U.S. patent application number 14/625575 was filed with the patent office on 2015-09-24 for immunobinders directed against sclerostin.
The applicant listed for this patent is AbbVie Inc.. Invention is credited to Lorenzo Benatuil, Jacqueline Bixby, Sahana Bose, Alyssa Brito, Emma Fung, Chung-Ming Hsieh, Alexander Ivanov, Yulia Kutskova, John Memmott, Wendy Waegell.
Application Number | 20150266977 14/625575 |
Document ID | / |
Family ID | 47144147 |
Filed Date | 2015-09-24 |
United States Patent
Application |
20150266977 |
Kind Code |
A1 |
Hsieh; Chung-Ming ; et
al. |
September 24, 2015 |
IMMUNOBINDERS DIRECTED AGAINST SCLEROSTIN
Abstract
Proteins that bind sclerostin or sclerostin and TNF are
described along with there use in composition and methods for
treating, preventing, and diagnosing sclerostin related diseases
and for detecting sclerostin or sclerostin and TNF in cells,
tissues, samples, and compositions.
Inventors: |
Hsieh; Chung-Ming; (Newton,
MA) ; Ivanov; Alexander; (Lexington, MA) ;
Waegell; Wendy; (Brookfield, MA) ; Kutskova;
Yulia; (Northborough, MA) ; Memmott; John;
(Framingham, MA) ; Benatuil; Lorenzo;
(Northborough, MA) ; Bixby; Jacqueline; (Auburn,
MA) ; Fung; Emma; (Northborough, MA) ; Bose;
Sahana; (Marlborough, MA) ; Brito; Alyssa;
(Hudson, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AbbVie Inc. |
North Chicago |
IL |
US |
|
|
Family ID: |
47144147 |
Appl. No.: |
14/625575 |
Filed: |
February 18, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13659647 |
Oct 24, 2012 |
8999331 |
|
|
14625575 |
|
|
|
|
61550724 |
Oct 24, 2011 |
|
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Current U.S.
Class: |
424/136.1 ;
424/142.1; 435/252.33; 435/254.21; 435/320.1; 435/328; 435/336;
435/69.6; 530/387.3; 530/391.1; 530/391.3; 530/391.7;
536/23.53 |
Current CPC
Class: |
A61P 11/00 20180101;
C07K 16/22 20130101; A61K 38/1793 20130101; A61P 37/06 20180101;
A61P 1/16 20180101; C07K 16/241 20130101; A61P 37/02 20180101; C07K
2317/31 20130101; A61K 38/2073 20130101; C07K 2317/76 20130101;
A61K 31/44 20130101; A61K 38/13 20130101; A61P 1/04 20180101; A61K
38/191 20130101; A61K 31/519 20130101; C07K 2317/60 20130101; A61K
38/18 20130101; A61P 35/00 20180101; A61P 37/00 20180101; C07K
2317/64 20130101; A61P 1/00 20180101; C07K 16/468 20130101; A61P
31/14 20180101; A61K 38/2026 20130101; A61K 31/56 20130101; A61K
2039/505 20130101; A61P 25/28 20180101; A61K 38/1808 20130101; C07K
2317/92 20130101; A61P 31/18 20180101; A61P 17/04 20180101; A61K
31/4439 20130101; A61P 25/00 20180101; C07K 2317/94 20130101; A61K
38/217 20130101; C07K 2317/21 20130101; A61K 38/1816 20130101; A61K
39/3955 20130101; A61P 11/06 20180101; A61P 19/02 20180101; A61P
29/00 20180101; A61P 11/02 20180101; A61K 38/2066 20130101; C07K
2317/90 20130101; A61K 31/439 20130101; A61K 38/19 20130101; A61K
31/353 20130101; A61P 37/08 20180101; C07K 2317/565 20130101; C07K
16/18 20130101 |
International
Class: |
C07K 16/46 20060101
C07K016/46; C07K 16/24 20060101 C07K016/24; C07K 16/22 20060101
C07K016/22 |
Claims
1-12. (canceled)
13. An isolated nucleic acid encoding a binding protein comprising
an antigen binding domain capable of binding human sclerostin, the
antigen binding domain comprising at least one CDR comprising an
amino acid consensus, wherein the at least one CDR comprises an
amino acid sequence selected from the group consisting of residues
31-35 of SEQ ID NO: 3; residues 50-66 of SEQ ID NO: 3; residues
99-108 of SEQ ID NO:3; residues 23-34 of SEQ ID NO: 4; residues
51-57 of SEQ ID NO: 4; residues 90-101 of SEQ ID NO:4; residues
31-35 of SEQ ID NO: 5; residues 50-66 of SEQ ID NO: 5; residues
99-115 of SEQ ID NO:5; residues 23-33 of SEQ ID NO: 6; residues
49-55 of SEQ ID NO: 6; residues 88-96 of SEQ ID NO:6; residues
31-35 of SEQ ID NO: 7; residues 50-66 of SEQ ID NO: 7; residues
99-107 of SEQ ID NO:7; residues 23-33 of SEQ ID NO: 8; residues
49-55 of SEQ ID NO: 8; residues 88-95 of SEQ ID NO:8; residues
31-35 of SEQ ID NO: 9; residues 50-66 of SEQ ID NO: 9; residues
99-107 of SEQ ID NO:9; residues 24-39 of SEQ ID NO: 10; residues
55-61 of SEQ ID NO: 10; residues 94-112 of SEQ ID NO:10; residues
31-35 of SEQ ID NO: 11; residues 50-66 of SEQ ID NO: 11; residues
99-111 of SEQ ID NO: 11; residues 24-39 of SEQ ID NO: 12; residues
55-61 of SEQ ID NO: 12; residues 94-113 of SEQ ID NO:12; residues
31-37 of SEQ ID NO: 13; residues 52-69 of SEQ ID NO: 13; residues
102-122 of SEQ ID NO:13; residues 24-34 of SEQ ID NO: 14; residues
50-56 of SEQ ID NO: 14; residues 89-97 of SEQ ID NO:14; Residues
31-35 of SEQ ID NO:1998; Residues 50-66 of SEQ ID NO:1998; Residues
99-110 of SEQ ID NO:1998; Residues 31-35 of SEQ ID NO.:1999;
Residues 50-66 of SEQ ID NO.:1999; Residues 99-110 of SEQ ID
NO.:1999; Residues 31-35 of SEQ ID NO.:2000; Residues 50-66 of SEQ
ID NO.:2000; Residues 99-110 of SEQ ID NO.:2000; Residues 31-35 of
SEQ ID NO.:2001; Residues 50-66 of SEQ ID NO.:2001; Residues 99-110
of SEQ ID NO.:2001; Residues 31-35 of SEQ ID NO.:2002; Residues
50-66 of SEQ ID NO.:2002; Residues 99-110 of SEQ ID NO.:2002;
Residues 31-35 of SEQ ID NO.:2003; Residues 50-66 of SEQ ID
NO.:2003; Residues 99-110 of SEQ ID NO.:2003; Residues 31-35 of SEQ
ID NO.:2004; Residues 50-66 of SEQ ID NO.:2004; Residues 99-110 of
SEQ ID NO.:2004; Residues 31-35 of SEQ ID NO.:2005; Residues 50-66
of SEQ ID NO.:2005; Residues 99-110 of SEQ ID NO.:2005; Residues
31-35 of SEQ ID NO.:2006; Residues 50-66 of SEQ ID NO.:2006;
Residues 99-110 of SEQ ID NO.:2006; Residues 31-35 of SEQ ID
NO.:2007; Residues 50-66 of SEQ ID NO.:2007; Residues 99-110 of SEQ
ID NO.:2007; Residues 23-36 of SEQ ID NO.:2008; Residues 52-58 of
SEQ ID NO.:2008; Residues 101-109 of SEQ ID NO.:2008; Residues
23-36 of SEQ ID NO.:2009; Residues 52-58 of SEQ ID NO.:2009;
Residues 101-109 of SEQ ID NO.:2009; Residues 23-36 of SEQ ID
NO.:2008; Residues 52-58 of SEQ ID NO.:2010; Residues 101-109 of
SEQ ID NO.:2010; Residues 23-36 of SEQ ID NO.:2011; Residues 52-58
of SEQ ID NO.:2011; Residues 101-109 of SEQ ID NO.:2011; Residues
23-36 of SEQ ID NO.:2012; Residues 52-58 of SEQ ID NO.:2012;
Residues 101-109 of SEQ ID NO.:2012; Residues 23-36 of SEQ ID
NO.:2013; Residues 52-58 of SEQ ID NO.:2013; Residues 101-109 of
SEQ ID NO.:2013; Residues 23-36 of SEQ ID NO.:2014; Residues 52-58
of SEQ ID NO.:2014; Residues 101-109 of SEQ ID NO.:2014; Residues
23-36 of SEQ ID NO.:2015; Residues 52-58 of SEQ ID NO.:2015;
Residues 101-109 of SEQ ID NO.:2015; Residues 23-36 of SEQ ID
NO.:2016; Residues 52-58 of SEQ ID NO.:2016; Residues 101-109 of
SEQ ID NO.:2016; Residues 23-36 of SEQ ID NO.:2017; Residues 52-58
of SEQ ID NO.:2017; Residues 101-109 of SEQ ID NO.:2017; Residues
31-35 of SEQ ID NO:2020; Residues 50-66 of SEQ ID NO:2020; Residues
99-108 of SEQ ID NO:2020; Residues 31-35 of SEQ ID NO:2021;
Residues 50-66 of SEQ ID NO:2021; Residues 99-108 of SEQ ID
NO:2021; Residues 31-35 of SEQ ID NO:2022; Residues 50-66 of SEQ ID
NO:2022; Residues 99-108 of SEQ ID NO:2022; Residues 31-35 of SEQ
ID NO:2023; Residues 50-66 of SEQ ID NO:2023; Residues 99-108 of
SEQ ID NO:2023; Residues 31-35 of SEQ ID NO:2024; Residues 50-66 of
SEQ ID NO:2024; Residues 99-108 of SEQ ID NO:2024; Residues 31-35
of SEQ ID NO:2025; Residues 50-66 of SEQ ID NO:2025; Residues
99-108 of SEQ ID NO:2025; Residues 31-35 of SEQ ID NO:2026;
Residues 50-66 of SEQ ID NO:2026; Residues 99-108 of SEQ ID
NO:2026; Residues 31-35 of SEQ ID NO:2027; Residues 50-66 of SEQ ID
NO:2027; Residues 99-108 of SEQ ID NO:2027; Residues 31-35 of SEQ
ID NO:2028; Residues 50-66 of SEQ ID NO:2028; Residues 99-108 of
SEQ ID NO:2028; Residues 31-35 of SEQ ID NO:2029; Residues 50-66 of
SEQ ID NO:2029; Residues 99-108 of SEQ ID NO:2029; Residues 31-35
of SEQ ID NO:2030; Residues 50-66 of SEQ ID NO:2030; Residues
99-108 of SEQ ID NO:2030; Residues 31-35 of SEQ ID NO:2031;
Residues 50-66 of SEQ ID NO:2031; Residues 99-108 of SEQ ID
NO:2031; Residues 31-35 of SEQ ID NO:2032; Residues 50-66 of SEQ ID
NO:2032; Residues 99-108 of SEQ ID NO:2032; Residues 31-35 of SEQ
ID NO:2033; Residues 50-66 of SEQ ID NO:2033; Residues 99-108 of
SEQ ID NO:2033; Residues 31-35 of SEQ ID NO:2034; Residues 50-66 of
SEQ ID NO:2034; Residues 99-110 of SEQ ID NO:2034; Residues 31-35
of SEQ ID NO.:2035; Residues 51-57 of SEQ ID NO.:2035; Residues
90-101 of SEQ ID NO.:2035; Residues 31-35 of SEQ ID NO.:2036;
Residues 51-57 of SEQ ID NO.:2036; Residues 90-101 of SEQ ID
NO.:2036; Residues 31-35 of SEQ ID NO.:2037; Residues 51-57 of SEQ
ID NO.:2035; Residues 90-101 of SEQ ID NO.:2037; Residues 31-35 of
SEQ ID NO.:2038; Residues 51-57 of SEQ ID NO.:2038; Residues 90-101
of SEQ ID NO.:2038; Residues 31-35 of SEQ ID NO.:2039; Residues
51-57 of SEQ ID NO.:2039; Residues 90-101 of SEQ ID NO.:2039;
Residues 31-35 of SEQ ID NO.:2040; Residues 51-57 of SEQ ID
NO.:2040; Residues 90-101 of SEQ ID NO.:2040; Residues 31-35 of SEQ
ID NO.:2041; Residues 51-57 of SEQ ID NO.:2041; Residues 90-101 of
SEQ ID NO.:2041; Residues 31-35 of SEQ ID NO.:2042; Residues 51-57
of SEQ ID NO.:2042; Residues 90-101 of SEQ ID NO.:2042; Residues
31-35 of SEQ ID NO.:2043; Residues 51-57 of SEQ ID NO.:2043;
Residues 90-101 of SEQ ID NO.:2043; Residues 31-35 of SEQ ID
NO.:2044; Residues 51-57 of SEQ ID NO.:2044; Residues 90-101 of SEQ
ID NO.:2044; Residues 31-35 of SEQ ID NO.:2045; Residues 51-57 of
SEQ ID NO.:2045; Residues 90-101 of SEQ ID NO.:2045; Residues 31-35
of SEQ ID NO.:2046; Residues 51-57 of SEQ ID NO.:2046; Residues
90-101 of SEQ ID NO.:2046; Residues 31-35 of SEQ ID NO.:2047;
Residues 51-57 of SEQ ID NO.:2047; Residues 90-101 of SEQ ID
NO.:2047; Residues 31-35 of SEQ ID NO.:2048; Residues 51-57 of SEQ
ID NO.:2048; Residues 90-101 of SEQ ID NO.:2048; Residues 31-35 of
SEQ ID NO.:2049; Residues 51-57 of SEQ ID NO.:2049; and Residues
90-101 of SEQ ID NO.:2049.
14. A vector comprising the isolated nucleic acid of claim 13.
15. A host cell comprising the vector of claim 15.
16. A method of producing a protein capable of binding sclerostin,
comprising culturing the host cell of claim 16 in culture medium
under conditions sufficient to produce a binding protein capable of
binding sclerostin.
17-23. (canceled)
24. A method for treating a subject for a disease or a disorder in
which sclerostin activity is detrimental by administering to the
subject a binding protein such that treatment is achieved, wherein
the binding protein comprises an antigen binding domain capable of
binding human sclerostin, the antigen binding domain comprising at
least one CDR comprising an amino acid consensus wherein the at
least one CDR comprises an amino acid sequence selected from the
group consisting of residues 31-35 of SEQ ID NO: 3; residues 50-66
of SEQ ID NO: 3; residues 99-108 of SEQ ID NO:3; residues 23-34 of
SEQ ID NO: 4; residues 51-57 of SEQ ID NO: 4; residues 90-101 of
SEQ ID NO:4; residues 31-35 of SEQ ID NO: 5; residues 50-66 of SEQ
ID NO: 5; residues 99-115 of SEQ ID NO:5; residues 23-33 of SEQ ID
NO: 6; residues 49-55 of SEQ ID NO: 6; residues 88-96 of SEQ ID
NO:6; residues 31-35 of SEQ ID NO: 7; residues 50-66 of SEQ ID NO:
7; residues 99-107 of SEQ ID NO:7; residues 23-33 of SEQ ID NO: 8;
residues 49-55 of SEQ ID NO: 8; residues 88-95 of SEQ ID NO:8;
residues 31-35 of SEQ ID NO: 9; residues 50-66 of SEQ ID NO: 9;
residues 99-107 of SEQ ID NO:9; residues 24-39 of SEQ ID NO: 10;
residues 55-61 of SEQ ID NO: 10; residues 94-112 of SEQ ID NO:10;
residues 31-35 of SEQ ID NO: 11; residues 50-66 of SEQ ID NO: 11;
residues 99-111 of SEQ ID NO: 11; residues 24-39 of SEQ ID NO: 12;
residues 55-61 of SEQ ID NO: 12; residues 94-113 of SEQ ID NO:12;
residues 31-37 of SEQ ID NO: 13; residues 52-69 of SEQ ID NO: 13;
residues 102-122 of SEQ ID NO:13; residues 24-34 of SEQ ID NO: 14;
residues 50-56 of SEQ ID NO: 14; residues 89-97 of SEQ ID NO:14;
Residues 31-35 of SEQ ID NO:1998; Residues 50-66 of SEQ ID NO:1998;
Residues 99-110 of SEQ ID NO:1998; Residues 31-35 of SEQ ID
NO.:1999; Residues 50-66 of SEQ ID NO.:1999; Residues 99-110 of SEQ
ID NO.:1999; Residues 31-35 of SEQ ID NO.:2000; Residues 50-66 of
SEQ ID NO.:2000; Residues 99-110 of SEQ ID NO.:2000; Residues 31-35
of SEQ ID NO.:2001; Residues 50-66 of SEQ ID NO.:2001; Residues
99-110 of SEQ ID NO.:2001; Residues 31-35 of SEQ ID NO.:2002;
Residues 50-66 of SEQ ID NO.:2002; Residues 99-110 of SEQ ID
NO.:2002; Residues 31-35 of SEQ ID NO.:2003; Residues 50-66 of SEQ
ID NO.:2003; Residues 99-110 of SEQ ID NO.:2003; Residues 31-35 of
SEQ ID NO.:2004; Residues 50-66 of SEQ ID NO.:2004; Residues 99-110
of SEQ ID NO.:2004; Residues 31-35 of SEQ ID NO.:2005; Residues
50-66 of SEQ ID NO.:2005; Residues 99-110 of SEQ ID NO.:2005;
Residues 31-35 of SEQ ID NO.:2006; Residues 50-66 of SEQ ID
NO.:2006; Residues 99-110 of SEQ ID NO.:2006; Residues 31-35 of SEQ
ID NO.:2007; Residues 50-66 of SEQ ID NO.:2007; Residues 99-110 of
SEQ ID NO.:2007; Residues 23-36 of SEQ ID NO.:2008; Residues 52-58
of SEQ ID NO.:2008; Residues 101-109 of SEQ ID NO.:2008; Residues
23-36 of SEQ ID NO.:2009; Residues 52-58 of SEQ ID NO.:2009;
Residues 101-109 of SEQ ID NO.:2009; Residues 23-36 of SEQ ID
NO.:2008; Residues 52-58 of SEQ ID NO.:2010; Residues 101-109 of
SEQ ID NO.:2010; Residues 23-36 of SEQ ID NO.:2011; Residues 52-58
of SEQ ID NO.:2011; Residues 101-109 of SEQ ID NO.:2011; Residues
23-36 of SEQ ID NO.:2012; Residues 52-58 of SEQ ID NO.:2012;
Residues 101-109 of SEQ ID NO.:2012; Residues 23-36 of SEQ ID
NO.:2013; Residues 52-58 of SEQ ID NO.:2013; Residues 101-109 of
SEQ ID NO.:2013; Residues 23-36 of SEQ ID NO.:2014; Residues 52-58
of SEQ ID NO.:2014; Residues 101-109 of SEQ ID NO.:2014; Residues
23-36 of SEQ ID NO.:2015; Residues 52-58 of SEQ ID NO.:2015;
Residues 101-109 of SEQ ID NO.:2015; Residues 23-36 of SEQ ID
NO.:2016; Residues 52-58 of SEQ ID NO.:2016; Residues 101-109 of
SEQ ID NO.:2016; Residues 23-36 of SEQ ID NO.:2017; Residues 52-58
of SEQ ID NO.:2017; Residues 101-109 of SEQ ID NO.:2017; Residues
31-35 of SEQ ID NO:2020; Residues 50-66 of SEQ ID NO:2020; Residues
99-108 of SEQ ID NO:2020; Residues 31-35 of SEQ ID NO:2021;
Residues 50-66 of SEQ ID NO:2021; Residues 99-108 of SEQ ID
NO:2021; Residues 31-35 of SEQ ID NO:2022; Residues 50-66 of SEQ ID
NO:2022; Residues 99-108 of SEQ ID NO:2022; Residues 31-35 of SEQ
ID NO:2023; Residues 50-66 of SEQ ID NO:2023; Residues 99-108 of
SEQ ID NO:2023; Residues 31-35 of SEQ ID NO:2024; Residues 50-66 of
SEQ ID NO:2024; Residues 99-108 of SEQ ID NO:2024; Residues 31-35
of SEQ ID NO:2025; Residues 50-66 of SEQ ID NO:2025; Residues
99-108 of SEQ ID NO:2025; Residues 31-35 of SEQ ID NO:2026;
Residues 50-66 of SEQ ID NO:2026; Residues 99-108 of SEQ ID
NO:2026; Residues 31-35 of SEQ ID NO:2027; Residues 50-66 of SEQ ID
NO:2027; Residues 99-108 of SEQ ID NO:2027; Residues 31-35 of SEQ
ID NO:2028; Residues 50-66 of SEQ ID NO:2028; Residues 99-108 of
SEQ ID NO:2028; Residues 31-35 of SEQ ID NO:2029; Residues 50-66 of
SEQ ID NO:2029; Residues 99-108 of SEQ ID NO:2029; Residues 31-35
of SEQ ID NO:2030; Residues 50-66 of SEQ ID NO:2030; Residues
99-108 of SEQ ID NO:2030; Residues 31-35 of SEQ ID NO:2031;
Residues 50-66 of SEQ ID NO:2031; Residues 99-108 of SEQ ID
NO:2031; Residues 31-35 of SEQ ID NO:2032; Residues 50-66 of SEQ ID
NO:2032; Residues 99-108 of SEQ ID NO:2032; Residues 31-35 of SEQ
ID NO:2033; Residues 50-66 of SEQ ID NO:2033; Residues 99-108 of
SEQ ID NO:2033; Residues 31-35 of SEQ ID NO:2034; Residues 50-66 of
SEQ ID NO:2034; Residues 99-110 of SEQ ID NO:2034; Residues 31-35
of SEQ ID NO.:2035; Residues 51-57 of SEQ ID NO.:2035; Residues
90-101 of SEQ ID NO.:2035; Residues 31-35 of SEQ ID NO.:2036;
Residues 51-57 of SEQ ID NO.:2036; Residues 90-101 of SEQ ID
NO.:2036; Residues 31-35 of SEQ ID NO.:2037; Residues 51-57 of SEQ
ID NO.:2035; Residues 90-101 of SEQ ID NO.:2037; Residues 31-35 of
SEQ ID NO.:2038; Residues 51-57 of SEQ ID NO.:2038; Residues 90-101
of SEQ ID NO.:2038; Residues 31-35 of SEQ ID NO.:2039; Residues
51-57 of SEQ ID NO.:2039; Residues 90-101 of SEQ ID NO.:2039;
Residues 31-35 of SEQ ID NO.:2040; Residues 51-57 of SEQ ID
NO.:2040; Residues 90-101 of SEQ ID NO.:2040; Residues 31-35 of SEQ
ID NO.:2041; Residues 51-57 of SEQ ID NO.:2041; Residues 90-101 of
SEQ ID NO.:2041; Residues 31-35 of SEQ ID NO.:2042; Residues 51-57
of SEQ ID NO.:2042; Residues 90-101 of SEQ ID NO.:2042; Residues
31-35 of SEQ ID NO.:2043; Residues 51-57 of SEQ ID NO.:2043;
Residues 90-101 of SEQ ID NO.:2043; Residues 31-35 of SEQ ID
NO.:2044; Residues 51-57 of SEQ ID NO.:2044; Residues 90-101 of SEQ
ID NO.:2044; Residues 31-35 of SEQ ID NO.:2045; Residues 51-57 of
SEQ ID NO.:2045; Residues 90-101 of SEQ ID NO.:2045; Residues 31-35
of SEQ ID NO.:2046; Residues 51-57 of SEQ ID NO.:2046; Residues
90-101 of SEQ ID NO.:2046; Residues 31-35 of SEQ ID NO.:2047;
Residues 51-57 of SEQ ID NO.:2047; Residues 90-101 of SEQ ID
NO.:2047; Residues 31-35 of SEQ ID NO.:2048; Residues 51-57 of SEQ
ID NO.:2048; Residues 90-101 of SEQ ID NO.:2048; Residues 31-35 of
SEQ ID NO.:2049; Residues 51-57 of SEQ ID NO.:2049; and Residues
90-101 of SEQ ID NO.:2049.
25. The method of claim 24, wherein the disorder is selected from
the group consisting of a respiratory disorder; asthma; allergic
and nonallergic asthma; asthma due to infection; asthma due to
infection with respiratory syncytial virus (RSV); chronic
obstructive pulmonary disease (COPD); a condition involving airway
inflammation; eosinophilia; fibrosis and/or excess mucus
production; cystic fibrosis; pulmonary fibrosis; an atopic
disorder; atopic dermatitis; urticaria; eczema; allergic rhinitis;
allergic enterogastritis; an inflammatory and/or autoimmune
condition of the skin; an inflammatory and/or autoimmune condition
of gastrointestinal organs; inflammatory bowel disease (IBD);
ulcerative colitis; Crohn's disease; an inflammatory and/or
autoimmune condition of the liver; liver cirrhosis; liver fibrosis;
liver fibrosis caused by hepatitis B and/or C virus; scleroderma; a
tumors; a cancer; hepatocellular carcinoma; glioblastoma; lymphoma;
Hodgkin's lymphoma; a viral infection; HTLV-1 infection (e.g., from
HTLV-1); a suppression of expression of protective type 1 immune
response, and a suppression of expression of protective type 1
immune responses during vaccination.
26-31. (canceled)
32. A binding protein comprising first and second polypeptide
chains, wherein the first polypeptide chain comprises a first
VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable
domain; VD2 is a second heavy chain variable domain; C is a heavy
chain constant domain; X1 is a first linker; X2 is an Fc region;
(X1)n is (X1)0 or (X1)1; (X2)n is (X2)0 or (X2)1; and wherein the
second polypeptide chain comprises a second VD1-(X1)n-VD2-C-(X2)n,
wherein VD1 is a first light chain variable domain; VD2 is a second
light chain variable domain; C is a light chain constant domain; X1
is a second linker; X2 does not comprise an Fc region; (X1)n is
(X1)0 or (X1)1; (X2)n is (X2)0 or (X2)1; and wherein the first and
second X1 linker are the same or different; wherein the first X1
linker is not CH1 and/or the second X1 linker is not CL and wherein
(a) the VD1 or VD2 heavy chain variable domain comprise three CDRs
selected from the group consisting of SEQ ID NO: 24, 34, 44, 54,
64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194,
204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324,
334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454,
464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584,
594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714,
724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824, 834, 844,
854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954, 964, 974,
984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084,
1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204,
1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314,
1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424,
1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534,
1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644,
1654, 1664, 1674, and 1684, the VD1 or VD2 light chain variable
domain comprises three CDRs selected from the group consisting of
SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129, 139, 149,
159, 169, 179, 189, 199, 209, 219, 229, 239, 249, 259, 269, 279,
289, 299, 309, 319, 329, 339, 349, 359, 369, 379, 389, 399, 409,
419, 429, 439, 449, 459, 469, 479, 489, 499, 509, 519, 529, 539,
549, 559, 569, 579, 589, 599, 609, 619, 629, 639, 649, 659, 669,
679, 689, 699, 709, 719, 729, 739, 749, 759, 769, 779, 789, 799,
809, 819, 829, 839, 849, 859, 869, 879, 889, 899, 909, 919, 929,
939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029, 1039, 1049,
1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159,
1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269,
1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379,
1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489,
1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599,
1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, and 1689, and the
binding protein is capable of binding sclerostin and another
target; (b) the VD1 and VD2 heavy chain variable domains
independently comprise three CDRs selected from the group
consisting of SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114,
124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244,
254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374,
384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504,
514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634,
644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764,
774, 784, 794, 804, 814, 824, 834, 844, 854, 864, 874, 884, 894,
904, 914, 924, 934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024,
1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144,
1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254,
1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364,
1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474,
1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584,
1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674, and 1684, the
VD1 or VD2 light chain variable domain comprises three CDRs
selected from the group consisting of SEQ ID NO: 29, 39, 49, 59,
69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199,
209, 219, 229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329,
339, 349, 359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459,
469, 479, 489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589,
599, 609, 619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719,
729, 739, 749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849,
859, 869, 879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979,
989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089,
1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199,
1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309,
1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419,
1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529,
1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639,
1649, 1659, 1669, 1679, and 1689, and the binding protein is
capable of binding sclerostin and sclerostin; (c) the VD1 heavy
chain variable domain comprises three CDRs selected from the group
consisting of SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114,
124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244,
254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374,
384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504,
514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634,
644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764,
774, 784, 794, 804, 814, 824, 834, 844, 854, 864, 874, 884, 894,
904, 914, 924, 934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024,
1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144,
1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254,
1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364,
1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474,
1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584,
1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674, and 1684, and
the VD2 heavy chain variable domain comprises three CDRs selected
from the group consisting of SEQ ID NO: 22, 32, 42, 52, 62, 72, 82,
92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212,
222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342,
352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472,
482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602,
612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732,
742, 752, 762, 772, 782, 792, 802, 812, 822, 832, 842, 852, 862,
872, 882, 892, 902, 912, 922, 932, 942, 952, 962, 972, 982, 992,
1002, 1012, 1022, 1032, 1042, 1052, 1062, 1072, 1082, 1092, 1102,
1112, 1122, 1132, 1142, 1152, 1162, 1172, 1182, 1192, 1202, 1212,
1222, 1232, 1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322,
1332, 1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322, 1332,
1342, 1352, 1362, 1372, 1382, 1392, 1402, 1412, 1422, 1432, 1442,
1452, 1462, 1472, 1482, 1492, 1502, 1512, 1522, 1532, 1542, 1552,
1562, 1572, 1582, 1592, 1602, 1612, 1622, 1632, 1642, 1652, 1662,
1672, and 1682; the VD1 light chain variable domain comprises three
CDRs selected from the group consisting of SEQ ID NO: 29, 39, 49,
59, 69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189,
199, 209, 219, 229, 239, 249, 259, 269, 279, 289, 299, 309, 319,
329, 339, 349, 359, 369, 379, 389, 399, 409, 419, 429, 439, 449,
459, 469, 479, 489, 499, 509, 519, 529, 539, 549, 559, 569, 579,
589, 599, 609, 619, 629, 639, 649, 659, 669, 679, 689, 699, 709,
719, 729, 739, 749, 759, 769, 779, 789, 799, 809, 819, 829, 839,
849, 859, 869, 879, 889, 899, 909, 919, 929, 939, 949, 959, 969,
979, 989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079,
1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189,
1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299,
1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409,
1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519,
1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629,
1639, 1649, 1659, 1669, 1679, and 1689, and the VD2 light chain
variable domain comprises three CDRs selected from the group
consisting of SEQ ID NO: 27, 37, 47, 57, 67, 77, 87, 97, 107, 117,
127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247,
257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377,
387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507,
517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637,
647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767,
777, 787, 797, 807, 817, 827, 837, 847, 857, 867, 877, 887, 897,
907, 917, 927, 937, 947, 957, 967, 977, 987, 997, 1007, 1017, 1027,
1037, 1047, 1057, 1067, 1077, 1087, 1097, 1107, 1117, 1127, 1137,
1147, 1157, 1167, 1177, 1187, 1197, 1207, 1217, 1227, 1237, 1247,
1257, 1267, 1277, 1287, 1297, 1307, 1317, 1327, 1337, 1347, 1357,
1367, 1377, 1387, 1397, 1407, 1417, 1427, 1437, 1447, 1457, 1467,
1477, 1487, 1497, 1507, 1517, 1527, 1537, 1547, 1557, 1567, 1577,
1587, 1597, 1607, 1617, 1627, 1637, 1647, 1657, 1667, 1677, and
1687, and the binding protein is capable of binding sclerostin and
TNF-.alpha.; or (d) the VD2 heavy chain variable domain comprises
three CDRs selected from the group consisting of SEQ ID NO: 24, 34,
44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174,
184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304,
314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434,
444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564,
574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694,
704, 714, 724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824,
834, 844, 854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954,
964, 974, 984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074,
1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194,
1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304,
1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414,
1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524,
1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634,
1644, 1654, 1664, 1674, and 1684, and the VD1 heavy chain variable
domain comprises three CDRs selected from the group consisting of
SEQ ID NO: 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142,
152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272,
282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402,
412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532,
542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662,
672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792,
802, 812, 822, 832, 842, 852, 862, 872, 882, 892, 902, 912, 922,
932, 942, 952, 962, 972, 982, 992, 1002, 1012, 1022, 1032, 1042,
1052, 1062, 1072, 1082, 1092, 1102, 1112, 1122, 1132, 1142, 1152,
1162, 1172, 1182, 1192, 1202, 1212, 1222, 1232, 1242, 1252, 1262,
1272, 1282, 1292, 1302, 1312, 1322, 1332, 1242, 1252, 1262, 1272,
1282, 1292, 1302, 1312, 1322, 1332, 1342, 1352, 1362, 1372, 1382,
1392, 1402, 1412, 1422, 1432, 1442, 1452, 1462, 1472, 1482, 1492,
1502, 1512, 1522, 1532, 1542, 1552, 1562, 1572, 1582, 1592, 1602,
1612, 1622, 1632, 1642, 1652, 1662, 1672, and 1682; the VD2 light
chain variable domain comprises three CDRs selected from the group
consisting of SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119,
129, 139, 149, 159, 169, 179, 189, 199, 209, 219, 229, 239, 249,
259, 269, 279, 289, 299, 309, 319, 329, 339, 349, 359, 369, 379,
389, 399, 409, 419, 429, 439, 449, 459, 469, 479, 489, 499, 509,
519, 529, 539, 549, 559, 569, 579, 589, 599, 609, 619, 629, 639,
649, 659, 669, 679, 689, 699, 709, 719, 729, 739, 749, 759, 769,
779, 789, 799, 809, 819, 829, 839, 849, 859, 869, 879, 889, 899,
909, 919, 929, 939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029,
1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139,
1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249,
1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359,
1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469,
1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579,
1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, and
1689, and the VD1 light chain variable domain comprises three CDRs
selected from the group consisting of SEQ ID NO: 27, 37, 47, 57,
67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197,
207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327,
337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457,
467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587,
597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717,
727, 737, 747, 757, 767, 777, 787, 797, 807, 817, 827, 837, 847,
857, 867, 877, 887, 897, 907, 917, 927, 937, 947, 957, 967, 977,
987, 997, 1007, 1017, 1027, 1037, 1047, 1057, 1067, 1077, 1087,
1097, 1107, 1117, 1127, 1137, 1147, 1157, 1167, 1177, 1187, 1197,
1207, 1217, 1227, 1237, 1247, 1257, 1267, 1277, 1287, 1297, 1307,
1317, 1327, 1337, 1347, 1357, 1367, 1377, 1387, 1397, 1407, 1417,
1427, 1437, 1447, 1457, 1467, 1477, 1487, 1497, 1507, 1517, 1527,
1537, 1547, 1557, 1567, 1577, 1587, 1597, 1607, 1617, 1627, 1637,
1647, 1657, 1667, 1677, and 1687, and the binding protein is
capable of binding TNF-.alpha., and sclerostin.
33-40. (canceled)
41. A binding protein capable of binding two antigens comprising
four polypeptide chains, wherein two polypeptide chains comprise
VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable
domain; VD2 is a second heavy chain variable domain; C is a heavy
chain constant domain; X1 is a first linker; X2 is an Fc region;
(X1)n is (X1)0 or (X1)1; (X2)n is (X2)0 or (X2)1; and wherein two
polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a
first light chain variable domain; VD2 is a second light chain
variable domain; C is a light chain constant domain; X1 is a second
linker; X2 does not comprise an Fc region; (X1)n is (X1)0 or (X1)1;
(X2)n is (X2)0 or (X2)1; and wherein the first and second X1 linker
are the same or different; wherein the first X1 linker is not CH1
and/or the second X1 linker is not CL and wherein (a) the VD1 or
VD2 heavy chain variable domain comprise three CDRs selected from
the group consisting of SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94,
104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224,
234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354,
364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484,
494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614,
624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744,
754, 764, 774, 784, 794, 804, 814, 824, 834, 844, 854, 864, 874,
884, 894, 904, 914, 924, 934, 944, 954, 964, 974, 984, 994, 1004,
1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124,
1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234,
1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344,
1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454,
1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564,
1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674,
and 1684, the VD1 or VD2 light chain variable domain comprises
three CDRs selected from the group consisting of SEQ ID NO: 29, 39,
49, 59, 69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169, 179,
189, 199, 209, 219, 229, 239, 249, 259, 269, 279, 289, 299, 309,
319, 329, 339, 349, 359, 369, 379, 389, 399, 409, 419, 429, 439,
449, 459, 469, 479, 489, 499, 509, 519, 529, 539, 549, 559, 569,
579, 589, 599, 609, 619, 629, 639, 649, 659, 669, 679, 689, 699,
709, 719, 729, 739, 749, 759, 769, 779, 789, 799, 809, 819, 829,
839, 849, 859, 869, 879, 889, 899, 909, 919, 929, 939, 949, 959,
969, 979, 989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079,
1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189,
1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299,
1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409,
1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519,
1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629,
1639, 1649, 1659, 1669, 1679, and 1689, and the binding protein is
capable of binding sclerostin and another target; (b) the VD1 and
VD2 heavy chain variable domains independently comprise three CDRs
selected from the group consisting of SEQ ID NO: 24, 34, 44, 54,
64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194,
204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324,
334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454,
464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584,
594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714,
724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824, 834, 844,
854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954, 964, 974,
984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084,
1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204,
1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314,
1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424,
1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534,
1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644,
1654, 1664, 1674, and 1684, the VD1 or VD2 light chain variable
domain comprises three CDRs selected from the group consisting of
SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129, 139, 149,
159, 169, 179, 189, 199, 209, 219, 229, 239, 249, 259, 269, 279,
289, 299, 309, 319, 329, 339, 349, 359, 369, 379, 389, 399, 409,
419, 429, 439, 449, 459, 469, 479, 489, 499, 509, 519, 529, 539,
549, 559, 569, 579, 589, 599, 609, 619, 629, 639, 649, 659, 669,
679, 689, 699, 709, 719, 729, 739, 749, 759, 769, 779, 789, 799,
809, 819, 829, 839, 849, 859, 869, 879, 889, 899, 909, 919, 929,
939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029, 1039, 1049,
1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159,
1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269,
1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379,
1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489,
1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599,
1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, and 1689, and the
binding protein is capable of binding sclerostin and sclerostin;
(c) the VD1 heavy chain variable domain comprises three CDRs
selected from the group consisting of SEQ ID NO: 24, 34, 44, 54,
64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194,
204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324,
334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454,
464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584,
594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714,
724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824, 834, 844,
854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954, 964, 974,
984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084,
1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204,
1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314,
1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424,
1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534,
1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644,
1654, 1664, 1674, and 1684, and the VD2 heavy chain variable domain
comprises three CDRs selected from the group consisting of SEQ ID
NO: 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152,
162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282,
292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412,
422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542,
552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672,
682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792, 802,
812, 822, 832, 842, 852, 862, 872, 882, 892, 902, 912, 922, 932,
942, 952, 962, 972, 982, 992, 1002, 1012, 1022, 1032, 1042, 1052,
1062, 1072, 1082, 1092, 1102, 1112, 1122, 1132, 1142, 1152, 1162,
1172, 1182, 1192, 1202, 1212, 1222, 1232, 1242, 1252, 1262, 1272,
1282, 1292, 1302, 1312, 1322, 1332, 1242, 1252, 1262, 1272, 1282,
1292, 1302, 1312, 1322, 1332, 1342, 1352, 1362, 1372, 1382, 1392,
1402, 1412, 1422, 1432, 1442, 1452, 1462, 1472, 1482, 1492, 1502,
1512, 1522, 1532, 1542, 1552, 1562, 1572, 1582, 1592, 1602, 1612,
1622, 1632, 1642, 1652, 1662, 1672, and 1682; the VD1 light chain
variable domain comprises three CDRs selected from the group
consisting of SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119,
129, 139, 149, 159, 169, 179, 189, 199, 209, 219, 229, 239, 249,
259, 269, 279, 289, 299, 309, 319, 329, 339, 349, 359, 369, 379,
389, 399, 409, 419, 429, 439, 449, 459, 469, 479, 489, 499, 509,
519, 529, 539, 549, 559, 569, 579, 589, 599, 609, 619, 629, 639,
649, 659, 669, 679, 689, 699, 709, 719, 729, 739, 749, 759, 769,
779, 789, 799, 809, 819, 829, 839, 849, 859, 869, 879, 889, 899,
909, 919, 929, 939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029,
1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139,
1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249,
1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359,
1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469,
1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579,
1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, and
1689, and the VD2 light chain variable domain comprises three CDRs
selected from the group consisting of SEQ ID NO: 27, 37, 47, 57,
67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197,
207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327,
337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457,
467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587,
597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717,
727, 737, 747, 757, 767, 777, 787, 797, 807, 817, 827, 837, 847,
857, 867, 877, 887, 897, 907, 917, 927, 937, 947, 957, 967, 977,
987, 997, 1007, 1017, 1027, 1037, 1047, 1057, 1067, 1077, 1087,
1097, 1107, 1117, 1127, 1137, 1147, 1157, 1167, 1177, 1187, 1197,
1207, 1217, 1227, 1237, 1247, 1257, 1267, 1277, 1287, 1297, 1307,
1317, 1327, 1337, 1347, 1357, 1367, 1377, 1387, 1397, 1407, 1417,
1427, 1437, 1447, 1457, 1467, 1477, 1487, 1497, 1507, 1517, 1527,
1537, 1547, 1557, 1567, 1577, 1587, 1597, 1607, 1617, 1627, 1637,
1647, 1657, 1667, 1677, and 1687, and the binding protein is
capable of binding sclerostin and TNF-.alpha.; or (d) the VD2 heavy
chain variable domain comprises three CDRs selected from the group
consisting of SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114,
124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244,
254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374,
384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504,
514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634,
644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764,
774, 784, 794, 804, 814, 824, 834, 844, 854, 864, 874, 884, 894,
904, 914, 924, 934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024,
1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144,
1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254,
1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364,
1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474,
1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584,
1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674, and 1684, and
the VD1 heavy chain variable domain comprises three CDRs selected
from the group consisting of SEQ ID NO: 22, 32, 42, 52, 62, 72, 82,
92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212,
222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342,
352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472,
482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602,
612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732,
742, 752, 762, 772, 782, 792, 802, 812, 822, 832, 842, 852, 862,
872, 882, 892, 902, 912, 922, 932, 942, 952, 962, 972, 982, 992,
1002, 1012, 1022, 1032, 1042, 1052, 1062, 1072, 1082, 1092, 1102,
1112, 1122, 1132, 1142, 1152, 1162, 1172, 1182, 1192, 1202, 1212,
1222, 1232, 1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322,
1332, 1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322, 1332,
1342, 1352, 1362, 1372, 1382, 1392, 1402, 1412, 1422, 1432, 1442,
1452, 1462, 1472, 1482, 1492, 1502, 1512, 1522, 1532, 1542, 1552,
1562, 1572, 1582, 1592, 1602, 1612, 1622, 1632, 1642, 1652, 1662,
1672, and 1682; the VD2 light chain variable domain comprises three
CDRs selected from the group consisting of SEQ ID NO: 29, 39, 49,
59, 69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189,
199, 209, 219, 229, 239, 249, 259, 269, 279, 289, 299, 309, 319,
329, 339, 349, 359, 369, 379, 389, 399, 409, 419, 429, 439, 449,
459, 469, 479, 489, 499, 509, 519, 529, 539, 549, 559, 569, 579,
589, 599, 609, 619, 629, 639, 649, 659, 669, 679, 689, 699, 709,
719, 729, 739, 749, 759, 769, 779, 789, 799, 809, 819, 829, 839,
849, 859, 869, 879, 889, 899, 909, 919, 929, 939, 949, 959, 969,
979, 989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079,
1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189,
1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299,
1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409,
1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519,
1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629,
1639, 1649, 1659, 1669, 1679, and 1689, and the VD1 light chain
variable domain comprises three CDRs selected from the group
consisting of SEQ ID NO: 27, 37, 47, 57, 67, 77, 87, 97, 107, 117,
127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247,
257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377,
387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507,
517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637,
647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767,
777, 787, 797, 807, 817, 827, 837, 847, 857, 867, 877, 887, 897,
907, 917, 927, 937, 947, 957, 967, 977, 987, 997, 1007, 1017, 1027,
1037, 1047, 1057, 1067, 1077, 1087, 1097, 1107, 1117, 1127, 1137,
1147, 1157, 1167, 1177, 1187, 1197, 1207, 1217, 1227, 1237, 1247,
1257, 1267, 1277, 1287, 1297, 1307, 1317, 1327, 1337, 1347, 1357,
1367, 1377, 1387, 1397, 1407, 1417, 1427, 1437, 1447, 1457, 1467,
1477, 1487, 1497, 1507, 1517, 1527, 1537, 1547, 1557, 1567, 1577,
1587, 1597, 1607, 1617, 1627, 1637, 1647, 1657, 1667, 1677, and
1687, and the binding protein is capable of binding TNF-.alpha. and
sclerostin.
42-44. (canceled)
45. A binding protein construct comprising a binding protein
sequence of claim 32 further comprising a linker polypeptide and/or
an immunoglobulin constant domain.
46-48. (canceled)
49. A binding protein conjugate comprising a binding protein
construct of claim 32, the binding protein conjugate further
comprising an immunoadhesion molecule, an imaging agent, a
therapeutic agent, or a cytotoxic agent.
50. An isolated nucleic acid encoding a binding protein amino acid
sequence of claim 32.
51. An isolated nucleic acid encoding a sclerostin binding protein
construct amino acid sequence of claim 50.
52. A vector comprising the isolated nucleic acid of claim 51.
53. A host cell comprising the vector of claim 52.
54. A method of producing a protein capable of binding sclerostin,
comprising culturing the host cell of claim 53 in culture medium
under conditions sufficient to produce a binding protein capable of
binding sclerostin.
55. (canceled)
56. A pharmaceutical composition comprising the binding protein of
claim 32, and a pharmaceutically acceptable carrier.
57. A method for treating a mammal comprising the step of
administering to the mammal an effective amount of the composition
of claim 56.
58-61. (canceled)
62. A method for treating a subject for a disease or a disorder in
which sclerostin activity is detrimental by administering to the
subject the binding protein of claim 32 such that treatment is
achieved.
63. The method of claim 62, wherein the disorder is selected from
the group consisting of a respiratory disorder; asthma; allergic
and nonallergic asthma; asthma due to infection; asthma due to
infection with respiratory syncytial virus (RSV); chronic
obstructive pulmonary disease (COPD); a condition involving airway
inflammation; eosinophilia; fibrosis and/or excess mucus
production; cystic fibrosis; pulmonary fibrosis; an atopic
disorder; atopic dermatitis; urticaria; eczema; allergic rhinitis;
allergic enterogastritis; an inflammatory and/or autoimmune
condition of the skin; an inflammatory and/or autoimmune condition
of gastrointestinal organs; inflammatory bowel disease (IBD);
ulcerative colitis; Crohn's disease; an inflammatory and/or
autoimmune condition of the liver; liver cirrhosis; liver fibrosis;
liver fibrosis caused by hepatitis B and/or C virus; scleroderma; a
tumors; a cancer; hepatocellular carcinoma; glioblastoma; lymphoma;
Hodgkin's lymphoma; a viral infection; HTLV-1 infection (e.g., from
HTLV-1); a suppression of expression of protective type 1 immune
response, and a suppression of expression of protective type 1
immune responses during vaccination.
64. (canceled)
65. A method for generating a DVD-binding protein comprising the
steps of a) obtaining a first parent antibody or antigen binding
portion thereof, capable of binding TNF-.alpha.; b) obtaining a
second parent antibody or antigen binding portion thereof, capable
of binding human sclerostin; c) constructing polypeptide chains
comprising the VD1-(X1)n-VD2-C-(X2)n of any one of claim 32; e)
expressing the polypeptide chains; such that a DVD-binding protein
is generated.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S. Utility
application Ser. No. 13/659,647, filed Oct. 24, 2012, which claims
priority to U.S. Provisional Application Ser. No. 61/550,724, filed
Oct. 24, 2011, each of which is incorporated herein by reference in
its entirety.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted in ASCII format via EFS-Web and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Dec. 19, 2012, is named 532352_Seq_List.txt and is 2,706,515
bytes in size.
FIELD OF THE INVENTION
[0003] Sclerostin binding proteins, and specifically their uses in
the prevention and/or treatment of acute and chronic immunological
diseases such as rheumatoid arthritis, osteoarthritis, psoriasis,
multiple sclerosis, and other autoimmune diseases are provided.
BACKGROUND OF THE INVENTION
[0004] The SOST gene encodes a 24 KD protein called sclerostin that
has been classified as a member of the DAN family of cysteine knot
containing glycoproteins based on sequence similarity
(Avasian-Kretchmer (2004) Mol. Endocrinol. 8(1):1-12). Sclerostin
is a negative regulator of bone formation that inhibits osteoblast
proliferation as well as differentiation and suppresses
mineralization of osteoblastic cells in vitro (Poole et al. (2005)
FASEB J. 19:1836-38; Winkler et al. (2005) J. Biol. Chem. 280(4):
2498-2502).
[0005] Sclerostin is an inhibitor of the canonical Wnt signaling
pathway. It binds to LRP4, LRP5 and/or LRP6 receptors leading to
stabilization of .beta.-catenin leading to regulation of gene
transcription through transcription regulators including lymphoid
enhancing factor-1 (LEF) and T cell factors (TCF). Sclerostin
inhibition allows signaling through the Wnt pathway resulting in
bone formation (van Bezooijen et al. (2007) J. Bone Min. Res.
22(1):19-28).
[0006] An increase in canonical Wnt signaling results in increased
bone mass (Li et al. (2005) J. Biol. Chem. 280(20):19883-7; Semenov
et al. (2005) J. Biol. Chem. 280(29):26770-775). Loss of function
mutants in LRP5 lead to the low bone mass phenotype seen in
osteoporosis-pseudoglioma syndrome in humans and LRP5 KO mice
demonstrate phenotypes similar to those seen in these patients
(Balemans et al. (2008) Calcif. Tissue Int. 82:445-53). Two human
mutations of the SOST gene have been identified that lead to
Sclerosteosis and Van Buchem's disease, both of which result in a
high bone mass phenotype (Brunkow et al. (2001) Am. J. Hum. Genet.
68:577-89; Balemans et al. (2001) Hum Mol Genet 10:537-43).
Additionally, sclerostin KO mice demonstrate a high bone mass
phenotype while sclerostin over-producing Tg mice have a low bone
mass phenotype (Li et al. (2008) J. Bone and Min. Res.
23(6):860-9).
[0007] UCB Celltech (formerly Celltech), in collaboration with
AMGEN, is developing a sclerostin neutralizing mAb for the
treatment of osteoporosis and fracture healing. Phase I clinical
trials have been completed. A Phase II trial has been completed in
osteoporosis and Phase III trials have been initiated. Multiple
Phase II trials are ongoing for the treatment of fracture healing.
The pathogenic role of TNF in arthritis is well established as
TNF-.alpha. antagonists reduce inflammation and limit progression
of cartilage damage and bone erosion in human disease (van den Berg
(2001) Arthritis Res. 3:18-26). Although TNF antagonists have
revolutionized RA therapy, a significant portion of patients do not
respond adequately to these drugs. Preclinical studies with
TNF-.alpha. and SOST point to both independent and overlapping
roles in arthritis pathophysiology. Whereas sclerostin or
TNF-.alpha. inhibition alone exert only modest effects on
proinflammatory gene expression, the combination of SOST inhibition
with TNF-.alpha. inhibition leads to strong synergistic responses.
In particular, the combination of inhibiting sclerostin and
TNF-.alpha. has the potential to both block inflammation and
promote bone healing providing greater clinical benefit to
patients.
[0008] Although a variety of antibodies to sclerostin have been
described since the discovery of this critical proinflammatory
cytokine, there remains a need for improved antibodies that can
effectively mediate or neutralize the activity of sclerostin during
an inflammatory response or autoimmune disorder, while protecting
or restoring bone mineral density, bone volume and bone
strength.
BRIEF SUMMARY OF THE INVENTION
[0009] Proteins that bind human sclerostin are provided. Binding
proteins are provided that include but are not limited to
antibodies, antigen binding portions thereof, and multivalent,
multispecific binding proteins such as DVD-binding proteins that
can bind human Sclerostin and another target, such as TNF-.alpha..
Methods of making and using the sclerostin binding proteins
described herein as well as various compositions that may be used
in methods of detecting sclerostin in a sample or in methods of
treating or preventing a disorder in an individual that is
associated with or suspected to be associated with sclerostin
activity are provided.
[0010] In one aspect, there is provided a binding protein
comprising an antigen binding domain capable of binding human
sclerostin, said antigen binding domain comprising at least one CDR
comprising:
TABLE-US-00001 CDR-H1. (SEQ ID NO: 15)
X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-X.sub.6-X.sub.7 CDR-H2.
(SEQ ID NO: 16)
X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-X.sub.6-X.sub.7-X.sub.8-X.sub.9-X-
.sub.10-X.sub.11-X.sub.12-X.sub.13-X.sub.14-
X.sub.15-X.sub.16-X.sub.17-X.sub.18 CDR-H3. (SEQ ID NO: 17)
X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-X.sub.6-X.sub.7-X.sub.8-X.sub.9-X-
.sub.10-X.sub.11-X.sub.12-X.sub.13-X.sub.14-
X.sub.15-X.sub.16-X.sub.17-X.sub.18-X.sub.19-X.sub.20-X.sub.21
CDR-L1. (SEQ ID NO: 18)
X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-X.sub.6-X.sub.7-X.sub.8-X.sub.9-X-
.sub.10-X.sub.11-X.sub.12-X.sub.13-X.sub.14- X.sub.15-X.sub.16
CDR-L2. (SEQ ID NO: 19)
X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-X.sub.6-X.sub.7 or CDR-L3.
(SEQ ID NO: 20
X.sub.1-X.sub.2-X.sub.3-X.sub.4-X.sub.5-X.sub.6-X.sub.7-X.sub.8-X.sub.9-X-
.sub.10-X.sub.11-X.sub.12.
[0011] In an embodiment, a binding protein is provided that
comprises at least one CDR comprising residues 31-35 of SEQ ID NO:
3; residues 50-66 of SEQ ID NO: 3; residues 99-108 of SEQ ID NO:3;
residues 23-34 of SEQ ID NO: 4; residues 51-57 of SEQ ID NO: 4;
residues 90-101 of SEQ ID NO:4; residues 31-35 of SEQ ID NO: 5;
residues 50-66 of SEQ ID NO: 5; residues 99-115 of SEQ ID NO:5;
residues 23-33 of SEQ ID NO: 6; residues 49-55 of SEQ ID NO: 6;
residues 88-96 of SEQ ID NO:6; residues 31-35 of SEQ ID NO: 7;
residues 50-66 of SEQ ID NO: 7; residues 99-107 of SEQ ID NO:7;
residues 23-33 of SEQ ID NO: 8; residues 49-55 of SEQ ID NO: 8;
residues 88-95 of SEQ ID NO:8; residues 31-35 of SEQ ID NO: 9;
residues 50-66 of SEQ ID NO: 9; residues 99-107 of SEQ ID NO:9;
residues 24-39 of SEQ ID NO: 10; residues 55-61 of SEQ ID NO: 10;
residues 94-112 of SEQ ID NO:10; residues 31-35 of SEQ ID NO: 11;
residues 50-66 of SEQ ID NO: 11; residues 99-111 of SEQ ID NO: 11;
residues 24-39 of SEQ ID NO: 12; residues 55-61 of SEQ ID NO: 12;
residues 94-113 of SEQ ID NO:12; residues 31-37 of SEQ ID NO: 13;
residues 52-69 of SEQ ID NO: 13; residues 102-122 of SEQ ID NO:13;
residues 24-34 of SEQ ID NO: 14; residues 50-56 of SEQ ID NO: 14;
residues 89-97 of SEQ ID NO:14; Residues 31-35 of SEQ ID NO:1998;
Residues 50-66 of SEQ ID NO:1998; Residues 99-110 of SEQ ID
NO:1998; Residues 31-35 of SEQ ID NO.:1999; Residues 50-66 of SEQ
ID NO.:1999; Residues 99-110 of SEQ ID NO.:1999; Residues 31-35 of
SEQ ID NO.:2000; Residues 50-66 of SEQ ID NO.:2000; Residues 99-110
of SEQ ID NO.:2000; Residues 31-35 of SEQ ID NO.:2001; Residues
50-66 of SEQ ID NO.:2001; Residues 99-110 of SEQ ID NO.:2001;
Residues 31-35 of SEQ ID NO.:2002; Residues 50-66 of SEQ ID
NO.:2002; Residues 99-110 of SEQ ID NO.:2002; Residues 31-35 of SEQ
ID NO.:2003; Residues 50-66 of SEQ ID NO.:2003; Residues 99-110 of
SEQ ID NO.:2003; Residues 31-35 of SEQ ID NO.:2004; Residues 50-66
of SEQ ID NO.:2004; Residues 99-110 of SEQ ID NO.:2004; Residues
31-35 of SEQ ID NO.:2005; Residues 50-66 of SEQ ID NO.:2005;
Residues 99-110 of SEQ ID NO.:2005; Residues 31-35 of SEQ ID
NO.:2006; Residues 50-66 of SEQ ID NO.:2006; Residues 99-110 of SEQ
ID NO.:2006; Residues 31-35 of SEQ ID NO.:2007; Residues 50-66 of
SEQ ID NO.:2007; Residues 99-110 of SEQ ID NO.:2007; Residues 23-36
of SEQ ID NO.:2008; Residues 52-58 of SEQ ID NO.:2008; Residues
101-109 of SEQ ID NO.:2008; Residues 23-36 of SEQ ID NO.:2009;
Residues 52-58 of SEQ ID NO.:2009; Residues 101-109 of SEQ ID
NO.:2009; Residues 23-36 of SEQ ID NO.:2008; Residues 52-58 of SEQ
ID NO.:2010; Residues 101-109 of SEQ ID NO.:2010; Residues 23-36 of
SEQ ID NO.:2011; Residues 52-58 of SEQ ID NO.:2011; Residues
101-109 of SEQ ID NO.:2011; Residues 23-36 of SEQ ID NO.:2012;
Residues 52-58 of SEQ ID NO.:2012; Residues 101-109 of SEQ ID
NO.:2012; Residues 23-36 of SEQ ID NO.:2013; Residues 52-58 of SEQ
ID NO.:2013; Residues 101-109 of SEQ ID NO.:2013; Residues 23-36 of
SEQ ID NO.:2014; Residues 52-58 of SEQ ID NO.:2014; Residues
101-109 of SEQ ID NO.:2014; Residues 23-36 of SEQ ID NO.:2015;
Residues 52-58 of SEQ ID NO.:2015; Residues 101-109 of SEQ ID
NO.:2015; Residues 23-36 of SEQ ID NO.:2016; Residues 52-58 of SEQ
ID NO.:2016; Residues 101-109 of SEQ ID NO.:2016; Residues 23-36 of
SEQ ID NO.:2017; Residues 52-58 of SEQ ID NO.:2017; Residues
101-109 of SEQ ID NO.:2017; Residues 31-35 of SEQ ID NO:2020;
Residues 50-66 of SEQ ID NO:2020; Residues 99-108 of SEQ ID
NO:2020; Residues 31-35 of SEQ ID NO:2021; Residues 50-66 of SEQ ID
NO:2021; Residues 99-108 of SEQ ID NO:2021; Residues 31-35 of SEQ
ID NO:2022; Residues 50-66 of SEQ ID NO:2022; Residues 99-108 of
SEQ ID NO:2022; Residues 31-35 of SEQ ID NO:2023; Residues 50-66 of
SEQ ID NO:2023; Residues 99-108 of SEQ ID NO:2023; Residues 31-35
of SEQ ID NO:2024; Residues 50-66 of SEQ ID NO:2024; Residues
99-108 of SEQ ID NO:2024; Residues 31-35 of SEQ ID NO:2025;
Residues 50-66 of SEQ ID NO:2025; Residues 99-108 of SEQ ID
NO:2025; Residues 31-35 of SEQ ID NO:2026; Residues 50-66 of SEQ ID
NO:2026; Residues 99-108 of SEQ ID NO:2026; Residues 31-35 of SEQ
ID NO:2027; Residues 50-66 of SEQ ID NO:2027; Residues 99-108 of
SEQ ID NO:2027; Residues 31-35 of SEQ ID NO:2028; Residues 50-66 of
SEQ ID NO:2028; Residues 99-108 of SEQ ID NO:2028; Residues 31-35
of SEQ ID NO:2029; Residues 50-66 of SEQ ID NO:2029; Residues
99-108 of SEQ ID NO:2029; Residues 31-35 of SEQ ID NO:2030;
Residues 50-66 of SEQ ID NO:2030; Residues 99-108 of SEQ ID
NO:2030; Residues 31-35 of SEQ ID NO:2031; Residues 50-66 of SEQ ID
NO:2031; Residues 99-108 of SEQ ID NO:2031; Residues 31-35 of SEQ
ID NO:2032; Residues 50-66 of SEQ ID NO:2032; Residues 99-108 of
SEQ ID NO:2032; Residues 31-35 of SEQ ID NO:2033; Residues 50-66 of
SEQ ID NO:2033; Residues 99-108 of SEQ ID NO:2033; Residues 31-35
of SEQ ID NO:2034; Residues 50-66 of SEQ ID NO:2034; Residues
99-110 of SEQ ID NO:2034; Residues 31-35 of SEQ ID NO.:2035;
Residues 51-57 of SEQ ID NO.:2035; Residues 90-101 of SEQ ID
NO.:2035; Residues 31-35 of SEQ ID NO.:2036; Residues 51-57 of SEQ
ID NO.:2036; Residues 90-101 of SEQ ID NO.:2036; Residues 31-35 of
SEQ ID NO.:2037; Residues 51-57 of SEQ ID NO.:2035; Residues 90-101
of SEQ ID NO.:2037; Residues 31-35 of SEQ ID NO.:2038; Residues
51-57 of SEQ ID NO.:2038; Residues 90-101 of SEQ ID NO.:2038;
Residues 31-35 of SEQ ID NO.:2039; Residues 51-57 of SEQ ID
NO.:2039; Residues 90-101 of SEQ ID NO.:2039; Residues 31-35 of SEQ
ID NO.:2040; Residues 51-57 of SEQ ID NO.:2040; Residues 90-101 of
SEQ ID NO.:2040; Residues 31-35 of SEQ ID NO.:2041; Residues 51-57
of SEQ ID NO.:2041; Residues 90-101 of SEQ ID NO.:2041; Residues
31-35 of SEQ ID NO.:2042; Residues 51-57 of SEQ ID NO.:2042;
Residues 90-101 of SEQ ID NO.:2042; Residues 31-35 of SEQ ID
NO.:2043; Residues 51-57 of SEQ ID NO.:2043; Residues 90-101 of SEQ
ID NO.:2043; Residues 31-35 of SEQ ID NO.:2044; Residues 51-57 of
SEQ ID NO.:2044; Residues 90-101 of SEQ ID NO.:2044; Residues 31-35
of SEQ ID NO.:2045; Residues 51-57 of SEQ ID NO.:2045; Residues
90-101 of SEQ ID NO.:2045; Residues 31-35 of SEQ ID NO.:2046;
Residues 51-57 of SEQ ID NO.:2046; Residues 90-101 of SEQ ID
NO.:2046; Residues 31-35 of SEQ ID NO.:2047; Residues 51-57 of SEQ
ID NO.:2047; Residues 90-101 of SEQ ID NO.:2047; Residues 31-35 of
SEQ ID NO.:2048; Residues 51-57 of SEQ ID NO.:2048; Residues 90-101
of SEQ ID NO.:2048; Residues 31-35 of SEQ ID NO.:2049; Residues
51-57 of SEQ ID NO.:2049; and Residues 90-101 of SEQ ID
NO.:2049.
[0012] In another embodiment, a sclerostinbinding protein
comprising at least 3 CDRs described above is provided.
[0013] In another embodiment, a sclerostin binding protein is
provided that comprises at least 3 CDRs of Table 1:
TABLE-US-00002 TABLE 1 VH MSL10 CDR Set VH MSL10 CDR-H1 Residues
31-35 of SEQ ID NO: 3 VH MSL10 CDR-H2 Residues 50-66 of SEQ ID NO:
3 VH MSL10 CDR-H3 Residues 99-108 of SEQ ID NO: 3 VL MSL10 CDR Set
VL MSL10 CDR-L1 Residues 23-34 of SEQ ID NO: 4 VL MSL10 CDR-L2
Residues 51-57 of SEQ ID NO: 4 VL MSL10 CDR-L3 Residues 90-101 of
SEQ ID NO: 4 VH MSL10 AM1 CDR Set VH MSL10 AM1 CDR-H1 Residues
31-35 of SEQ ID NO: 2020 VH MSL10 AM1 CDR-H2 Residues 50-66 of SEQ
ID NO: 2020 VH MSL10 AM1 CDR-H3 Residues 99-108 of SEQ ID NO: 2020
VL MSL10 AM1 CDR Set VL MSL10 AM1 CDR-L1 Residues 23-34 of SEQ ID
NO: 2035 VL MSL10 AM1 CDR-L2 Residues 51-57 of SEQ ID NO: 2035 VL
MSL10 AM1 CDR-L3 Residues 90-101 of SEQ ID NO: 2035 VH MSL10 AM2
CDR Set VH MSL10 AM2 CDR-H1 Residues 31-35 of SEQ ID NO: 2021 VH
MSL10 AM2 CDR-H2 Residues 50-66 of SEQ ID NO: 2021 VH MSL10 AM2
CDR-H3 Residues 99-108 of SEQ ID NO: 2021 VL MSL10 AM2 CDR Set VL
MSL10 AM2 CDR-L1 Residues 23-34 of SEQ ID NO: 2036 VL MSL10 AM2
CDR-L2 Residues 51-57 of SEQ ID NO: 2036 VL MSL10 AM2 CDR-L3
Residues 90-101 of SEQ ID NO: 2036 VH MSL10 AM3 CDR Set VH MSL10
AM3 CDR-H1 Residues 31-35 of SEQ ID NO: 2022 VH MSL10 AM3 CDR-H2
Residues 50-66 of SEQ ID NO: 2022 VH MSL10 AM3 CDR-H3 Residues
99-108 of SEQ ID NO: 2022 VL MSL10 AM3 CDR Set VL MSL10 AM3 CDR-L1
Residues 23-34 of SEQ ID NO: 2037 VL MSL10 AM3 CDR-L2 Residues
51-57 of SEQ ID NO: 2037 VL MSL10 AM3 CDR-L3 Residues 90-101 of SEQ
ID NO: 2037 VH MSL10 AM4 CDR Set VH MSL10 AM4 CDR-H1 Residues 31-35
of SEQ ID NO: 2023 VH MSL10 AM4 CDR-H2 Residues 50-66 of SEQ ID NO:
2023 VH MSL10 AM4 CDR-H3 Residues 99-108 of SEQ ID NO: 2023 VL
MSL10 AM4 CDR Set VL MSL10 AM4 CDR-L1 Residues 23-34 of SEQ ID NO:
2038 VL MSL10 AM4 CDR-L2 Residues 51-57 of SEQ ID NO: 2038 VL MSL10
AM4 CDR-L3 Residues 90-101 of SEQ ID NO: 2038 VH MSL10 AM5 CDR Set
VH MSL10 AM5 CDR-H1 Residues 31-35 of SEQ ID NO: 2024 VH MSL10 AM5
CDR-H2 Residues 50-66 of SEQ ID NO: 2024 VH MSL10 AM5 CDR-H3
Residues 99-108 of SEQ ID NO: 2024 VL MSL10 AM5 CDR Set VL MSL10
AM5 CDR-L1 Residues 23-34 of SEQ ID NO: 2039 VL MSL10 AM5 CDR-L2
Residues 51-57 of SEQ ID NO: 2039 VL MSL10 AM5 CDR-L3 Residues
90-101 of SEQ ID NO: 2039 VH MSL10 AM6 CDR Set VH MSL10 AM6 CDR-H1
Residues 31-35 of SEQ ID NO: 2025 VH MSL10 AM6 CDR-H2 Residues
50-66 of SEQ ID NO: 2025 VH MSL10 AM6 CDR-H3 Residues 99-108 of SEQ
ID NO: 2025 VL MSL10 AM6 CDR Set VL MSL10 AM6 CDR-L1 Residues 23-34
of SEQ ID NO: 2040 VL MSL10 AM6 CDR-L2 Residues 51-57 of SEQ ID NO:
2040 VL MSL10 AM6 CDR-L3 Residues 90-101 of SEQ ID NO: 2040 VH
MSL10 AM7 CDR Set VH MSL10 AM7 CDR-H1 Residues 31-35 of SEQ ID NO:
2026 VH MSL10 AM7 CDR-H2 Residues 50-66 of SEQ ID NO: 2026 VH MSL10
AM7 CDR-H3 Residues 99-108 of SEQ ID NO: 2026 VL MSL10 AM7 CDR Set
VL MSL10 AM7 CDR-L1 Residues 23-34 of SEQ ID NO: 2041 VL MSL10 AM7
CDR-L2 Residues 51-57 of SEQ ID NO: 2041 VL MSL10 AM7 CDR-L3
Residues 90-101 of SEQ ID NO: 2041 VH MSL10 AM8 CDR Set VH MSL10
AM8 CDR-H1 Residues 31-35 of SEQ ID NO: 2027 VH MSL10 AM8 CDR-H2
Residues 50-66 of SEQ ID NO: 2027 VH MSL10 AM8 CDR-H3 Residues
99-108 of SEQ ID NO: 2027 VL MSL10 AM8 CDR Set VL MSL10 AM8 CDR-L1
Residues 23-34 of SEQ ID NO: 2042 VL MSL10 AM8 CDR-L2 Residues
51-57 of SEQ ID NO: 2042 VL MSL10 AM8 CDR-L3 Residues 90-101 of SEQ
ID NO: 2042 VH MSL10 AM9 CDR Set VH MSL10 AM9 CDR-H1 Residues 31-35
of SEQ ID NO: 2028 VH MSL10 AM9 CDR-H2 Residues 50-66 of SEQ ID NO:
2028 VH MSL10 AM9 CDR-H3 Residues 99-108 of SEQ ID NO: 2028 VL
MSL10 AM9 CDR Set VL MSL10 AM9 CDR-L1 Residues 23-34 of SEQ ID NO:
2043 VL MSL10 AM9 CDR-L2 Residues 51-57 of SEQ ID NO: 2043 VL MSL10
AM9 CDR-L3 Residues 90-101 of SEQ ID NO: 2043 VH MSL10 AM10 CDR Set
VH MSL10 AM10 CDR-H1 Residues 31-35 of SEQ ID NO: 2029 VH MSL10
AM10 CDR-H2 Residues 50-66 of SEQ ID NO: 2029 VH MSL10 AM10 CDR-H3
Residues 99-108 of SEQ ID NO: 2029 VL MSL10 AM10 CDR Set VL MSL10
AM10 CDR-L1 Residues 23-34 of SEQ ID NO: 2044 VL MSL10 AM10 CDR-L2
Residues 51-57 of SEQ ID NO: 2044 VL MSL10 AM10 CDR-L3 Residues
90-101 of SEQ ID NO: 2044 VH MSL10 AM10 CDR Set VH MSL10 AM11
CDR-H1 Residues 31-35 of SEQ ID NO: 2030 VH MSL10 AM11 CDR-H2
Residues 50-66 of SEQ ID NO: 2030 VH MSL10 AM11 CDR-H3 Residues
99-108 of SEQ ID NO: 2030 VL MSL10 AM10 CDR Set VL MSL10 AM11
CDR-L1 Residues 23-34 of SEQ ID NO: 2045 VL MSL10 AM11 CDR-L2
Residues 51-57 of SEQ ID NO: 2045 VL MSL10 AM11 CDR-L3 Residues
90-101 of SEQ ID NO: 2045 VH MSL10 AM1.2 CDR Set VH MSL10 AM1.2
CDR-H1 Residues 31-35 of SEQ ID NO: 2031 VH MSL10 AM1.2 CDR-H2
Residues 50-66 of SEQ ID NO: 2031 VH MSL10 AM1.2 CDR-H3 Residues
99-108 of SEQ ID NO: 2031 VL MSL10 AM1.2 CDR Set VL MSL10 AM1.2
CDR-L1 Residues 23-34 of SEQ ID NO: 2046 VL MSL10 AM1.2 CDR-L2
Residues 51-57 of SEQ ID NO: 2046 VL MSL10 AM1.2 CDR-L3 Residues
90-101 of SEQ ID NO: 2046 VH MSL10 AM2.2 CDR Set VH MSL10 AM2.2
CDR-H1 Residues 31-35 of SEQ ID NO: 2032 VH MSL10 AM2.2 CDR-H2
Residues 50-66 of SEQ ID NO: 2032 VH MSL10 AM2.2 CDR-H3 Residues
99-108 of SEQ ID NO: 2032 VL MSL10 AM2.2 CDR Set VL MSL10 AM2.2
CDR-L1 Residues 23-34 of SEQ ID NO: 2047 VL MSL10 AM2.2 CDR-L2
Residues 51-57 of SEQ ID NO: 2047 VL MSL10 AM2.2 CDR-L3 Residues
90-101 of SEQ ID NO: 2047 VH MSL10 AM3.2 CDR Set VH MSL10 AM3.2
CDR-H1 Residues 31-35 of SEQ ID NO: 2033 VH MSL10 AM3.2 CDR-H2
Residues 50-66 of SEQ ID NO: 2033 VH MSL10 AM3.2 CDR-H3 Residues
99-108 of SEQ ID NO: 2033 VL MSL10 AM3.2 CDR Set VL MSL10 AM3.2
CDR-L1 Residues 23-34 of SEQ ID NO: 2048 VL MSL10 AM3.2 CDR-L2
Residues 51-57 of SEQ ID NO: 2048 VL MSL10 AM3.2 CDR-L3 Residues
90-101 of SEQ ID NO: 2048 VH MSL10 AM4.2 CDR Set VH MSL10 AM4.2
CDR-H1 Residues 31-35 of SEQ ID NO: 2034 VH MSL10 AM4.2 CDR-H2
Residues 50-66 of SEQ ID NO: 2034 VH MSL10 AM4.2 CDR-H3 Residues
99-108 of SEQ ID NO: 2034 VL MSL10 AM4.2 CDR Set VL MSL10 AM4.2
CDR-L1 Residues 23-34 of SEQ ID NO: 2049 VL MSL10 AM4.2 CDR-L2
Residues 51-57 of SEQ ID NO: 2049 VL MSL10 AM4.2 CDR-L3 Residues
90-101 of SEQ ID NO: 2049 VH MSL17 CDR Set VH MSL17 CDR-H1 Residues
31-35 of SEQ ID NO: 5 VH MSL17 CDR-H2 Residues 50-66 of SEQ ID NO:
5 VH MSL17 CDR-H3 Residues 99-115 of SEQ ID NO: 5 VL MSL17 CDR Set
VL MSL17 CDR-L1 Residues 23-33 of SEQ ID NO: 6 VL MSL17 CDR-L2
Residues 49-55 of SEQ ID NO: 6 VL MSL17 CDR-L3 Residues 88-96 of
SEQ ID NO: 6 VH MSL9-8 CDR Set VH MSL9-8 CDR-H1 Residues 31-35 of
SEQ ID NO: 7 VH MSL9-8 CDR-H2 Residues 50-66 of SEQ ID NO: 7 VH
MSL9-8 CDR-H3 Residues 99-107 of SEQ ID NO: 7 VL MSL9-8 CDR Set VL
MSL9-8 CDR-L1 Residues 23-33 of SEQ ID NO: 8 VL MSL9-8 CDR-L2
Residues 49-55 of SEQ ID NO: 8 VL MSL9-8 CDR-L3 Residues 88-95 of
SEQ ID NO: 8 VH MSK9 CDR Set VH MSK9 CDR-H1 Residues 31-35 of SEQ
ID NO: 9 VH MSK9 CDR-H2 Residues 50-66 of SEQ ID NO: 9 VH MSK9
CDR-H3 Residues 99-107 of SEQ ID NO: 9 VL MSK9 CDR Set VL MSK9
CDR-L1 Residues 24-39 of SEQ ID NO: 10 VL MSK9 CDR-L2 Residues
55-61 of SEQ ID NO: 10 VL MSK9 CDR-L3 Residues 94-112 of SEQ ID NO:
10 VH MSK13 CDR Set VH MSK13 CDR-H1 Residues 31-35 of SEQ ID NO: 11
VH MSK13 CDR-H2 Residues 50-66 of SEQ ID NO: 11 VH MSK13 CDR-H3
Residues 99-111 of SEQ ID NO: 11 VL MSK13 CDR Set VL MSK13 CDR-L1
Residues 24-39 of SEQ ID NO: 12 VL MSK13 CDR-L2 Residues 55-61 of
SEQ ID NO: 12 VL MSK13 CDR-L3 Residues 94-113 of SEQ ID NO: 12 VH
MSK21 CDR Set VH MSK21 CDR-H1 Residues 31-37 of SEQ ID NO: 13 VH
MSK21 CDR-H2 Residues 52-69 of SEQ ID NO: 13 VH MSK21 CDR-H3
Residues 102-122 of SEQ ID NO: 13 VL MSK21 CDR Set VL MSK21 CDR-L1
Residues 24-34 of SEQ ID NO: 14 VL MSK21 CDR-L2 Residues 50-56 of
SEQ ID NO: 14 VL MSK21 CDR-L3 Residues 89-97 of SEQ ID NO: 14 VH
AE10-6 AM1 CDR Set VH AE10-6 AM1 CDR-H1 Residues 31-37 of SEQ ID
NO: 1998 VH AE10-6 AM1 CDR-H2 Residues 52-69 of SEQ ID NO: 1998 VH
AE10-6 AM1 CDR-H3 Residue 102-122 of SEQ ID NO: 1998 VL AE10-6 AM1
Set VL AE10-6 AM1 CDR-L1 Residues 24-34 of SEQ ID NO: 2008 VL
AE10-6 AM1 CDR-L2 Residues 50-56 of SEQ ID NO: 2008 VL AE10-6 AM1
CDR-L3 Residues 89-97 of SEQ ID NO: 2008 VH AE10-6 AM2 CDR Set VH
AE10-6 AM2 CDR-H1 Residues 31-37 of SEQ ID NO: 1999 VH AE10-6 AM2
CDR-H2 Residues 52-69 of SEQ ID NO: 1999 VH AE10-6 AM2 CDR-H3
Residue 102-122 of SEQ ID NO: 1999 VL AE10-6 AM2 Set VL AE10-6 AM2
CDR-L1 Residues 24-34 of SEQ ID NO: 2009 VL AE10-6 AM2 CDR-L2
Residues 50-56 of SEQ ID NO: 2009 VL AE10-6 AM2 CDR-L3 Residues
89-97 of SEQ ID NO: 2009 VH AE10-6 AM3 CDR Set VH AE10-6 AM3 CDR-H1
Residues 31-37 of SEQ ID NO: 2000 VH AE10-6 AM3 CDR-H2 Residues
52-69 of SEQ ID NO: 2000 VH AE10-6 AM3 CDR-H3 Residue 102-122 of
SEQ ID NO: 2000 VL AE10-6 AM3 Set VL AE10-6 AM3 CDR-L1 Residues
24-34 of SEQ ID NO: 2010 VL AE10-6 AM3 CDR-L2 Residues 50-56 of SEQ
ID NO: 2010 VL AE10-6 AM3 CDR-L3 Residues 89-97 of SEQ ID NO: 2010
VH AE10-6 AM4 CDR Set VH AE10-6 AM4 CDR-H1 Residues 31-37 of SEQ ID
NO: 2001 VH AE10-6 AM4 CDR-H2 Residues 52-69 of SEQ ID NO: 2001 VH
AE10-6 AM4 CDR-H3 Residue 102-122 of SEQ ID NO: 2001 VL AE10-6 AM4
Set VL AE10-6 AM4 CDR-L1 Residues 24-34 of SEQ ID NO: 2011 VL
AE10-6 AM4 CDR-L2 Residues 50-56 of SEQ ID NO: 2011 VL AE10-6 AM4
CDR-L3 Residues 89-97 of SEQ ID NO: 2011 VH AE10-6 AM5 CDR Set VH
AE10-6 AM5 CDR-H1 Residues 31-37 of SEQ ID NO: 2002 VH AE10-6 AM5
CDR-H2 Residues 52-69 of SEQ ID NO: 2002 VH AE10-6 AM5 CDR-H3
Residue 102-122 of SEQ ID NO: 2002 VL AE10-6 AM5 Set VL AE10-6 AM5
CDR-L1 Residues 24-34 of SEQ ID NO: 2012 VL AE10-6 AM5 CDR-L2
Residues 50-56 of SEQ ID NO: 2012 VL AE10-6 AM5 CDR-L3 Residues
89-97 of SEQ ID NO: 2012 VH AE10-6 AM6 CDR Set VH AE10-6 AM6 CDR-H1
Residues 31-37 of SEQ ID NO: 2003 VH AE10-6 AM6 CDR-H2 Residues
52-69 of SEQ ID NO: 2003 VH AE10-6 AM6 CDR-H3 Residue 102-122 of
SEQ ID NO: 2003 VL AE10-6 AM6 Set VL AE10-6 AM6 CDR-L1 Residues
24-34 of SEQ ID NO: 2013 VL AE10-6 AM6 CDR-L2 Residues 50-56 of SEQ
ID NO: 2013 VL AE10-6 AM6 CDR-L3 Residues 89-97 of SEQ ID NO: 2013
VH AE10-6 AM7 CDR Set VH AE10-6 AM7 CDR-H1 Residues 31-37 of SEQ ID
NO: 2004 VH AE10-6 AM7 CDR-H2 Residues 52-69 of SEQ ID NO: 2004 VH
AE10-6 AM7 CDR-H3 Residue 102-122 of SEQ ID NO: 2004 VL AE10-6 AM7
Set VL AE10-6 AM7 CDR-L1 Residues 24-34 of SEQ ID NO: 2014 VL
AE10-6 AM7 CDR-L2 Residues 50-56 of SEQ ID NO: 2014 VL AE10-6 AM7
CDR-L3 Residues 89-97 of SEQ ID NO: 2014 VH AE10-6 AM8 CDR Set VH
AE10-6 AM8 CDR-H1 Residues 31-37 of SEQ ID NO: 2005 VH AE10-6 AM8
CDR-H2 Residues 52-69 of SEQ ID NO: 2005 VH AE10-6 AM8 CDR-H3
Residue 102-122 of SEQ ID NO: 2005 VL AE10-6 AM8 Set VL AE10-6 AM8
CDR-L1 Residues 24-34 of SEQ ID NO: 2015 VL AE10-6 AM8 CDR-L2
Residues 50-56 of SEQ ID NO: 2015 VL AE10-6 AM8 CDR-L3 Residues
89-97 of SEQ ID NO: 2015 VH AE10-6 AM9 CDR Set VH AE10-6 AM9 CDR-H1
Residues 31-37 of SEQ ID NO: 2006 VH AE10-6 AM9 CDR-H2 Residues
52-69 of SEQ ID NO: 2006 VH AE10-6 AM9 CDR-H3 Residue 102-122 of
SEQ ID NO: 2006 VL AE10-6 AM9 Set VL AE10-6 AM9 CDR-L1 Residues
24-34 of SEQ ID NO: 2016 VL AE10-6 AM9 CDR-L2 Residues 50-56 of SEQ
ID NO: 2016 VL AE10-6 AM9 CDR-L3 Residues 89-97 of SEQ ID NO: 2016
VH AE10-6 AM10 CDR Set VH AE10-6 AM10 CDR-H1 Residues 31-37 of SEQ
ID NO: 2007 VH AE10-6 AM10 CDR-H2 Residues 52-69 of SEQ ID NO: 2007
VH AE10-6 AM10 CDR-H3 Residue 102-122 of SEQ ID NO: 2007 VL AE10-6
AM10 Set VL AE10-6 AM10 CDR-L1 Residues 24-34 of SEQ ID NO: 2017 VL
AE10-6 AM10 CDR-L2 Residues 50-56 of SEQ ID NO: 2017 VL AE10-6 AM10
CDR-L3 Residues 89-97 of SEQ ID NO: 2017
[0014] In another embodiment, a sclerostin binding protein may
comprise at least two variable domain CDR sets described above. In
an embodiment, the two variable domain CDR sets are VH MSL10 CDR
Set and VL MSL10 CDR Set; VH MSL17 CDR Set and VL MSL17 CDR Set; VH
MSL9-8 CDR Set and VL MSL9-8 CDR Set; VH MSK9 CDR Set and VL MSK9
CDR Set; VH MSK13 CDR Set and VL MSK13 CDR Set; VH MSK21 CDR Set or
VL MSK21 CDR Set; VH AE10-6 AM1 CDR Set and VL AE10-6 AM1 CDR Set;
VH AE10-6 AM2 CDR Set and VL AE10-6 AM2 CDR Set; VH AE10-6 AM3 CDR
Set and VL AE10-6 AM3 CDR Set; VH AE10-6 AM4 CDR Set and VL AE10-6
AM4 CDR Set; VH AE10-6 AM5 CDR Set and VL AE10-6 AM5 CDR Set; VH
AE10-6 AM6 CDR Set and VL AE10-6 AM6 CDR Set; VH AE10-6 AM7 CDR Set
and VL AE10-6 AM7 CDR Set; VH AE10-6 AM8 CDR Set and VL AE10-6
AM8CDR Set; VH AE10-6 AM9CDR Set and VL AE10-6 AM9CDR Set; VH
AE10-6 AM10CDR Set and VL AE10-6 AM10 CDR Set; VH MSL10 AM1 CDR Set
and VL MSL10 AM1 CDR Set; VH MSL10 AM2 CDR Set and VL MSL10 AM2 CDR
Set; VH MSL10 AM3 CDR Set and VL MSL10 AM3 CDR Set; VH MSL10 AM4
CDR Set and VL MSL10 AM4 CDR Set; VH MSL10 AM5 CDR Set and VL MSL10
AM5 CDR Set; VH MSL10 AM6 CDR Set and VL MSL10 AM6 CDR Set; VH
MSL10 AM7 CDR Set and VL MSL10 AM7 CDR Set; VH MSL10 AM8 CDR Set
and VL MSL10 AM8 CDR Set; VH MSL10 AM9 CDR Set and VL MSL10 AM9 CDR
Set; VH MSL10 AM10 CDR Set and VL MSL10 AM10 CDR Set; VH MSL10
AM1.2 CDR Set and VL MSL10 AM1.2 CDR Set; VH MSL10 AM2.2 CDR Set
and VL MSL10 AM2.2 CDR Set; VH MSL10 AM3.2 CDR Set and VL MSL10
AM3.2 CDR Set; and VH MSL10 AM4.2 CDR Set and VL MSL10 AM4.2 CDR
Set.
[0015] In another embodiment, a sclerostinbinding protein described
herein comprises two variable domains, wherein first variable
domain comprises a sequence selected from the group consisting of
SEQ ID NOs 3, 5, 7, 9, 11, 13, 1719-1866, 1998-2007, 2018, and
2020-2034 and wherein the second variable domain comprises a
sequence selected from the group consisting of SEQ ID NOs 4, 6, 8,
10, 12, 1867-1997, 2007-2017, 2019, and 2035-2049.
[0016] In another embodiment, a sclerostinbinding protein
comprising an antigen binding domain that comprises a V.sub.H is
provided. In an embodiment, the V.sub.H comprises any one of SEQ ID
NOs 3, 5, 7, 9, 11, 13, 1719-1866, 1998-2007, 2018, or 2020-2034.
In another embodiment, the sclerostinbinding protein comprising an
antigen binding domain that comprises a V.sub.L is provided. In an
embodiment, the V.sub.L comprises any one of SEQ ID NOs 4, 6, 8,
10, 12, 14, 1867-1997, 2008-2017, 2019, or 2035-2049.
[0017] In another embodiment, the sclerostin binding protein
comprising an antigen binding domain that comprises a V.sub.H and a
V.sub.L is provided. In an embodiment, the V.sub.H comprises SEQ ID
NO: 3, 5, 7, 9, 11, or 13 and the V.sub.L comprises SEQ ID NO: 4,
6, 8, 10, 12, 14, 1867-1997, 2008-2017, 2019, or 2035-2049.
[0018] An sclerostinbinding protein may comprise an alternative
human acceptor framework comprising at least one framework region
amino acid substitution, wherein the amino acid sequence of the
framework is at least 65% identical to the sequence of said human
acceptor framework and comprises at least 70 amino acid residues
identical to said human acceptor framework.
[0019] In another embodiment, an sclerostin binding protein
comprises an alternative human acceptor framework, wherein said
acceptor framework comprises at least one framework region amino
acid substitution at a key residue, said key residue
comprising:
[0020] a residue adjacent to a CDR;
[0021] a glycosylation site residue;
[0022] a rare residue;
[0023] a residue capable of interacting with human SOST;
[0024] a residue capable of interacting with a CDR;
[0025] a canonical residue;
[0026] a contact residue between heavy chain variable region and
light chain variable region;
[0027] a residue within a Vernier zone; or
[0028] a residue in a region that overlaps between a
Chothia-defined variable heavy chain CDR1
[0029] and a Kabat-defined first heavy chain framework.
[0030] In another embodiment, a sclerostin binding protein
described herein, further comprises a heavy chain immunoglobulin
constant domain of: a human IgM constant domain; a human IgG1
constant domain; a human IgG2 constant domain; a human IgG3
constant domain; a human IgG4 constant domain; a human IgE constant
domain; or a human IgA constant domain. In an embodiment, the heavy
chain immunoglobulin constant region is a human IgG1 constant
domain. In an embodiment, the human IgG1 constant domain comprises
SEQ ID NO: 2060, SEQ ID NO: 2061, SEQ ID NO: 2062 or SEQ ID NO:
2063.
[0031] In another embodiment, a Sclerostin binding protein
described herein comprises a light chain immunoglobulin constant
domain is a human Ig kappa constant domain or a human Ig lambda
constant domain. An exemplary human Ig kappa constant domain
comprises amino acid sequence SEQ ID NO: 2064. An exemplary human
Ig lambda constant domain comprises amino acid sequence SEQ ID NO:
2065.
[0032] In another embodiment, a sclerostinbinding protein described
herein is an immunoglobulin molecule; an scFv; a monoclonal
antibody; a human antibody; a chimeric antibody; a humanized
antibody; a single domain antibody; an Fab fragment; an Fab'
fragment; an F(ab')2; an Fv; or a disulfide linked Fv. In a
particular embodiment, the sclerostin binding protein is a human
antibody.
[0033] Another aspect provides a binding protein capable of binding
human sclerostin, wherein the binding protein comprises: [0034] an
Ig constant heavy region having an amino acid sequence of SEQ ID
NO: 2060, SEQ ID NO: 2061, SEQ ID NO: 2062 or SEQ ID NO: 2063;
[0035] an Ig constant light region having an amino acid sequence of
SEQ ID NO: 2064 or SEQ ID NO: 2065; [0036] an Ig variable heavy
region having an amino acid sequence of SEQ ID NO: 3, 5, 7, 9, 11,
13, 1719-1866, 1998-2007, 2018, or 2020-2034; and [0037] an Ig
variable light region having an amino acid sequence of SEQ ID NO:
4, 6, 8, 10, 12, 14, 1867-1997, 2008-2017, 2019, or 2035-2049.
[0038] In an embodiment, a binding protein capable of binding human
sclerostin is provided and comprises: [0039] an Ig constant heavy
region having an amino acid sequence of SEQ ID NO:3; [0040] an Ig
constant light region having an amino acid sequence of SEQ ID NO:5;
[0041] an Ig variable heavy region having an amino acid sequence of
a VH in Table 18; [0042] an amino acid sequence of a VH of SEQ ID
NO: 3, 5, 7, 9, 11, 13, 1719-1866, 1998-2007, 2018, or 2020-2034;
and [0043] an Ig variable light region having an amino acid
sequence of a VL of SEQ ID NO: 4, 6, 8, 10, 12, 14, 1867-1997,
2008-2017, 2019, or 2035-2049.
[0044] Another aspect provides a multivalent, multispecific
DVD-binding protein comprising a polypeptide chain, wherein the
polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein [0045]
VD1 is a first heavy chain variable domain; [0046] VD2 is a second
heavy chain variable domain; [0047] C is a heavy chain constant
domain; [0048] X1 is a linker with the proviso that it is not CH1;
[0049] X2 is an Fc region; [0050] (X1)n is (X1)0 or (X1)1; (X2)n is
(X2)0 or (X2)1; and [0051] wherein [0052] (a) VD1 or VD2 comprise
three CDRs from SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104,
114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234,
244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364,
374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494,
504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624,
634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754,
764, 774, 784, 794, 804, 814, 824, 834, 844, 854, 864, 874, 884,
894, 904, 914, 924, 934, 944, 954, 964, 974, 984, 994, 1004, 1014,
1024, 1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134,
1144, 1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244,
1254, 1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354,
1364, 1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464,
1474, 1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574,
1584, 1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674, or
1684, and the binding protein is capable of binding sclerostin and
another target; [0053] (b) VD1 and VD2 independently comprise three
CDRs from SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124,
134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254,
264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384,
394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514,
524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644,
654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774,
784, 794, 804, 814, 824, 834, 844, 854, 864, 874, 884, 894, 904,
914, 924, 934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024,
1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144,
1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254,
1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364,
1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474,
1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584,
1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674, or 1684, and
the binding protein is capable of binding sclerostin and
sclerostin; [0054] (c) VD1 comprises three CDRs from SEQ ID NO: 24,
34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174,
184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304,
314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434,
444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564,
574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694,
704, 714, 724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824,
834, 844, 854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954,
964, 974, 984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074,
1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194,
1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304,
1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414,
1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524,
1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634,
1644, 1654, 1664, 1674, or 1684, and VD2 comprises three CDRs from
SEQ ID NO: 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142,
152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272,
282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402,
412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532,
542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662,
672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792,
802, 812, 822, 832, 842, 852, 862, 872, 882, 892, 902, 912, 922,
932, 942, 952, 962, 972, 982, 992, 1002, 1012, 1022, 1032, 1042,
1052, 1062, 1072, 1082, 1092, 1102, 1112, 1122, 1132, 1142, 1152,
1162, 1172, 1182, 1192, 1202, 1212, 1222, 1232, 1242, 1252, 1262,
1272, 1282, 1292, 1302, 1312, 1322, 1332, 1242, 1252, 1262, 1272,
1282, 1292, 1302, 1312, 1322, 1332, 1342, 1352, 1362, 1372, 1382,
1392, 1402, 1412, 1422, 1432, 1442, 1452, 1462, 1472, 1482, 1492,
1502, 1512, 1522, 1532, 1542, 1552, 1562, 1572, 1582, 1592, 1602,
1612, 1622, 1632, 1642, 1652, 1662, 1672, or 1682, and the binding
protein is capable of binding sclerostin and TNF-.alpha.; or [0055]
(d) VD2 comprises three CDRs from SEQ ID NO: 24, 34, 44, 54, 64,
74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204,
214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334,
344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464,
474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594,
604, 614, 624, or 634, and VD1 comprises three CDRs from SEQ ID NO:
22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162,
172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292,
302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422,
432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552,
562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682,
692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792, 802, 812,
822, 832, 842, 852, 862, 872, 882, 892, 902, 912, 922, 932, 942,
952, 962, 972, 982, 992, 1002, 1012, 1022, 1032, 1042, 1052, 1062,
1072, 1082, 1092, 1102, 1112, 1122, 1132, 1142, 1152, 1162, 1172,
1182, 1192, 1202, 1212, 1222, 1232, 1242, 1252, 1262, 1272, 1282,
1292, 1302, 1312, 1322, 1332, 1242, 1252, 1262, 1272, 1282, 1292,
1302, 1312, 1322, 1332, 1342, 1352, 1362, 1372, 1382, 1392, 1402,
1412, 1422, 1432, 1442, 1452, 1462, 1472, 1482, 1492, 1502, 1512,
1522, 1532, 1542, 1552, 1562, 1572, 1582, 1592, 1602, 1612, 1622,
1632, 1642, 1652, 1662, 1672, or 1682, and the binding protein is
capable of binding TNF-.alpha. and sclerostin.
[0056] In an embodiment of the DVD-binding protein described above,
VD1-(X1)n-VD2 comprises SEQ ID NO: 21, 31, 41, 51, 61, 71, 81, 91,
101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221,
231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351,
361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481,
491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611,
621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741,
751, 761, 771, 781, 791, 801, 811, 821, 831, 841, 851, 861, 871,
881, 891, 901, 911, 921, 931, 941, 951, 961, 971, 981, 991, 1001,
1011, 1021, 1031, 1041, 1051, 1061, 1071, 1081, 1091, 1101, 1111,
1121, 1131, 1141, 1151, 1161, 1171, 1181, 1191, 1201, 1211, 1221,
1231, 1241, 1251, 1261, 1271, 1281, 1291, 1301, 1311, 1321, 1331,
1341, 1351, 1361, 1371, 1381, 1391, 1401, 1411, 1421, 1431, 1441,
1451, 1461, 1471, 1481, 1491, 1501, 1511, 1521, 1531, 1541, 1551,
1561, 1571, 1581, 1591, 1601, 1611, 1621, 1631, 1641, 1651, 1661,
1671, or 1681.
[0057] Another aspect provides a multivalent, multispecific
DVD-binding protein comprising a polypeptide chain, wherein the
polypeptide chain comprises VD1-(X1)n-VD2-C-(X2)n, wherein [0058]
VD1 is a first light chain variable domain; [0059] VD2 is a second
light chain variable domain; [0060] C is a light chain constant
domain; [0061] X1 is a linker with the proviso that it is not CL;
[0062] X2 does not comprise an Fc region; [0063] (X1)n is (X1)0 or
(X1)1; [0064] (X2)n is (X2)0 or (X2)1; and [0065] wherein [0066]
(a) VD1 or VD2 comprise three CDRs from SEQ ID NO: 29, 39, 49, 59,
69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199,
209, 219, 229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329,
339, 349, 359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459,
469, 479, 489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589,
599, 609, 619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719,
729, 739, 749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849,
859, 869, 879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979,
989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089,
1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199,
1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309,
1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419,
1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529,
1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639,
1649, 1659, 1669, 1679, or 1689, and the binding protein is capable
of binding sclerostin and another target; [0067] (b) VD1 and VD2
independently comprise three CDRs from SEQ ID NO: 29, 39, 49, 59,
69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199,
209, 219, 229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329,
339, 349, 359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459,
469, 479, 489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589,
599, 609, 619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719,
729, 739, 749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849,
859, 869, 879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979,
989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089,
1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199,
1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309,
1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419,
1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529,
1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639,
1649, 1659, 1669, 1679, or 1689, and the binding protein is capable
of binding sclerostin and sclerostin; [0068] (c) VD1 comprises
three CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109,
119, 129, 139, 149, 159, 169, 179, 189, 199, 209, 219, 229, 239,
249, 259, 269, 279, 289, 299, 309, 319, 329, 339, 349, 359, 369,
379, 389, 399, 409, 419, 429, 439, 449, 459, 469, 479, 489, 499,
509, 519, 529, 539, 549, 559, 569, 579, 589, 599, 609, 619, 629,
639, 649, 659, 669, 679, 689, 699, 709, 719, 729, 739, 749, 759,
769, 779, 789, 799, 809, 819, 829, 839, 849, 859, 869, 879, 889,
899, 909, 919, 929, 939, 949, 959, 969, 979, 989, 999, 1009, 1019,
1029, 1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129,
1139, 1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239,
1249, 1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349,
1359, 1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459,
1469, 1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569,
1579, 1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679,
or 1689, and VD2 comprises three CDRs from SEQ ID NO: 27, 37, 47,
57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187,
197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317,
327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447,
457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577,
587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707,
717, 727, 737, 747, 757, 767, 777, 787, 797, 807, 817, 827, 837,
847, 857, 867, 877, 887, 897, 907, 917, 927, 937, 947, 957, 967,
977, 987, 997, 1007, 1017, 1027, 1037, 1047, 1057, 1067, 1077,
1087, 1097, 1107, 1117, 1127, 1137, 1147, 1157, 1167, 1177, 1187,
1197, 1207, 1217, 1227, 1237, 1247, 1257, 1267, 1277, 1287, 1297,
1307, 1317, 1327, 1337, 1347, 1357, 1367, 1377, 1387, 1397, 1407,
1417, 1427, 1437, 1447, 1457, 1467, 1477, 1487, 1497, 1507, 1517,
1527, 1537, 1547, 1557, 1567, 1577, 1587, 1597, 1607, 1617, 1627,
1637, 1647, 1657, 1667, 1677, or 1687, and the binding protein is
capable of binding sclerostin and TNF-.alpha.; or [0069] (d) VD2
comprises three CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79, 89,
99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199, 209, 219,
229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329, 339, 349,
359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459, 469, 479,
489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589, 599, 609,
619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719, 729, 739,
749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849, 859, 869,
879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979, 989, 999,
1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109,
1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219,
1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329,
1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439,
1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549,
1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659,
1669, 1679, or 1689, and VD1 comprises three CDRs from SEQ ID NO:
27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167,
177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297,
307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427,
437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557,
567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687,
697, 707, 717, 727, 737, 747, 757, 767, 777, 787, 797, 807, 817,
827, 837, 847, 857, 867, 877, 887, 897, 907, 917, 927, 937, 947,
957, 967, 977, 987, 997, 1007, 1017, 1027, 1037, 1047, 1057, 1067,
1077, 1087, 1097, 1107, 1117, 1127, 1137, 1147, 1157, 1167, 1177,
1187, 1197, 1207, 1217, 1227, 1237, 1247, 1257, 1267, 1277, 1287,
1297, 1307, 1317, 1327, 1337, 1347, 1357, 1367, 1377, 1387, 1397,
1407, 1417, 1427, 1437, 1447, 1457, 1467, 1477, 1487, 1497, 1507,
1517, 1527, 1537, 1547, 1557, 1567, 1577, 1587, 1597, 1607, 1617,
1627, 1637, 1647, 1657, 1667, 1677, or 1687, and the binding
protein is capable of binding TNF-.alpha. and sclerostin. [0070] In
an embodiment of the DVD-binding protein described above,
VD1-(X1)n-VD2 comprises SEQ ID NO: 26, 36, 46, 56, 66, 76, 86, 96,
106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226,
236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356,
366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486,
496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616,
626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746,
756, 766, 776, 786, 796, 806, 816, 826, 836, 846, 856, 866, 876,
886, 896, 906, 916, 926, 936, 946, 956, 966, 976, 986, 996, 1006,
1116, 1126, 1136, 1146, 1156, 1166, 1176, 1186, 1196, 1206, 1216,
1226, 1236, 1246, 1256, 1266, 1276, 1286, 1296, 1306, 1316, 1326,
1336, 1346, 1356, 1366, 1376, 1386, 1396, 1406, 1416, 1426, 1436,
1446, 1456, 1466, 1476, 1486, 1496, 1506, 1516, 1526, 1536, 1546,
1556, 1566, 1576, 1586, 1596, 1606, 1616, 1626, 1636, 1646, 1656,
1666, 1676, or 1686.
[0071] Another embodiment provides a multivalent, multispecific
DVD-A binding protein comprising first and second polypeptide
chains, wherein the first polypeptide chain comprises a first
VD1-(X1)n-VD2-C-(X2)n, wherein [0072] VD1 is a first heavy chain
variable domain; [0073] VD2 is a second heavy chain variable
domain; [0074] C is a heavy chain constant domain; [0075] X1 is a
first linker; [0076] X2 is an Fc region; [0077] (X1)n is (X1)0 or
(X1)1; [0078] (X2)n is (X2)0 or (X2)1; and
[0079] wherein the second polypeptide chain comprises a second
VD1-(X1)n-VD2-C-(X2)n, wherein [0080] VD1 is a first light chain
variable domain; [0081] VD2 is a second light chain variable
domain; [0082] C is a light chain constant domain; [0083] X1 is a
second linker; [0084] X2 does not comprise an Fc region; (X1)n is
(X1)0 or (X1)1; [0085] (X2)n is (X2)0 or (X2)1; and [0086] wherein
the first and second X1 linker are the same or different; [0087]
wherein the first X1 linker is not CH1 and/or the second X1 linker
is not CL and wherein [0088] (a) the VD1 or VD2 heavy chain
variable domain comprise three CDRs from SEQ ID NO: 24, 34, 44, 54,
64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194,
204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324,
334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454,
464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584,
594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714,
724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824, 834, 844,
854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954, 964, 974,
984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084,
1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204,
1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314,
1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424,
1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534,
1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644,
1654, 1664, 1674, or 1684, the VD1 or VD2 light chain variable
domain comprises three CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79,
89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199, 209, 219,
229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329, 339, 349,
359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459, 469, 479,
489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589, 599, 609,
619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719, 729, 739,
749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849, 859, 869,
879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979, 989, 999,
1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109,
1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219,
1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329,
1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439,
1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549,
1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659,
1669, 1679, or1689, and the binding protein is capable of binding
sclerostin and another target; [0089] (b) the VD1 and VD2 heavy
chain variable domains independently comprise three CDRs from SEQ
ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144,
154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274,
284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404,
414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534,
544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664,
674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794,
804, 814, 824, 834, 844, 854, 864, 874, 884, 894, 904, 914, 924,
934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024, 1034, 1044,
1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164,
1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274,
1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384,
1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494,
1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604,
1614, 1624, 1634, 1644, 1654, 1664, 1674, or 1684, the VD1 or VD2
light chain variable domain comprises three CDRs from SEQ ID NO:
29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169,
179, 189, 199, 209, 219, 229, 239, 249, 259, 269, 279, 289, 299,
309, 319, 329, 339, 349, 359, 369, 379, 389, 399, 409, 419, 429,
439, 449, 459, 469, 479, 489, 499, 509, 519, 529, 539, 549, 559,
569, 579, 589, 599, 609, 619, 629, 639, 649, 659, 669, 679, 689,
699, 709, 719, 729, 739, 749, 759, 769, 779, 789, 799, 809, 819,
829, 839, 849, 859, 869, 879, 889, 899, 909, 919, 929, 939, 949,
959, 969, 979, 989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069,
1079, 1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179,
1189, 1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289,
1299, 1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399,
1409, 1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509,
1519, 1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619,
1629, 1639, 1649, 1659, 1669, 1679, or1689, and the binding protein
is capable of binding sclerostin and sclerostin; [0090] (c) the VD1
heavy chain variable domain comrpises three CDRs from SEQ ID NO:
24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164,
174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294,
304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424,
434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554,
564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684,
694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794, 804, 814,
824, 834, 844, 854, 864, 874, 884, 894, 904, 914, 924, 934, 944,
954, 964, 974, 984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064,
1074, 1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184,
1194, 1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294,
1304, 1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404,
1414, 1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514,
1524, 1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624,
1634, 1644, 1654, 1664, 1674, or 1684, and the VD2 heavy chain
variable domain comprises three CDRs from SEQ ID NO: 22, 32, 42,
52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182,
192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312,
322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442,
452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572,
582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702,
712, 722, 732, 742, 752, 762, 772, 782, 792, 802, 812, 822, 832,
842, 852, 862, 872, 882, 892, 902, 912, 922, 932, 942, 952, 962,
972, 982, 992, 1002, 1012, 1022, 1032, 1042, 1052, 1062, 1072,
1082, 1092, 1102, 1112, 1122, 1132, 1142, 1152, 1162, 1172, 1182,
1192, 1202, 1212, 1222, 1232, 1242, 1252, 1262, 1272, 1282, 1292,
1302, 1312, 1322, 1332, 1242, 1252, 1262, 1272, 1282, 1292, 1302,
1312, 1322, 1332, 1342, 1352, 1362, 1372, 1382, 1392, 1402, 1412,
1422, 1432, 1442, 1452, 1462, 1472, 1482, 1492, 1502, 1512, 1522,
1532, 1542, 1552, 1562, 1572, 1582, 1592, 1602, 1612, 1622, 1632,
1642, 1652, 1662, 1672, or 1682; the VD1 light chain variable
domain comprises three CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79,
89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199, 209, 219,
229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329, 339, 349,
359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459, 469, 479,
489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589, 599, 609,
619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719, 729, 739,
749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849, 859, 869,
879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979, 989, 999,
1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109,
1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219,
1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329,
1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439,
1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549,
1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659,
1669, 1679, or 1689, and the VD2 light chain variable domain
comprises three CDRs from SEQ ID NO: 27, 37, 47, 57, 67, 77, 87,
97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217,
227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347,
357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477,
487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607,
617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737,
747, 757, 767, 777, 787, 797, 807, 817, 827, 837, 847, 857, 867,
877, 887, 897, 907, 917, 927, 937, 947, 957, 967, 977, 987, 997,
1007, 1017, 1027, 1037, 1047, 1057, 1067, 1077, 1087, 1097, 1107,
1117, 1127, 1137, 1147, 1157, 1167, 1177, 1187, 1197, 1207, 1217,
1227, 1237, 1247, 1257, 1267, 1277, 1287, 1297, 1307, 1317, 1327,
1337, 1347, 1357, 1367, 1377, 1387, 1397, 1407, 1417, 1427, 1437,
1447, 1457, 1467, 1477, 1487, 1497, 1507, 1517, 1527, 1537, 1547,
1557, 1567, 1577, 1587, 1597, 1607, 1617, 1627, 1637, 1647, 1657,
1667, 1677, or 1687, and the binding protein is capable of binding
sclerostin and TNF-.alpha.; or [0091] (d) the VD2 heavy chain
variable domain comrpises three CDRs from SEQ ID NO: 24, 34, 44,
54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184,
194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314,
324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444,
454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574,
584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704,
714, 724, 734, 744, 754, 764, 774, 784, 794, 804, 814, 824, 834,
844, 854, 864, 874, 884, 894, 904, 914, 924, 934, 944, 954, 964,
974, 984, 994, 1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074,
1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194,
1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304,
1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414,
1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524,
1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634,
1644, 1654, 1664, 1674, or 1684, and the VD1 heavy chain variable
domain comprises three CDRs from SEQ ID NO: 22, 32, 42, 52, 62, 72,
82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212,
222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342,
352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472,
482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602,
612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732,
742, 752, 762, 772, 782, 792, 802, 812, 822, 832, 842, 852, 862,
872, 882, 892, 902, 912, 922, 932, 942, 952, 962, 972, 982, 992,
1002, 1012, 1022, 1032, 1042, 1052, 1062, 1072, 1082, 1092, 1102,
1112, 1122, 1132, 1142, 1152, 1162, 1172, 1182, 1192, 1202, 1212,
1222, 1232, 1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322,
1332, 1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322, 1332,
1342, 1352, 1362, 1372, 1382, 1392, 1402, 1412, 1422, 1432, 1442,
1452, 1462, 1472, 1482, 1492, 1502, 1512, 1522, 1532, 1542, 1552,
1562, 1572, 1582, 1592, 1602, 1612, 1622, 1632, 1642, 1652, 1662,
1672, or 1682; the VD2 light chain variable domain comprises three
CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129,
139, 149, 159, 169, 179, 189, 199, 209, 219, 229, 239, 249, 259,
269, 279, 289, 299, 309, 319, 329, 339, 349, 359, 369, 379, 389,
399, 409, 419, 429, 439, 449, 459, 469, 479, 489, 499, 509, 519,
529, 539, 549, 559, 569, 579, 589, 599, 609, 619, 629, 639, 649,
659, 669, 679, 689, 699, 709, 719, 729, 739, 749, 759, 769, 779,
789, 799, 809, 819, 829, 839, 849, 859, 869, 879, 889, 899, 909,
919, 929, 939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029,
1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139,
1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249,
1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359,
1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469,
1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579,
1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, or
1689, and the VD1 light chain variable domain comprises three CDRs
from SEQ ID NO: 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137,
147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267,
277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397,
407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527,
537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657,
667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787,
797, 807, 817, 827, 837, 847, 857, 867, 877, 887, 897, 907, 917,
927, 937, 947, 957, 967, 977, 987, 997, 1007, 1017, 1027, 1037,
1047, 1057, 1067, 1077, 1087, 1097, 1107, 1117, 1127, 1137, 1147,
1157, 1167, 1177, 1187, 1197, 1207, 1217, 1227, 1237, 1247, 1257,
1267, 1277, 1287, 1297, 1307, 1317, 1327, 1337, 1347, 1357, 1367,
1377, 1387, 1397, 1407, 1417, 1427, 1437, 1447, 1457, 1467, 1477,
1487, 1497, 1507, 1517, 1527, 1537, 1547, 1557, 1567, 1577, 1587,
1597, 1607, 1617, 1627, 1637, 1647, 1657, 1667, 1677, or 1687, and
the binding protein is capable of binding TNF-.alpha. and
sclerostin. [0092] In an embodiment of the DVD-binding protein
described above, wherein X1 or X2 is SEQ ID NO: 1695, 1696, 1697,
1698, 1699, 1700, 1701, 1702, 1703, 1704, 1705, 1706, 1707, 1708,
1709, 1710, 1711, 1712, 1713, 1714, 1715, 1716, 2050, 2051, 2052,
2053, 2054, 2055, 2056, 2057, 2058, and 2059.
[0093] Another embodiment provides a multivalent, multispecific
DVD-A binding protein capable of binding two antigens comprising
four polypeptide chains, wherein two polypeptide chains comprise
VD1-(X1)n-VD2-C-(X2)n, wherein [0094] VD1 is a first heavy chain
variable domain; [0095] VD2 is a second heavy chain variable
domain; [0096] C is a heavy chain constant domain; [0097] X1 is a
first linker; [0098] X2 is an Fc region; [0099] (X1)n is (X1)0 or
(X1)1; [0100] (X2)n is (X2)0 or (X2)1; and [0101] wherein two
polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein [0102]
VD1 is a first light chain variable domain; [0103] VD2 is a second
light chain variable domain; [0104] C is a light chain constant
domain; [0105] X1 is a second linker; [0106] X2 does not comprise
an Fc region; [0107] (X1)n is (X1)0 or (X1)1; [0108] (X2)n is (X2)0
or (X2)1; and [0109] wherein the first and second X1 linker are the
same or different; [0110] wherein the first X1 linker is not CH1
and/or the second X1 linker is not CL and wherein [0111] (a) the
VD1 or VD2 heavy chain variable domain comprise three CDRs from SEQ
ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144,
154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274,
284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404,
414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534,
544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664,
674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794,
804, 814, 824, 834, 844, 854, 864, 874, 884, 894, 904, 914, 924,
934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024, 1034, 1044,
1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164,
1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274,
1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384,
1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494,
1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604,
1614, 1624, 1634, 1644, 1654, 1664, 1674, or 1684, the VD1 or VD2
light chain variable domain comprises three CDRs from SEQ ID NO:
29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129, 139, 149, 159, 169,
179, 189, 199, 209, 219, 229, 239, 249, 259, 269, 279, 289, 299,
309, 319, 329, 339, 349, 359, 369, 379, 389, 399, 409, 419, 429,
439, 449, 459, 469, 479, 489, 499, 509, 519, 529, 539, 549, 559,
569, 579, 589, 599, 609, 619, 629, 639, 649, 659, 669, 679, 689,
699, 709, 719, 729, 739, 749, 759, 769, 779, 789, 799, 809, 819,
829, 839, 849, 859, 869, 879, 889, 899, 909, 919, 929, 939, 949,
959, 969, 979, 989, 999, 1009, 1019, 1029, 1039, 1049, 1059, 1069,
1079, 1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159, 1169, 1179,
1189, 1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269, 1279, 1289,
1299, 1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379, 1389, 1399,
1409, 1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489, 1499, 1509,
1519, 1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599, 1609, 1619,
1629, 1639, 1649, 1659, 1669, 1679, or 1689, and the binding
protein is capable of binding sclerostin and another target; [0112]
(b) the VD1 and VD2 heavy chain variable domains independently
comprise three CDRs from SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94,
104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224,
234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354,
364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484,
494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614,
624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744,
754, 764, 774, 784, 794, 804, 814, 824, 834, 844, 854, 864, 874,
884, 894, 904, 914, 924, 934, 944, 954, 964, 974, 984, 994, 1004,
1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114, 1124,
1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224, 1234,
1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334, 1344,
1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444, 1454,
1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554, 1564,
1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664, 1674,
or 1684, the VD1 or VD2 light chain variable domain comprises three
CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129,
139, 149, 159, 169, 179, 189, 199, 209, 219, 229, 239, 249, 259,
269, 279, 289, 299, 309, 319, 329, 339, 349, 359, 369, 379, 389,
399, 409, 419, 429, 439, 449, 459, 469, 479, 489, 499, 509, 519,
529, 539, 549, 559, 569, 579, 589, 599, 609, 619, 629, 639, 649,
659, 669, 679, 689, 699, 709, 719, 729, 739, 749, 759, 769, 779,
789, 799, 809, 819, 829, 839, 849, 859, 869, 879, 889, 899, 909,
919, 929, 939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029,
1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139,
1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249,
1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359,
1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469,
1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579,
1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, or
1689, and the binding protein is capable of binding sclerostin and
sclerostin; [0113] (c) the VD1 heavy chain variable domain
comrpises three CDRs from SEQ ID NO: 24, 34, 44, 54, 64, 74, 84,
94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214,
224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344,
354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474,
484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604,
614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734,
744, 754, 764, 774, 784, 794, 804, 814, 824, 834, 844, 854, 864,
874, 884, 894, 904, 914, 924, 934, 944, 954, 964, 974, 984, 994,
1004, 1014, 1024, 1034, 1044, 1054, 1064, 1074, 1084, 1094, 1114,
1124, 1134, 1144, 1154, 1164, 1174, 1184, 1194, 1204, 1214, 1224,
1234, 1244, 1254, 1264, 1274, 1284, 1294, 1304, 1314, 1324, 1334,
1344, 1354, 1364, 1374, 1384, 1394, 1404, 1414, 1424, 1434, 1444,
1454, 1464, 1474, 1484, 1494, 1504, 1514, 1524, 1534, 1544, 1554,
1564, 1574, 1584, 1594, 1604, 1614, 1624, 1634, 1644, 1654, 1664,
1674, or 1684, and the VD2 heavy chain variable domain comprises
three CDRs from SEQ ID NO: 22, 32, 42, 52, 62, 72, 82, 92, 102,
112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232,
242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362,
372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492,
502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622,
632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752,
762, 772, 782, 792, 802, 812, 822, 832, 842, 852, 862, 872, 882,
892, 902, 912, 922, 932, 942, 952, 962, 972, 982, 992, 1002, 1012,
1022, 1032, 1042, 1052, 1062, 1072, 1082, 1092, 1102, 1112, 1122,
1132, 1142, 1152, 1162, 1172, 1182, 1192, 1202, 1212, 1222, 1232,
1242, 1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322, 1332, 1242,
1252, 1262, 1272, 1282, 1292, 1302, 1312, 1322, 1332, 1342, 1352,
1362, 1372, 1382, 1392, 1402, 1412, 1422, 1432, 1442, 1452, 1462,
1472, 1482, 1492, 1502, 1512, 1522, 1532, 1542, 1552, 1562, 1572,
1582, 1592, 1602, 1612, 1622, 1632, 1642, 1652, 1662, 1672, or
1682; the VD1 light chain variable domain comrpises three CDRs from
SEQ ID NO: 29, 39, 49, 59, 69, 79, 89, 99, 109, 119, 129, 139, 149,
159, 169, 179, 189, 199, 209, 219, 229, 239, 249, 259, 269, 279,
289, 299, 309, 319, 329, 339, 349, 359, 369, 379, 389, 399, 409,
419, 429, 439, 449, 459, 469, 479, 489, 499, 509, 519, 529, 539,
549, 559, 569, 579, 589, 599, 609, 619, 629, 639, 649, 659, 669,
679, 689, 699, 709, 719, 729, 739, 749, 759, 769, 779, 789, 799,
809, 819, 829, 839, 849, 859, 869, 879, 889, 899, 909, 919, 929,
939, 949, 959, 969, 979, 989, 999, 1009, 1019, 1029, 1039, 1049,
1059, 1069, 1079, 1089, 1099, 1109, 1119, 1129, 1139, 1149, 1159,
1169, 1179, 1189, 1199, 1209, 1219, 1229, 1239, 1249, 1259, 1269,
1279, 1289, 1299, 1309, 1319, 1329, 1339, 1349, 1359, 1369, 1379,
1389, 1399, 1409, 1419, 1429, 1439, 1449, 1459, 1469, 1479. 1489,
1499, 1509, 1519, 1529, 1539, 1549, 1559, 1569, 1579, 1589, 1599,
1609, 1619, 1629, 1639, 1649, 1659, 1669, 1679, or1689, and the VD2
light chain variable domain comprises three CDRs from SEQ ID NO:
27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167,
177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297,
307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427,
437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557,
567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687,
697, 707, 717, 727, 737, 747, 757, 767, 777, 787, 797, 807, 817,
827, 837, 847, 857, 867, 877, 887, 897, 907, 917, 927, 937, 947,
957, 967, 977, 987, 997, 1007, 1017, 1027, 1037, 1047, 1057, 1067,
1077, 1087, 1097, 1107, 1117, 1127, 1137, 1147, 1157, 1167, 1177,
1187, 1197, 1207, 1217, 1227, 1237, 1247, 1257, 1267, 1277, 1287,
1297, 1307, 1317, 1327, 1337, 1347, 1357, 1367, 1377, 1387, 1397,
1407, 1417, 1427, 1437, 1447, 1457, 1467, 1477, 1487, 1497, 1507,
1517, 1527, 1537, 1547, 1557, 1567, 1577, 1587, 1597, 1607, 1617,
1627, 1637, 1647, 1657, 1667, 1677, or 1687, and the binding
protein is capable of binding sclerostin and TNF-.alpha.; or [0114]
(d) the VD2 heavy chain variable domain comprises three CDRs from
SEQ ID NO: 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144,
154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274,
284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404,
414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534,
544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664,
674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794,
804, 814, 824, 834, 844, 854, 864, 874, 884, 894, 904, 914, 924,
934, 944, 954, 964, 974, 984, 994, 1004, 1014, 1024, 1034, 1044,
1054, 1064, 1074, 1084, 1094, 1114, 1124, 1134, 1144, 1154, 1164,
1174, 1184, 1194, 1204, 1214, 1224, 1234, 1244, 1254, 1264, 1274,
1284, 1294, 1304, 1314, 1324, 1334, 1344, 1354, 1364, 1374, 1384,
1394, 1404, 1414, 1424, 1434, 1444, 1454, 1464, 1474, 1484, 1494,
1504, 1514, 1524, 1534, 1544, 1554, 1564, 1574, 1584, 1594, 1604,
1614, 1624, 1634, 1644, 1654, 1664, 1674, or 1684, and the VD1
heavy chain variable domain comprises three CDRs from SEQ ID NO:
22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162,
172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292,
302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422,
432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552,
562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682,
692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792, 802, 812,
822, 832, 842, 852, 862, 872, 882, 892, 902, 912, 922, 932, 942,
952, 962, 972, 982, 992, 1002, 1012, 1022, 1032, 1042, 1052, 1062,
1072, 1082, 1092, 1102, 1112, 1122, 1132, 1142, 1152, 1162, 1172,
1182, 1192, 1202, 1212, 1222, 1232, 1242, 1252, 1262, 1272, 1282,
1292, 1302, 1312, 1322, 1332, 1242, 1252, 1262, 1272, 1282, 1292,
1302, 1312, 1322, 1332, 1342, 1352, 1362, 1372, 1382, 1392, 1402,
1412, 1422, 1432, 1442, 1452, 1462, 1472, 1482, 1492, 1502, 1512,
1522, 1532, 1542, 1552, 1562, 1572, 1582, 1592, 1602, 1612, 1622,
1632, 1642, 1652, 1662, 1672, or 1682; the VD2 light chain variable
domain comrpises three CDRs from SEQ ID NO: 29, 39, 49, 59, 69, 79,
89, 99, 109, 119, 129, 139, 149, 159, 169, 179, 189, 199, 209, 219,
229, 239, 249, 259, 269, 279, 289, 299, 309, 319, 329, 339, 349,
359, 369, 379, 389, 399, 409, 419, 429, 439, 449, 459, 469, 479,
489, 499, 509, 519, 529, 539, 549, 559, 569, 579, 589, 599, 609,
619, 629, 639, 649, 659, 669, 679, 689, 699, 709, 719, 729, 739,
749, 759, 769, 779, 789, 799, 809, 819, 829, 839, 849, 859, 869,
879, 889, 899, 909, 919, 929, 939, 949, 959, 969, 979, 989, 999,
1009, 1019, 1029, 1039, 1049, 1059, 1069, 1079, 1089, 1099, 1109,
1119, 1129, 1139, 1149, 1159, 1169, 1179, 1189, 1199, 1209, 1219,
1229, 1239, 1249, 1259, 1269, 1279, 1289, 1299, 1309, 1319, 1329,
1339, 1349, 1359, 1369, 1379, 1389, 1399, 1409, 1419, 1429, 1439,
1449, 1459, 1469, 1479. 1489, 1499, 1509, 1519, 1529, 1539, 1549,
1559, 1569, 1579, 1589, 1599, 1609, 1619, 1629, 1639, 1649, 1659,
1669, 1679, or 1689, and the VD1 light chain variable domain
comprises three CDRs from SEQ ID NO: 27, 37, 47, 57, 67, 77, 87,
97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217,
227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347,
357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477,
487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607,
617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737,
747, 757, 767, 777, 787, 797, 807, 817, 827, 837, 847, 857, 867,
877, 887, 897, 907, 917, 927, 937, 947, 957, 967, 977, 987, 997,
1007, 1017, 1027, 1037, 1047, 1057, 1067, 1077, 1087, 1097, 1107,
1117, 1127, 1137, 1147, 1157, 1167, 1177, 1187, 1197, 1207, 1217,
1227, 1237, 1247, 1257, 1267, 1277, 1287, 1297, 1307, 1317, 1327,
1337, 1347, 1357, 1367, 1377, 1387, 1397, 1407, 1417, 1427, 1437,
1447, 1457, 1467, 1477, 1487, 1497, 1507, 1517, 1527, 1537, 1547,
1557, 1567, 1577, 1587, 1597, 1607, 1617, 1627, 1637, 1647, 1657,
1667, 1677, or 1687, and the binding protein is capable of binding
TNF-.alpha. and sclerostin.
[0115] In an embodiment of a multivalent, multispecific DVD-binding
protein described herein, n is 0. [0116] An embodiment of a
multivalent, multispecific DVD-binding protein is provided, wherein
X1 or X2 is SEQ ID NO: 1695, 1696, 1697, 1698, 1699, 1700, 1701,
1702, 1703, 1704, 1705, 1706, 1707, 1708, 1709, 1710, 1711, 1712,
1713, 1714, 1715, 1716, 2050, 2051, 2052, 2053, 2054, 2055, 2056,
2057, 2058, and 2059.
[0117] In another embodiment, a multivalent, multispecific
DVD-binding protein described herein comprises two first
polypeptide chains and two second polypeptide chains.
[0118] In an embodiment of a multivalent, multispecific DVD-binding
protein described herein, the Fc region is a native sequence Fc
region or a variant sequence Fc region.
[0119] In an embodiment, the Fc region is an Fc region from an
IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD.
[0120] In another embodiment of a multivalent, multispecif
DVD-binding protein described herein comprising a first and second
polypeptide chains, VD1 of the first polypeptide chain and said VD1
of the second polypeptide chain are obtained from the same first
and second parent antibody, respectively, or antigen binding
portion thereof.
[0121] In an embodiment of a TNF-.alpha. and sclerostin DVD-binding
protein described herein, a parental anti-TNF-.alpha. antibody
binds TNF-.alpha. with a potency different from the potency with
which a parental anti-sclerostin antibody binds human
sclerostin.
[0122] In another embodiment of a TNF-.alpha. and SOST DVD-binding
protein described herein, a parental anti-TNF-.alpha. antibody
binds TNF-.alpha. with an affinity different from the affinity with
which said anti-sclerostin antibody binds human sclerostin.
[0123] In another embodiment of a TNF-.alpha. and SOST DVD-binding
protein described herein, an anti-TNF-.alpha. antibody and said
anti-sclerostin antibody are a human antibody, a CDR grafted
antibody, or a humanized antibody.
[0124] In another embodiment, a TNF-.alpha. and SOST DVD-binding
protein described herein possesses at least one desired property
exhibited by said anti-TNF-.alpha. antibody or said anti-sclerostin
antibody. In an embodiment, the desired property is one or more
antibody parameters. In an embodiment, the antibody parameters are
antigen specificity, affinity to antigen, potency, biological
function, epitope recognition, stability, solubility, production
efficiency, immunogenicity, pharmacokinetics, bioavailability,
tissue cross reactivity, or orthologous antigen binding.
[0125] Another embodiment provides a method of producing a
multivalent, multispecific DVD-binding protein described herein,
comprising culturing a host cell carrying a vector comprising a
nucleic acid described herein in culture medium under conditions
sufficient to produce the binding protein. In an embodiment,
50%-75% of the binding protein produced according the method is a
dual specific tetravalent DVD-binding protein described herein. In
an embodiment, 75%-90% of the binding protein produced according to
this method is a dual specific tetravalent binding protein. In an
embodiment, 90%-95% of the binding protein produced is a dual
specific tetravalent binding protein.
[0126] Another embodiment provides a protein produced according to
the described method.
[0127] In another embodiment, a pharmaceutical composition
comprising a multivalent, multispecific DVD-binding protein
described herein and a pharmaceutically acceptable carrier is
provided.
[0128] In another embodiment, a pharmaceutical composition
comprising a multivalent, multispecific DVD-binding protein further
comprises at least one additional agent. In an embodiment, the
additional agent is a therapeutic agent; an imaging agent; a
cytotoxic agent; an angiogenesis inhibitor; a kinase inhibitor; a
co-stimulation molecule blocker; an adhesion molecule blocker; an
anti-cytokine antibody or functional fragment thereof;
methotrexate; cyclosporin; rapamycin; FK506; a detectable label or
reporter; a TNF antagonist; an antirheumatic; a muscle relaxant, a
narcotic, a non-steroid anti-inflammatory drug (NSAID); an
analgesic; an anesthetic; a sedative; a local anesthetic; a
neuromuscular blocker; an antimicrobial; an antipsoriatic; a
corticosteroid; an anabolic steroid; an erythropoietin; an
immunization; an immunoglobulin; an immunosuppressive; a growth
hormone; a hormone replacement drug; a radiopharmaceutical; an
antidepressant; an antipsychotic; a stimulant; an asthma
medication; a beta agonist; an inhaled steroid; an epinephrine or
analog; a cytokine; or a cytokine antagonist.
[0129] Another embodiment provides a method for treating a subject
for a disease or a disorder by administering to the subject a
multivalent, multispecific DVD-binding protein described herein
that binds TNF-.alpha. and sclerostin such that treatment is
achieved.
[0130] A method for generating a multivalent, multispecific
DVD-binding protein described herein is provided, comprising the
steps of:
[0131] a) obtaining a first parent antibody or antigen binding
portion thereof
[0132] b) obtaining a second parent antibody or antigen binding
portion thereof;
[0133] c) constructing polypeptide chains described herein;
[0134] e) expressing said polypeptide chains;
such that a DVD-binding protein is generated.
[0135] In another embodiment of the method described above, said
first parent antibody or antigen binding portion thereof, and said
second parent antibody or antigen binding portion thereof, are a
human antibody, a CDR grafted antibody, or a humanized
antibody.
[0136] In another embodiment of the method described above, said
first parent antibody or antigen binding portion thereof, and said
second parent antibody or antigen binding portion thereof, are an
Fab fragment, an F(ab').sub.2 fragment, a bivalent fragment
comprising two Fab fragments linked by a disulfide bridge at the
hinge region; an Fd fragment consisting of the VH and CH1 domains;
an Fv fragment consisting of the VL and VH domains of a single arm
of an antibody, a dAb fragment, an isolated complementarity
determining region (CDR), a single chain antibody, or
diabodies.
[0137] In another embodiment of the method, the first parent
antibody or antigen binding portion thereof possesses at least one
desired property exhibited by the DVD-binding protein.
[0138] In another embodiment of the method described above the
second parent antibody or antigen binding portion thereof possesses
at least one desired property exhibited by the DVD-binding
protein.
[0139] In an embodiment, in the method described above, the Fc
region is a native sequence Fc region or a variant sequence Fc
region. In an embodiment, the Fc region is an Fc region from an
IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD.
[0140] In another embodiment, in a method described above, a
desired property is one or more antibody parameters of the first
parent antibody or antigen binding portion thereof.
[0141] In another embodiment, in a method described above, a
desired property is one or more antibody parameters of the second
parent antibody.
[0142] In an embodiment, said antibody parameters are antigen
specificity, affinity to antigen, potency, biological function,
epitope recognition, stability, solubility, production efficiency,
immunogenicity, pharmacokinetics, bioavailability, tissue cross
reactivity, or orthologous antigen binding.
[0143] In another embodiment of the method described above, the
first parent antibody or antigen binding portion thereof, binds
said first antigen with a different affinity than the affinity with
which said second parent antibody or antigen binding portion
thereof, binds said second antigen.
[0144] In another embodiment, the first parent antibody or antigen
binding portion thereof, binds said first antigen with a different
potency than the potency with which said second parent antibody or
antigen binding portion thereof, binds said second antigen.
[0145] In another embodiment, an sclerostin binding protein
described herein binds human sclerostin and is capable of
modulating a biological function of SOST.
[0146] A neutralizing binding protein is provided, wherein the
neutralizing binding protein comprises a Sclerostin binding protein
as described above, and wherein said neutralizing binding protein
is capable of neutralizing sclerostin.
[0147] In another embodiment, a neutralizing sclerostin binding
protein that binds pro-human sclerostin, mature-human sclerostin,
or truncated-human sclerostin is provided.
[0148] In an embodiment, a neutralizing sclerostin binding protein
described herein diminishes the ability of sclerostin to bind to
its receptor. In an embodiment, a neutralizing sclerostin binding
protein diminishes the ability of pro-human sclerostin, mature
human sclerostin, or a truncated human sclerostin to bind to the
sclerostinreceptor.
[0149] In another embodiment, a neutralizing sclerostin binding
protein described herein is capable of reducing one or more of
sclerostin biological activities, including: including inhibition
of osteoblast differentiation, and osteoblast function leading to
inhibition of bone formation.
[0150] In an embodiment, an sclerostin binding protein having an on
rate constant (K.sub.on) to said target of: at least about 10.sup.2
M.sup.-1 s.sup.-1; at least about 10.sup.3 M.sup.-1 s.sup.-1; at
least about 10.sup.4 M.sup.-1 s.sup.-1; at least about 10.sup.5
M.sup.-1 s.sup.-1; or at least about 10.sup.6 M.sup.-1 s.sup.-1; as
measured by surface plasmon resonance, is provided.
[0151] In another embodiment, an sclerostin binding protein having
an off rate constant (K.sub.off) to said target of: at most about
10.sup.-3 s.sup.-1; at most about 10.sup.-4 s.sup.-1; at most about
10.sup.-5 s.sup.-1; or at most about 10.sup.-6 s.sup.-1, as
measured by surface plasmon resonance, is provided.
[0152] In another embodiment, an sclerostinbinding protein having a
dissociation constant (K.sub.D) to said target of: at most about
10.sup.-7 M; at most about 10.sup.-8 M; at most about 10.sup.-9 M;
at most about 10.sup.-10 M; at most about 10.sup.-11 M; at most
about 10.sup.-12 M; or at most 10.sup.-13 M, is provided.
[0153] Another aspect provides an sclerostinbinding protein
construct that comprises an sclerostin binding protein described
herein and further comprises a linker polypeptide or an
immunoglobulin constant domain. In an embodiment, the sclerostin
binding protein construct is provided, wherein said construct
comprises an sclerostin binding protein of an immunoglobulin
molecule, a disulfide linked Fv, a monoclonal antibody, an scFv, a
chimeric antibody, a single domain antibody, a CDR-grafted
antibody, a diabody, a humanized antibody, a multispecific
antibody, an Fab, a dual specific antibody, an Fab', a bispecific
antibody, an F(ab')2, an Fv, or a DVD-binding protein.
[0154] In an embodiment, an sclerostin binding protein construct is
provided, wherein said construct comprises a heavy chain
immunoglobulin constant domain of a human IgM constant domain, a
human IgG4 constant domain, a human IgG1 constant domain, a human
IgE constant domain, a human IgG2 constant domain, a human IgG3
constant domain, or a human IgA constant domain.
[0155] In yet another embodiment, asclerostin binding protein
construct comprises an immunoglobulin constant domain having an
amino acid sequence SEQ ID NO: 2060, SEQ ID NO: 2061, SEQ ID
NO:2062; SEQ ID NO:2063; SEQ ID NO:2063; and SEQ ID NO:2065.
[0156] In another embodiment, a sclerostin binding protein
construct described herein has a greater half life in vivo than the
soluble counterpart of said sclerostin binding protein
construct.
[0157] Another aspect provides asclerostin binding protein
conjugate comprising asclerostin binding protein construct, wherein
the sclerostin binding protein conjugate further comprises an
immunoadhesion molecule, an imaging agent, a therapeutic agent, or
a cytotoxic agent.
[0158] Exemplary imaging agents useful in making sclerostin binding
protein are provided and include, but are not limited to, a
radiolabel, an enzyme, a fluorescent label, a luminescent label, a
bioluminescent label, a magnetic label, and biotin.
[0159] Exemplary radiolabels are provided and include, but are not
limited, to .sup.3H, .sup.14C .sup.35S, .sup.90Y, .sup.99Tc,
.sup.111In, .sup.125I, .sup.131I, .sup.177Lu, .sup.166Ho, or
.sup.153Sm.
[0160] In another embodiment, asclerostin binding protein conjugate
comprising a therapeutic or cytotoxic agent are provided, said
agent further comprising an anti-metabolite, an alkylating agent,
an antibiotic, a growth factor, a cytokine, an anti-angiogenic
agent, an anti-mitotic agent, an anthracycline, toxin, or an
apoptotic agent.
[0161] In another embodiment, binding proteins described herein
possesses a human glycosylation pattern.
[0162] In another embodiment, asclerostin binding protein described
herein, including sclerostin binding protein constructs and
sclerostin binding protein conjugates, may be in the form of a
crystallized binding protein. Exemplary crystalline forms retain at
least some of the biologically activity of the uncrystallized form
of asclerostin binding protein described herein. Such crystalline
forms may also be used as a carrier-free pharmaceutical controlled
release crystallized sclerostin binding proteins.
[0163] Another embodiment provides isolated nucleic acids encoding
sclerostin binding proteins, including binding protein constructs,
described herein. Such nucleic acids may be inserted into a vector
for carrying out various genetic analyses and recombinant
techniques for expressing, characterizing, or improving one or more
properties of asclerostin binding protein described herein.
Exemplary vectors for cloning nucleic acids encoding binding
proteins described herein include, but are not limited, pcDNA, pTT,
pTT3, pEFBOS, pBV, pJV, and pBJ.
[0164] A host cell comprising a vector comprising a nucleic acid
encoding a binding protein are provided and described herein. Host
cells are provided and may be prokaryotic or eukaryotic. An
exemplary prokaryotic host cell is Escherichia coli. Eukaryotic
cells useful as host cells are provided and include protist cell,
animal cell, plant cell, and fungal cell.
[0165] An exemplary fungal cell is a yeast cell, including
Saccharomyces cerevisiae. An exemplary animal cell useful as a host
cell is provided and includes, but is not limited to, a mammalian
cell, an avian cell, and an insect cell. Exemplary mammalian cells
include CHO and COS cells. An insect cell useful as a host cell is
provided and is an insect Sf9 cell.
[0166] A vector may comprise a nucleic acid encoding a sclerostin
binding protein described herein in which the nucleic acid is
operably linked to appropriate transcriptional and/or translational
sequences that permit expression of the binding protein in a
particular host cell carrying the vector.
[0167] Another aspect provides a method of producing asclerostin
binding protein comprising culturing a host cell comprising a
vector encoding the sclerostin binding protein in culture medium
under conditions sufficient to produce the binding protein capable
of binding sclerostin. The protein so produced can be isolated and
used in various compositions and methods described herein.
[0168] Compositions are provided and include a composition for the
release of a binding protein, wherein said composition comprises:
(a) a formulation, wherein said formulation comprises a
crystallized binding protein, described herein, and an ingredient;
and (b) at least one polymeric carrier.
[0169] Exemplary polymeric carriers useful in compositions are
provided and include, without limitation, one or more of the group
consisting of: poly (acrylic acid), poly (cyanoacrylates), poly
(amino acids), poly (anhydrides), poly (depsipeptide), poly
(esters), poly (lactic acid), poly (lactic-co-glycolic acid) or
PLGA, poly (b-hydroxybutryate), poly (caprolactone), poly
(dioxanone); poly (ethylene glycol), poly ((hydroxypropyl)
methacrylamide, poly [(organo) phosphazene], poly (ortho esters),
poly (vinyl alcohol), poly (vinylpyrrolidone), maleic
anhydride-alkyl vinyl ether copolymers, pluronic polyols, albumin,
alginate, cellulose and cellulose derivatives, collagen, fibrin,
gelatin, hyaluronic acid, oligosaccharides, glycaminoglycans,
sulfated polysaccharides, blends and copolymers thereof.
[0170] In another aspect, an ingredient of a composition is
provided, wherein the ingredient is albumin, sucrose, trehalose,
lactitol, gelatin, hydroxypropyl-.beta.-cyclodextrin,
methoxypolyethylene glycol or polyethylene glycol.
[0171] Another embodiment provides a method for treating a mammal
comprising the step of administering to the mammal an effective
amount of a composition described herein.
[0172] Pharmaceutical compositions comprising a sclerostin binding
protein described herein and a pharmaceutically acceptable carrier
are provided. A pharmaceutically acceptable carrier may also serve
as an adjuvant to increase the absorption or dispersion of the
sclerostin binding protein in a composition. An exemplary adjuvant
is hyaluronidase.
[0173] In another embodiment, a pharmaceutical composition further
comprises at least one additional therapeutic agent for treating a
disorder in which SOST activity is detrimental.
[0174] Another embodiment provides a method for reducing human SOST
activity comprising contacting human SOST with a sclerostin binding
protein herein such that human SOST activity is reduced.
[0175] In another embodiment, a pharmaceutical composition
comprising a sclerostin binding protein described herein comprises
at least one additional agent. In an embodiment, the additional
agent is a therapeutic agent; an imaging agent; a cytotoxic agent;
an angiogenesis inhibitors; a kinase inhibitors; a co-stimulation
molecule blockers; an adhesion molecule blockers; a anti-cytokine
antibody or functional fragment thereof; methotrexate; cyclosporin;
rapamycin; FK506; a detectable label or reporter; a TNF antagonist;
an antirheumatic; a muscle relaxant; a narcotic; a non-steroid
anti-inflammatory drug (NSAID); an analgesic; an anesthetic; a
sedative; a local anesthetic; a neuromuscular blocker; an
antimicrobial; an antipsoriatic; a corticosteroid; an anabolic
steroid; an erythropoietin; an immunization; an immunoglobulin; an
immunosuppressive; a growth hormone; a hormone replacement drug; a
radiopharmaceutical; an antidepressant; an antipsychotic; a
stimulant; an asthma medication; a beta agonist; an inhaled
steroid; an epinephrine or analog; a cytokine; or a cytokine
antagonist. Another embodiment provides a method for treating a
subject for a disease or a disorder by administering to the subject
a multivalent, multispecific DVD-binding protein described herein
that binds TNF-.alpha. and sclerostin such that treatment is
achieved.
[0176] In another embodiment, a disorder that may treated by a
method of administering to a subject a sclerostin binding protein
described herein is provided, wherein the disorder is rheumatoid
arthritis, osteoarthritis, juvenile chronic arthritis, septic
arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis,
spondyloarthropathy, systemic lupus erythematosus, Crohn's disease,
ulcerative colitis, inflammatory bowel disease, insulin dependent
diabetes mellitus, thyroiditis, asthma, allergic diseases,
psoriasis, dermatitis scleroderma, graft versus host disease, organ
transplant rejection, acute or chronic immune disease associated
with organ transplantation, sarcoidosis, atherosclerosis,
disseminated intravascular coagulation, Kawasaki's disease, Grave's
disease, nephrotic syndrome, chronic fatigue syndrome, Wegener's
granulomatosis, Henoch-Schoenlein purpurea, microscopic vasculitis
of the kidneys, chronic active hepatitis, uveitis, septic shock,
toxic shock syndrome, sepsis syndrome, cachexia, infectious
diseases, parasitic diseases, acquired immunodeficiency syndrome,
acute transverse myelitis, Huntington's chorea, Parkinson's
disease, Alzheimer's disease, stroke, primary biliary cirrhosis,
hemolytic anemia, malignancies, heart failure, myocardial
infarction, Addison's disease, sporadic, polyglandular deficiency
type I and polyglandular deficiency type II, Schmidt's syndrome,
adult (acute) respiratory distress syndrome, alopecia, alopecia
areata, seronegative arthropathy, arthropathy, Reiter's disease,
psoriatic arthropathy, ulcerative colitic arthropathy, enteropathic
synovitis, chlamydia, yersinia and salmonella associated
arthropathy, spondyloarthropathy, atheromatous
disease/arteriosclerosis, atopic allergy, autoimmune bullous
disease, pemphigus vulgaris, pemphigus foliaceus, pemphigoid,
linear IgA disease, autoimmune haemolytic anaemia, Coombs positive
haemolytic anaemia, acquired pernicious anaemia, juvenile
pernicious anaemia, myalgic encephalitis/Royal Free Disease,
chronic mucocutaneous candidiasis, giant cell arteritis, primary
sclerosing hepatitis, cryptogenic autoimmune hepatitis, Acquired
Immunodeficiency Disease Syndrome, Acquired Immunodeficiency
Related Diseases, Hepatitis B, Hepatitis C, common varied
immunodeficiency (common variable hypogammaglobulinaemia), dilated
cardiomyopathy, female infertility, ovarian failure, premature
ovarian failure, fibrotic lung disease, cryptogenic fibrosing
alveolitis, post-inflammatory interstitial lung disease,
interstitial pneumonitis, connective tissue disease associated
interstitial lung disease, mixed connective tissue disease
associated lung disease, systemic sclerosis associated interstitial
lung disease, rheumatoid arthritis associated interstitial lung
disease, systemic lupus erythematosus associated lung disease,
dermatomyositis/polymyositis associated lung disease, Sjogren's
disease associated lung disease, ankylosing spondylitis associated
lung disease, vasculitic diffuse lung disease, haemosiderosis
associated lung disease, drug-induced interstitial lung disease,
fibrosis, radiation fibrosis, bronchiolitis obliterans, chronic
eosinophilic pneumonia, lymphocytic infiltrative lung disease,
postinfectious interstitial lung disease, gouty arthritis,
autoimmune hepatitis, type-1 autoimmune hepatitis (classical
autoimmune or lupoid hepatitis), type-2 autoimmune hepatitis
(anti-LKM antibody hepatitis), autoimmune mediated hypoglycaemia,
type B insulin resistance with acanthosis nigricans,
hypoparathyroidism, acute immune disease associated with organ
transplantation, chronic immune disease associated with organ
transplantation, osteoarthrosis, primary sclerosing cholangitis,
psoriasis type 1, psoriasis type 2, idiopathic leucopaenia,
autoimmune neutropaenia, renal disease NOS, glomerulonephritides,
microscopic vasculitis of the kidneys, Lyme disease, discoid lupus
erythematosus, male infertility idiopathic or NOS, sperm
autoimmunity, multiple sclerosis (all subtypes), sympathetic
ophthalmia, pulmonary hypertension secondary to connective tissue
disease, Goodpasture's syndrome, pulmonary manifestation of
polyarteritis nodosa, acute rheumatic fever, rheumatoid
spondylitis, Still's disease, systemic sclerosis, Sjorgren's
syndrome, Takayasu's disease/arteritis, autoimmune
thrombocytopaenia, idiopathic thrombocytopaenia, autoimmune thyroid
disease, hyperthyroidism, goitrous autoimmune hypothyroidism
(Hashimoto's disease), atrophic autoimmune hypothyroidism, primary
myxoedema, phacogenic uveitis, primary vasculitis, vitiligo acute
liver disease, chronic liver diseases, alcoholic cirrhosis,
alcohol-induced liver injury, choleosatatis, idiosyncratic liver
disease, Drug-Induced hepatitis, Non-alcoholic Steatohepatitis,
allergy and asthma, group B streptococci (GBS) infection, mental
disorders (e.g., depression and schizophrenia), Th2 Type and Th1
Type mediated diseases, acute and chronic pain (different forms of
pain), and cancers such as lung, breast, stomach, bladder, colon,
pancreas, ovarian, prostate and rectal cancer and hematopoietic
malignancies (leukemia and lymphoma) Abetalipoproteinemia,
Acrocyanosis, acute and chronic parasitic or infectious processes,
acute leukemia, acute lymphoblastic leukemia (ALL), acute myeloid
leukemia (AML), acute or chronic bacterial infection, acute
pancreatitis, acute renal failure, adenocarcinomas, aerial ectopic
beats, AIDS dementia complex, alcohol-induced hepatitis, allergic
conjunctivitis, allergic contact dermatitis, allergic rhinitis,
allograft rejection, alpha-1-antitrypsin deficiency, amyotrophic
lateral sclerosis, anemia, angina pectoris, anterior horn cell
degeneration, anti-CD3 therapy, antiphospholipid syndrome,
anti-receptor hypersensitivity reactions, aortic and peripheral
aneurysms, aortic dissection, arterial hypertension,
arteriosclerosis, arteriovenous fistula, ataxia, atrial
fibrillation (sustained or paroxysmal), atrial flutter,
atrioventricular block, B cell lymphoma, bone graft rejection, bone
marrow transplant (BMT) rejection, bundle branch block, Burkitt's
lymphoma, Burns, cardiac arrhythmias, cardiac stun syndrome,
cardiac tumors, cardiomyopathy, cardiopulmonary bypass inflammation
response, cartilage transplant rejection, cerebellar cortical
degenerations, cerebellar disorders, chaotic or multifocal atrial
tachycardia, chemotherapy associated disorders, chronic myelocytic
leukemia (CML), chronic alcoholism, chronic inflammatory
pathologies, chronic lymphocytic leukemia (CLL), chronic
obstructive pulmonary disease (COPD), chronic salicylate
intoxication, colorectal carcinoma, congestive heart failure,
conjunctivitis, contact dermatitis, cor pulmonale, coronary artery
disease, Creutzfeldt-Jakob disease, culture negative sepsis, cystic
fibrosis, cytokine therapy associated disorders, Dementia
pugilistica, demyelinating diseases, dengue hemorrhagic fever,
dermatitis, dermatologic conditions, diabetes, diabetes mellitus,
diabetic arteriosclerotic disease, Diffuse Lewy body disease,
dilated congestive cardiomyopathy, disorders of the basal ganglia,
Down's Syndrome in middle age, drug-induced movement disorders
induced by drugs which block CNS dopamine receptors, drug
sensitivity, eczema, encephalomyelitis, endocarditis,
endocrinopathy, epiglottitis, Epstein-Barr virus infection,
erythromelalgia, extrapyramidal and cerebellar disorders, familial
hematophagocytic lymphohistiocytosis, fetal thymus implant
rejection, Friedreich's ataxia, functional peripheral arterial
disorders, fungal sepsis, gas gangrene, gastric ulcer, glomerular
nephritis, graft rejection of any organ or tissue, gram negative
sepsis, gram positive sepsis, granulomas due to intracellular
organisms, hairy cell leukemia, Hallervorden-Spatz disease,
Hashimoto's thyroiditis, hay fever, heart transplant rejection,
hemachromatosis, hemodialysis, hemolytic uremic
syndrome/thrombolytic thrombocytopenic purpura, hemorrhage,
hepatitis (A), His bundle arrhythmias, HIV infection/HIV
neuropathy, Hodgkin's disease, hyperkinetic movement disorders,
hypersensitivity reactions, hypersensitivity pneumonitis,
hypertension, hypokinetic movement disorders,
hypothalamic-pituitary-adrenal axis evaluation, idiopathic
Addison's disease, idiopathic pulmonary fibrosis, antibody mediated
cytotoxicity, Asthenia, infantile spinal muscular atrophy,
inflammation of the aorta, influenza a, ionizing radiation
exposure, iridocyclitis/uveitis/optic neuritis,
ischemia-reperfusion injury, ischemic stroke, juvenile rheumatoid
arthritis, juvenile spinal muscular atrophy, Kaposi's sarcoma,
kidney transplant rejection, legionella, leishmaniasis, leprosy,
lesions of the corticospinal system, lipedema, liver transplant
rejection, lymphedema, malaria, malignant Lymphoma, malignant
histiocytosis, malignant melanoma, meningitis, meningococcemia,
metabolic/idiopathic, migraine headache, mitochondrial multi-system
disorder, mixed connective tissue disease, monoclonal gammopathy,
multiple myeloma, multiple systems degenerations (Mencel
Dejerine-Thomas Shi-Drager and Machado-Joseph), myasthenia gravis,
mycobacterium avium intracellulare, mycobacterium tuberculosis,
myelodysplastic syndrome, myocardial infarction, myocardial
ischemic disorders, nasopharyngeal carcinoma, neonatal chronic lung
disease, nephritis, nephrosis, neurodegenerative diseases,
neurogenic I muscular atrophies, neutropenic fever, non-Hodgkins
lymphoma, occlusion of the abdominal aorta and its branches,
occlusive arterial disorders, OKT3.RTM. therapy,
orchitis/epidydimitis, orchitis/vasectomy reversal procedures,
organomegaly, osteoporosis, pancreas transplant rejection,
pancreatic carcinoma, paraneoplastic syndrome/hypercalcemia of
malignancy, parathyroid transplant rejection, pelvic inflammatory
disease, perennial rhinitis, pericardial disease, peripheral
atherosclerotic disease, peripheral vascular disorders,
peritonitis, pernicious anemia, pneumocystis carinii pneumonia,
pneumonia, POEMS syndrome (polyneuropathy, organomegaly,
endocrinopathy, monoclonal gammopathy, and skin changes syndrome),
post perfusion syndrome, post pump syndrome, post-MI cardiotomy
syndrome, preeclampsia, Progressive supranucleo Palsy, primary
pulmonary hypertension, radiation therapy, Raynaud's phenomenon and
disease, Raynaud's disease, Refsum's disease, regular narrow QRS
tachycardia, renovascular hypertension, reperfusion injury,
restrictive cardiomyopathy, sarcomas, scleroderma, senile chorea,
Senile Dementia of Lewy body type, seronegative arthropathies,
shock, sickle cell anemia, skin allograft rejection, skin changes
syndrome, small bowel transplant rejection, solid tumors, specific
arrhythmias, spinal ataxia, spinocerebellar degenerations,
streptococcal myositis, structural lesions of the cerebellum,
Subacute sclerosing panencephalitis, Syncope, syphilis of the
cardiovascular system, systemic anaphylaxis, systemic inflammatory
response syndrome, systemic onset juvenile rheumatoid arthritis,
T-cell or FAB ALL, Telangiectasia, thromboangitis obliterans,
thrombocytopenia, toxicity, transplants, trauma/hemorrhage, type
III hypersensitivity reactions, type IV hypersensitivity, unstable
angina, uremia, urosepsis, urticaria, valvular heart diseases,
varicose veins, vasculitis, venous diseases, venous thrombosis,
ventricular fibrillation, viral and fungal infections, viral
encephalitis/aseptic meningitis, viral-associated hemaphagocytic
syndrome, Wernicke-Korsakoff syndrome, Wilson's disease, xenograft
rejection of any organ or tissue, acute coronary syndromes, acute
idiopathic polyneuritis, acute inflammatory demyelinating
polyradiculoneuropathy, acute ischemia, adult Still's disease,
alopecia areata, anaphylaxis, anti-phospholipid antibody syndrome,
aplastic anemia, arteriosclerosis, atopic eczema, atopic
dermatitis, autoimmune dermatitis, autoimmune disorder associated
with streptococcus infection, autoimmune enteropathy, autoimmune
hearing loss, autoimmune lymphoproliferative syndrome (ALPS),
autoimmune myocarditis, autoimmune premature ovarian failure,
blepharitis, bronchiectasis, bullous pemphigoid, cardiovascular
disease, catastrophic antiphospholipid syndrome, celiac disease,
cervical spondylosis, chronic ischemia, cicatricial pemphigoid,
clinically isolated syndrome (cis) with risk for multiple
sclerosis, conjunctivitis, childhood onset psychiatric disorder,
chronic obstructive pulmonary disease (COPD), dacryocystitis,
dermatomyositis, diabetic retinopathy, diabetes mellitus, disk
herniation, disk prolapse, drug induced immune hemolytic anemia,
endocarditis, endometriosis, endophthalmitis, episcleritis,
erythema multiforme, erythema multiforme major, gestational
pemphigoid, Guillain-Barre syndrome (GBS), hay fever, Hughes
syndrome, idiopathic Parkinson's disease, idiopathic interstitial
pneumonia, IgE-mediated allergy, immune hemolytic anemia, inclusion
body myositis, infectious ocular inflammatory disease, inflammatory
demyelinating disease, inflammatory heart disease, inflammatory
kidney disease, IPF/UIP, iritis, keratitis, keratojunctivitis
sicca, Kussmaul disease or Kussmaul-Meier disease, Landry's
paralysis, Langerhan's cell histiocytosis, livedo reticularis,
macular degeneration, microscopic polyangiitis, morbus bechterev,
motor neuron disorders, mucous membrane pemphigoid, multiple organ
failure, myasthenia gravis, myelodysplastic syndrome, myocarditis,
nerve root disorders, neuropathy, non-A non-B hepatitis, optic
neuritis, osteolysis, ovarian cancer, pauciarticular JRA,
peripheral artery occlusive disease (PAOD), peripheral vascular
disease (PVD), peripheral artery, disease (PAD), phlebitis,
polyarteritis nodosa (or periarteritis nodosa), polychondritis,
polymyalgia rheumatica, poliosis, polyarticular JRA, polyendocrine
deficiency syndrome, polymyositis, polymyalgia rheumatica (PMR),
post-pump syndrome, primary Parkinsonism, prostate and rectal
cancer and hematopoietic malignancies (leukemia and lymphoma),
prostatitis, pure red cell aplasia, primary adrenal insufficiency,
recurrent neuromyelitis optica, restenosis, rheumatic heart
disease, sapho (synovitis, acne, pustulosis, hyperostosis, and
osteitis), scleroderma, secondary amyloidosis, shock lung,
scleritis, sciatica, secondary adrenal insufficiency, silicone
associated connective tissue disease, sneddon-wilkinson dermatosis,
spondylitis ankylosans, Stevens-Johnson syndrome (SJS), systemic
inflammatory response syndrome, temporal arteritis, toxoplasmic
retinitis, toxic epidermal necrolysis, transverse myelitis, TRAPS
(tumor necrosis factor receptor, type 1 allergic reaction, type II
diabetes, urticaria, usual interstitial pneumonia (UIP),
vasculitis, vernal conjunctivitis, viral retinitis,
Vogt-Koyanagi-Harada syndrome (VKH syndrome), wet macular
degeneration, wound healing, or yersinia and salmonella associated
arthropathy.
[0177] Another embodiment provides a method for treating a subject
for a disease or a disorder in which SOST activity is detrimental
by administering to the subject a sclerostin binding protein
described herein such that treatment is achieved. The method can be
used to a treat respiratory disorders; asthma; allergic and
nonallergic asthma; asthma due to infection; asthma due to
infection with respiratory syncytial virus (RSV); chronic
obstructive pulmonary disease (COPD); other conditions involving
airway inflammation; eosinophilia; fibrosis and excess mucus
production; cystic fibrosis; pulmonary fibrosis; atopic disorders;
atopic dermatitis; urticaria; eczema; allergic rhinitis; and
allergic enterogastritis; inflammatory and/or autoimmune conditions
of the skin; inflammatory and/or autoimmune conditions of
gastrointestinal organs; inflammatory bowel diseases (IBD);
ulcerative colitis; Crohn's disease; inflammatory and/or autoimmune
conditions of the liver; liver cirrhosis; liver fibrosis; liver
fibrosis caused by hepatitis B and/or C virus; scleroderma; tumors
or cancers; hepatocellular carcinoma; glioblastoma; lymphoma;
Hodgkin's lymphoma; viral infections; HTLV-1 infection (e.g., from
HTLV-1); suppression of expression of protective type 1 immune
responses, or suppression of expression of protective type 1 immune
responses during vaccination.
[0178] In a further embodiment of the above method, the
administering to the subject is by parenteral, subcutaneous,
intramuscular, intravenous, intra-articular, intrabronchial,
intraabdominal, intracapsular, intracartilaginous, intracavitary,
intracelial, intracerebellar, intracerebroventricular, intracolic,
intracervical, intragastric, intrahepatic, intramyocardial,
intraosteal, intrapelvic, intrapericardiac, intraperitoneal,
intrapleural, intraprostatic, intrapulmonary, intrarectal,
intrarenal, intraretinal, intraspinal, intrasynovial,
intrathoracic, intrauterine, intravesical, bolus, vaginal, rectal,
buccal, sublingual, intranasal, or transdermal.
[0179] A method of treating a patient suffering from a disorder in
which sclerostin is detrimental comprising the step of
administering asclerostin binding protein described herein before,
concurrent with, or after the administration of a second agent is
provided, wherein the second agent is inhaled steroids;
beta-agonists; short-acting or long-acting beta-agonists;
antagonists of leukotrienes or leukotriene receptors; ADVAIR; IgE
inhibitors; anti-IgE antibodies; XOLAIR; phosphodiesterase
inhibitors; PDE4 inhibitors; xanthines; anticholinergic drugs; mast
cell-stabilizing agents; Cromolyn; IL-4 inhibitors; IL-5
inhibitors; eotaxin/CCR3 inhibitors; antagonists of histamine or
its receptors including H1, H2, H3, and H4; antagonists of
prostaglandin D or its receptors DP1 and CRTH2; TNF antagonists; a
soluble fragment of a TNF receptor; ENBREL.RTM.; TNF enzyme
antagonists; TNF converting enzyme (TACE) inhibitors; muscarinic
receptor antagonists; TGF-beta antagonists; interferon gamma;
perfenidone; chemotherapeutic agents, methotrexate; leflunomide;
sirolimus (rapamycin) or an analog thereof, CCI-779; COX2 or cPLA2
inhibitors; NSAIDs; immunomodulators; p38 inhibitors; TPL-2, MK-2
and NFkB inhibitors; budenoside; epidermal growth factor;
corticosteroids; cyclosporine; sulfasalazine; aminosalicylates;
6-mercaptopurine; azathioprine; metronidazole; lipoxygenase
inhibitors; mesalamine; olsalazine; balsalazide; antioxidants;
thromboxane inhibitors; IL-1 receptor antagonists; anti-IL-1.beta.
antibodies; anti-IL-6 antibodies; growth factors; elastase
inhibitors; pyridinyl-imidazole compounds; antibodies or agonists
of TNF, LT, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9,
IL-10, IL-11, IL-12, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19,
IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28,
IL-29, IL-30, IL-31, IL-32, IL-33, EMAP-II, GM-CSF, FGF, or PDGF;
antibodies of CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45,
CD69, CD90 or their ligands; FK506; rapamycin; mycophenolate
mofetil; ibuprofen; prednisolone; phosphodiesterase inhibitors;
adenosine agonists; antithrombotic agents; complement inhibitors;
adrenergic agents; IRAK, NIK, IKK, p38, or MAP kinase inhibitors;
IL-1.beta. converting enzyme inhibitors; TNF-.alpha. converting
enzyme inhibitors; T-cell signaling inhibitors; metalloproteinase
inhibitors; 6-mercaptopurines; angiotensin converting enzyme
inhibitors; soluble cytokine receptors; soluble p55 TNF receptor;
soluble p75 TNF receptor; sIL-1RI; sIL-1RII; sIL-6R;
anti-inflammatory cytokines; IL-4; IL-10; IL-11; SOST; or
TGF-.beta..
BRIEF DESCRIPTION OF THE DRAWINGS
[0180] FIG. 1 contains panels A and B. Panel A is a schematic
representation of Dual Variable Domain (DVD) constructs and shows
the strategy for generation of a DVD-binding protein from two
parent antibodies.
[0181] FIG. 1 also includes panel B, which is a schematic
representation of constructs DVD1-Ig, DVD2-Ig, and two chimeric
mono-specific antibody clones.
[0182] FIG. 2A, FIG. 2B and FIG. 2C demonstrate the effect of
anti-TNF, anti-sclerostin, or combined therapies on paw swelling in
a mouse model of induced arthritis. FIG. 2A shows the change in paw
thickness when dosed with anti-mouse TNF mAb, anti-sclerostin mAb,
or a combination of both anti-mouse TNF mAb and anti-sclerostin
mAb. FIG. 2B shows the arthritic ankle bone volume when dosed with
anti-mouse TNF mAb, anti-sclerostin mAb, or a combination of both
anti-mouse TNF mAb and anti-sclerostin mAb. FIG. 2C shows the bone
density of trabecular bone in the lumbar spine (L5) when dosed with
anti-mouse TNF mAb, anti-sclerostin mAb, or a combination of both
anti-mouse TNF mAb and anti-sclerostin mAb.
[0183] FIG. 3 demonstrates the alignment of anti-SOST antibodies
(SEQ ID NOS 2066-2093).
[0184] FIG. 4A, FIG. 4B and FIG. 4C show the combined
neutralization of TNF and SOST in a late therapeutic mouse collagen
induced arthritis (CIA) model in which therapy began five days
after the onset of inflammation. FIG. 4A shows the change in paw
thickness when dosed with anti-mouse TNF mAb, anti-sclerostin mAb,
or a combination of both anti-mouse TNF mAb and anti-sclerostin
mAb. FIG. 4B shows the arthritic ankle bone volume when dosed with
anti-mouse TNF mAb, anti-sclerostin mAb, or a combination of both
anti-mouse TNF mAb and anti-sclerostin mAb. FIG. 4C shows the bone
density of trabecular bone in the lumbar spine (L5) when dosed with
anti-mouse TNF mAb, anti-sclerostin mAb, or a combination of both
anti-mouse TNF mAb and anti-sclerostin mAb.
[0185] FIG. 5A, FIG. 5B and FIG. 5C show the ability of sclerostin
inhibition to restore bone in the arthritic joint in mice in which
therapy began five days after the onset of inflammation. FIG. 5A
shows the change in paw thickness when dosed with an
anti-IL1.alpha. therapeutic and an anti-IL1.beta. therapeutic,
anti-sclerostin, or a combination of an anti-IL1.alpha. therapeutic
and an anti-IL1.beta. therapeutic and anti-sclerostin. FIG. 5B
shows the arthritic ankle bone volume when dosed with an
anti-IL1.alpha. therapeutic and an anti-IL1.beta. therapeutic,
anti-sclerostin, or a combination of an anti-IL1.alpha. therapeutic
and an anti-IL1.beta. therapeutic and anti-sclerostin. FIG. 5C
shows the bone density of trabecular bone in the lumbar spine (L5)
when dosed with an anti-IL1.alpha. therapeutic and an
anti-IL1.beta. therapeutic, anti-sclerostin, or a combination of an
anti-IL1.alpha. therapeutic and an anti-IL1.beta. therapeutic and
anti-sclerostin.
[0186] FIG. 6A and FIG. 6B show data from a mouse model of Crohn's
disease. FIG. 6A shows the colon inflammation score when mice are
dosed with anti-TNF mAb, anti-sclerostin mAb, or a combination of
anti-TNF mAb and anti-sclerostin mAb. FIG. 6B shows the bone
density of trabecular bone in the lumbar spine (L5) when mice are
dosed with anti-TNF mAb, anti-sclerostin mAb, or a combination of
anti-TNF mAb and anti-sclerostin mAb.
[0187] FIG. 7A-1, FIG. 7A-2, FIG. 7B-1, and FIG. 7B-2 show binding
profile data which demonstrate that exemplary Sclerostin/TNF
DVD-Igs bind sclerostin once saturated with TNF and vice versa.
DETAILED DESCRIPTION OF THE INVENTION
[0188] Sclerostin binding proteins, including, but not limited to,
anti-sclerostin antibodies, or antigen-binding portions thereof,
that bind sclerostin and multivalent, multispecific binding
proteins such as DVD-binding proteins that bind SOST and another
target are provided. Various aspects relating to antibodies and
antibody fragments, DVD-binding proteins, and pharmaceutical
compositions thereof, as well as nucleic acids, recombinant
expression vectors and host cells for making such sclerostin
binding proteins, including antibodies, DVD-binding proteins, and
fragments thereof are provided. Methods of using the sclerostin
binding proteins to detect human sclerostin, either in vitro or in
vivo; and to regulate gene expression are also provided.
[0189] Any binding protein or antibody capable of competing with a
sclerostin binding protein described herein are also provided.
[0190] Unless otherwise defined herein, scientific and technical
terms shall have the meanings that are commonly understood by those
of ordinary skill in the art. The meaning and scope of the terms
should be clear, however, in the event of any latent ambiguity,
definitions provided herein take precedent over any dictionary or
extrinsic definition. Further, unless otherwise required by
context, singular terms shall include pluralities and plural terms
shall include the singular. In this application, the use of "or"
means "and/or" unless stated otherwise. Furthermore, the use of the
term "including", as well as other forms, such as "includes" and
"included", is not limiting. Also, terms such as "element" or
"component" encompass both elements and components comprising one
unit and elements and components that comprise more than one
subunit unless specifically stated otherwise.
[0191] Generally, nomenclatures used in connection with, and
techniques of, cell and tissue culture, molecular biology,
immunology, microbiology, genetics and protein and nucleic acid
chemistry and hybridization described herein are those well known
and commonly used in the art. The methods and techniques provided
are generally performed according to conventional methods well
known in the art and as described in various general and more
specific references that are cited and discussed throughout the
present specification unless otherwise indicated. Enzymatic
reactions and purification techniques are performed according to
manufacturer's specifications, as commonly accomplished in the art
or as described herein. The nomenclatures used in connection with,
and the laboratory procedures and techniques of, analytical
chemistry, synthetic organic chemistry, and medicinal and
pharmaceutical chemistry described herein are those well known and
commonly used in the art. Standard techniques are used for chemical
syntheses, chemical analyses, pharmaceutical preparation,
formulation, and delivery, and treatment of patients.
[0192] That what is provided may be more readily understood, select
terms are defined below.
[0193] The term "polypeptide" refers to any polymeric chain of
amino acids. The terms "peptide" and "protein" are used
interchangeably with the term polypeptide and also refer to a
polymeric chain of amino acids. The term "polypeptide" encompasses
native or artificial proteins, protein fragments and polypeptide
analogs of a protein sequence. A polypeptide may be monomeric or
polymeric. Use of "polypeptide" herein is intended to encompass
polypeptide and fragments and variants (including fragments of
variants) thereof, unless otherwise contradicted by context. For an
antigenic polypeptide, a fragment of polypeptide optionally
contains at least one contiguous or nonlinear epitope of
polypeptide. The precise boundaries of the at least one epitope
fragment can be confirmed using ordinary skill in the art. The
fragment comprises at least about 5 contiguous amino acids, such as
at least about 10 contiguous amino acids, at least about 15
contiguous amino acids, or at least about 20 contiguous amino
acids. A variant of polypeptide is as described herein.
[0194] The term "isolated protein" or "isolated polypeptide" refers
to a protein or polypeptide that by virtue of its origin or source
of derivation is not associated with naturally associated
components that accompany it in its native state; is substantially
free of other proteins from the same species; is expressed by a
cell from a different species; or does not occur in nature. Thus, a
polypeptide that is chemically synthesized or synthesized in a
cellular system different from the cell from which it naturally
originates will be "isolated" from its naturally associated
components. A protein may also be rendered substantially free of
naturally associated components by isolation, using protein
purification techniques well known in the art.
[0195] The term "recovering" refers to the process of rendering a
chemical species such as a polypeptide substantially free of
naturally associated components by isolation, e.g., using protein
purification techniques well known in the art.
[0196] The terms "human sclerostin" or "human SOST" (abbreviated
herein as "hSOST") refer to a 24 KD protein, or active fragments
thereof, called sclerostin that has been classified as a member of
the DAN family of cysteine knot containing glycoproteins based on
sequence similarity (Avasian-Kretchmer (2004) Mol. Endocrinol.
8(1):1-12). Sclerostin is a negative regulator of bone formation
that inhibits osteoblast proliferation as well as differentiation
and suppresses mineralization of osteoblastic cells in vitro (Poole
et al. (2005) FASEB J. 19:1836-38; Winkler et al. (2005) J. Biol.
Chem. 280(4): 2498-2502). The term human "SOST" is intended to
include recombinant human sclerostin (rhSOST) which can be prepared
by standard recombinant expression methods. The sequence of human
SOST is shown in Table 2.
TABLE-US-00003 TABLE 2 Sequence of Human Sclerostin Sequence
Sequence Protein Identifier 123456789012345678901234567890 Human
Sclerostin SEQ ID QGWQAFKNDATEIIPELGEYPEPPPELENNKTMNRAE NO.: 1694
NGGRPPHHPFETKDVSEYSCREL HFTRYVTDGPCRSAKPVTELVCSGQCGPARLLPNAIG
RGKWWRPSGPDFRCIPDRYRAQR VQLLCPGGEAPRARKVRLVASCKCKRLTRFHNQSELK
DFGTEAARPQKGRKPRPRARSAK ANQAELENAY
[0197] "Biological activity" refers to all inherent biological
properties of the cytokine. Biological properties of sclerostin
include, but are not limited to, binding to an sclerostin
receptor.
[0198] The terms "specific binding" or "specifically binding" in
reference to the interaction of an antibody, a protein, or a
peptide with a second chemical species, mean that the interaction
is dependent upon the presence of a particular structure (e.g., an
antigenic determinant or epitope) on the chemical species; for
example, an antibody recognizes and binds to a specific protein
structure rather than to proteins generally. If an antibody is
specific for epitope "A", the presence of a molecule containing
epitope A (or free, unlabeled A), in a reaction containing labeled
"A" and the antibody, will reduce the amount of labeled A bound to
the antibody.
[0199] The term "antibody" broadly refers to any immunoglobulin
(Ig) molecule comprised of four polypeptide chains, two heavy (H)
chains and two light (L) chains, or any functional fragment,
mutant, variant, or derivation thereof, which retains the essential
epitope binding features of an Ig molecule. Such mutant, variant,
or derivative antibody formats are known in the art. Nonlimiting
embodiments of which are discussed below.
[0200] In a full-length antibody, each heavy chain is comprised of
a heavy chain variable region (abbreviated herein as HCVR or VH)
and a heavy chain constant region. The heavy chain constant region
is comprised of three domains: CH1, CH2, and CH3. Each light chain
is comprised of a light chain variable region (abbreviated herein
as LCVR or VL) and a light chain constant region. The light chain
constant region is comprised of one domain, CL. The VH and VL
regions can be further subdivided into regions of hypervariability,
termed complementarity determining regions (CDR), interspersed with
regions that are more conserved, termed framework regions (FR).
Each VH and VL is composed of three CDRs and four FRs, arranged
from amino-terminus to carboxy-terminus in the following order:
FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Immunoglobulin molecules can
be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class
(e.g., IgG 1, IgG2, IgG 3, IgG4, IgA1 and IgA2) or subclass.
[0201] The term "Fc region" is used to define the C-terminal region
of an immunoglobulin heavy chain, which may be generated by papain
digestion of an intact antibody. The Fc region may be a native
sequence Fc region or a variant Fc region. The Fc region of an
immunoglobulin generally comprises two constant domains, a CH2
domain, and a CH3 domain, and optionally comprises a CH4 domain.
Replacements of amino acid residues in the Fc portion to alter
antibody effector function are known in the art (U.S. Pat. Nos.
5,648,260 and 5,624,821). The Fc portion of an antibody mediates
several important effector functions, for example, cytokine
induction, ADCC, phagocytosis, complement dependent cytotoxicity
(CDC), and half-life/clearance rate of antibody and
antigen-antibody complexes. In some cases these effector functions
are desirable for therapeutic antibody but in other cases might be
unnecessary or even deleterious, depending on the therapeutic
objectives. Certain human IgG isotypes, particularly IgG1 and IgG3,
mediate ADCC and CDC via binding to Fc.gamma.Rs and complement C1q,
respectively. Neonatal Fc receptors (FcRn) are the critical
components determining the circulating half-life of antibodies. In
still another embodiment at least one amino acid residue is
replaced in the constant region of the antibody, for example the Fc
region of the antibody, such that effector functions of the
antibody are altered. The dimerization of two identical heavy
chains of an immunoglobulin is mediated by the dimerization of CH3
domains and is stabilized by the disulfide bonds within the hinge
region (Huber et al. Nature 264:415-20; Thies et al. (1999) J. Mol.
Biol. 293:67-79.). Mutation of cysteine residues within the hinge
regions to prevent heavy chain-heavy chain disulfide bonds will
destabilize dimeration of CH3 domains. Residues responsible for CH3
dimerization have been identified (Dall'Acqua (1998) Biochem.
37:9266-9273.). Therefore, it is possible to generate a monovalent
half-Ig. Interestingly, these monovalent half Ig molecules have
been found in nature for both IgG and IgA subclasses (Seligman
(1978) Ann Immunol. 129:855-70; Biewenga et al. (1983) Clin. Exp.
Immunol. 51: 395-400). The stoichiometry of FcRn: Ig Fc region has
been determined to be 2:1 (West et al. (2000) Biochem. 39:
9698-708), and half Fc is sufficient for mediating FcRn binding
(Kim et al. (1994) Eur. J. Immunol. 24: 542-548). Mutations to
disrupt the dimerization of CH3 domain may not have greater adverse
effect on its FcRn binding as the residues important for CH3
dimerization are located on the inner interface of CH3 b sheet
structure, whereas the region responsible for FcRn binding is
located on the outside interface of CH2-CH3 domains. However, the
half Ig molecule may have certain advantage in tissue penetration
due to its smaller size than that of a regular antibody. In one
provided embodiment, at least one amino acid residue is replaced in
the constant region of the binding protein, for example the Fc
region, such that the dimerization of the heavy chains is
disrupted, resulting in half DVD Ig molecules. The
anti-inflammatory activity of IgG is completely dependent on
sialylation of the N-linked glycan of the IgG Fc fragment. The
precise glycan requirements for anti-inflammatory activity has been
determined, such that an appropriate IgG1 Fc fragment can be
created, thereby generating a fully recombinant, sialylated IgG1 Fc
with greatly enhanced potency (Anthony et al. (2008) Science
320:373-376).
[0202] The term "antigen-binding portion" of an antibody (or simply
"antibody portion") refers to one or more fragments of an antibody
that retain the ability to specifically bind to an antigen (e.g.,
human sclerostin (hSOST)). It has been shown that the
antigen-binding function of an antibody can be performed by
fragments of a full-length antibody. Such antibody embodiments may
also be bispecific, dual specific, or multi-specific formats;
specifically binding to two or more different antigens. Examples of
binding fragments encompassed within the term "antigen-binding
portion" of an antibody include (i) a Fab fragment, a monovalent
fragment consisting of the VL, VH, CL, and CH1 domains; (ii) a
F(ab').sub.2 fragment, a bivalent fragment comprising two Fab
fragments linked by a disulfide bridge at the hinge region; (iii) a
Fd fragment consisting of the VH and CH1 domains; (iv) a Fv
fragment consisting of the VL and VH domains of a single arm of an
antibody, (v) a dAb fragment (Ward et al. (1989) Nature
341:544-546, PCT Publication No. WO 90/05144), which comprises a
single variable domain; and (vi) an isolated complementarity
determining region (CDR). Furthermore, although the two domains of
the Fv fragment, VL and VH, are coded for by separate genes, they
can be joined, using recombinant methods, by a synthetic linker
that enables them to be made as a single protein chain in which the
VL and VH regions pair to form monovalent molecules (known as
single chain Fv (scFv); see, for example, Bird et al. (1988)
Science 242: 423-426 and Huston et al. (1988) Proc. Natl. Acad.
Sci. USA 85: 5879-5883). Such single chain antibodies are also
intended to be encompassed within the term "antigen-binding
portion" of an antibody. Other forms of single chain antibodies,
such as diabodies are also encompassed. Diabodies are bivalent,
bispecific antibodies in which VH and VL domains are expressed on a
single polypeptide chain, but using a linker that is too short to
allow for pairing between the two domains on the same chain,
thereby forcing the domains to pair with complementary domains of
another chain and creating two antigen binding sites (see, for
example, Holliger, et al. (1993) Proc. Natl. Acad. Sci. USA 90:
6444-6448; Poljak et al. (1994) Structure 2: 1121-1123). Such
antibody binding portions are known in the art (Kontermann and
Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York.
790 pp. (ISBN 3-540-41354-5)). In addition single chain antibodies
also include "linear antibodies" comprising a pair of tandem Fv
segments (VH-CH1-VH-CH1) which, together with complementary light
chain polypeptides, form a pair of antigen binding regions (Zapata
et al. (1995) Protein Eng. 8(10):1057-1062; and U.S. Pat. No.
5,641,870).
[0203] An immunoglobulin constant (C) domain refers to a heavy (CH)
or light (CL) chain constant domain. Murine and human IgG heavy
chain and light chain constant domain amino acid sequences are
known in the art.
[0204] The term "binding protein construct" refers to a polypeptide
comprising one or more of the antigen binding portions linked to a
linker polypeptide or an immunoglobulin constant domain. Linker
polypeptides comprise two or more amino acid residues joined by
peptide bonds and are used to link one or more antigen binding
portions. Such linker polypeptides are well known in the art (see
e.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:
6444-6448; Poljak et al. (1994) Structure 2: 1121-1123). An
immunoglobulin constant domain refers to a heavy or light chain
constant domain. Human IgG heavy chain and light chain constant
domain amino acid sequences are provided in Table 3.
[0205] The VH and VL domain sequences provided below comprise
complementarity determining region (CDR) and framework sequences
that are either known in the art or readily discernable using
methods known in the art. In some embodiments, one or more of these
CDR and/or framework sequences are replaced, without loss or
function, by other CDR and/or framework sequences from binding
proteins that are known in the art to bind to the same antigen.
TABLE-US-00004 TABLE 3 Sequence of Human IgG Heavy Chain Constant
Domain and Light Chain Constant Domain Sequence Sequence Protein
Identifier 123456789012345678901234567890 Ig gamma-1 SEQ ID
ASTKGPSVFFLAPSSKSTSGGTAALGCLVKDYFPEPV constant region NO.: 2060
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD
VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Ig gamma-1 SEQ ID NO.:
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV constant region 2061
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS QL
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDQLMISRTPEVTCVVVD
VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK Ig gamma-1 SEQ ID NO.:
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV constant region 2062
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS mutant
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD
VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Ig gamma-1 SEQ ID NO.:
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV constant region 2063
TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS QL mutant
SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVD
VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV
VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA
KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD
KSRWQQGNVFSCSVLHEALHNHYTQKSLSLSPGK Ig Kappa constant SEQ ID NO.:
TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK region 2064
VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS
KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Ig Lambda SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAV constant region NO.: 2065
TVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT
PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS
Still further, a sclerostin binding protein, such as an antibody or
antigen-binding portion thereof, may be part of a larger
immunoadhesion molecule, formed by covalent or noncovalent
association of the antibody or antibody portion with one or more
other proteins or peptides. Examples of such immunoadhesion
molecules include use of the streptavidin core region to make a
tetrameric scFv molecule (Kipriyanov et al. (1995) Human Antibod.
Hybridomas 6: 93-101) and use of a cysteine residue, a marker
peptide and a C-terminal polyhistidine tag to make bivalent and
biotinylated scFv molecules (Kipriyanov et al. (1994) Mol. Immunol.
31: 1047-1058). Antibody portions, such as Fab and F(ab').sub.2
fragments, can be prepared from whole antibodies using conventional
techniques, such as papain or pepsin digestion, respectively, of
whole antibodies. Moreover, antibodies, antibody portions and
immunoadhesion molecules can be obtained using standard recombinant
DNA techniques, as described herein.
[0206] The term "isolated antibody" refers to an antibody that is
substantially free of other antibodies having different antigenic
specificities (e.g., an isolated antibody that specifically binds
hSOST is substantially free of antibodies that specifically bind
antigens other than hSOST). An isolated antibody that specifically
binds hSOST may, however, have cross-reactivity to other antigens,
such as sclerostin molecules from other species. Moreover, an
isolated antibody may be substantially free of other cellular
material and/or chemicals.
[0207] The term "monoclonal antibody" or "mAb" refers to an
antibody obtained from a population of substantially homogeneous
antibodies, i.e., the individual antibodies comprising the
population are identical except for possible naturally occurring
mutations that may be present in minor amounts. Monoclonal
antibodies are highly specific, being directed against a single
antigen. Furthermore, in contrast to polyclonal antibody
preparations that typically include different antibodies directed
against different determinants (epitopes), each mAb is directed
against a single determinant on the antigen. The modifier
"monoclonal" is not to be construed as requiring production of the
antibody by any particular method.
[0208] The term "human antibody" is intended to include antibodies
having variable and constant regions derived from human germline
immunoglobulin sequences. Human antibodies are provided and may
include amino acid residues not encoded by human germline
immunoglobulin sequences (e.g., mutations introduced by random or
site-specific mutagenesis in vitro or by somatic mutation in vivo),
for example in the CDRs and in particular CDR3. However, the term
"human antibody" is not intended to include antibodies in which CDR
sequences derived from the germline of another mammalian species,
such as a mouse, have been grafted onto human framework
sequences.
[0209] The term "recombinant human antibody" is intended to include
all human antibodies that are prepared, expressed, created or
isolated by recombinant means, such as antibodies expressed using a
recombinant expression vector transfected into a host cell
(described further in Section II C, below), antibodies isolated
from a recombinant, combinatorial human antibody library
(Hoogenboom (1997) TIB Tech. 15:62-70; Azzazy and Highsmith (2002)
Clin. Biochem. 35: 425-445; Gavilondo and Larrick (2002)
BioTechniques 29: 128-145; Hoogenboom and Chames (2000) Immunol.
Today 21: 371-378), antibodies isolated from an animal (e.g., a
mouse) that is transgenic for human immunoglobulin genes (see,
e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295;
Kellermann and Green (2002) Curr. Opin. Biotechnol. 13:593-597;
Little et al (2000) Immunol. Today 21: 364-370) or antibodies
prepared, expressed, created or isolated by any other means that
involves splicing of human immunoglobulin gene sequences to other
DNA sequences. Such recombinant human antibodies have variable and
constant regions derived from human germline immunoglobulin
sequences. In certain embodiments, however, such recombinant human
antibodies are subjected to in vitro mutagenesis (or, when an
animal transgenic for human Ig sequences is used, in vivo somatic
mutagenesis) and thus the amino acid sequences of the VH and VL
regions of the recombinant antibodies are sequences that, while
derived from and related to human germline VH and VL sequences, may
not naturally exist within the human antibody germline repertoire
in vivo.
[0210] The term "chimeric antibody" refers to antibodies that
comprise heavy and light chain variable region sequences from one
species and constant region sequences from another species, such as
antibodies having murine heavy and light chain variable regions
linked to human constant regions.
[0211] The term "CDR-grafted antibody" refers to antibodies that
comprise heavy and light chain variable region sequences from one
species but in which the sequences of one or more of the CDR
regions of VH and/or VL are replaced with CDR sequences of another
species, such as antibodies having murine heavy and light chain
variable regions in which one or more of the murine CDRs (e.g.,
CDR3) has been replaced with human CDR sequences.
[0212] The terms "Kabat numbering", "Kabat definitions", and "Kabat
labeling" are used interchangeably herein. These terms, which are
recognized in the art, refer to a system of numbering amino acid
residues which are more variable (i.e., hypervariable) than other
amino acid residues in the heavy and light chain variable regions
of an antibody, or an antigen binding portion thereof (Kabat et al.
(1971) Ann. NY Acad. Sci. 190:382-391 and Kabat, E. A., et al.
(1991) Sequences of Proteins of Immunological Interest, Fifth
Edition, U.S. Department of Health and Human Services, NIH
Publication No. 91-3242). For the heavy chain variable region, the
hypervariable region ranges from amino acid positions 31 to 35 for
CDR1, amino acid positions 50 to 65 for CDR2, and amino acid
positions 95 to 102 for CDR3. For the light chain variable region,
the hypervariable region ranges from amino acid positions 24 to 34
for CDR1, amino acid positions 50 to 56 for CDR2, and amino acid
positions 89 to 97 for CDR3.
[0213] The term "CDR" refers to the complementarity determining
region within antibody variable sequences. There are three CDRs in
each of the variable regions of the heavy chain and the light
chain, which are designated CDR1, CDR2 and CDR3, for each of the
variable regions. The term "CDR set" refers to a group of three
CDRs that occur in a single variable region capable of binding the
antigen. The exact boundaries of these CDRs have been defined
differently according to different systems. The system described by
Kabat (Kabat et al., Sequences of Proteins of Immunological
Interest (National Institutes of Health, Bethesda, Md. (1987) and
(1991)) not only provides an unambiguous residue numbering system
applicable to any variable region of an antibody, but also provides
precise residue boundaries defining the three CDRs. These CDRs may
be referred to as Kabat CDRs. Chothia and coworkers (Chothia and
Lesk (1987) J. Mol. Biol. 196:901-917 and Chothia et al. (1989)
Nature 342:877-883) found that certain sub-portions within Kabat
CDRs adopt nearly identical peptide backbone conformations, despite
having great diversity at the level of amino acid sequence. These
sub-portions were designated as L1, L2, and L3 or H1, H2, and H3
where the "L" and the "H" designates the light chain and the heavy
chains regions, respectively. These regions may be referred to as
Chothia CDRs, which have boundaries that overlap with Kabat CDRs.
Other boundaries defining CDRs overlapping with the Kabat CDRs have
been described by Padlan (1995). The methods used herein may
utilize CDRs defined according to any of these systems, although
certain embodiments use Kabat or Chothia defined CDRs.
[0214] The term "canonical residue" refers to a residue in a CDR or
framework that defines a particular canonical CDR structure as
defined by Chothia et al. (1987) J. Mol. Biol. 196:901-907; Chothia
et al. (1992) J. Mol. Biol. 227:799). According to Chothia et al.,
critical portions of the CDRs of many antibodies have nearly
identical peptide backbone confirmations despite great diversity at
the level of amino acid sequence. Each canonical structure
specifies primarily a set of peptide backbone torsion angles for a
contiguous segment of amino acid residues forming a loop.
[0215] An "affinity matured" antibody is an antibody with one or
more alterations in one or more CDRs thereof which result in an
improvement in the affinity of the antibody for a target antigen,
compared to a parent antibody which does not possess the
alteration(s). Exemplary affinity matured antibodies will have
nanomolar or even picomolar affinities for the target antigen. A
variety of procedures for producing affinity matured antibodies are
known in the art. For example, Marks et al. (1992) Bio/Technol. 10:
779-783 describes affinity maturation by VH and VL domain
shuffling. Random mutagenesis of CDR and/or framework residues is
described by Barbas et al. (1994) Proc. Nat. Acad. Sci. USA
91:3809-3813; Schier et al. (1995) Gene 169:147-155; Yelton et al.
(1995) J. Immunol. 155:1994-2004; Jackson et al. (1995) J. Immunol.
154(7):3310-3319; Hawkins et al. (1992) J. Mol. Biol. 226:889-896.
Selective mutation at selective mutagenesis positions and at
contact or hypermutation positions with an activity enhancing amino
acid residue is described in U.S. Pat. No. 6,914,128.
[0216] The term "multivalent binding protein" denotes a binding
protein comprising two or more antigen binding sites. In an
embodiment, multivalent binding protein is engineered to have three
or more antigen binding sites, and is generally not a naturally
occurring antibody. The term "multispecific binding protein" refers
to a binding protein capable of binding two or more related or
unrelated targets. "Dual variable domain" ("DVD") binding proteins
are provided and comprise two or more antigen binding sites and are
tetravalent or multivalent binding proteins. DVDs may be
monospecific, i.e., capable of binding one antigen, or
multispecific, i.e., capable of binding two or more antigens. A
"DVD binding protein" comprises two heavy chain DVD polypeptides
and two light chain DVD polypeptides. Each half of a DVD-binding
protein comprises a heavy chain DVD polypeptide and a light chain
DVD polypeptide, and two or more antigen binding sites. Each
binding site comprises a heavy chain variable domain and a light
chain variable domain with a total of six CDRs involved in antigen
binding per antigen binding site. DVD binding proteins are also
known as DVD-Ig.TM. molecules.
[0217] A description of the design, expression, and
characterization of DVD-binding proteins is provided in PCT
Publication No. WO 2007/024715, U.S. Pat. No. 7,612,181, and Wu et
al. (2007) Nature Biotech. 25:1290-1297. An example of such
DVD-binding proteins comprises a heavy chain that comprises the
structural formula VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first
heavy chain variable domain, VD2 is a second heavy chain variable
domain, C is a heavy chain constant domain, X1 is a linker with the
proviso that it is not CH1, X2 is an Fc region, and n is 0 or 1,
but, in an embodiment, 1; and a light chain that comprises the
structural formula VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first
light chain variable domain, VD2 is a second light chain variable
domain, C is a light chain constant domain, X1 is a linker with the
proviso that it is not CL, and X2 does not comprise an Fc region;
and n is 0 or 1, but, in an embodiment, 1. Such a DVD-binding
protein may comprise two such heavy chains and two such light
chains, wherein each chain comprises variable domains linked in
tandem without an intervening constant region between variable
regions, wherein a heavy chain and a light chain associate to form
tandem functional antigen binding sites, and a pair of heavy and
light chains may associate with another pair of heavy and light
chains to form a tetrameric binding protein with four functional
antigen binding sites. In another example, a DVD-binding protein
may comprise heavy and light chains that each comprise three
variable domains (VD1, VD2, VD3) linked in tandem without an
intervening constant region between variable domains, wherein a
pair of heavy and light chains may associate to form three antigen
binding sites, and wherein a pair of heavy and light chains may
associate with another pair of heavy and light chains to form a
tetrameric binding protein with six antigen binding sites.
[0218] A DVD-binding protein may bind one or more epitopes of
sclerostin. A DVD-binding protein may also bind an epitope of
sclerostin and an epitope of a second target antigen other than a
sclerostin polypeptide.
[0219] The term "bispecific antibody" refers to full-length
antibodies that are generated by quadroma technology (see Milstein
and Cuello (1983) Nature 305(5934):537-40), by chemical conjugation
of two different monoclonal antibodies (see Staerz et al. (1985)
Nature 314(6012): 628-31), or by knob-into-hole or similar
approaches which introduces mutations in the Fc region (see
Holliger et al. (1993) Proc. Natl. Acad. Sci. USA
90(14):6444-6448), resulting in multiple different immunoglobulin
species of which only one is the functional bispecific antibody. By
molecular function, a bispecific antibody binds one antigen (or
epitope) on one of its two binding arms (one pair of HC/LC), and
binds a different antigen (or epitope) on its second arm (a
different pair of HC/LC). By this definition, a bispecific antibody
has two distinct antigen binding arms (in both specificity and CDR
sequences), and is monovalent for each antigen to which it
binds.
[0220] The term "dual-specific antibody" refers to full-length
antibodies that can bind two different antigens (or epitopes) in
each of its two binding arms (a pair of HC/LC) (see PCT Publication
No. WO 02/02773). Accordingly a dual-specific binding protein has
two identical antigen binding arms, with identical specificity and
identical CDR sequences, and is bivalent for each antigen to which
it binds.
[0221] A "functional antigen binding site" of a binding protein is
one that is capable of binding a target antigen. The antigen
binding affinity of the antigen binding site is not necessarily as
strong as the parent antibody from which the antigen binding site
is derived, but the ability to bind antigen must be measurable
using any one of a variety of methods known for evaluating antibody
binding to an antigen. Moreover, the antigen binding affinity of
each of the antigen binding sites of a multivalent antibody herein
need not be quantitatively the same.
[0222] The term "cytokine" is a generic term for proteins that are
released by one cell population and that act on another cell
population as intercellular mediators. Examples of such cytokines
are lymphokines, monokines, and traditional polypeptide hormones.
Included among the cytokines are growth hormones, such as human
growth hormone, N-methionyl human growth hormone, and bovine growth
hormone; parathyroid hormone; thyroxine; insulin; proinsulin;
relaxin; prorelaxin; glycoprotein hormones, such as follicle
stimulating hormone (FSH), thyroid stimulating hormone (TSH), and
luteinizing hormone (LH); hepatic growth factor; fibroblast growth
factor; prolactin; placental lactogen; a tumor necrosis factor such
as tumor necrosis factor-alpha (TNF-.alpha.) and tumor necrosis
factor-beta (TNF-.beta.); mullerian-inhibiting substance; mouse
gonadotropin-associated peptide; inhibin; activin; vascular
endothelial growth factor; integrin; thrombopoietin (TPO); nerve
growth factors such as NGF-alpha (NGF-.alpha.); platelet-growth
factor; placental growth factor; transforming growth factors (TGFs)
such as TGF-alpha (TGF-.alpha.) and TGF-beta (TGF-.beta.);
insulin-like growth factor-1 and -11; erythropoietin (EPO);
osteoinductive factors; interferons such as interferon-alpha
(IFN-.alpha.), interferon-beta (IFN-.beta.), and interferon-gamma
(IFN-.gamma.); colony stimulating factors (CSFs) such as
macrophage-CSF (M-CSF); granulocyte macrophage-CSF (GM-CSF); and
granulocyte-CSF (G-CSF); interleukins (ILs) such as IL-1, IL-2,
IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12,
IL-13, IL-15, IL-17, IL-18, IL-21, IL-22, IL-23, IL-33; and other
polypeptide factors including LIF and kit ligand (KL). The term
cytokine includes proteins from natural sources or from recombinant
cell culture and biologically active equivalents of the native
sequence cytokines.
[0223] The terms "donor" and "donor antibody" refer to an antibody
providing one or more CDRs. In an embodiment, the donor antibody is
an antibody from a species different from the antibody from which
the framework regions are obtained or derived. In the context of a
humanized antibody, the term "donor antibody" refers to a non-human
antibody providing one or more CDRs.
[0224] The terms "framework" and "framework sequence" refers to the
remaining sequences of a variable region minus the CDRs. Because
the exact definition of a CDR sequence can be determined by
different systems, the meaning of a framework sequence is subject
to correspondingly different interpretations. The six CDRs (CDR-L1,
-L2, and -L3 of light chain and CDR-H1, -H2, and -H3 of heavy
chain) also divide the framework regions on the light chain and the
heavy chain into four sub-regions (FR1, FR2, FR3 and FR4) on each
chain, in which CDR1 is positioned between FR1 and FR2, CDR2
between FR2 and FR3, and CDR3 between FR3 and FR4. Without
specifying the particular sub-regions as FR1, FR2, FR3 or FR4, a
framework region, as referred by others, represents the combined
FR's within the variable region of a single, naturally occurring
immunoglobulin chain. An FR represents one of the four sub-regions,
and FRs represents two or more of the four sub-regions constituting
a framework region.
[0225] The terms "acceptor" and "acceptor antibody" refer to the
antibody providing or nucleic acid sequence encoding at least 80%,
at least 85%, at least 90%, at least 95%, at least 98%, or 100% of
the amino acid sequences of one or more of the framework regions.
In some embodiments, the term "acceptor" refers to the antibody
amino acid providing or nucleic acid sequence encoding the constant
region(s). In yet another embodiment, the term "acceptor" refers to
the antibody amino acid providing or nucleic acid sequence encoding
one or more of the framework regions and the constant region(s). In
a specific embodiment, the term "acceptor" refers to a human
antibody amino acid or nucleic acid sequence that provides or
encodes at least 80%, in an embodiment, at least 85%, at least 90%,
at least 95%, at least 98%, or 100% of the amino acid sequences of
one or more of the framework regions. In accordance with this
embodiment, an acceptor may contain at least 1, at least 2, at
least 3, least 4, at least 5, or at least 10 amino acid residues
that does (do) not occur at one or more specific positions of a
human antibody. An acceptor framework region and/or acceptor
constant region(s) may be, e.g., derived or obtained from a
germline antibody gene, a mature antibody gene, a functional
antibody (e.g., antibodies well known in the art, antibodies in
development, or antibodies commercially available).
[0226] Human heavy chain and light chain acceptor sequences are
known in the art. In one embodiment, human heavy chain and light
chain acceptor sequences from V-base (hvbase.mrc-cpe.cam.ac.uk/) or
from IMGT.RTM., the international ImMunoGeneTics information
System.RTM. (himgt.cines.fr/textes/IMGTrepertoire/LocusGenes/) are
provided. The terms "germline antibody gene" or "gene fragment"
refers to an immunoglobulin sequence encoded by non-lymphoid cells
that have not undergone the maturation process that leads to
genetic rearrangement and mutation for expression of a particular
immunoglobulin. (See, e.g., Shapiro et al. (2002) Crit. Rev.
Immunol. 22(3): 183-200; Marchalonis et al. (2001) Adv. Exp. Med.
Biol. 484:13-30). One of the advantages provided by various
embodiments stems from the recognition that germline antibody genes
are more likely than mature antibody genes to conserve essential
amino acid sequence structures characteristic of individuals in the
species, hence less likely to be recognized as from a foreign
source when used therapeutically in that species.
[0227] The terms "key" residues refer to certain residues within
the variable region that have more impact on the binding
specificity and/or affinity of an antibody, in particular a
humanized antibody. A key residue includes, but is not limited to,
one or more of the following: a residue that is adjacent to a CDR,
a potential glycosylation site (can be either N- or O-glycosylation
site), a rare residue, a residue capable of interacting with the
antigen, a residue capable of interacting with a CDR, a canonical
residue, a contact residue between heavy chain variable region and
light chain variable region, a residue within the Vernier zone, and
a residue in the region that overlaps between the Chothia
definition of a variable heavy chain CDR1 and the Kabat definition
of the first heavy chain framework.
[0228] The terms "humanized antibody" refers to antibodies that
comprise heavy and light chain variable region sequences from a
non-human species (e.g., a mouse) but in which at least a portion
of the VH and/or VL sequence has been altered to be more
"human-like", i.e., more similar to human germline variable
sequences. One type of humanized antibody is a CDR-grafted
antibody, in which human CDR sequences are introduced into
non-human VH and VL sequences to replace the corresponding nonhuman
CDR sequences. Also "humanized antibody" is an antibody or a
variant, derivative, analog or fragment thereof which
immunospecifically binds to an antigen of interest and which
comprises a framework (FR) region having substantially the amino
acid sequence of a human antibody and a complementary determining
region (CDR) having substantially the amino acid sequence of a
non-human antibody. The term "substantially" in the context of a
CDR refers to a CDR having an amino acid sequence at least 80%, at
least 85%, at least 90%, at least 95%, at least 98% or at least 99%
identical to the amino acid sequence of a non-human antibody CDR. A
humanized antibody comprises substantially all of at least one, and
typically two, variable domains (Fab, Fab', F(ab')2, FabC, Fv) in
which all or substantially all of the CDR regions correspond to
those of a non-human immunoglobulin (i.e., donor antibody) and all
or substantially all of the framework regions are those of a human
immunoglobulin consensus sequence. In an embodiment, a humanized
antibody also comprises at least a portion of an immunoglobulin
constant region (Fc), typically that of a human immunoglobulin. In
some embodiments, a humanized antibody contains both the light
chain as well as at least the variable domain of a heavy chain. The
antibody also may include the CH1, hinge, CH2, CH3, and CH4 regions
of the heavy chain. In some embodiments, a humanized antibody only
contains a humanized light chain. In some embodiments, a humanized
antibody only contains a humanized heavy chain. In specific
embodiments, a humanized antibody only contains a humanized
variable domain of a light chain and/or humanized heavy chain.
[0229] A humanized antibody may be from any class of
immunoglobulins, including IgM, IgG, IgD, IgA or IgE, or any
isotype including without limitation IgG1, IgG2, IgG3, or IgG4. The
humanized antibody may comprise sequences from more than one class
or isotype, and particular constant domains may be selected to
optimize desired effector functions using techniques well known in
the art.
[0230] The framework and CDR regions of a humanized antibody need
not correspond precisely to the parental sequences, e.g., the donor
antibody CDR or the consensus framework may be mutagenized by
substitution, insertion and/or deletion of at least one amino acid
residue so that the CDR or framework residue at that site does not
correspond to either the donor antibody or the consensus framework.
In an embodiment, such mutations, however, will not be extensive.
Usually, at least 80%, at least 85%, more at least 90%, and at
least 95% of the humanized antibody residues will correspond to
those of the parental FR and CDR sequences. The term "consensus
framework" refers to the framework region in the consensus
immunoglobulin sequence. The term "consensus immunoglobulin
sequence" refers to the sequence formed from the most frequently
occurring amino acids (or nucleotides) in a family of related
immunoglobulin sequences (see e.g., Winnaker, From Genes to Clones
(Verlagsgesellschaft, Weinheim, Germany 1987)). In a family of
immunoglobulins, each position in the consensus sequence is
occupied by the amino acid occurring most frequently at that
position in the family. If two amino acids occur equally
frequently, either can be included in the consensus sequence.
[0231] With respect to constructing DVD-binding protein or other
binding protein molecules, a "linker" is used to denote
polypeptides comprising two or more amino acid residues joined by
peptide bonds and are used to link one or more antigen binding
portions. Such linker polypeptides are well known in the art (see,
e.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA, 90:
6444-6448; Poljak et al. (1994) Structure, 2: 1121-1123). Exemplary
linkers include, but are not limited to, GGGGSG (SEQ ID NO:1695),
GGSGG (SEQ ID NO:1696), GGGGSGGGGS (SEQ ID NO:1697), GGSGGGGSGS
(SEQ ID NO:1698), GGSGGGGSGGGGS (SEQ ID NO:1699), GGGGSGGGGSGGGG
(SEQ ID NO:1700), GGGGSGGGGSGGGGS (SEQ ID NO:1701), ASTKGP (SEQ ID
NO:1702), ASTKGPSVFPLAP (SEQ ID NO:1703), TVAAP (SEQ ID NO:1704),
TVAAPSVFIFPP (SEQ ID NO:1705), AKTTPKLEEGEFSEAR (SEQ ID NO:1706),
AKTTPKLEEGEFSEARV (SEQ ID NO:1707), AKTTPKLGG (SEQ ID NO:1710),
SAKTTPKLGG (SEQ ID NO:1709), SAKTTP (SEQ ID NO:1702), RADAAP (SEQ
ID NO:1711), RADAAPTVS (SEQ ID NO:1712), RADAAAAGGPGS (SEQ ID
NO:1713), RADAAAAGGGGSGGGGSGGGGSGGGGS (SEQ ID NO:1714),
SAKTTPKLEEGEFSEARV (SEQ ID NO:1715), ADAAP (SEQ ID NO:1716),
ADAAPTVSIFPP (SEQ ID NO:2050), QPKAAP (SEQ ID NO:2051),
QPKAAPSVTLFPP (SEQ ID NO:2052), AKTTPP (SEQ ID NO:2053),
AKTTPPSVTPLAP (SEQ ID NO:2054), AKTTAP (SEQ ID NO:2055),
AKTTAPSVYPLAP (SEQ ID NO:2056), GENKVEYAPALMALS (SEQ ID NO:2057),
GPAKELTPLKEAKVS (SEQ ID NO:2058), and GHEAAAVMQVQYPAS (SEQ ID
NO:2059).
[0232] The term "Vernier" zone refers to a subset of framework
residues that may adjust CDR structure and fine-tune the fit to
antigen as described by Foote and Winter (1992) J. Mol. Biol.
224:487-499). Vernier zone residues form a layer underlying the
CDRs and may impact on the structure of CDRs and the affinity of
the antibody.
[0233] The term "neutralizing" refers to neutralization of the
biological activity of an antigen (e.g., SOST) when a binding
protein specifically binds the antigen. In an embodiment, a
neutralizing binding protein described herein binds to hSOST
resulting in the inhibition of a biological activity of hSOST. In
an embodiment, the neutralizing binding protein binds hSOST and
reduces a biologically activity of hSOST by at least about 20%,
40%, 60%, 80%, 85%, or more Inhibition of a biological activity of
hSOST by a neutralizing binding protein can be assessed by
measuring one or more indicators of hSOST biological activity well
known in the art.
[0234] The term "activity" includes activities such as the binding
specificity/affinity of an antibody for an antigen, for example, an
anti-hSOST antibody that binds to an SOST antigen and/or the
neutralizing potency of an antibody, for example, an anti-hSOST
antibody whose binding to hSOST inhibits the biological activity of
hSOST.
[0235] The term "epitope" includes any polypeptide determinant
capable of specific binding to an immunoglobulin or T-cell
receptor. In certain embodiments, epitope determinants include
chemically active surface groupings of molecules such as amino
acids, sugar side chains, phosphoryl, or sulfonyl, and, in certain
embodiments, may have specific three dimensional structural
characteristics, and/or specific charge characteristics. An epitope
is a region of an antigen that is bound by an antibody. In certain
embodiments, an antibody is said to specifically bind an antigen
when it preferentially recognizes its target antigen in a complex
mixture of proteins and/or macromolecules. Antibodies are said to
"bind to the same epitope" if the antibodies cross-compete (one
prevents the binding or modulating effect of the other). In
addition, structural definitions of epitopes (overlapping, similar,
identical) are informative, but functional definitions are often
more relevant as they encompass structural (binding) and functional
(modulation, competition) parameters.
[0236] The term "surface plasmon resonance" refers to an optical
phenomenon that allows for the analysis of real-time biospecific
interactions by detection of alterations in protein concentrations
within a biosensor matrix, for example using the BIAcore system
(Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, N.J.). For
further descriptions, see Jonsson et al. (1993) Ann. Biol. Clin.
51:19-26; Jonsson et al. (1991) BioTechniques, 11:620-627; Johnsson
et al. (1995) J. Mol. Recognit. 8:125-131; and Johnnson et al.
(1991) Anal. Biochem. 198:268-277.
[0237] The term "K.sub.on" (also "Kon", "kon") is intended to refer
to the on rate constant for association of a binding protein (e.g.,
an antibody) to an antigen to form an association complex, e.g.,
antibody/antigen complex, as is known in the art. The "Kon" also is
known by the terms "association rate constant", or "k.sub.a", as
used interchangeably herein. This value indicates the binding rate
of an antibody to its target antigen or the rate of complex
formation between an antibody and antigen as is shown by the
equation below:
Antibody("Ab")+Antigen("Ag").fwdarw.Ab-Ag.
[0238] The term "K.sub.off" (also "Koff", "koff") is intended to
refer to the off rate constant for dissociation, of a binding
protein (e.g., an antibody) from an association complex (e.g., an
antibody/antigen complex) as is known in the art. The "koff" also
is known by the terms "dissociaciation rate constant", or
"k.sub.d", as used interchangeably herein. This value indicates the
dissociation rate of an antibody from its target antigen or
separation of Ab-Ag complex over time into free antibody and
antigen as shown by the equation below:
Ab+Ag.rarw.Ab-Ag.
[0239] The term "K.sub.D" (also "K.sub.d") is intended to refer to
the "equilibrium dissociation constant", and refers to the value
obtained in a titration measurement at equilibrium, or by dividing
the dissociation rate constant (Koff) by the association rate
constant (Kon). The association rate constant (Kon), the
dissociation rate constant (Koff), and the equilibrium dissociation
constant (K are used to represent the binding affinity of an
antibody to an antigen. Methods for determining association and
dissociation rate constants are well known in the art. Using
fluorescence-based techniques offers high sensitivity and the
ability to examine samples in physiological buffers at equilibrium.
Other experimental approaches and instruments such as a
BIAcore.RTM. (biomolecular interaction analysis) assay can be used
(e.g., instrument available from BIAcore International AB, a GE
Healthcare company, Uppsala, Sweden). Additionally, a KinExA.RTM.
(Kinetic Exclusion Assay) assay, available from Sapidyne
Instruments (Boise, Id.) can also be used.
[0240] The terms "label" and "detectable label" mean a moiety
attached to a specific binding partner, such as an antibody or an
analyte, e.g., to render the reaction between members of a specific
binding pair, such as an antibody and an analyte, detectable. The
specific binding partner, e.g., antibody or analyte, so labeled is
referred to as "detectably labeled". Thus, the term "labeled
binding protein" refers to a protein with a label incorporated that
provides for the identification of the binding protein. In an
embodiment, the label is a detectable marker that can produce a
signal that is detectable by visual or instrumental means, e.g.,
incorporation of a radiolabeled amino acid or attachment to a
polypeptide of biotinyl moieties that can be detected by marked
avidin or streptavidin (e.g., streptavidin containing a fluorescent
marker or enzymatic activity that can be detected by optical or
colorimetric methods). Examples of labels for polypeptides include,
but are not limited to, the following: radioisotopes or
radionuclides (e.g., .sup.3H, .sup.14C, .sup.35S, .sup.90Y,
.sup.99Tc, .sup.111In, .sup.125I, .sup.131I .sup.177Lu, .sup.166Ho,
or .sup.153Sm), chromogens, fluorescent labels (e.g., FITC,
rhodamine, lanthanide phosphors), enzymatic labels (e.g.,
horseradish peroxidase, luciferase, alkaline phosphatase),
chemiluminescent markers, biotinyl groups, predetermined
polypeptide epitopes recognized by a secondary reporter (e.g.,
leucine zipper pair sequences, binding sites for secondary
antibodies, metal binding domains, epitope tags), and magnetic
agents (e.g., gadolinium chelates). Representative examples of
labels commonly employed for immunoassays include moieties that
produce light, e.g., acridinium compounds, and moieties that
produce fluorescence, e.g., fluorescein. Other labels are described
herein. In this regard, the moiety itself may not be detectably
labeled but may become detectable upon reaction with yet another
moiety. Use of the term "detectably labeled" is intended to
encompass the latter type of detectable labeling.
[0241] The term "SOST binding protein conjugate" or "sclerostin
binding protein conjugate" refers to a sclerostin binding protein
described herein chemically linked to a second chemical moiety,
such as a therapeutic or cytotoxic agent. The term "agent" is used
herein to denote a chemical compound, a mixture of chemical
compounds, a biological macromolecule, or an extract made from
biological materials. In an embodiment, the therapeutic or
cytotoxic agents include, but are not limited to, pertussis toxin,
taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine,
mitomycin, etoposide, tenoposide, vincristine, vinblastine,
colchicine, doxorubicin, daunorubicin, dihydroxy anthracin dione,
mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone,
glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and
puromycin and analogs or homologs thereof. When employed in the
context of an immunoassay, a sclerostin binding protein conjugate
may be a detectably labeled antibody, which is used as the
detection antibody.
[0242] The terms "crystal" and "crystallized" refer to a binding
protein (e.g., an antibody), or antigen binding portion thereof,
that exists in the form of a crystal. Crystals are one form of the
solid state of matter that is distinct from other forms such as the
amorphous solid state or the liquid crystalline state. Crystals are
composed of regular, repeating, three-dimensional arrays of atoms,
ions, molecules (e.g., proteins such as antibodies), or molecular
assemblies (e.g., antigen/antibody complexes). These
three-dimensional arrays are arranged according to specific
mathematical relationships that are well-understood in the field.
The fundamental unit, or building block, that is repeated in a
crystal is called the asymmetric unit. Repetition of the asymmetric
unit in an arrangement that conforms to a given, well-defined
crystallographic symmetry provides the "unit cell" of the crystal.
Repetition of the unit cell by regular translations in all three
dimensions provides the crystal. See Giege et al., Chapter 1, In
Crystallization of Nucleic Acids and Proteins, a Practical
Approach, 2nd ed., (Ducruix and Giege, eds.) (Oxford University
Press, New York, 1999) pp. 1-16.
[0243] The term "polynucleotide" means a polymeric form of two or
more nucleotides, either ribonucleotides or deoxynucleotides or a
modified form of either type of nucleotide. The term includes
single and double stranded forms of DNA.
[0244] The term "isolated polynucleotide" shall mean a
polynucleotide (e.g., of genomic, cDNA, or synthetic origin, or
some combination thereof) that, by virtue of its origin, the
"isolated polynucleotide" is not associated with all or a portion
of a polynucleotide with which the "isolated polynucleotide" is
found in nature; is operably linked to a polynucleotide that it is
not linked to in nature; or does not occur in nature as part of a
larger sequence.
[0245] The term "vector" is intended to refer to a nucleic acid
molecule capable of transporting another nucleic acid to which it
has been linked One type of vector is a "plasmid", which refers to
a circular double stranded DNA loop into which additional DNA
segments may be ligated. Another type of vector is a viral vector,
wherein additional DNA segments may be ligated into the viral
genome. Certain vectors are capable of autonomous replication in a
host cell into which they are introduced (e.g., bacterial vectors
having a bacterial origin of replication and episomal mammalian
vectors). Other vectors (e.g., non-episomal mammalian vectors) can
be integrated into the genome of a host cell upon introduction into
the host cell, and thereby are replicated along with the host
genome. Moreover, certain vectors are capable of directing the
expression of genes to which they are operatively linked Such
vectors are referred to herein as "recombinant expression vectors"
(or simply, "expression vectors"). In general, expression vectors
of utility in recombinant DNA techniques are often in the form of
plasmids. In the present specification, "plasmid" and "vector" may
be used interchangeably as the plasmid is the most commonly used
form of vector. However, other forms of expression vectors, such as
viral vectors (e.g., replication defective retroviruses,
adenoviruses and adeno-associated viruses), which serve equivalent
functions are provided.
[0246] The term "operably linked" refers to a juxtaposition wherein
the components described are in a relationship permitting them to
function in their intended manner. A control sequence "operably
linked" to a coding sequence is ligated in such a way that
expression of the coding sequence is achieved under conditions
compatible with the control sequences. "Operably linked" sequences
include both expression control sequences that are contiguous with
the gene of interest and expression control sequences that act in
trans or at a distance to control the gene of interest. The term
"expression control sequence" refers to polynucleotide sequences
that are necessary to effect the expression and processing of
coding sequences to which they are ligated. Expression control
sequences include appropriate transcription initiation,
termination, promoter and enhancer sequences; efficient RNA
processing signals such as splicing and polyadenylation signals;
sequences that stabilize cytoplasmic mRNA; sequences that enhance
translation efficiency (i.e., Kozak consensus sequence); sequences
that enhance protein stability; and when desired, sequences that
enhance protein secretion. The nature of such control sequences
differs depending upon the host organism; in prokaryotes, such
control sequences generally include promoter, ribosomal binding
site, and transcription termination sequence; in eukaryotes,
generally, such control sequences include promoters and
transcription termination sequence. The term "control sequences" is
intended to include components whose presence is essential for
expression and processing, and can also include additional
components whose presence is advantageous, for example, leader
sequences and fusion partner sequences.
[0247] "Transformation", as defined herein, refers to any process
by which exogenous DNA enters a host cell. Transformation may occur
under natural or artificial conditions using various methods well
known in the art. Transformation may rely on any known method for
the insertion of foreign nucleic acid sequences into a prokaryotic
or eukaryotic host cell. The method is selected based on the host
cell being transformed and may include, but is not limited to,
viral infection, electroporation, lipofection, and particle
bombardment. Such "transformed" cells include stably transformed
cells in which the inserted DNA is capable of replication either as
an autonomously replicating plasmid or as part of the host
chromosome. They also include cells which transiently express the
inserted DNA or RNA for limited periods of time.
[0248] The term "recombinant host cell" (or simply "host cell"), is
intended to refer to a cell into which exogenous DNA has been
introduced. In an embodiment, the host cell comprises two or more
(e.g., multiple) nucleic acids encoding antibodies, such as the
host cells described in U.S. Pat. No. 7,262,028, for example. Such
terms are intended to refer not only to the particular subject
cell, but also to the progeny of such a cell. Because certain
modifications may occur in succeeding generations due to either
mutation or environmental influences, such progeny may not, in
fact, be identical to the parent cell, but are still included
within the scope of the term "host cell". In an embodiment, host
cells include prokaryotic and eukaryotic cells from any of the
Kingdoms of life. In another embodiment, eukaryotic cells include
protist, fungal, plant or animal cells. In another embodiment, host
cells include but are not limited to the prokaryotic cell line
Escherichia coli; mammalian cell lines CHO, HEK 293, COS, NS0, SP2
and PER.C6; the insect cell line Sf9; and the fungal cell
Saccharomyces cerevisiae.
[0249] Standard techniques may be used for recombinant DNA,
oligonucleotide synthesis, and tissue culture and transformation
(e.g., electroporation, lipofection). Enzymatic reactions and
purification techniques may be performed according to
manufacturer's specifications or as commonly accomplished in the
art or as described herein. The foregoing techniques and procedures
may be generally performed according to conventional methods well
known in the art and as described in various general and more
specific references that are cited and discussed throughout the
present specification. See e.g., Sambrook et al., Molecular
Cloning: A Laboratory Manual, 2nd ed. (Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, N.Y., 1989).
[0250] "Transgenic organism", as known in the art, refers to an
organism having cells that contain a transgene, wherein the
transgene introduced into the organism (or an ancestor of the
organism) expresses a polypeptide not naturally expressed in the
organism. A "transgene" is a DNA construct, which is stably and
operably integrated into the genome of a cell from which a
transgenic organism develops, directing the expression of an
encoded gene product in one or more cell types or tissues of the
transgenic organism.
[0251] The terms "regulate" and "modulate" are used
interchangeably, and refers to a change or an alteration in the
activity of a molecule of interest (e.g., the biological activity
of hSOST). Modulation may be an increase or a decrease in the
magnitude of a certain activity or function of the molecule of
interest. Exemplary activities and functions of a molecule include,
but are not limited to, binding characteristics, enzymatic
activity, cell receptor activation, and signal transduction.
[0252] Correspondingly, the term "modulator" is a compound capable
of changing or altering an activity or function of a molecule of
interest (e.g., the biological activity of hSOST). For example, a
modulator may cause an increase or decrease in the magnitude of a
certain activity or function of a molecule compared to the
magnitude of the activity or function observed in the absence of
the modulator. In certain embodiments, a modulator is an inhibitor,
which decreases the magnitude of at least one activity or function
of a molecule. Exemplary inhibitors include, but are not limited
to, proteins, peptides, antibodies, peptibodies, carbohydrates or
small organic molecules. Peptibodies are described, e.g., in PCT
Publication No. WO01/83525.
[0253] The term "agonist" refers to a modulator that, when
contacted with a molecule of interest, causes an increase in the
magnitude of a certain activity or function of the molecule
compared to the magnitude of the activity or function observed in
the absence of the agonist. Particular agonists of interest may
include, but are not limited to, sclerostin polypeptides, nucleic
acids, carbohydrates, or any other molecule that binds to human
sclerostin (hSOST).
[0254] The terms "antagonist" and "inhibitor" refer to a modulator
that, when contacted with a molecule of interest causes a decrease
in the magnitude of a certain activity or function of the molecule
compared to the magnitude of the activity or function observed in
the absence of the antagonist. Particular antagonists of interest
include those that block or modulate the biological or
immunological activity of human sclerostin. Antagonists and
inhibitors of human sclerostin may include, but are not limited to,
proteins, nucleic acids, carbohydrates, or any other molecules,
which bind to human sclerostin.
[0255] The term "effective amount" refers to the amount of a
therapy that is sufficient to reduce or ameliorate the severity
and/or duration of a disorder or one or more symptoms thereof;
prevent the advancement of a disorder; cause regression of a
disorder; prevent the recurrence, development, onset, or
progression of one or more symptoms associated with a disorder;
detect a disorder; or enhance or improve the prophylactic or
therapeutic effect(s) of another therapy (e.g., prophylactic or
therapeutic agent).
[0256] "Patient" and "subject" may be used interchangeably herein
to refer to an animal, such as a mammal, including a primate (for
example, a human, a monkey, and a chimpanzee), a non-primate (for
example, a cow, a pig, a camel, a llama, a horse, a goat, a rabbit,
a sheep, a hamster, a guinea pig, a cat, a dog, a rat, a mouse, a
whale), a bird (e.g., a duck or a goose), and a shark. In an
embodiment, a patient or subject is a human, such as a human being
treated or assessed for a disease, disorder or condition, a human
at risk for a disease, disorder or condition, a human having a
disease, disorder or condition, and/or human being treated for a
disease, disorder or condition.
[0257] The term "sample" is used in its broadest sense. A
"biological sample" includes, but is not limited to, any quantity
of a substance from a living thing or formerly living thing. Such
living things include, but are not limited to, humans, non-human
primates, mice, rats, monkeys, dogs, rabbits and other animals.
Such substances include, but are not limited to, blood (e.g., whole
blood), plasma, serum, urine, amniotic fluid, synovial fluid,
endothelial cells, leukocytes, monocytes, other cells, organs,
tissues, bone marrow, lymph nodes and spleen.
[0258] "Component", "components," and "at least one component,"
refer generally to a capture antibody, a detection or conjugate
antibody, a control, a calibrator, a series of calibrators, a
sensitivity panel, a container, a buffer, a diluent, a salt, an
enzyme, a co-factor for an enzyme, a detection reagent, a
pretreatment reagent/solution, a substrate (e.g., as a solution), a
stop solution, and the like that can be included in a kit for assay
of a test sample, such as a patient urine, serum or plasma sample,
in accordance with the methods described herein and other methods
known in the art. Thus, in the context of the present disclosure,
"at least one component," "component," and "components" can include
a polypeptide or other analyte as above, such as a composition
comprising an analyte such as polypeptide, which is optionally
immobilized on a solid support, such as by binding to an
anti-analyte (e.g., anti-polypeptide) antibody. Some components can
be in solution or lyophilized for reconstitution for use in an
assay.
[0259] "Control" refers to a composition known to not analyte
("negative control") or to contain analyte ("positive control"). A
positive control can comprise a known concentration of analyte.
"Control," "positive control," and "calibrator" may be used
interchangeably herein to refer to a composition comprising a known
concentration of analyte. A "positive control" can be used to
establish assay performance characteristics and is a useful
indicator of the integrity of reagents (e.g., analytes).
[0260] "Predetermined cutoff" and "predetermined level" refer
generally to an assay cutoff value that is used to assess
diagnostic/prognostic/therapeutic efficacy results by comparing the
assay results against the predetermined cutoff/level, where the
predetermined cutoff/level already has been linked or associated
with various clinical parameters (e.g., severity of disease,
progression/nonprogression/improvement, etc.). While the present
disclosure may provide exemplary predetermined levels, it is
well-known that cutoff values may vary depending on the nature of
the immunoassay (e.g., antibodies employed, etc.). It further is
well within the ordinary skill of one in the art to adapt the
disclosure herein for other immunoassays to obtain
immunoassay-specific cutoff values for those other immunoassays
based on this disclosure. Whereas the precise value of the
predetermined cutoff/level may vary between assays, correlations as
described herein (if any) should be generally applicable.
[0261] "Pretreatment reagent," e.g., lysis, precipitation and/or
solubilization reagent, as used in a diagnostic assay as described
herein is one that lyses any cells and/or solubilizes any analyte
that is/are present in a test sample. Pretreatment is not necessary
for all samples, as described further herein. Among other things,
solubilizing the analyte (e.g., polypeptide of interest) may entail
release of the analyte from any endogenous binding proteins present
in the sample. A pretreatment reagent may be homogeneous (not
requiring a separation step) or heterogeneous (requiring a
separation step). With use of a heterogeneous pretreatment reagent
there is removal of any precipitated analyte binding proteins from
the test sample prior to proceeding to the next step of the
assay.
[0262] "Quality control reagents" in the context of immunoassays
and kits described herein, include, but are not limited to,
calibrators, controls, and sensitivity panels. A "calibrator" or
"standard" typically is used (e.g., one or more, such as a
plurality) in order to establish calibration (standard) curves for
interpolation of the concentration of an analyte, such as an
antibody or an analyte. Alternatively, a single calibrator, which
is near a predetermined positive/negative cutoff, can be used.
Multiple calibrators (i.e., more than one calibrator or a varying
amount of calibrator(s)) can be used in conjunction so as to
comprise a "sensitivity panel." "Risk" refers to the possibility or
probability of a particular event occurring either presently or at
some point in the future. "Risk stratification" refers to an array
of known clinical risk factors that allows physicians to classify
patients into a low, moderate, high or highest risk of developing a
particular disease, disorder or condition.
[0263] "Specific" and "specificity" in the context of an
interaction between members of a specific binding pair (e.g., an
antigen (or fragment thereof) and an antibody (or antigenically
reactive fragment thereof)) refer to the selective reactivity of
the interaction. The phrase "specifically binds to" and analogous
phrases refer to the ability of antibodies (or antigenically
reactive fragments thereof) to bind specifically to analyte (or a
fragment thereof) and not bind specifically to other entities.
[0264] "Specific binding partner" is a member of a specific binding
pair. A specific binding pair comprises two different molecules,
which specifically bind to each other through chemical or physical
means. Therefore, in addition to antigen and antibody specific
binding pairs of common immunoassays, other specific binding pairs
can include biotin and avidin (or streptavidin), carbohydrates and
lectins, complementary nucleotide sequences, effector and receptor
molecules, cofactors and enzymes, enzyme inhibitors and enzymes,
and the like. Furthermore, specific binding pairs can include
members that are analogs of the original specific binding members,
for example, an analyte-analog. Immunoreactive specific binding
members include antigens, antigen fragments, and antibodies,
including monoclonal and polyclonal antibodies as well as
complexes, fragments, and variants (including fragments of
variants) thereof, whether isolated or recombinantly produced.
[0265] The term "variant" means a polypeptide that differs from a
given polypeptide (e.g., sclerostin, BNP, NGAL, or HIV polypeptide,
or anti-polypeptide antibody) in amino acid sequence by the
addition (e.g., insertion), deletion, or conservative substitution
of amino acids, but that retains the biological activity of the
given polypeptide (e.g., a variant sclerostin can compete with
anti-sclerostin antibody for binding to sclerostin). A conservative
substitution of an amino acid, i.e., replacing an amino acid with a
different amino acid of similar properties (e.g., hydrophilicity
and degree and distribution of charged regions) is recognized in
the art as typically involving a minor change. These minor changes
can be identified, in part, by considering the hydropathic index of
amino acids, as understood in the art (see, e.g., Kyte et al.
(1982) J. Mol. Biol. 157:105-132). The hydropathic index of an
amino acid is based on a consideration of its hydrophobicity and
charge. It is known in the art that amino acids of similar
hydropathic indexes can be substituted and still retain protein
function. In one aspect, amino acids having hydropathic indexes of
.+-.2 are substituted. The hydrophilicity of amino acids also can
be used to reveal substitutions that would result in proteins
retaining biological function. A consideration of the
hydrophilicity of amino acids in the context of a peptide permits
calculation of the greatest local average hydrophilicity of that
peptide, a useful measure that has been reported to correlate well
with antigenicity and immunogenicity (see, e.g., U.S. Pat. No.
4,554,101). Substitution of amino acids having similar
hydrophilicity values can result in peptides retaining biological
activity, for example immunogenicity, as is understood in the art.
In one aspect, substitutions are performed with amino acids having
hydrophilicity values within .+-.2 of each other. Both the
hydrophobicity index and the hydrophilicity value of amino acids
are influenced by the particular side chain of that amino acid.
Consistent with that observation, amino acid substitutions that are
compatible with biological function are understood to depend on the
relative similarity of the amino acids, and particularly the side
chains of those amino acids, as revealed by the hydrophobicity,
hydrophilicity, charge, size, and other properties. "Variant" also
can be used to describe a polypeptide or fragment thereof that has
been differentially processed, such as by proteolysis,
phosphorylation, or other post-translational modification, yet
retains its biological activity or antigen reactivity, e.g., the
ability to bind to sclerostin. Use of "variant" herein is intended
to encompass fragments of a variant unless otherwise contradicted
by context.
I. Antibodies that Bind Human SOST.
[0266] One aspect provides isolated murine monoclonal antibodies,
or antigen-binding portions thereof, that bind to sclerostin with
high affinity, a slow off rate and high neutralizing capacity. A
second aspect provides chimeric antibodies that bind sclerostin. A
third aspect provides CDR grafted antibodies, or antigen-binding
portions thereof, that bind sclerostin. A fourth aspect provides
humanized antibodies, or antigen-binding portions thereof, that
bind sclerostin. A fifth aspect provides dual variable domain
binding proteins (DVD-binding proteins) that bind sclerostin and
one other target. In an embodiment, the antibodies, or portions
thereof, are isolated antibodies. In an embodiment, the antibodies
neutralizing human anti-sclerostin are provided.
I.A. Method of Making Anti Sclerostin Antibodies
[0267] Anti sclerostin antibodies made by any of a number of
techniques known in the art are provided.
I.A.1. Anti Sclerostin Monoclonal Antibodies Using Hybridoma
Technology
[0268] Monoclonal antibodies can be prepared using a wide variety
of techniques known in the art including the use of hybridoma,
recombinant, and phage display technologies, or a combination
thereof. For example, monoclonal antibodies can be produced using
hybridoma techniques including those known in the art and taught,
for example, in Harlow et al., Antibodies: A Laboratory Manual, 2nd
ed. (Cold Spring Harbor Laboratory Press, 1988); Hammerling, et
al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681
(Elsevier, N.Y., 1981). The term "monoclonal antibody" is not
limited to antibodies produced through hybridoma technology. The
term "monoclonal antibody" refers to an antibody that is derived
from a single clone, including any eukaryotic, prokaryotic, or
phage clone, and not the method by which it is produced.
[0269] Methods for producing and screening for specific
anti-sclerostin antibodies using hybridoma technology are routine
and well known in the art. One embodiment provides methods of
generating monoclonal antibodies as well as antibodies produced by
the method comprising culturing a hybridoma cell secreting an
antibody wherein, in an embodiment, the hybridoma is generated by
fusing splenocytes isolated from a mouse immunized with an antigen
with myeloma cells and then screening the hybridomas resulting from
the fusion for hybridoma clones that secrete an antibody able to
bind a polypeptide. Briefly, mice can be immunized with an
sclerostin antigen. In an embodiment, the sclerostin antigen is
administered with an adjuvant to stimulate the immune response.
Such adjuvants include complete or incomplete Freund's adjuvant,
RIBI (muramyl dipeptides) or ISCOM (immunostimulating complexes).
Such adjuvants may protect the polypeptide from rapid dispersal by
sequestering it in a local deposit, or they may contain substances
that stimulate the host to secrete factors that are chemotactic for
macrophages and other components of the immune system. In an
embodiment, if a polypeptide is being administered, the
immunization schedule will involve two or more administrations of
the polypeptide, spread out over several weeks.
[0270] After immunization of an animal with an sclerostin antigen,
antibodies and/or antibody-producing cells may be obtained from the
animal. An anti-sclerostin antibody-containing serum is obtained
from the animal by bleeding or sacrificing the animal. The serum
may be used as it is obtained from the animal, an immunoglobulin
fraction may be obtained from the serum, or the anti-sclerostin
antibodies may be purified from the serum. Serum or immunoglobulins
obtained in this manner are polyclonal, thus having a heterogeneous
array of properties.
[0271] Once an immune response is detected, e.g., antibodies
specific for the antigen SOST are detected in the mouse serum, the
mouse spleen is harvested and splenocytes isolated. The splenocytes
are then fused by well-known techniques to any suitable myeloma
cells, for example cells from cell line SP20 available from the
American Type Culture Collection (ATCC, Manassas, Va., US).
Hybridomas are selected and cloned by limited dilution. The
hybridoma clones are then assayed by methods known in the art for
cells that secrete antibodies capable of binding SOST. Ascites
fluid, which generally contains high levels of antibodies, can be
generated by immunizing mice with positive hybridoma clones.
[0272] In another embodiment, antibody-producing immortalized
hybridomas may be prepared from the immunized animal. After
immunization, the animal is sacrificed and the splenic B cells are
fused to immortalized myeloma cells as is well known in the art.
See, e.g., Harlow and Lane, supra. In an embodiment, the myeloma
cells do not secrete immunoglobulin polypeptides (a non-secretory
cell line). After fusion and antibiotic selection, the hybridomas
are screened using SOST, or a portion thereof, or a cell expressing
SOST. In an embodiment, the initial screening is performed using an
enzyme-linked immunoassay (ELISA) or a radioimmunoassay (RIA), in
an embodiment, an ELISA. An example of ELISA screening is provided
in PCT Publication No. WO 00/37504.
[0273] Anti-sclerostin antibody-producing hybridomas are selected,
cloned, and further screened for desirable characteristics,
including robust hybridoma growth, high antibody production and
desirable antibody characteristics, as discussed further below.
Hybridomas may be cultured and expanded in vivo in syngeneic
animals, in animals that lack an immune system, e.g., nude mice, or
in cell culture in vitro. Methods of selecting, cloning and
expanding hybridomas are well known to those of ordinary skill in
the art.
[0274] In an embodiment, the hybridomas are mouse hybridomas, as
described above. In another embodiment, the hybridomas are produced
in a non-human, non-mouse species such as rats, sheep, pigs, goats,
cattle or horses. In another embodiment, the hybridomas are human
hybridomas, in which a human non-secretory myeloma is fused with a
human cell expressing an anti-sclerostin antibody.
[0275] Antibody fragments that recognize specific epitopes may be
generated by known techniques. For example, Fab and F(ab')2
fragments produced by proteolytic cleavage of immunoglobulin
molecules, using enzymes such as papain (to produce Fab fragments)
or pepsin (to produce F(ab')2 fragments), are provided. F(ab')2
fragments contain the variable region, the light chain constant
region and the CHI domain of the heavy chain.
I.A.2. Anti-Sclerostin Monoclonal Antibodies Using SLAM
[0276] In another embodiment, recombinant antibodies generated from
single, isolated lymphocytes using a procedure referred to in the
art as the selected lymphocyte antibody method (SLAM) are provided,
as described in U.S. Pat. No. 5,627,052; PCT Publication No. WO
92/02551; and Babcook et al. (1996) Proc. Natl. Acad. Sci. USA 93:
7843-7848. In this method, single cells secreting antibodies of
interest, e.g., lymphocytes derived from any one of the immunized
animals described in Section 1, are screened using an
antigen-specific hemolytic plaque assay, wherein the antigen SOST,
a subunit of SOST, or a fragment thereof, is coupled to sheep red
blood cells using a linker, such as biotin, and used to identify
single cells that secrete antibodies with specificity for SOST.
Following identification of antibody-secreting cells of interest,
heavy and light chain variable region (VH and VL) cDNAs are rescued
from the cells by reverse transcriptase-PCR, and these variable
regions can then be expressed, in the context of appropriate
immunoglobulin constant regions (e.g., human constant regions), in
mammalian host cells, such as COS or CHO cells. The host cells
transfected with the amplified immunoglobulin sequences, derived
from in vivo selected lymphocytes, can then undergo further
analysis and selection in vitro, for example, by panning the
transfected cells to isolate cells expressing antibodies to SOST.
The amplified immunoglobulin sequences further can be manipulated
in vitro, such as by in vitro affinity maturation methods such as
those described in PCT Publication Nos. WO 97/29131 and WO
00/56772.
I.A.3. Anti-Sclerostin Monoclonal Antibodies Using Transgenic
Animals
[0277] In another embodiment, antibodies produced by immunizing a
non-human animal comprising some, or all, of the human
immunoglobulin locus with an SOST antigen are provided. In an
embodiment, the non-human animal is a XENOMOUSE transgenic mouse,
an engineered mouse strain that comprises large fragments of the
human immunoglobulin loci and is deficient in mouse antibody
production. See, e.g., Green et al. (1994) Nature Genet. 7:13-21
and U.S. Pat. Nos. 5,916,771; 5,939,598; 5,985,615; 5,998,209;
6,075,181; 6,091,001; 6,114,598 and 6,130,364. See also PCT
Publication Nos. WO 91/10741; WO 94/02602; WO 96/34096 and WO
96/33735; WO 98/16654; WO 98/24893; WO 98/50433; WO 99/45031; WO
99/53049; WO 00/09560; and WO 00/037504. The XENOMOUSE.RTM.
transgenic mouse produces an adult-like human repertoire of fully
human antibodies, and generates antigen-specific human Mabs. The
XENOMOUSE.RTM. transgenic mouse contains approximately 80% of the
human antibody repertoire through introduction of megabase sized,
germline configuration YAC fragments of the human heavy chain loci
and x light chain loci. See, Mendez et al., Nature Genetics,
15:146-156 (1997); and Green and Jakobovits, J. Exp. Med., 188:
483-495 (1998).
I.A.4. Anti-Sclerostin Monoclonal Antibodies Using Recombinant
Antibody Libraries
[0278] In vitro methods to make antibodies are provided, wherein an
antibody library is screened to identify an antibody having the
desired binding specificity. Methods for such screening of
recombinant antibody libraries are well known in the art and
include methods described in, for example, Ladner et al., U.S. Pat.
No. 5,223,409; Kang et al., PCT Publication No. WO 92/18619; Dower
et al., PCT Publication No. WO 91/17271; Winter et al., PCT
Publication No. WO 92/20791; Markland et al., PCT Publication No.
WO 92/15679; Breitling et al., PCT Publication No. WO 93/01288;
McCafferty et al., PCT Publication No. WO 92/01047; Garrard et al.,
PCT Publication No. WO 92/09690; Fuchs et al., Bio/Technology, 9:
1369-1372 (1991); Hay et al., Hum. Antibod. Hybridomas, 3: 81-85
(1992); Huse et al., Science, 246: 1275-1281 (1989); McCafferty et
al., Nature, 348: 552-554 (1990); Griffiths et al., EMBO J., 12:
725-734 (1993); Hawkins et al., J. Mol. Biol., 226: 889-896 (1992);
Clackson et al., Nature, 352: 624-628 (1991); Gram et al., Proc.
Natl. Acad. Sci. USA, 89: 3576-3580 (1992); Garrard et al.,
Bio/Technology, 9: 1373-1377 (1991); Hoogenboom et al., Nucl. Acid
Res., 19: 4133-4137 (1991); and Barbas et al., Proc. Natl. Acad.
Sci. USA, 88: 7978-7982 (1991); US patent application publication
No. 2003/0186374; and PCT Publication No. WO 97/29131.
[0279] The recombinant antibody library may be from a subject
immunized with sclerostin, or a portion of sclerostin.
Alternatively, the recombinant antibody library may be from a naive
subject, i.e., one who has not been immunized with sclerostin, such
as a human antibody library from a human subject who has not been
immunized with human sclerostin. Antibodies of the invention are
selected by screening the recombinant antibody library with the
peptide comprising human sclerostin to thereby select those
antibodies that recognize sclerostin. Methods for conducting such
screening and selection are well known in the art, such as
described in the references in the preceding paragraph. To select
antibodies having particular binding affinities for human
sclerostin, such as those that dissociate from human sclerostin
with a particular K.sub.off rate constant, the art-known method of
surface plasmon resonance can be used to select antibodies having
the desired K.sub.off rate constant. To select antibodies having a
particular neutralizing activity for human sclerostin, such as
those with a particular an IC.sub.50, standard methods known in the
art for assessing the inhibition of human sclerostin activity may
be used.
[0280] In one aspect, an isolated antibody, or an antigen-binding
portion thereof, that binds human sclerostin is provided. In an
embodiment, the antibody is a neutralizing antibody. In various
embodiments, the antibody is a recombinant antibody or a monoclonal
antibody.
[0281] For example, the antibodies that are provided can also be
generated using various phage display methods known in the art. In
phage display methods, functional antibody domains are displayed on
the surface of phage particles which carry the polynucleotide
sequences encoding them. In a particular, such phage can be
utilized to display antigen-binding domains expressed from a
repertoire or combinatorial antibody library (e.g., human or
murine). Phage expressing an antigen binding domain that binds the
antigen of interest can be selected or identified with antigen,
e.g., using labeled antigen or antigen bound or captured to a solid
surface or bead. Phage used in these methods are typically
filamentous phage including fd and M13 binding domains expressed
from phage with Fab, Fv, or disulfide stabilized Fv antibody
domains recombinantly fused to either the phage gene III or gene
VIII protein. Phage display methods that can be used to make the
antibodies invention are provided and include those disclosed in
Brinkmann et al., J. Immunol. Methods, 182: 41-50 (1995); Ames et
al., J. Immunol. Methods, 184: 177-186 (1995); Kettleborough et
al., Eur. J. Immunol., 24: 952-958 (1994); Persic et al., Gene,
187: 9-18 (1997); Burton et al., Advances in Immunology, 57:191-280
(1994); PCT Publications Nos. WO 90/02809; WO 91/10737; WO 92/01047
(PCT/GB91/01134); WO 92/18619; WO 93/11236; WO 95/15982; WO
95/20401; and U.S. Pat. Nos. 5,698,426; 5,223,409; 5,403,484;
5,580,717; 5,427,908; 5,750,753; 5,821,047; 5,571,698; 5,427,908;
5,516,637; 5,780, 225; 5,658,727; 5,733,743 and 5,969,108.
[0282] As described in the above references, after phage selection,
the antibody coding regions from the phage can be isolated and used
to generate whole antibodies including human antibodies or any
other desired antigen binding fragment, and expressed in any
desired host, including mammalian cells, insect cells, plant cells,
yeast, and bacteria, e.g., as described in detail below. For
example, techniques to recombinantly produce Fab, Fab' and F(ab')2
fragments can also be employed using methods known in the art such
as those disclosed in PCT publication WO 92/22324; Mullinax et al.,
BioTechniques, 12(6): 864-869 (1992); and Sawai et al., Am. J.
Reprod. Immunol., 34: 26-34 (1995); and Better et al., Science,
240: 1041-1043 (1988). Examples of techniques which can be used to
produce single-chain Fvs and antibodies include those described in
U.S. Pat. Nos. 4,946,778 and 5,258,498; Huston et al., Methods in
Enzymology, 203: 46-88 (1991); Shu et al., Proc. Natl. Acad. Sci.
USA, 90: 7995-7999 (1993); and Skerra et al., Science, 240:
1038-1041 (1988).
[0283] Alternative to screening of recombinant antibody libraries
by phage display, other methodologies known in the art for
screening large combinatorial libraries are provided and can be
applied to the identification of dual specificity antibodies. One
type of alternative expression system is one in which the
recombinant antibody library is expressed as RNA-protein fusions,
as described in PCT Publication No. WO 98/31700 by Szostak and
Roberts, and in Roberts, R. W. and Szostak, J. W. (1997) Proc.
Natl. Acad. Sci. USA, 94: 12297-12302. In this system, a covalent
fusion is created between an mRNA and the peptide or protein that
it encodes by in vitro translation of synthetic mRNAs that carry
puromycin, a peptidyl acceptor antibiotic, at their 3' end. Thus, a
specific mRNA can be enriched from a complex mixture of mRNAs
(e.g., a combinatorial library) based on the properties of the
encoded peptide or protein, e.g., antibody, or portion thereof,
such as binding of the antibody, or portion thereof, to the dual
specificity antigen. Nucleic acid sequences encoding antibodies, or
portions thereof, recovered from screening of such libraries can be
expressed by recombinant means as described above (e.g., in
mammalian host cells) and, moreover, can be subjected to further
affinity maturation by either additional rounds of screening of
mRNA-peptide fusions in which mutations have been introduced into
the originally selected sequence(s), or by other methods for
affinity maturation in vitro of recombinant antibodies, as
described above.
[0284] In another approach, antibodies generated using yeast
display methods known in the art are provided. In yeast display
methods, genetic methods are used to tether antibody domains to the
yeast cell wall and display them on the surface of yeast. In
particular, such yeast can be utilized to display antigen-binding
domains expressed from a repertoire or combinatorial antibody
library (e. g., human or murine). Examples of yeast display methods
that can be used to make the antibodies are provided, and include
those disclosed by Wittrup et al. in U.S. Pat. No. 6,699,658.
I.B. Production of Recombinant Sclerostin Antibodies
[0285] Antibodies produced by any of a number of techniques known
in the art are provided. For example, expression from host cells,
wherein expression vector(s) encoding the heavy and light chains is
(are) transfected into a host cell by standard techniques. The
various forms of the term "transfection" are intended to encompass
a wide variety of techniques commonly used for the introduction of
exogenous DNA into a prokaryotic or eukaryotic host cell, e.g.,
electroporation, calcium-phosphate precipitation, DEAE-dextran
transfection, and the like. Although it is possible to express the
antibodies that are provided in either prokaryotic or eukaryotic
host cells, expression of antibodies in eukaryotic cells is
preferable, and most preferable in mammalian host cells, because
such eukaryotic cells (and in particular mammalian cells) are more
likely than prokaryotic cells to assemble and secrete a properly
folded and immunologically active antibody.
[0286] Exemplary mammalian host cells for expressing the
recombinant antibodies that are provided include Chinese Hamster
Ovary (CHO cells) (including dhfr-CHO cells, described in Urlaub
and Chasin, (1980) Proc. Natl. Acad. Sci. USA, 77: 4216-4220, used
with a DHFR selectable marker, e.g., as described in R. J. Kaufman
and P. A. Sharp (1982) Mol. Biol., 159: 601-621), NS0 myeloma
cells, COS cells and SP2 cells. When recombinant expression vectors
encoding antibody genes are introduced into mammalian host cells,
the antibodies are produced by culturing the host cells for a
period of time sufficient to allow for expression of the antibody
in the host cells or, in an embodiment, secretion of the antibody
into the culture medium in which the host cells are grown.
Antibodies can be recovered from the culture medium using standard
protein purification methods.
[0287] Host cells can also be used to produce functional antibody
fragments, such as Fab fragments or scFv molecules. It will be
understood that variations on the above procedure are within the
scope of what is provided. For example, it may be desirable to
transfect a host cell with DNA encoding functional fragments of
either the light chain and/or the heavy chain of an antibody, as
provided. Recombinant DNA technology may also be used to remove
some, or all, of the DNA encoding either or both of the light and
heavy chains that is not necessary for binding to the antigens of
interest. The molecules expressed from such truncated DNA molecules
are also encompassed by the antibodies, as provided. In addition,
bifunctional antibodies are provided and may be produced in which
one heavy and one light chain are an antibody and the other heavy
and light chain are specific for an antigen other than the antigens
of interest by crosslinking an antibody to a second antibody by
standard chemical crosslinking methods.
[0288] In an exemplary system for recombinant expression of an
antibody, or antigen-binding portion thereof, a recombinant
expression vector encoding both the antibody heavy chain and the
antibody light chain is introduced into dhfr-CHO cells by calcium
phosphate-mediated transfection is provided. Within the recombinant
expression vector, the antibody heavy and light chain genes are
each operatively linked to CMV enhancer/AdMLP promoter regulatory
elements to drive high levels of transcription of the genes. The
recombinant expression vector also carries a DHFR gene, which
allows for selection of CHO cells that have been transfected with
the vector using methotrexate selection/amplification. The selected
transformant host cells are cultured to allow for expression of the
antibody heavy and light chains and intact antibody is recovered
from the culture medium. Standard molecular biology techniques are
used to prepare the recombinant expression vector, transfect the
host cells, select for transformants, culture the host cells and
recover the antibody from the culture medium. Still further, a
method of synthesizing a recombinant antibody by culturing a host
cell in a suitable culture medium until a recombinant antibody is
synthesized is provided. The method can further comprise isolating
the recombinant antibody from the culture medium.
I.B.1. Anti Human Sclerostin Antibodies
[0289] Tables herein provide a list of amino acid sequences of VH
and VL regions of exemplary human anti-human sclerostin
antibodies.
[0290] Table 6 provides a sclerostin binding protein comprising an
antigen binding domain capable of binding human sclerostin, said
antigen binding domain comprising at least one CDR comprising an
amino acid sequence provided therein.
I.B.2. Anti Human Sclerostin Chimeric Antibodies
[0291] A chimeric antibody is a molecule in which different
portions of the antibody are derived from different animal species,
such as antibodies having a variable region derived from a murine
monoclonal antibody and a human immunoglobulin constant region.
Methods for producing chimeric antibodies are known in the art and
discussed in detail in the Examples section. See e.g., Morrison,
Science, 229: 1202-1207 (1985); Oi et al., BioTechniques, 4:
214-221 (1986); Gillies et al., J. Immunol. Methods, 125: 191-202
(1989); U.S. Pat. Nos. 5,807,715; 4,816,567; and 4,816,397. In
addition, techniques developed for the production of "chimeric
antibodies" (Morrison et al., Proc. Natl. Acad. Sci. USA, 81:
6851-6855 (1984); Neuberger et al., Nature, 312: 604-608 (1984);
Takeda et al., Nature, 314: 452-454 (1985) by splicing genes from a
mouse antibody molecule of appropriate antigen specificity together
with genes from a human antibody molecule of appropriate biological
activity can be used.
[0292] In one embodiment, chimeric antibodies produced by replacing
the heavy chain constant region of the murine monoclonal anti human
sclerostin antibodies described in section 1 with a human IgG1
constant region are provided.
I.B.3. Anti SOST CDR-Grafted Antibodies
[0293] CDR-grafted antibodies comprising heavy and light chain
variable region sequences from a human antibody wherein one or more
of the CDR regions of V.sub.H and/or V.sub.L are replaced with CDR
sequences of the murine antibodies are provided. A framework
sequence from any human antibody may serve as the template for CDR
grafting. However, straight chain replacement onto such a framework
often leads to some loss of binding affinity to the antigen. The
more homologous a human antibody is to the original murine
antibody, the less likely the possibility that combining the murine
CDRs with the human framework will introduce distortions in the
CDRs that could reduce affinity. Therefore, it is preferable that
the human variable framework that is chosen to replace the murine
variable framework apart from the CDRs have at least a 65% sequence
identity with the murine antibody variable region framework. It is
more preferable that the human and murine variable regions apart
from the CDRs have at least 70% sequence identify. It is even more
preferable that the human and murine variable regions apart from
the CDRs have at least 75% sequence identity. It is most preferable
that the human and murine variable regions apart from the CDRs have
at least 80% sequence identity. Methods for producing chimeric
antibodies are known in the art. (also see EP 0 239 400; PCT
Publication No. WO 91/09967; U.S. Pat. Nos. 5,225,539; 5,530,101;
and 5,585,089); veneering or resurfacing (see, e.g., EP 0 592 106;
EP 0 519 596; Padlan, Molecular Immunology, 28(4/5): 489-498
(1991); Studnicka et al., Protein Engineering, 7(6): 805-814
(1994); Roguska et al., Proc. Natl. Acad. Sci. USA, 91: 969-973
(1994)); and chain shuffling (see, e.g., U.S. Pat. No.
5,565,352).
I.B.4. Anti-Human Sclerostin Humanized Antibodies
[0294] Humanized antibodies are antibody molecules derived from
non-human species antibody that binds the desired antigen having
one or more complementarity determining regions (CDRs) from the
non-human species antibody and framework regions from a human
immunoglobulin molecule. Known human Ig sequences are disclosed,
e.g., at worldwide web sites:
www.ncbi.nlm.nih.gov/entrez-/query.fcgi;
www.atcc.org/phage/hdb.html; www.sciquest.com/; www.abcam.com/;
www.antibodyresource.com/onlinecomp.html;
www.public.iastate.edu/.about.pedro/research_tools.html;
www.mgen.uni-heidelberg.de/SD/IT/IT.html;
www.whfreeman.com/immunology/CH-05/kuby05.htm;
www.library.thinkquest.org/12429/Immune/Antibody.html; www hhmi
org/grants/lectures/1996/vlab/;
www.path.cam.ac.uk/.about.mrc7/m-ikeimages.html;
www.antibodyresource.com/;
mcb.harvard.edu/BioLinks/Immunology.html.www.immunologylink.com/;
pathbox.wustl.edu/.about.hcenter/index.-html;
www.biotech.ufl.edu/.about.hcl/;
www.pebio.com/pa/340913/340913.html-;
www.nal.usda.gov/awic/pubs/antibody/;
www.m.ehime-u.acjp/.about.yasuhito-/Elisa.html;
www.biodesign.com/table.asp;
www.icnet.uk/axp/facs/davies/lin-ks.html;
www.biotech.ufl.edu/.about.fccl/protocol.html;
www.isac-net.org/sites_geo.html;
aximtl.imt.uni-marburg.de/.about.rek/AEP-Start.html;
baserv.uci.kun.nl.about.jraats/linksl.html;
www.recab.uni-hd.de/immuno.bme.nwu.edu/;
www.mrc-cpe.cam.ac.uk/imt-doc/public/INTRO.html;
www.ibt.unam.mx/vir/V_mice.html; imgt.cnusc.fr:8104/;
www.biochem.ucl.ac.uk/.about.martin/abs/index.html;
antibody.bath.ac.uk/; abgen.cvm.tamu.edu/lab/wwwabgen.html;
www.unizh.chLabouthonegger/AHOseminar/Slide01.html;
www.cryst.bbk.ac.uk/.aboutubcg07s/; www.nimr
mrc.ac.uk/CC/ccaewg/ccaewg.htm;
www.path.cam.ac.uk/.about.mrc7/humanisation/TAHHP.html; www.ibt
unam.mx/vir/structure/stat_aim.html;
www.biosci.missouri.edu/smithgp/index.html;
www.cryst.bioc.cam.ac.uk/.abo-ut.fmolina/Web-pages/Pept/spottech.html;
www.jerini.de/frroducts.htm; www.patents.ibm.com/ibm.html. Kabat et
al., Sequences of Proteins of Immunological Interest, U.S. Dept.
Health (1983). Such imported sequences can be used to reduce
immunogenicity or reduce, enhance or modify binding, affinity,
on-rate, off-rate, avidity, specificity, half-life, or any other
suitable characteristic, as known in the art.
[0295] Framework (FR) residues in the human framework regions may
be substituted with the corresponding residue from the CDR donor
antibody to alter, in an embodiment, improve, antigen binding.
These framework substitutions are identified by methods well known
in the art, e.g., by modeling of the interactions of the CDR and
framework residues to identify framework residues important for
antigen binding and sequence comparison to identify unusual
framework residues at particular positions. See, e.g., Queen et
al., U.S. Pat. No. 5,585,089; Riechmann et al., Nature, 332:
323-327 (1988). Three-dimensional immunoglobulin models are
commonly available and are familiar to those skilled in the art.
Computer programs are available which illustrate and display
probable three-dimensional conformational structures of selected
candidate immunoglobulin sequences. Inspection of these displays
permits analysis of the likely role of the residues in the
functioning of the candidate immunoglobulin sequence, i.e., the
analysis of residues that influence the ability of the candidate
immunoglobulin to bind its antigen. In this way, FR residues can be
selected and combined from the consensus and import sequences so
that the desired antibody characteristic, such as increased
affinity for the target antigen(s), is achieved. In general, the
CDR residues are directly and most substantially involved in
influencing antigen binding. Antibodies can be humanized using a
variety of techniques known in the art, such as but not limited to
those described in Jones et al., Nature, 321:522-525 (1986);
Verhoeyen et al., Science, 239:1534-1536 (1988); Sims et al., J.
Immunol., 151: 2296-2308 (1993); Chothia and Lesk, J. Mol. Biol.,
196: 901-917 (1987), Carter et al., Proc. Natl. Acad. Sci. USA, 89:
4285-4289 (1992); Presta et al., J. Immunol., 151: 2623-2632
(1993); Padlan, Molecular Immunology, 28(4/5): 489-498 (1991);
Studnicka et al., Protein Engineering, 7(6): 805-814 (1994);
Roguska et al., Proc. Natl. Acad. Sci. USA, 91: 969-973 (1994); PCT
Publication Nos. WO 91/09967; WO 90/14443; WO 90/14424; WO
90/14430; WO 99/06834 (PCT/US98/16280); WO 97/20032
(PCT/US96/18978); WO 92/11272 (PCT/US91/09630); WO 92/03461
(PCT/US91/05939); WO 94/18219 (PCT/US94/01234); WO 92/01047
(PCT/GB91/01134); and WO 93/06213 (PCT/GB92/01755); EP Patent Nos.
EP 0 592 106; EP 0 519 596 and EP 0 239 400; U.S. Pat. Nos.
5,565,332; 5,723,323; 5,976,862; 5,824,514; 5,817,483; 5,814,476;
5,763,192; 5,723,323; 5,766,886; 5,714,352; 6,204,023; 6,180,370;
5,693,762; 5,530,101; 5,585,089; 5,225,539 and 4,816,567.
I.B.5. Anti-Sclerostin DVD-Binding Proteins
[0296] Also provided are dual variable domain binding proteins
(DVD-binding proteins) that bind one or more epitopes of
sclerostin. A DVD-binding protein may also bind an epitope of
sclerostin and an epitope of a second target antigen other than an
sclerostin polypeptide. An embodiment of such DVD-binding proteins
comprises a heavy chain that comprises the structural formula
VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable
domain, VD2 is a second heavy chain variable domain, C is a heavy
chain constant domain, X1 is a linker with the proviso that it is
not CH1, X2 is an Fc region, and n is 0 or 1, and, in an
embodiment, 1; and a light chain that comprises the structural
formula VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain
variable domain, VD2 is a second light chain variable domain, C is
a light chain constant domain, X1 is a linker with the proviso that
it is not CL, and X2 does not comprise an Fc region; and n is 0 or
1, and, in an embodiment, 1. Such a DVD-binding protein may
comprise two such heavy chains and two such light chains, wherein
each chain comprises variable domains linked in tandem without an
intervening constant region between variable regions, wherein a
heavy chain and a light chain associate to form two tandem antigen
binding sites, and a pair of heavy and light chains may associate
with another pair of heavy and light chains to form a tetrameric
binding protein with four antigen binding sites. In another
embodiment, a DVD-binding protein may comprise heavy and light
chains that each comprise three variable domains, e.g., VD1, VD2,
VD3, linked in tandem without an intervening constant region
between variable domains, wherein a pair of heavy and light chains
may associate to form three antigen binding sites, and wherein a
pair of heavy and light chains may associate with another pair of
heavy and light chains to form a tetrameric binding protein with
six antigen binding sites.
[0297] Each variable domain (VD) in a DVD-binding protein may be
obtained from one or more "parent" monoclonal antibodies that bind
one or more desired antigens or epitopes, such as sclerostin and/or
non-sclerostin antigens or epitopes (e.g., TNF-.alpha.).
III.A. Generation of Parent Monoclonal Antibodies
[0298] The variable domains of the DVD-binding protein can be
obtained from parent antibodies, including monoclonal antibodies
(mAb), capable of binding antigens of interest. These antibodies
may be naturally occurring or may be generated by recombinant
technology. It is understood that if an antibody that binds a
desired target antigen or epitope is polyclonal then it is still
necessary to obtain the variable domains of an antigen binding site
of a single antibody from the polyclonal population, i.e., of a
single monoclonal member of the polyclonal population, for use in
generating a DVD-binding protein. Monoclonal antibodies may be
generated by any of variety of methods known in the art, including
those described herein (see, sections A.1.-A.4., above).
III.B. Criteria for Selecting Parent Monoclonal Antibodies
[0299] An embodiment pertaining to selecting parent antibodies with
at least one or more properties desired in the DVD-binding protein
is provided. In an embodiment, the desired property is one or more
antibody parameters. In another embodiment, the antibody parameters
are antigen specificity, affinity to antigen, potency, biological
function, epitope recognition, stability, solubility, production
efficiency, immunogenicity, pharmacokinetics, bioavailability,
tissue cross reactivity, and orthologous antigen binding.
III.B.1. Affinity to Antigen
[0300] The desired affinity of a therapeutic mAb may depend upon
the nature of the antigen, and the desired therapeutic end-point.
In an embodiment, monoclonal antibodies have higher affinities
(Kd=0.01-0.50 pM) when blocking a cytokine-cytokine receptor
interaction as such interaction are usually high affinity
interactions (e.g., <pM-<nM ranges). In such instances, the
mAb affinity for its target should be equal to or better than the
affinity of the cytokine (ligand) for its receptor. On the other
hand, mAb with lesser affinity (>nM range) could be
therapeutically effective, e.g., in clearing circulating
potentially pathogenic proteins e.g., monoclonal antibodies that
bind to, sequester, and clear circulating species of a target
antigen, such as A-.beta. amyloid. In other instances, reducing the
affinity of an existing high affinity mAb by site-directed
mutagenesis or using a mAb with lower affinity for its target could
be used to avoid potential side-effects, e.g., a high affinity mAb
may sequester or neutralize all of its intended target, thereby
completely depleting/eliminating the function(s) of the targeted
protein. In this scenario, a low affinity mAb may
sequester/neutralize a fraction of the target that may be
responsible for the disease symptoms (the pathological or
over-produced levels), thus allowing a fraction of the target to
continue to perform its normal physiological function(s).
Therefore, it may be possible to reduce the Kd to adjust dose
and/or reduce side-effects. The affinity of the parental mAb might
play a role in appropriately targeting cell surface molecules to
achieve desired therapeutic out-come. For example, if a target is
expressed on cancer cells with high density and on normal cells
with low density, a lower affinity mAb will bind a greater number
of targets on tumor cells than normal cells, resulting in tumor
cell elimination via ADCC or CDC, and therefore might have
therapeutically desirable effects. Thus, selecting a mAb with
desired affinity may be relevant for both soluble and surface
targets.
[0301] Signaling through a receptor upon interaction with its
ligand may depend upon the affinity of the receptor-ligand
interaction. Similarly, it is conceivable that the affinity of a
mAb for a surface receptor could determine the nature of
intracellular signaling and whether the mAb may deliver an agonist
or an antagonist signal. The affinity-based nature of mAb-mediated
signaling may have an impact of its side-effect profile. Therefore,
the desired affinity and desired functions of therapeutic
monoclonal antibodies need to be determined carefully by in vitro
and in vivo experimentation.
[0302] The desired Kd of a binding protein (e.g., an antibody) may
be determined experimentally depending on the desired therapeutic
outcome. In an embodiment, parent antibodies with affinity (Kd) for
a particular antigen equal to, or better than, the desired affinity
of the DVD-binding protein for the same antigen are selected. The
antigen binding affinity and kinetics are assessed by Biacore or
another similar technique. In one embodiment, each parent antibody
has a dissociation constant (Kd) to its antigen of: at most about
10.sup.-7 M; at most about 10.sup.-8 M; at most about 10.sup.-9 M;
at most about 10.sup.-10 M; at most about 10.sup.-11 M; at most
about 10.sup.-12 M; or at most 10.sup.-13 M. First parent antibody
from which VD1 is obtained and second parent antibody from which
VD2 is obtained may have similar or different affinity (K.sub.D)
for the respective antigen. Each parent antibody has an on rate
constant (Kon) to the antigen of: at least about 10.sup.2 M.sup.-1
s.sup.-1; at least about 10.sup.3 M.sup.-1 s.sup.-1; at least about
10.sup.4 M.sup.-1 s.sup.-1; at least about 10.sup.5 M.sup.-1
s.sup.-1; or at least about 10.sup.6 M.sup.-1 s.sup.-1, as measured
by surface plasmon resonance. The first parent antibody from which,
for example, a VD1 is obtained and the second parent antibody from
which a VD2 is obtained may have similar or different on rate
constant (Kon) for the respective antigen. In one embodiment, each
parent antibody has an off rate constant (Koff) to the antigen of:
at most about 10.sup.-3 s.sup.-1; at most about 10.sup.-4 s.sup.-1;
at most about 10.sup.-5 s.sup.-1; or at most about 10.sup.-6
s.sup.-1, as measured by surface plasmon resonance. The first
parent antibody from which VD1 is obtained and the second parent
antibody from which VD2 is obtained may have similar or different
off rate constants (Koff) for the respective antigen.
III.B.2. Potency
[0303] The desired affinity/potency of parental monoclonal
antibodies will depend on the desired therapeutic outcome. For
example, for receptor-ligand (R-L) interactions the affinity (kd)
is equal to or better than the R-L kd (pM range). For simple
clearance of a pathologic circulating proteins, the Kd could be in
low nM range, e.g., clearance of various species of circulating
A-.beta. peptide. In addition, the Kd will also depend on whether
the target expresses multiple copies of the same epitope, e.g., an
mAb targeting conformational epitope in A.beta. oligomers.
[0304] Where VD1 and VD2 bind the same antigen, but distinct
epitopes, the DVD-binding protein will contain binding sites for
the same antigen, thus increasing avidity and thereby the apparent
Kd of the DVD-binding protein. In an embodiment, parent antibodies
with equal or lower Kd than that desired in the DVD-binding protein
are chosen. The affinity considerations of a parental mAb may also
depend upon whether the DVD-binding protein contains four or more
identical antigen binding sites (i.e., a DVD-binding protein from a
single mAb). In this case, the apparent Kd would be greater than
the mAb due to avidity. Such DVD-binding proteins can be employed
for cross-linking surface receptor, increased neutralization
potency, enhanced clearance of pathological proteins, etc.
[0305] In another embodiment, parent antibodies with neutralization
potency for specific antigen equal to or better than the desired
neutralization potential of the DVD-binding protein for the same
antigen are selected. The neutralization potency can be assessed by
a target-dependent bioassay where cells of appropriate type produce
a measurable signal (i.e., proliferation or cytokine production) in
response to target stimulation, and target neutralization by the
mAb can reduce the signal in a dose-dependent manner.
III.B.3. Biological Functions
[0306] Monoclonal antibodies can perform potentially several
functions. Some of these functions are listed in Table 4. These
functions can be assessed by both in vitro assays (e.g., cell-based
and biochemical assays) and in vivo animal models.
TABLE-US-00005 TABLE 4 Some Potential Applications For Therapeutic
Antibodies. Target (Class) Mechanism of Action (target) Soluble
Neutralization of activity (e.g., a cytokine, such SOST)
(cytokines, other) Enhance clearance (e.g., A.beta. oligomers)
Increase half-life (e.g., GLP 1) Cell Surface Agonist (e.g., GLP1
R, EPO R, etc.) (Receptors, other) Antagonist (e.g., integrins,
etc.) Cytotoxic (CD 20, etc.) Protein deposits Enhance
clearance/degradation (e.g., A.beta. plaques, amyloid deposits)
[0307] MAbs with distinct functions described in the examples
herein and in Table 8 can be selected to achieve desired
therapeutic outcomes. Two or more selected parent monoclonal
antibodies can then be used in DVD-binding protein format to
achieve two distinct functions in a single DVD-binding protein. For
example, a DVD-binding protein can be generated by selecting a
parent mAb that neutralizes function of a specific cytokine, such
as sclerostin, and selecting a parent mAb that enhances clearance
of a pathological protein. Similarly, two parent mAbs may be
selected that recognize two different cell surface receptors, one
mAb with an agonist function on one receptor and the other mAb with
an antagonist function on a different receptor. These two selected
mAbs, each with a distinct function, can be used to construct a
single DVD-binding protein that will possess the two distinct
functions (agonist and antagonist) of the selected monoclonal
antibodies in a single molecule. Similarly, two antagonistic mAbs
to cell surface receptors, each blocking binding of respective
receptor ligands (e.g., EGF and IGF), may be used in a DVD-binding
protein format. Conversely, an antagonistic anti-receptor mAb
(e.g., anti-EGFR) and a neutralizing anti-soluble mediator (e.g.,
anti-IGF1/2) mAb can be selected to make a DVD-binding protein.
III.B.4. Epitope Recognition:
[0308] Different regions of proteins may perform different
functions. For example, specific regions of a cytokine, such as
sclerostin, interact with the cytokine receptor to bring about
receptor activation whereas other regions of the protein may be
required for stabilizing the cytokine. In this instance, one may
select a mAb that binds specifically to the receptor interacting
region(s) on the cytokine and thereby block cytokine-receptor
interaction. In some cases, for example certain chemokine receptors
that bind multiple ligands, a mAb that binds to the epitope (region
on chemokine receptor) that interacts with only one ligand can be
selected. In other instances, monoclonal antibodies can bind to
epitopes on a target that are not directly responsible for
physiological functions of the protein, but binding of a mAb to
these regions could either interfere with physiological functions
(steric hindrance) or alter the conformation of the protein such
that the protein cannot function (mAb to receptors with multiple
ligand which alter the receptor conformation such that none of the
ligand can bind). Anti-cytokine monoclonal antibodies that do not
block binding of the cytokine to its receptor, but block signal
transduction have also been identified (e.g., 125-2H, an anti-IL-18
mAb).
[0309] Examples of epitopes and mAb functions include, but are not
limited to, blocking Receptor-Ligand (R-L) interaction
(neutralizing mAb that binds R-interacting site); steric hindrance
resulting in diminished or no R-binding. An antibody can bind the
target at a site other than a receptor binding site, but still
interfere with receptor binding and functions of the target by
inducing conformational change and eliminate function (e.g.,
XOLAIR.RTM. omalizumab, Genetech/Novartis), binding to R but block
signaling (125-2H mAb).
[0310] In an embodiment, the parental mAb needs to target the
appropriate epitope for maximum efficacy. Such epitope should be
conserved in the DVD-binding protein. The binding epitope of a mAb
can be determined by several approaches, including
co-crystallography, limited proteolysis of mAb-antigen complex plus
mass spectrometric peptide mapping (Legros V. et al 2000 Protein
Sci. 9:1002-10), phage displayed peptide libraries (O'Connor K H et
al 2005 J Immunol Methods. 299:21-35), as well as mutagenesis (Wu
C. et al. 2003 J Immunol 170:5571-7).
III.B.5. Physicochemical and Pharmaceutical Properties:
[0311] Therapeutic treatment with antibodies often requires
administration of high doses, often several mg/kg (due to a low
potency on a mass basis as a consequence of a typically large
molecular weight). In order to accommodate patient compliance and
to adequately address chronic disease therapies and outpatient
treatment, subcutaneous (s.c.) or intramuscular (i.m.)
administration of therapeutic mAbs is desirable. For example, the
maximum desirable volume for s.c administration is .about.1.0 mL,
and therefore, concentrations of >100 mg/mL are desirable to
limit the number of injections per dose. In an embodiment, the
therapeutic antibody is administered in one dose. The development
of such formulations is constrained, however, by protein-protein
interactions (e.g., aggregation, which potentially increases
immunogenicity risks) and by limitations during processing and
delivery (e.g., viscosity). Consequently, the large quantities
required for clinical efficacy and the associated development
constraints limit full exploitation of the potential of antibody
formulation and s.c administration in high-dose regimens. It is
apparent that the physicochemical and pharmaceutical properties of
a protein molecule and the protein solution are of utmost
importance, e.g., stability, solubility and viscosity features.
III.B.5.i. Stability
[0312] A "stable" antibody formulation is one in which the antibody
therein essentially retains its physical stability and/or chemical
stability and/or biological activity upon storage. Stability can be
measured at a selected temperature for a selected time period. In
an embodiment, the antibody in the formulation is stable at room
temperature (about 30.degree. C.) or at 40.degree. C. for at least
1 month and/or stable at about 2-8.degree. C. for at least 1 year
for at least 2 years. Furthermore, in an embodiment, the
formulation is stable following freezing (to, e.g., -70.degree. C.)
and thawing of the formulation, hereinafter referred to as a
"freeze/thaw cycle." In another example, a "stable" formulation may
be one wherein less than about 10% and less than about 5% of the
protein is present as an aggregate in the formulation.
[0313] A DVD-binding protein stable in vitro at various
temperatures for an extended time period is desirable. One can
achieve this by rapid screening of parental mAbs stable in vitro at
elevated temperature, e.g., at 40.degree. C. for 2-4 weeks, and
then assess stability. During storage at 2-8.degree. C., the
protein reveals stability for at least 12 months, e.g., at least 24
months. Stability (% of monomeric, intact molecule) can be assessed
using various techniques such as cation exchange chromatography,
size exclusion chromatography, SDS-PAGE, as well as bioactivity
testing. For a more comprehensive list of analytical techniques
that may be employed to analyze covalent and conformational
modifications, see, Jones, A. J. S. (1993) "Analytical methods for
the assessment of protein formulations and delivery systems," In
Cleland, J. L.; Langer, R., editors. Formulation and delivery of
peptides and proteins, 1st edition (Washington, ACS), pages 22-45;
and Pearlman, R.; Nguyen, T. H. (1990) "Analysis of protein drugs,"
In Lee, V. H., editor. Peptide and protein drug delivery, 1st
edition (New York, Marcel Dekker, Inc.), pages 247-301.
[0314] Heterogeneity and aggregate formation: stability of the
antibody may be such that the formulation may reveal less than
about 10%, and, in an embodiment, less than about 5%, in another
embodiment, less than about 2%, or, in an embodiment, within the
range of 0.5% to 1.5% or less in the GMP antibody material that is
present as aggregate. Size exclusion chromatography is a method
that is sensitive, reproducible, and very robust in the detection
of protein aggregates.
[0315] In addition to low aggregate levels, the antibody must, in
an embodiment, be chemically stable. Chemical stability may be
determined by ion exchange chromatography (e.g., cation or anion
exchange chromatography), hydrophobic interaction chromatography,
or other methods such as isoelectric focusing or capillary
electrophoresis. For instance, chemical stability of the antibody
may be such that after storage of at least 12 months at 2-8.degree.
C. the peak representing unmodified antibody in a cation exchange
chromatography may increase not more than 20%, in an embodiment,
not more than 10%, or, in another embodiment, not more than 5% as
compared to the antibody solution prior to storage testing.
[0316] In an embodiment, the parent antibodies display structural
integrity; correct disulfide bond formation, and correct folding:
Chemical instability due to changes in secondary or tertiary
structure of an antibody may impact antibody activity. For
instance, stability as indicated by activity of the antibody may be
such that after storage of at least 12 months at 2-8.degree. C. the
activity of the antibody may decrease not more than 50%, in an
embodiment not more than 30%, or even not more than 10%, or in an
embodiment not more than 5% or 1% as compared to the antibody
solution prior to storage testing. Suitable antigen-binding assays
can be employed to determine antibody activity.
III.B.5.ii. Solubility:
[0317] The "solubility" of a mAb correlates with the production of
correctly folded, monomeric IgG. The solubility of the IgG may
therefore be assessed by HPLC. For example, soluble (monomeric) IgG
will give rise to a single peak on the HPLC chromatograph, whereas
insoluble (e.g., multimeric and aggregated) will give rise to a
plurality of peaks. A person skilled in the art will therefore be
able to detect an increase or decrease in solubility of an IgG
using routine HPLC techniques. For a more comprehensive list of
analytical techniques that may be employed to analyze solubility
(see, Jones, A. G. Dep. Chem. Biochem. Eng., Univ. Coll. London,
London, UK. Editor(s): Shamlou, P. Ayazi, Process. Solid-Liquid
Suspensions (1993), 93-117. (Butterworth-Heinemann, Oxford, UK) and
Pearlman, Rodney; Nguyen, Tue H, Advances in Parenteral Sciences
(1990), 4 (Pept. Protein Drug Delivery), 247-301. Solubility of a
therapeutic mAb is critical for formulating to high concentration
often required for adequate dosing. As outlined herein,
solubilities of >100 mg/mL may be required to accommodate
efficient antibody dosing. For instance, antibody solubility may be
not less than about 5 mg/mL in early research phase, in an
embodiment not less than about 25 mg/mL in advanced process science
stages, or in an embodiment not less than about 100 mg/mL, or in an
embodiment not less than about 150 mg/mL. The intrinsic properties
of a protein molecule are important to the physico-chemical
properties of the protein solution, e.g., stability, solubility,
viscosity. However, a person skilled in the art will appreciate
that a broad variety of excipients exist that may be used as
additives to beneficially impact the characteristics of the final
protein formulation. These excipients may include: (i) liquid
solvents, cosolvents (e.g., alcohols such as ethanol); (ii)
buffering agents (e.g., phosphate, acetate, citrate, amino acid
buffers); (iii) sugars or sugar alcohols (e.g., sucrose, trehalose,
fructose, raffinose, mannitol, sorbitol, dextrans); (iv)
surfactants (e.g., polysorbate 20, 40, 60, 80, poloxamers); (v)
isotonicity modifiers (e.g., salts such as NaCl, sugars, sugar
alcohols); and (vi) others (e.g., preservatives, chelating agents,
antioxidants, chelating substances (e.g., EDTA), biodegradable
polymers, carrier molecules (e.g., HSA, PEGs)
[0318] Viscosity is a parameter of high importance with regard to
antibody manufacture and antibody processing (e.g.,
diafiltration/ultrafiltration), fill-finish processes (pumping
aspects, filtration aspects) and delivery aspects (syringeability,
sophisticated device delivery). Low viscosities enable the liquid
solution of the antibody having a higher concentration. This
enables the same dose to be administered in smaller volumes. Small
injection volumes inhere the advantage of lower pain on injection
sensations, and the solutions do not necessarily have to be
isotonic to reduce pain on injection in the patient. The viscosity
of the antibody solution may be such that at shear rates of 100
(1/s) antibody solution viscosity is below 200 mPas, in an
embodiment below 125 mPas, in another embodiment below 70 mPas, and
in yet another embodiment below 25 mPas or even below 10 mPas.
III.B.5.iii. Production Efficiency
[0319] The generation of a DVD-binding protein that is efficiently
expressed in mammalian cells, such as Chinese hamster ovary cells
(CHO), will in an embodiment require two parental monoclonal
antibodies which are themselves expressed efficiently in mammalian
cells. The production yield from a stable mammalian line (i.e.,
CHO) should be above about 0.5 g/L, in an embodiment above about 1
g/L, and in another embodiment in the range of about 2 to about 5
g/L or more (Kipriyanov S M, Little M. 1999 Mol Biotechnol.
12:173-201; Carroll S, Al-Rubeai M. 2004 Expert Opin Biol Ther.
4:1821-9).
[0320] Production of antibodies and Ig fusion proteins in mammalian
cells is influenced by several factors. Engineering of the
expression vector via incorporation of strong promoters, enhancers
and selection markers can maximize transcription of the gene of
interest from an integrated vector copy. The identification of
vector integration sites that are permissive for high levels of
gene transcription can augment protein expression from a vector
(Wurm et al, 2004, Nature Biotechnology, 2004, 22(11): 1393-1398).
Furthermore, levels of production are affected by the ratio of
antibody heavy and light chains and various steps in the process of
protein assembly and secretion (Jiang et al. 2006, Biotechnology
Progress, January-February 2006, vol. 22, no. 1, pp. 313-318).
III.B.6. Immunogenicity
[0321] Administration of a therapeutic mAb may result in certain
incidence of an immune response (i.e., the formation of endogenous
antibodies directed against the therapeutic mAb). Potential
elements that might induce immunogenicity should be analyzed during
selection of the parental monoclonal antibodies, and steps to
reduce such risk can be taken to optimize the parental monoclonal
antibodies prior to DVD-binding protein construction. Mouse-derived
antibodies have been found to be highly immunogenic in patients.
The generation of chimeric antibodies comprised of mouse variable
and human constant regions presents a logical next step to reduce
the immunogenicity of therapeutic antibodies (Morrison and Schlom,
1990). Alternatively, immunogenicity can be reduced by transferring
murine CDR sequences into a human antibody framework (reshaping/CDR
grafting/humanization), as described for a therapeutic antibody by
Riechmann et al., 1988. Another method is referred to as
"resurfacing" or "veneering", starting with the rodent variable
light and heavy domains, only surface-accessible framework amino
acids are altered to human ones, while the CDR and buried amino
acids remain from the parental rodent antibody (Roguska et al.,
1996). In another type of humanization, instead of grafting the
entire CDRs, one technique grafts only the "specificity-determining
regions" (SDRs), defined as the subset of CDR residues that are
involved in binding of the antibody to its target (Kashmiri et al.,
2005). This necessitates identification of the SDRs either through
analysis of available three-dimensional structures of
antibody-target complexes or mutational analysis of the antibody
CDR residues to determine which interact with the target.
Alternatively, fully human antibodies may have reduced
immunogenicity compared to murine, chimeric, or humanized
antibodies.
[0322] Another approach to reduce the immunogenicity of therapeutic
antibodies is the elimination of certain specific sequences that
are predicted to be immunogenic. In one approach, after a first
generation biologic has been tested in humans and found to be
unacceptably immunogenic, the B-cell epitopes can be mapped and
then altered to avoid immune detection. Another approach uses
methods to predict and remove potential T-cell epitopes.
Computational methods have been developed to scan and to identify
the peptide sequences of biologic therapeutics with the potential
to bind to MHC proteins (Desmet et al., 2005). Alternatively a
human dendritic cell-based method can be used to identify CD4.sup.+
T-cell epitopes in potential protein allergens (Stickler et al.,
2005; S. L. Morrison and J. Schlom, Important Adv. Oncol. (1990),
pp. 3-18; Riechmann, L., Clark, M., Waldmann, H. and Winter, G.
"Reshaping human antibodies for therapy," Nature (1988) 332:
323-327; Roguska-M-A, Pedersen-J-T, Henry-A-H, Searle-S-M,
Roja-C-M, Avery-B, Hoffee-M, Cook-S, Lambert-J-M, Blattler-W-A,
Rees-A-R, Guild-B-C, "A comparison of two murine mAbs humanized by
CDR-grafting and variable domain resurfacing," Protein Engineering,
(1996), 9: 895-904; Kashmiri-Syed-V-S, De-Pascalis-Roberto,
Gonzales-Noreen-R, Schlom-Jeffrey, "SDR grafting--a new approach to
antibody humanization," Methods (San Diego Calif.), May 2005,
36(1): 25-34; Desmet-Johan, Meersseman-Geert, Boutonnet-Nathalie,
Pletinckx-Jurgen, De-Clercq-Krista, Debulpaep-Maja,
Braeckman-Tessa, Lasters-Ignace, "Anchor profiles of HLA-specific
peptides: analysis by a novel affinity scoring method and
experimental validation," Proteins (2005) 58: 53-69; Stickler-M-M,
Estell-D-A, Harding-F-A., "CD4+ T-cell epitope determination using
unexposed human donor peripheral blood mononuclear cells," J.
Immunother. (2000) 23: 654-60.)
III.B.7. In Vivo Efficacy
[0323] To generate a DVD-binding protein with desired in vivo
efficacy, it is important to generate and select mAbs with
similarly desired in vivo efficacy when given in combination.
However, in some instances the DVD-binding protein may exhibit in
vivo efficacy that cannot be achieved with the combination of two
separate mAbs. For instance, a DVD-binding protein may bring two
targets in close proximity leading to an activity that cannot be
achieved with the combination of two separate mAbs. Additional
desirable biological functions are described herein in section B3.
Parent antibodies with characteristics desirable in the DVD-binding
protein may be selected based on factors such as pharmacokinetic
half-life (t1/2); tissue distribution; soluble versus cell surface
targets; and target concentration-soluble/density-surface.
III.B.8. In Vivo Tissue Distribution
[0324] To generate a DVD-binding protein with desired in vivo
tissue distribution, in an embodiment, parent mAbs with similar
desired in vivo tissue distribution profile must be selected.
Alternatively, based on the mechanism of the dual-specific
targeting strategy, it may at other times not be required to select
parent mAbs with the similarly desired in vivo tissue distribution
when given in combination. For instance, in the case of a
DVD-binding protein in which one binding component targets the
DVD-binding protein to a specific site thereby bringing the second
binding component to the same target site. For example, one binding
specificity of a DVD-binding protein could target pancreas (islet
cells) and the other specificity could bring GLP1 to the pancreas
to induce insulin.
III.B.9. Isotype
[0325] To generate a DVD-binding protein with desired properties
including, but not limited to, isotype, effector functions, and the
circulating half-life, parent mAbs are selected that possess
appropriate Fc-effector functions depending on the therapeutic
utility and the desired therapeutic end-point. There are five main
heavy chain classes or isotypes, some of which have several
sub-types and these determine the effector functions of an antibody
molecule. These effector functions reside in the hinge region, CH2,
and CH3 domains of the antibody molecule. However, residues in
other parts of an antibody molecule may have effects on effector
functions as well. The hinge region Fc-effector functions include:
(i) antibody-dependent cellular cytotoxicity (ADCC), (ii)
complement (C1q) binding, activation, and complement-dependent
cytotoxicity (CDC), (iii) phagocytosis/clearance of
antigen-antibody complexes, and (iv) cytokine release in some
instances. These Fc-effector functions of an antibody molecule are
mediated through the interaction of the Fc-region with a set of
class-specific cell surface receptors. Antibodies of the IgG1
isotype are most active while IgG2 and IgG4 having minimal or no
effector functions. The effector functions of the IgG antibodies
are mediated through interactions with three structurally
homologous cellular Fc receptor types (and sub-types) (FcgR1,
FcgRII, and FcgRIII). These effector functions of an IgG1 can be
eliminated by mutating specific amino acid residues in the lower
hinge region (e.g., L234A, L235A) that are required for FcgR and
C1q binding Amino acid residues in the Fc region, in particular the
CH2-CH3 domains, also determine the circulating half-life of the
antibody molecule. This Fc function is mediated through the binding
of the Fc-region to the neonatal Fc receptor (FcRn), which is
responsible for recycling of antibody molecules from the acidic
lysosomes back to the general circulation.
[0326] Whether a mAb should have an active or an inactive isotype
will depend on the desired therapeutic end-point for an antibody.
Some examples of usage of isotypes and desired therapeutic outcome
are listed below: [0327] 1. If the desired end-point is functional
neutralization of a soluble cytokine then an inactive isotype may
be used; [0328] 2. If the desired out-come is clearance of a
pathological protein an active isotype may be used; [0329] 3. If
the desired out-come is clearance of protein aggregates an active
isotype may be used; [0330] 4. If the desired outcome is to
antagonize a surface receptor an inactive isotype is used (Tysabri,
IgG4; OKT3.RTM., mutated IgG1); [0331] 5. If the desired outcome is
to eliminate target cells an active isotype is used (Herceptin,
IgG1 (and with enhanced effector functions); and [0332] 6. If the
desired outcome is to clear proteins from circulation without
entering the CNS an IgM isotype may be used (e.g., clearing
circulating Ab peptide species). The Fc effector functions of a
parental mAb can be determined by various in vitro methods well
known in the art.
[0333] As discussed, the selection of isotype, and thereby the
effector functions will depend upon the desired therapeutic
end-point. In cases where simple neutralization of a circulating
target is desired, for example blocking receptor-ligand
interactions, the effector functions may not be required. In such
instances, isotypes or mutations in the Fc-region of an antibody
that eliminate effector functions are desirable. In other instances
where elimination of target cells is the therapeutic end-point, for
example elimination of tumor cells, isotypes or mutations or
de-fucosylation in the Fc-region that enhance effector functions
are desirable (Presta GL, Adv. Drug Delivery Rev., 58: 640-656,
2006; Satoh M., Iida S., Shitara K., Expert Opinion Biol. Ther., 6:
1161-1173, 2006). Similarly, depending up on the therapeutic
utility, the circulating half-life of an antibody molecule can be
reduced/prolonged by modulating antibody-FcRn interactions by
introducing specific mutations in the Fc region (Dall'Acqua W F,
Kiener P A, Wu H., J. Biol. Chem., 281: 23514-23524 (2006); Petkova
S B., Akilesh S., Sproule T J. et al., Internat. Immunol., 18:
1759-1769 (2006); Vaccaro C., Bawdon R., Wanjie S et al., Proc.
Natl. Acad. Sci. USA, 103: 18709-18714 (2007).
[0334] The published information on the various residues that
influence the different effector functions of a normal therapeutic
mAb may need to be confirmed for a DVD-binding protein. It may be
possible that in a DVD-binding protein format additional
(different) Fc-region residues, other than those identified for the
modulation of monoclonal antibody effector functions, may be
important.
[0335] Overall, the decision as to which Fc-effector functions
(isotype) will be critical in the final DVD-binding protein format
will depend up on the disease indication, therapeutic target,
desired therapeutic end-point, and safety considerations. Listed
below are exemplary appropriate heavy chain and light chain
constant regions including, but not limited to: [0336]
IgG1--allotype: Glmz [0337] IgG1 mutant--A234, A235 [0338]
IgG2--allotype: G2m(n-) [0339] Kappa--Km3 [0340] Lambda
[0341] Fc Receptor and C1q Studies:
[0342] The possibility of unwanted antibody-dependent cell-mediated
cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) by
antibody complexing to any overexpressed target on cell membranes
can be abrogated by (for example, L234A, L235A) hinge-region
mutations. These substituted amino acids, present in the IgG1 hinge
region of mAb, are expected to result in diminished binding of mAb
to human Fc receptors (but not FcRn), as FcgR binding is thought to
occur within overlapping sites on the IgG1 hinge region. This
feature of mAb may lead to an improved safety profile over
antibodies containing a wild-type IgG. Binding of mAb to human Fc
receptors can be determined by flow cytometry experiments using
cell lines (e.g., THP-1, K562) and an engineered CHO cell line that
expresses FcgRIIb (or other FcgRs). Compared to IgG1 control
monoclonal antibodies, mAb show reduced binding to FcgRI and
FcgRIIa whereas binding to FcgRIIb is unaffected. The binding and
activation of C1q by antigen/IgG immune complexes triggers the
classical complement cascade with consequent inflammatory and/or
immunoregulatory responses. The C1q binding site on IgGs has been
localized to residues within the IgG hinge region. C1q binding to
increasing concentrations of mAb was assessed by C1q ELISA. The
results demonstrate that mAb is unable to bind to C1q, as expected
when compared to the binding of a wildtype control IgG1. Overall,
the L234A, L235A hinge region mutation abolishes binding of mAb to
FcgRI, FcgRIIa, and C1q, but does not impact the interaction of mAb
with FcgRIIb. These data suggest that in vivo mAb with mutant Fc
will interact normally with the inhibitory FcgRIIb but will likely
fail to interact with the activating FcgRI and FcgRIIa receptors or
C1q.
[0343] Human FcRn Binding:
[0344] The neonatal receptor (FcRn) is responsible for transport of
IgG across the placenta and to control the catabolic half-life of
the IgG molecules. It might be desirable to increase the terminal
half-life of an antibody to improve efficacy, to reduce the dose or
frequency of administration, or to improve localization to the
target. Alternatively, it might be advantageous to do the converse
that is, to decrease the terminal half-life of an antibody to
reduce whole body exposure or to improve the target-to-non-target
binding ratios. Tailoring the interaction between IgG and its
salvage receptor, FcRn, offers a way to increase or decrease the
terminal half-life of IgG. Proteins in the circulation, including
IgG, are taken up in the fluid phase through micropinocytosis by
certain cells, such as those of the vascular endothelia. IgG can
bind FcRn in endosomes under slightly acidic conditions (pH
6.0-6.5) and can recycle to the cell surface, where it is released
under almost neutral conditions (pH 7.0-7.4). Mapping of the
Fc-region-binding site on FcRn80, 16, 17 showed that two histidine
residues that are conserved across species, His310 and His435, are
responsible for the pH dependence of this interaction. Using
phage-display technology, a mouse Fc-region mutation that increases
binding to FcRn and extends the half-life of mouse IgG was
identified (see Victor, G. et al., Nature Biotechnology, 15(7):
637-640 (1997)). Fc-region mutations that increase the binding
affinity of human IgG for FcRn at pH 6.0, but not at pH 7.4, have
also been identified (see, Dall'Acqua et al., J. Immunol., 169(9):
5171-80 (2002)). Moreover, in one case, a similar pH-dependent
increase in binding (up to 27-fold) was also observed for rhesus
FcRn, and this resulted in a twofold increase in serum half-life in
rhesus monkeys compared with the parent IgG (see, Hinton et al., J.
Biol. Chem., 279(8): 6213-6216 (2004)). These findings indicate
that it is feasible to extend the plasma half-life of antibody
therapeutics by tailoring the interaction of the Fc region with
FcRn. Conversely, Fc-region mutations that attenuate interaction
with FcRn can reduce antibody half-life.
III.B.10. Pharmacokinetics (PK)
[0345] To generate a DVD-binding protein with desired
pharmacokinetic profile, in an embodiment, parent mAbs with the
similarly desired pharmacokinetic profile are selected. One
consideration is that immunogenic response to monoclonal antibodies
(i.e., "HAHA", human anti-human antibody response; "HACA", human
anti-chimeric antibody response) further complicates the
pharmacokinetics of these therapeutic agents. In an embodiment,
monoclonal antibodies with minimal or no immunogenicity are used
for constructing DVD-binding proteins such that the resulting
DVD-binding proteins will also have minimal or no immunogenicity.
Some of the factors that determine the PK of a mAb include, but are
not limited to, intrinsic properties of the mAb (VH amino acid
sequence); immunogenicity; FcRn binding and Fc functions.
[0346] The PK profile of selected parental monoclonal antibodies
can be easily determined in rodents as the PK profile in rodents
correlates well with (or closely predicts) the PK profile of
monoclonal antibodies in cynomolgus monkey and humans.
[0347] After the parental monoclonal antibodies with desired PK
characteristics (and other desired functional properties as
discussed herein) are selected, the DVD-binding protein is
constructed. As the DVD-binding proteins contain two
antigen-binding domains from two parental monoclonal antibodies,
the PK properties of the DVD-binding protein are assessed as well.
Therefore, while determining the PK properties of the DVD-binding
protein, PK assays may be employed that determine the PK profile
based on functionality of both antigen-binding domains derived from
the 2 parent monoclonal antibodies. The PK profile of a DVD-binding
protein can be determined Additional factors that may impact the PK
profile of DVD-binding protein include the antigen-binding domain
(CDR) orientation, linker size, and Fc/FcRn interactions. PK
characteristics of parent antibodies can be evaluated by assessing
the following parameters: absorption, distribution, metabolism and
excretion.
[0348] Absorption:
[0349] To date, administration of therapeutic monoclonal antibodies
is via parenteral routes (e.g., intravenous [IV], subcutaneous
[SC], or intramuscular [IM]). Absorption of a mAb into the systemic
circulation following either SC or IM administration from the
interstitial space is primarily through the lymphatic pathway.
Saturable, presystemic, proteolytic degradation may result in
variable absolute bioavailability following extravascular
administration. Usually, increases in absolute bioavailability with
increasing doses of monoclonal antibodies may be observed due to
saturated proteolytic capacity at higher doses. The absorption
process for a mAb is usually quite slow as the lymph fluid drains
slowly into the vascular system, and the duration of absorption may
occur over hours to several days. The absolute bioavailability of
monoclonal antibodies following SC administration generally ranges
from 50% to 100%. In the case of a transport-mediating structure at
the blood-brain barrier (BBB) targeted by the DVD-binding protein
construct, circulation times in plasma may be reduced due to
enhanced trans-cellular transport at the blood brain barrier (BBB)
into the CNS compartment, where the DVD-binding protein is
liberated to enable interaction via its second antigen recognition
site.
[0350] Distribution:
[0351] Following IV administration, monoclonal antibodies usually
follow a biphasic serum (or plasma) concentration-time profile,
beginning with a rapid distribution phase, followed by a slow
elimination phase. In general, a biexponential pharmacokinetic
model best describes this kind of pharmacokinetic profile. The
volume of distribution in the central compartment (Vc) for a mAb is
usually equal to or slightly larger than the plasma volume (2-3
liters). A distinct biphasic pattern in serum (plasma)
concentration versus time profile may not be apparent with other
parenteral routes of administration, such as IM or SC, because the
distribution phase of the serum (plasma) concentration-time curve
is masked by the long absorption portion. Many factors, including
physicochemical properties, site-specific and target-oriented
receptor mediated uptake, binding capacity of tissue, and mAb dose
can influence biodistribution of a mAb. Some of these factors can
contribute to nonlinearity in biodistribution for a mAb.
[0352] Metabolism and Excretion:
[0353] Due to the molecular size, intact monoclonal antibodies are
not excreted into the urine via kidney. They are primarily
inactivated by metabolism (e.g., catabolism). For IgG-based
therapeutic monoclonal antibodies, half-lives typically ranges from
hours or 1-2 days to over 20 days. The elimination of a mAb can be
affected by many factors, including, but not limited to, affinity
for the FcRn receptor, immunogenicity of the mAb, the degree of
glycosylation of the mAb, the susceptibility for the mAb to
proteolysis, and receptor-mediated elimination.
III.B.11. Tissue Cross-Reactivity Pattern on Human and Tox
Species
[0354] Identical staining pattern suggests that potential human
toxicity can be evaluated in tox species. Tox species are those
animal in which unrelated toxicity is studied.
[0355] The individual antibodies are selected to meet two criteria:
(1) tissue staining appropriate for the known expression of the
antibody target and (2) similar staining pattern between human and
tox species tissues from the same organ.
[0356] Criterion 1: Immunizations and/or antibody selections
typically employ recombinant or synthesized antigens (proteins,
carbohydrates or other molecules). Binding to the natural
counterpart and counterscreen against unrelated antigens are often
part of the screening funnel for therapeutic antibodies. However,
screening against a multitude of antigens is often unpractical.
Therefore, tissue cross-reactivity studies with human tissues from
all major organs serve to rule out unwanted binding of the antibody
to any unrelated antigens.
[0357] Criterion 2: Comparative tissue cross reactivity studies
with human and tox species tissues (cynomolgus monkey, dog,
possibly rodents, and others, the same 36 or 37 tissues being
tested as in the human study) help to validate the selection of a
tox species. In the typical tissue cross-reactivity studies on
frozen tissue sections, therapeutic antibodies may demonstrate the
expected binding to the known antigen and/or to a lesser degree
binding to tissues based either on low level interactions
(unspecific binding, low level binding to similar antigens, low
level charge based interactions, etc.). In any case, the most
relevant toxicology animal species is the one with the highest
degree of coincidence of binding to human and animal tissue.
[0358] Tissue cross-reactivity studies follow the appropriate
regulatory guidelines including EC CPMP Guideline III/5271/94
"Production and quality control of mAbs" and the 1997 US FDA/CBER
"Points to Consider in the Manufacture and Testing of Monoclonal
Antibody Products for Human Use". Cryosections (5 .mu.m) of human
tissues obtained at autopsy or biopsy were fixed and dried on
object glass. The peroxidase staining of tissue sections are
performed, using the avidin-biotin system. FDA's Guidance "Points
to Consider in the Manufacture and Testing of Monoclonal Antibody
Products for Human Use". Relevant references include Clarke, J.
(2004), Boon, L. (2002a), Boon, L. (2002b), Ryan, A. (1999).
[0359] Tissue-cross reactivity studies are often done in two
stages, with the first stage including cryosections of 32 tissues
(typically: Adrenal Gland, Gastrointestinal Tract, Prostate,
Bladder, Heart, Skeletal Muscle, Blood Cells, Kidney, Skin, Bone
Marrow, Liver, Spinal Cord, Breast, Lung, Spleen, Cerebellum, Lymph
Node, Testes, Cerebral Cortex, Ovary, Thymus, Colon, Pancreas,
Thyroid, Endothelium, Parathyroid, Ureter, Eye, Pituitary, Uterus,
Fallopian Tube and Placenta) from one human donor. In the second
phase, a full cross reactivity study is performed with up to 38
tissues (including adrenal, blood, blood vessel, bone marrow,
cerebellum, cerebrum, cervix, esophagus, eye, heart, kidney, large
intestine, liver, lung, lymph node, breast mammary gland, ovary,
oviduct, pancreas, parathyroid, peripheral nerve, pituitary,
placenta, prostate, salivary gland, skin, small intestine, spinal
cord, spleen, stomach, striated muscle, testis, thymus, thyroid,
tonsil, ureter, urinary bladder, and uterus) from three unrelated
adults. Studies are done typically at minimally two dose
levels.
[0360] The therapeutic antibody (i.e., test article) and isotype
matched control antibody may be biotinylated for avidin-biotin
complex (ABC) detection; other detection methods may include
tertiary antibody detection for a FITC (or otherwise) labeled test
article, or precomplexing with a labeled anti-human IgG for an
unlabeled test article.
[0361] Briefly, cryosections (about 5 .mu.m) of human tissues
obtained at autopsy or biopsy are fixed and dried on object glass.
The peroxidase staining of tissue sections is performed, using the
avidin-biotin system. First (in case of a precomplexing detection
system), the test article is incubated with the secondary
biotinylated anti-human IgG and developed into immune complex. The
immune complex at the final concentrations of 2 and 10 .mu.g/mL of
test article is added onto tissue sections on object glass and then
the tissue sections were reacted for 30 minutes with a
avidin-biotin-peroxidase kit. Subsequently, DAB
(3,3'-diaminobenzidine), a substrate for the peroxidase reaction,
was applied for 4 minutes for tissue staining. Antigen-Sepharose
beads are used as positive control tissue sections.
[0362] Any specific staining is judged to be either an expected
(e.g., consistent with antigen expression) or unexpected reactivity
based upon known expression of the target antigen in question. Any
staining judged specific is scored for intensity and frequency.
Antigen or serum competition or blocking studies can assist further
in determining whether observed staining is specific or
nonspecific.
[0363] If two selected antibodies are found to meet the selection
criteria--appropriate tissue staining, matching staining between
human and toxicology animal specific tissue--they can be selected
for DVD-binding protein generation.
[0364] The tissue cross-reactivity study has to be repeated with
the final DVD-binding protein construct, but while these studies
follow the same protocol as outline herein, they are more complex
to evaluate because any binding can come from any of the two parent
antibodies, and any unexplained binding needs to be confirmed with
complex antigen competition studies.
[0365] It is readily apparent that the complex undertaking of
tissue cross-reactivity studies with a multispecific molecule like
a DVD-binding protein is greatly simplified if the two parental
antibodies are selected for: (1) lack of unexpected tissue
cross-reactivity findings and (2) appropriate similarity of tissue
cross-reactivity findings between the corresponding human and
toxicology animal species tissues.
III.B.12. Specificity and Selectivity
[0366] To generate a DVD-binding protein with desired specificity
and selectivity, one needs to generate and select parent mAbs with
the similarly desired specificity and selectivity profile.
[0367] Binding studies for specificity and selectivity with a
DVD-binding protein can be complex due to the four or more binding
sites, two each for each antigen. Briefly, binding studies using
ELISA, BIAcore, KinExA, or other interaction studies with a
DVD-binding protein need to monitor the binding of one, two, or
more antigens to the DVD-binding protein. While BIAcore technology
can resolve the sequential, independent binding of multiple
antigens, more traditional methods including ELISA or more modern
techniques like KinExA cannot. Therefore careful characterization
of each parent antibody is critical. After each individual antibody
has been characterized for specificity, confirmation of specificity
retention of the individual binding sites in the DVD-binding
protein is greatly simplified.
[0368] It is readily apparent that the complex undertaking of
determining the specificity of a DVD-binding protein is greatly
simplified if the two parental antibodies are selected for
specificity prior to being combined into a DVD-binding protein.
[0369] Antigen-antibody interaction studies can take many forms,
including many classical protein protein interaction studies,
including ELISA (enzyme linked immunosorbent assay), mass
spectrometry, chemical cross linking, SEC with light scattering,
equilibrium dialysis, gel permeation, ultrafiltration, gel
chromatography, large-zone analytical SEC, micropreparative
ultracentrifugation (sedimentation equilibrium), spectroscopic
methods, titration microcalorimetry, sedimentation equilibrium (in
analytical ultracentrifuge), sedimentation velocity (in analytical
centrifuge), surface plasmon resonance (including BIAcore).
Relevant references include "Current Protocols in Protein Science,"
John E. Coligan, Ben M. Dunn, David W. Speicher, Paul T, Wingfield
(eds.) Volume 3, chapters 19 and 20, published by John Wiley &
Sons Inc., and references included therein and "Current Protocols
in Immunology," John E. Coligan, Barbara E. Bierer, David H.
Margulies, Ethan M. Shevach, Warren Strober (eds.) published by
John Wiley & Sons Inc and relevant references included
therein.
[0370] Cytokine Release in Whole Blood:
[0371] The interaction of mAb with human blood cells can be
investigated by a cytokine release assay (Wing, M. G., Therapeutic
Immunology (1995), 2(4): 183-190; "Current Protocols in
Pharmacology," S. J. Enna, Michael Williams, John W. Ferkany, Terry
Kenakin, Paul Moser, (eds.) published by John Wiley & Sons Inc;
Madhusudan, S., Clinical Cancer Research (2004), 10(19): 6528-6534;
Cox, J. Methods (2006), 38(4): 274-282; Choi, I., Eur. J. Immunol.,
(2001), 31(1): 94-106). Briefly, various concentrations of mAb are
incubated with human whole blood for 24 hours. The concentration
tested should cover a wide range including final concentrations
mimicking typical blood levels in patients (including but not
limited to 100 ng/ml-100 .mu.g/ml). Following the incubation,
supernatants and cell lysates are analyzed for the presence of
IL-1R.alpha., TNF-.alpha., IL-lb, IL-6 and IL-8. Cytokine
concentration profiles generated for mAb are compared to profiles
produced by a negative human IgG control and a positive LPS or PHA
control. The cytokine profile displayed by mAb from both cell
supernatants and cell lysates are compared to that using control
human IgG. In an embodiment, the monoclonal antibody does not
interact with human blood cells to spontaneously release
inflammatory cytokines.
[0372] Cytokine release studies for a DVD-binding protein are
complex due to the four or more binding sites, two each for each
antigen. Briefly, cytokine release studies as described herein
measure the effect of the whole DVD-binding protein on whole blood
or other cell systems, but cannot resolve which portion of the
molecule causes cytokine release. Once cytokine release has been
detected, the purity of the DVD-binding protein preparation has to
be ascertained, because some co-purifying cellular components can
cause cytokine release on their own. If purity is not the issue,
fragmentation of DVD-binding protein (including but not limited to
removal of Fc portion, separation of binding sites etc.), binding
site mutagenesis or other methods may need to be employed to
deconvolute any observations. It is readily apparent that this
complex undertaking is greatly simplified if the two parental
antibodies are selected for lack of cytokine release prior to being
combined into a DVD-binding protein.
III.B.13. Cross Reactivity to Other Species for Toxicological
Studies
[0373] In an embodiment, the individual antibodies selected with
sufficient cross-reactivity to appropriate tox species, for
example, cynomolgus monkey. Parental antibodies need to bind to
orthologous species target (i.e. cynomolgus monkey) and elicit
appropriate response (modulation, neutralization, activation). In
an embodiment, the cross-reactivity (affinity/potency) to
orthologous species target should be within 10-fold of the human
target. In practice, the parental antibodies are evaluated for
multiple species, including mouse, rat, dog, monkey (and other
non-human primates), as well as disease model species (i.e. sheep
for asthma model). The acceptable cross-reactivity to tox species
from the parental monoclonal antibodies allows future toxicology
studies of DVD-binding protein in the same species. For that
reason, the two parental monoclonal antibodies should have
acceptable cross-reactivity for a common tox species therefore
allowing toxicology studies of DVD-binding protein in the same
species.
[0374] Parent mAbs may be selected from various mAbs capable of
binding specific targets and well known in the art. These include,
but are not limited to anti-sclerostin, anti-SOSTF, anti-TNF
antibody (U.S. Pat. No. 6,258,562), anti-IL-12 and/or anti-IL-12p40
antibody (U.S. Pat. No. 6,914,128); anti-IL-18 antibody (US patent
application publication No. 2005/0147610 A1), anti-C5, anti-CBL,
anti-CD147, anti-gp120, anti-VLA-4, anti-CD11a, anti-CD18,
anti-VEGF, anti-CD40L, anti CD-40 (e.g., see WO2007124299) anti-Id,
anti-ICAM-1, anti-CXCL13, anti-CD2, anti-EGFR, anti-TGF-beta 2,
anti-HGF, anti-cMet, anti DLL-4, anti-NPR1, anti-PLGF, anti-ErbB3,
anti-E-selectin, anti-Fact VII, anti-Her2/neu, anti-F gp,
anti-CD11/18, anti-CD14, anti-ICAM-3, anti-RON, anti CD-19,
anti-CD80 (e.g., see PCT Publication No. WO 2003/039486), anti-CD4,
anti-CD3, anti-CD23, anti-beta2-integrin, anti-alpha4beta7,
anti-CD52, anti-HLA DR, anti-CD22 (see, e.g., U.S. Pat. No.
5,789,554), anti-CD20, anti-MIF, anti-CD64 (FcR), anti-TCR alpha
beta, anti-CD2, anti-Hep B, anti-CA 125, anti-EpCAM, anti-gp120,
anti-CMV, anti-gpIIbIIIa, anti-IgE, anti-CD25, anti-CD33, anti-HLA,
anti-IGF1,2, anti IGFR, anti-VNRintegrin, anti-IL-1alpha,
anti-IL-1beta, anti-IL-1 receptor, anti-IL-2 receptor, anti-IL-4,
anti-IL-4 receptor, anti-IL5, anti-IL-5 receptor, anti-IL-6,
anti-IL-6R, RANKL, NGF, DKK, alphaVbeta3, anti-IL-8, anti-IL-9,
anti-IL-13, anti-IL-13 receptor, and anti-IL-23; IL-23p19; (see,
Presta, "Selection, design, and engineering of therapeutic
antibodies," J. Allergy Clin. Immunol., 116: 731-736 (2005) and at
worldwide website
hwww.path.cam.ac.uk/.about.mrc7/humanisation/antibodies.html).
[0375] Parent mAbs may also be selected from various therapeutic
antibodies approved for use, in clinical trials, or in development
for clinical use. Such therapeutic antibodies include, but are not
limited to, rituximab (Rituxan.RTM., IDEC/Genentech/Roche) (see for
example U.S. Pat. No. 5,736,137), a chimeric anti-CD20 antibody
approved to treat Non-Hodgkin's lymphoma; HuMax-CD20, an anti-CD20
currently being developed by Genmab, an anti-CD20 antibody
described in U.S. Pat. No. 5,500,362, AME-133 (Applied Molecular
Evolution), hA20 (Immunomedics, Inc.), HumaLYM (Intracel), and
PRO70769 (PCT/US2003/040426, entitled "Immunoglobulin Variants and
Uses Thereof"), trastuzumab (Herceptin.RTM., Genentech) (see for
example U.S. Pat. No. 5,677,171), a humanized anti-Her2/neu
antibody approved to treat breast cancer; pertuzumab (rhuMab-2C4,
Omnitarg.RTM.), currently being developed by Genentech; an
anti-Her2 antibody described in U.S. Pat. No. 4,753,894; cetuximab
(Erbitux.RTM., Imclone) (U.S. Pat. No. 4,943,533; PCT WO 96/40210),
a chimeric anti-EGFR antibody in clinical trials for a variety of
cancers; ABX-EGF (U.S. Pat. No. 6,235,883), currently being
developed by Abgenix-Immunex-Amgen; HuMax-EGFr (U.S. Ser. No.
10/172,317), currently being developed by Genmab; 425, EMD55900,
EMD62000, and EMD72000 (Merck KGaA) (U.S. Pat. No. 5,558,864;
Murthy et al. 1987, Arch Biochem Biophys. 252(2):549-60; Rodeck et
al., 1987, J Cell Biochem. 35(4):315-20; Kettleborough et al.,
1991, Protein Eng. 4(7):773-83); ICR62 (Institute of Cancer
Research) (PCT WO 95/20045; Modjtahedi et al., 1993, J. Cell
Biophys. 1993, 22(1-3):129-46; Modjtahedi et al., 1993, Br J
Cancer. 1993, 67(2):247-53; Modjtahedi et al, 1996, Br J Cancer,
73(2):228-35; Modjtahedi et al, 2003, Int J Cancer, 105(2):273-80);
TheraCIM hR3 (YM Biosciences, Canada and Centro de Immunologia
Molecular, Cuba (U.S. Pat. No. 5,891,996; U.S. Pat. No. 6,506,883;
Mateo et al, 1997, Immunotechnology, 3(1):71-81); mAb-806 (Ludwig
Institute for Cancer Research, Memorial Sloan-Kettering) (Jungbluth
et al. 2003, Proc Natl Acad Sci USA. 100(2):639-44); KSB-102 (KS
Biomedix); MR1-1 (IVAX, National Cancer Institute) (PCT WO
0162931A2); and SC100 (Scancell) (PCT WO 01/88138); alemtuzumab
(Campath.RTM., Millenium), a humanized mAb currently approved for
treatment of B-cell chronic lymphocytic leukemia; muromonab-CD3
(Orthoclone OKT3.RTM.), an anti-CD3 antibody developed by Ortho
Biotech/Johnson & Johnson, ibritumomab tiuxetan (Zevalin.RTM.),
an anti-CD20 antibody developed by IDEC/Schering AG, gemtuzumab
ozogamicin (Mylotarg.RTM.), an anti-CD33 (p67 protein) antibody
developed by Celltech/Wyeth, alefacept (Amevive.RTM.), an
anti-LFA-3 Fc fusion developed by Biogen), abciximab (ReoPro.RTM.),
developed by Centocor/Lilly, basiliximab (Simulect.RTM.), developed
by Novartis, palivizumab (Synagis.RTM.), developed by Medimmune,
infliximab (Remicade.RTM.), an anti-TNFalpha antibody developed by
Centocor, adalimumab (Humira.RTM.), an anti-TNFalpha antibody
developed by Abbott, Humicade.RTM., an anti-TNFalpha antibody
developed by Celltech, golimumab (CNTO-148), a fully human TNF
antibody developed by Centocor, etanercept (Enbrel.RTM.), an p75
TNF receptor Fc fusion developed by Immunex/Amgen, lenercept, an
p55TNF receptor Fc fusion previously developed by Roche, ABX-CBL,
an anti-CD147 antibody being developed by Abgenix, ABX-IL8, an
anti-IL8 antibody being developed by Abgenix, ABX-MA1, an
anti-MUC18 antibody being developed by Abgenix, Pemtumomab (R1549,
90Y-muHMFG1), an anti-MUC1 in development by Antisoma, Therex
(R1550), an anti-MUC 1 antibody being developed by Antisoma,
AngioMab (AS1405), being developed by Antisoma, HuBC-1, being
developed by Antisoma, Thioplatin (AS1407) being developed by
Antisoma, Antegren.RTM. (natalizumab), an anti-alpha-4-beta-1
(VLA-4) and alpha-4-beta-7 antibody being developed by Biogen,
VLA-1 mAb, an anti-VLA-1 integrin antibody being developed by
Biogen, LTBR mAb, an anti-lymphotoxin beta receptor (LTBR) antibody
being developed by Biogen, CAT-152, an anti-TGF-.beta.2 antibody
being developed by Cambridge Antibody Technology, ABT 874 (J695),
an anti-IL-12 p40 antibody being developed by Abbott, CAT-192, an
anti-TGF.beta.1 antibody being developed by Cambridge Antibody
Technology and Genzyme, CAT-213, an anti-Eotaxinl antibody being
developed by Cambridge Antibody Technology, LymphoStat-B.RTM. an
anti-Blys antibody being developed by Cambridge Antibody Technology
and Human Genome Sciences Inc., TRAIL-R1mAb, an anti-TRAIL-R1
antibody being developed by Cambridge Antibody Technology and Human
Genome Sciences, Inc., Avastin.RTM. bevacizumab, rhuMAb-VEGF), an
anti-VEGF antibody being developed by Genentech, an anti-HER
receptor family antibody being developed by Genentech, Anti-Tissue
Factor (ATF), an anti-Tissue Factor antibody being developed by
Genentech, Xolair.RTM. (Omalizumab), an anti-IgE antibody being
developed by Genentech, Raptiva.RTM. (Efalizumab), an anti-CD11a
antibody being developed by Genentech and Xoma, MLN-02 Antibody
(formerly LDP-02), being developed by Genentech and Millenium
Pharmaceuticals, HuMax CD4, an anti-CD4 antibody being developed by
Genmab, HuMax-IL15, an anti-IL15 antibody being developed by Genmab
and Amgen, HuMax-Inflam, being developed by Genmab and Medarex,
HuMax-Cancer, an anti-Heparanase I antibody being developed by
Genmab and Medarex and Oxford GcoSciences, HuMax-Lymphoma, being
developed by Genmab and Amgen, HuMax-TAC, being developed by
Genmab, IDEC-131, and anti-CD40L antibody being developed by IDEC
Pharmaceuticals, IDEC-151 (Clenoliximab), an anti-CD4 antibody
being developed by IDEC Pharmaceuticals, IDEC-114, an anti-CD80
antibody being developed by IDEC Pharmaceuticals, IDEC-152, an
anti-CD23 being developed by IDEC Pharmaceuticals, anti-macrophage
migration factor (MIF) antibodies being developed by IDEC
Pharmaceuticals, BEC2, an anti-idiotypic antibody being developed
by Imclone, IMC-1C11, an anti-KDR antibody being developed by
Imclone, DC101, an anti-flk-1 antibody being developed by Imclone,
anti-VE cadherin antibodies being developed by Imclone,
CEA-Cide.RTM. (labetuzumab), an anti-carcinoembryonic antigen (CEA)
antibody being developed by Immunomedics, LymphoCide.RTM.
(Epratuzumab), an anti-CD22 antibody being developed by
Immunomedics, AFP-Cide, being developed by Immunomedics,
MyelomaCide, being developed by Immunomedics, LkoCide, being
developed by Immunomedics, ProstaCide, being developed by
Immunomedics, MDX-010, an anti-CTLA4 antibody being developed by
Medarex, MDX-060, an anti-CD30 antibody being developed by Medarex,
MDX-070 being developed by Medarex, MDX-018 being developed by
Medarex, Osidem.RTM. (IDM-1), and anti-Her2 antibody being
developed by Medarex and Immuno-Designed Molecules, HuMax.RTM.-CD4,
an anti-CD4 antibody being developed by Medarex and Genmab,
HuMax-IL15, an anti-IL15 antibody being developed by Medarex and
Genmab, CNTO 148, an anti-TNF.alpha. antibody being developed by
Medarex and Centocor/Johnson & Johnson, CNTO 1275, an
anti-cytokine antibody being developed by Centocor/Johnson &
Johnson, MOR101 and MOR102, anti-intercellular adhesion molecule-1
(ICAM-1) (CD54) antibodies being developed by MorphoSys, MOR201, an
anti-fibroblast growth factor receptor 3 (FGFR-3) antibody being
developed by MorphoSys, Nuvion.RTM. (visilizumab), an anti-CD3
antibody being developed by Protein Design Labs, HuZAF.RTM., an
anti-gamma interferon antibody being developed by Protein Design
Labs, Anti-.alpha. 5.beta.1 Integrin, being developed by Protein
Design Labs, anti-IL-12, being developed by Protein Design Labs,
ING-1, an anti-Ep-CAM antibody being developed by Xoma, Xolair.RTM.
(Omalizumab) a humanized anti-IgE antibody developed by Genentech
and Novartis, and MLN01, an anti-Beta2 integrin antibody being
developed by Xoma. In another embodiment, the therapeutics include
KRN330 (Kirin); huA33 antibody (A33, Ludwig Institute for Cancer
Research); CNTO 95 (alpha V integrins, Centocor); MEDI-522 (alpha
V.beta.3 integrin, Medimmune); volociximab (alpha V.beta.1
integrin, Biogen/PDL); Human mAb 216 (B cell glycosolated epitope,
NCI); BiTE MT103 (bispecific CD19.times.CD3, Medimmune);
4G7.times.H22 (Bispecific Bcell.times.FcgammaR1, Medarex/Merck
KGa); rM28 (Bispecific CD28.times.MAPG, US Patent No. EP1444268);
MDX447 (EMD 82633) (Bispecific CD64.times.EGFR, Medarex);
Catumaxomab (removab) (Bispecific EpCAM.times.anti-CD3,
Trion/Fres); Ertumaxomab (bispecific HER2/CD3, Fresenius Biotech);
oregovomab (OvaRex) (CA-125, ViRexx); Rencarex.RTM. (WX G250)
(carbonic anhydrase IX, Wilex); CNTO 888 (CCL2, Centocor); TRC105
(CD105 (endoglin), Tracon); BMS-663513 (CD137 agonist, Brystol
Myers Squibb); MDX-1342 (CD19, Medarex); Siplizumab (MEDI-507)
(CD2, Medimmune); Ofatumumab (Humax-CD20) (CD20, Genmab); Rituximab
(Rituxan) (CD20, Genentech); veltuzumab (hA20) (CD20,
Immunomedics); Epratuzumab (CD22, Amgen); lumiliximab (IDEC 152)
(CD23, Biogen); muromonab-CD3 (CD3, Ortho); HuM291 (CD3 fc
receptor, PDL Biopharma); HeFi-1, CD30, NCI); MDX-060 (CD30,
Medarex); MDX-1401 (CD30, Medarex); SGN-30 (CD30, Seattle
Genentics); SGN-33 (Lintuzumab) (CD33, Seattle Genentics);
Zanolimumab (HuMax-CD4) (CD4, Genmab); HCD122 (CD40, Novartis);
SGN-40 (CD40, Seattle Genentics); Campath1h (Alemtuzumab) (CD52,
Genzyme); MDX-1411 (CD70, Medarex); hLL1 (EPB-1) (CD74.38,
Immunomedics); Galiximab (IDEC-144) (CD80, Biogen); MT293
(TRC093/D93) (cleaved collagen, Tracon); HuLuc63 (CS1, PDL Pharma);
ipilimumab (MDX-010) (CTLA4, Brystol Myers Squibb); Tremelimumab
(Ticilimumab, CP-675,2) (CTLA4, Pfizer); HGS-ETR1 (Mapatumumab)
(DR4 TRAIL-R1 agonist, Human Genome Science/Glaxo Smith Kline);
AMG-655 (DR5, Amgen); Apomab (DR5, Genentech); CS-1008 (DR5,
Daiichi Sankyo); HGS-ETR2 (lexatumumab) (DR5 TRAIL-R2 agonist,
HGS); Cetuximab (Erbitux) (EGFR, Imclone); IMC-11F8, (EGFR,
Imclone); Nimotuzumab (EGFR, YM Bio); Panitumumab (Vectabix) (EGFR,
Amgen); Zalutumumab (HuMaxEGFr) (EGFR, Genmab); CDX-110 (EGFRvIII,
AVANT Immunotherapeutics); adecatumumab (MT201) (Epcam, Merck);
edrecolomab (Panorex, 17-1A) (Epcam, Glaxo/Centocor); MORAb-003
(folate receptor a, Morphotech); KW-2871 (ganglioside GD3, Kyowa);
MORAb-009 (GP-9, Morphotech); CDX-1307 (MDX-1307) (hCGb, Celldex);
Trastuzumab (Herceptin) (HER2, Celldex); Pertuzumab (rhuMAb 2C4)
(HER2 (DI), Genentech); apolizumab (HLA-DR beta chain, PDL Pharma);
AMG-479 (IGF-1R, Amgen); anti-IGF-1R R1507 (IGF1-R, Roche); CP
751871 (IGF1-R, Pfizer); IMC-A12 (IGF1-R, Imclone); BIIB022
(IGF-1R, Biogen); Mik-beta-1 (IL-2Rb (CD122), Hoffman LaRoche);
CNTO 328 (IL6, Centocor); Anti-KIR (1-7F9) (Killer cell Ig-like
Receptor (KIR), Novo); Hu3S193 (Lewis (y), Wyeth, Ludwig Institute
of Cancer Research); hCBE-11 (LTBR, Biogen); HuHMFG1 (MUC1,
Antisoma/NCI); RAV12 (N-linked carbohydrate epitope, Raven); CAL
(parathyroid hormone-related protein (PTH-rP), University of
California); CT-011 (PD1, CureTech); MDX-1106 (ono-4538) (PD1,
Medarex/Ono); MAb CT-011 (PD1, Curetech); IMC-3G3 (PDGFRa,
Imclone); bavituximab (phosphatidylserine, Peregrine); huJ591
(PSMA, Cornell Research Foundation); muJ591 (PSMA, Cornell Research
Foundation); GC1008 (TGFb (pan) inhibitor (IgG4), Genzyme);
Infliximab (Remicade) (TNFa, Centocor); A27.15 (transferrin
receptor, Salk Institute, INSERN WO 2005/111082); E2.3 (transferrin
receptor, Salk Institute); Bevacizumab (Avastin) (VEGF, Genentech);
HuMV833 (VEGF, Tsukuba Research Lab-WO/2000/034337, University of
Texas); IMC-18F1 (VEGFR1, Imclone); IMC-1121 (VEGFR2, Imclone).
III.C. Construction of DVD-Binding Proteins
[0376] A multivalent multispecific dual variable domain binding
protein (DVD-binding protein) is designed such that two different
light chain variable domains (VL) from two different parent
monoclonal antibodies are linked in tandem directly or via a short
linker by recombinant DNA techniques, followed by the light chain
constant domain. Similarly, the heavy chain comprises two different
heavy chain variable domains (VH) linked in tandem, followed by the
constant domain CH1 and Fc region.
[0377] The variable domains can be obtained using recombinant DNA
techniques from a parent antibody generated by any one of the
methods described herein. In an embodiment, the variable domain is
a murine heavy or light chain variable domain. In another
embodiment, the variable domain is a CDR-grafted or a humanized
variable heavy or light chain domain. In an embodiment, the
variable domain is a human heavy or light chain variable
domain.
[0378] In one embodiment, the first and second variable domains are
linked directly to each other using recombinant DNA techniques. In
another embodiment the variable domains are linked via a linker
sequence. In an embodiment, two variable domains are linked Three
or more variable domains may also be linked directly or via a
linker sequence. The variable domains may bind the same antigen or
may bind different antigens. DVD-binding proteinswhich may include
one immunoglobulin variable domain and one non-immunoglobulin
variable domain, such as ligand binding domain of a receptor or
active domain of an enzyme, are provided. DVD-binding proteins may
also comprise two or more non-Ig domains.
[0379] The linker sequence may be a single amino acid or a
polypeptide sequence. In an embodiment, the linker sequences are
GGGGSG (SEQ ID NO:1695), GGSGG (SEQ ID NO:1696), GGGGSGGGGS (SEQ ID
NO:1697), GGSGGGGSGS (SEQ ID NO:1698), GGSGGGGSGGGGS (SEQ ID
NO:1699), GGGGSGGGGSGGGG (SEQ ID NO:1700), GGGGSGGGGSGGGGS (SEQ ID
NO:1701), ASTKGP (SEQ ID NO:1702), ASTKGPSVFPLAP (SEQ ID NO:1703),
TVAAP (SEQ ID NO:1704), TVAAPSVFIFPP (SEQ ID NO:1705),
AKTTPKLEEGEFSEAR (SEQ ID NO:1706), AKTTPKLEEGEFSEARV (SEQ ID
NO:1707), AKTTPKLGG (SEQ ID NO:1710), SAKTTPKLGG (SEQ ID NO:1709),
SAKTTP (SEQ ID NO:1702), RADAAP (SEQ ID NO:1711), RADAAPTVS (SEQ ID
NO:1712), RADAAAAGGPGS (SEQ ID NO:1713),
RADAAAAGGGGSGGGGSGGGGSGGGGS (SEQ ID NO:1714), SAKTTPKLEEGEFSEARV
(SEQ ID NO:1715), ADAAP (SEQ ID NO:1716), ADAAPTVSIFPP (SEQ ID
NO:2050), QPKAAP (SEQ ID NO:2051), QPKAAPSVTLFPP (SEQ ID NO:2052),
AKTTPP (SEQ ID NO:2053), AKTTPPSVTPLAP (SEQ ID NO:2054), AKTTAP
(SEQ ID NO:2055), AKTTAPSVYPLAP (SEQ ID NO:2056), GENKVEYAPALMALS
(SEQ ID NO:2057), GPAKELTPLKEAKVS (SEQ ID NO:2058), and
GHEAAAVMQVQYPAS (SEQ ID NO:2059). The choice of linker sequences is
based on crystal structure analysis of several Fab molecules. There
is a natural flexible linkage between the variable domain and the
CH1/CL constant domain in Fab or antibody molecular structure. This
natural linkage comprises approximately 10-12 amino acid residues,
contributed by 4-6 residues from C-terminus of V domain and 4-6
residues from the N-terminus of CL/CH1 domain. DVD-binding proteins
described herein can be generated using N-terminal 5-6 amino acid
residues, or 11-12 amino acid residues, of CL or CH1 as linker in
light chain and heavy chain of DVD-binding protein, respectively.
The N-terminal residues of CL or CH1 domains, particularly the
first 5-6 amino acid residues, adopt a loop conformation without
strong secondary structures, and therefore can act as flexible
linkers between the two variable domains. The N-terminal residues
of CL or CH1 domains are natural extension of the variable domains,
as they are part of the Ig sequences, and therefore minimize to a
large extent any immunogenicity potentially arising from the
linkers and junctions.
[0380] Other linker sequences may include any sequence of any
length of CL/CH1 domain but not all residues of CL/CH1 domain; for
example the first 5-12 amino acid residues of the CL/CH1 domains;
the light chain linkers can be from C.kappa. or C.lamda.; and the
heavy chain linkers can be derived from CH1 of any isotypes,
including C.gamma.1, C.gamma.2, C.gamma.3, C.gamma.4, C.alpha.1,
C.alpha.2, C.delta., C.epsilon., and C.mu.. Linker sequences may
also be derived from other proteins such as Ig-like proteins,
(e.g., TCR, FcR, KIR); G/S based sequences; hinge region-derived
sequences; and other natural sequences from other proteins.
[0381] In an embodiment a constant domain is linked to the two
linked variable domains using recombinant DNA techniques. In an
embodiment, a sequence comprising tandemly linked heavy chain
variable domains is linked to a heavy chain constant domain and a
sequence comprising tandemly linked light chain variable domains is
linked to a light chain constant domain. In an embodiment, the
constant domains are human heavy chain constant domain and human
light chain constant domain, respectively. In an embodiment, the
DVD heavy chain is further linked to an Fc region. The Fc region
may be a native sequence Fc region, or a variant Fc region. In
another embodiment, the Fc region is a human Fc region. In another
embodiment the Fc region includes Fc region from IgG1, IgG2, IgG3,
IgG4, IgA, IgM, IgE, or IgD.
[0382] In an embodiment, two heavy chain DVD polypeptides and two
light chain DVD polypeptides are combined to form a DVD-binding
protein. Detailed description of specific DVD-binding proteins
capable of binding specific target antigens, such as SOST, and
methods of making the same are provided in the Examples section
below.
III.D. Production of DVD-Binding Proteins
[0383] DVD-binding proteins produced by any of a number of
techniques known in the art are provided, including for example,
expression from host cells, wherein expression vector(s) encoding
the DVD-binding protein heavy and DVD-binding protein light chains
is (are) transfected into a host cell by standard techniques. The
various forms of the term "transfection" are intended to encompass
a wide variety of techniques commonly used for the introduction of
exogenous DNA into a prokaryotic or eukaryotic host cell, e.g.,
electroporation, calcium-phosphate precipitation, DEAE-dextran
transfection and the like. Although it is possible to express the
provided DVD-binding proteins in either prokaryotic or eukaryotic
host cells, DVD-binding proteins are expressed in eukaryotic cells,
for example, mammalian host cells, because such eukaryotic cells
(and in particular mammalian cells) are more likely than
prokaryotic cells to assemble and secrete a properly folded and
immunologically active DVD-binding protein.
[0384] Exemplary mammalian host cells for expressing the provided
recombinant antibodies include Chinese Hamster Ovary (CHO cells)
(including dhfr-CHO cells, described in Urlaub and Chasin, (1980)
Proc. Natl. Acad. Sci. USA, 77: 4216-4220, used with a DHFR
selectable marker, e.g., as described in R. J. Kaufman and P.A.
Sharp (1982) Mol. Biol., 159: 601-621), NS0 myeloma cells, COS
cells, SP2 and PER.C6 cells. When recombinant expression vectors
encoding DVD-binding proteins are introduced into mammalian host
cells, the DVD-binding proteins are produced by culturing the host
cells for a period of time sufficient to allow for expression of
the DVD-binding proteins in the host cells or secretion of the DVD
proteins into the culture medium in which the host cells are grown.
DVD-binding proteins can be recovered from the culture medium using
standard protein purification methods.
[0385] In an exemplary system for recombinant expression of the
provided DVD-binding proteins, a recombinant expression vector
encoding both the DVD-binding protein heavy chain and the
DVD-binding protein light chain is introduced into dhfr-CHO cells
by calcium phosphate-mediated transfection. Within the recombinant
expression vector, the DVD-binding protein heavy and light chain
genes are each operatively linked to CMV enhancer/AdMLP promoter
regulatory elements to drive high levels of transcription of the
genes. The recombinant expression vector also carries a DHFR gene,
which allows for selection of CHO cells that have been transfected
with the vector using methotrexate selection/amplification. The
selected transformant host cells are cultured to allow for
expression of the DVD-binding protein heavy and light chains and
intact DVD-binding protein is recovered from the culture medium.
Standard molecular biology techniques are used to prepare the
recombinant expression vector, transfect the host cells, select for
transformants, culture the host cells and recover the DVD-binding
protein from the culture medium. Still further, a method is
provided of synthesizing a DVD-binding protein by culturing a host
cell in a suitable culture medium until a DVD-binding protein is
synthesized. The method can further comprise isolating the
DVD-binding protein from the culture medium.
[0386] An important feature of DVD-binding protein is that it can
be produced and purified in a similar way as a conventional
antibody. The production of DVD-binding protein results in a
homogeneous, single major product with desired dual-specific
activity, without any sequence modification of the constant region
or chemical modifications of any kind. Other previously described
methods to generate "bi-specific", "multi-specific", and
"multi-specific multivalent" full length binding proteins do not
lead to a single primary product but instead lead to the
intracellular or secreted production of a mixture of assembled
inactive, mono-specific, multi-specific, multivalent, full length
binding proteins, and multivalent full length binding proteins with
combination of different binding sites. As an example, based on the
design described by Miller and Presta (PCT Publication No. WO
2001/077342(A1), there are 16 possible combinations of heavy and
light chains. Consequently only 6.25% of protein is likely to be in
the desired active form, and not as a single major product or
single primary product compared to the other 15 possible
combinations. Separation of the desired, fully active forms of the
protein from inactive and partially active forms of the protein
using standard chromatography techniques, typically used in large
scale manufacturing, is yet to be demonstrated.
[0387] Surprisingly, the provided design of the "dual-specific
multivalent full length binding proteins" of the leads to a dual
variable domain light chain and a dual variable domain heavy chain
which assemble primarily to the desired "dual-specific multivalent
full length binding proteins".
[0388] At least 50%, at least 75%, and at least 90% of the
assembled, and expressed DVD-binding proteins are the desired
dual-specific tetravalent protein. This aspect particularly
enhances the commercial utility of the invention provided.
Therefore, a method to express a dual variable domain light chain
and a dual variable domain heavy chain in a single cell leading to
a single primary product of a "dual-specific tetravalent full
length binding protein" is provided.
[0389] Methods of expressing a dual variable domain light chain and
a dual variable domain heavy chain in a single cell leading to a
"primary product" of a "dual-specific, tetravalent, full length
binding protein", where the "primary product" is more than 50% of
all assembled protein, comprising a dual variable domain light
chain and a dual variable domain heavy chain are provided.
[0390] Methods of expressing a dual variable domain light chain and
a dual variable domain heavy chain in a single cell leading to a
single "primary product" of a "dual-specific, tetravalent, full
length binding protein", where the "primary product" is more than
75% of all assembled protein, comprising a dual variable domain
light chain and a dual variable domain heavy chain are
provided.
[0391] Methods of expressing a dual variable domain light chain and
a dual variable domain heavy chain in a single cell leading to a
single "primary product" of a "dual-specific tetravalent full
length binding protein", where the "primary product" is more than
90% of all assembled protein, comprising a dual variable domain
light chain and a dual variable domain heavy chain are
provided.
IV. Production of Sclerostin Binding Proteins and Binding
Protein-Producing Cell Lines
[0392] In a provided embodiment, sclerostin binding proteins,
including anti-sclerostin antibodies, exhibit a high capacity to
reduce or to neutralize SOST activity, e.g., as assessed by any one
of several in vitro and in vivo assays known in the art.
Preferrably, sclerostin binding proteins also exhibit a high
capacity to reduce or to neutralize SOST activity
[0393] In embodiments, a binding protein, or antigen-binding
portion thereof, binds human sclerostin, wherein the binding
protein, or antigen-binding portion thereof, dissociates from human
SOST with a k.sub.off rate constant of about 0.1 s.sup.-1 or less,
as determined by surface plasmon resonance, or which inhibits human
SOST activity with an IC.sub.50 of about 1.times.10.sup.-6 M or
less. Alternatively, the binding protein, or an antigen-binding
portion thereof, may dissociate from human sclerostin with a
k.sub.off rate constant of about 1.times.10.sup.-2 s.sup.-1 or
less, as determined by surface plasmon resonance, or may inhibit
human sclerostin activity with an IC.sub.50 of about
1.times.10.sup.-7 M or less. Alternatively, the binding protein, or
an antigen-binding portion thereof, may dissociate from human
sclerostin with a k.sub.off rate constant of about
1.times.10.sup.-3 s.sup.-1 or less, as determined by surface
plasmon resonance, or may inhibit human sclerostin with an
IC.sub.50 of about 1.times.10.sup.-8 M or less. Alternatively, the
binding protein, or an antigen-binding portion thereof, may
dissociate from human sclerostin with a k.sub.off rate constant of
about 1.times.10.sup.-4 s.sup.-1 or less, as determined by surface
plasmon resonance, or may inhibit human sclerostin activity with an
IC.sub.50 of about 1.times.10.sup.-9 M or less. Alternatively, the
binding protein, or an antigen-binding portion thereof, may
dissociate from human sclerostin with a k.sub.off rate constant of
about 1.times.10.sup.-5 s.sup.-1 or less, as determined by surface
plasmon resonance, or may inhibit human sclerostin activity with an
IC.sub.50 of about 1.times.10.sup.10 M or less. Alternatively, the
binding protein, or an antigen-binding portion thereof, may
dissociate from human sclerostin with a k.sub.off rate constant of
about 1.times.10.sup.-5 s.sup.-1 or less, as determined by surface
plasmon resonance, or may inhibit human sclerostin activity with an
IC.sub.50 of about 1.times.10.sup.-11 M or less.
[0394] In certain embodiments, the binding protein comprises a
heavy chain constant region, such as an IgG1, IgG2, IgG3, IgG4,
IgA, IgE, IgM or IgD constant region. In an embodiment, the heavy
chain constant region is an IgG1 heavy chain constant region or an
IgG4 heavy chain constant region. Furthermore, the antibody can
comprise a light chain constant region, either a kappa light chain
constant region or a lambda light chain constant region. In an
embodiment, the antibody comprises a kappa light chain constant
region. Alternatively, the antibody portion can be, for example, a
Fab fragment or a single chain Fv fragment.
[0395] Replacements of amino acid residues in the Fc portion to
alter antibody effector function are known in the art (Winter et
al., U.S. Pat. Nos. 5,648,260 and 5,624,821). The Fc portion of an
antibody mediates several important effector functions e.g.
cytokine induction, ADCC, phagocytosis, complement dependent
cytotoxicity (CDC) and half-life/clearance rate of antibody and
antigen-antibody complexes. In some cases these effector functions
are desirable for therapeutic antibody but in other cases might be
unnecessary or even deleterious, depending on the therapeutic
objectives. Certain human IgG isotypes, particularly IgG1 and IgG3,
mediate ADCC and CDC via binding to Fc.gamma.Rs and complement C1q,
respectively. Neonatal Fc receptors (FcRn) are the critical
components determining the circulating half-life of antibodies. In
still another embodiment at least one amino acid residue is
replaced in the constant region of the antibody, for example the Fc
region of the antibody, such that effector functions of the
antibody are altered.
[0396] One embodiment provides a labeled binding protein wherein an
antibody or antibody portion is derivatized or linked to another
functional molecule (e.g., another peptide or protein). For
example, a labeled binding protein can be derived by functionally
linking an antibody or antibody portion (by chemical coupling,
genetic fusion, noncovalent association or otherwise) to one or
more other molecular entities, such as another antibody (e.g., a
bispecific antibody or a diabody), a detectable agent, a cytotoxic
agent, a pharmaceutical agent, and/or a protein or peptide that can
mediate associate of the antibody or antibody portion with another
molecule (such as a streptavidin core region or a polyhistidine
tag).
[0397] Useful detectable agents are provided with which a binding
protein, such as an antibody or antibody portion of the may be
derivatized include fluorescent compounds. Exemplary fluorescent
detectable agents include fluorescein, fluorescein isothiocyanate,
rhodamine, 5-dimethylamine-1-napthalenesulfonyl chloride,
phycoerythrin and the like. An antibody may also be derivatized
with detectable enzymes, such as alkaline phosphatase, horseradish
peroxidase, glucose oxidase and the like. When an antibody is
derivatized with a detectable enzyme, it is detected by adding
additional reagents that the enzyme uses to produce a detectable
reaction product. For example, when the detectable agent
horseradish peroxidase is present, the addition of hydrogen
peroxide and diaminobenzidine leads to a colored reaction product,
which is detectable. An antibody may also be derivatized with
biotin, and detected through indirect measurement of avidin or
streptavidin binding.
[0398] Another embodiment provides a crystallized binding protein.
In an embodiment, crystals of whole anti-sclerostin antibodies and
fragments thereof as disclosed herein, and formulations and
compositions comprising such crystals are provided. In one
embodiment the crystallized binding protein has a greater half-life
in vivo than the soluble counterpart of the binding protein. In
another embodiment the binding protein retains biological activity
after crystallization.
[0399] Crystallized binding protein are provided and may be
produced according methods known in the art and as disclosed in PCT
Publication No. WO 02072636.
[0400] Another embodiment provides a glycosylated binding protein
wherein the antibody or antigen-binding portion thereof comprises
one or more carbohydrate residues. Nascent in vivo protein
production may undergo further processing, known as
post-translational modification. In particular, sugar (glycosyl)
residues may be added enzymatically, a process known as
glycosylation. The resulting proteins bearing covalently linked
oligosaccharide side chains are known as glycosylated proteins or
glycoproteins.
[0401] Naturally occurring antibodies are glycoproteins with one or
more carbohydrate residues in the Fc domain, as well as the
variable domain. Carbohydrate residues in the Fc domain have
important effect on the effector function of the Fc domain, with
minimal effect on antigen binding or half-life of the antibody (R.
Jefferis, Biotechnol. Prog., 21: 11-16 (2005)). In contrast,
glycosylation of the variable domain may have an effect on the
antigen binding activity of the antibody. Glycosylation in the
variable domain may have a negative effect on antibody binding
affinity, likely due to steric hindrance (Co, M. S., et al., Mol.
Immunol., 30: 1361-1367 (1993)), or result in increased affinity
for the antigen (Wallick, S. C., et al., Exp. Med., 168:1099-1109
(1988); Wright, A., et al., EMBO J., 10: 2717-2723 (1991)).
[0402] One aspect of the provided is directed to generating
glycosylation site mutants in which the O- or N-linked
glycosylation site of the binding protein has been mutated. One
skilled in the art can generate such mutants using standard
well-known technologies. Glycosylation site mutants that retain the
biological activity but have increased or decreased binding
activity are provided.
[0403] In still another provided embodiment, the glycosylation of
the antibody or antigen-binding portion is modified. For example,
an aglycoslated antibody can be made (i.e., the antibody lacks
glycosylation). Glycosylation can be altered to, for example,
increase the affinity of the antibody for antigen. Such
carbohydrate modifications can be accomplished by, for example,
altering one or more sites of glycosylation within the antibody
sequence. For example, one or more amino acid substitutions can be
made that result in elimination of one or more variable region
glycosylation sites to thereby eliminate glycosylation at that
site. Such aglycosylation may increase the affinity of the antibody
for antigen. Such an approach is described in further detail in PCT
Publication WO 2003/016466A2, and U.S. Pat. Nos. 5,714,350 and
6,350,861.
[0404] Additionally or alternatively, a modified binding protein is
provided and can be made to have an altered type of glycosylation,
such as a hypofucosylated antibody having reduced amounts of
fucosyl residues (see Kanda, Yutaka et al., Journal of
Biotechnology (2007), 130(3), 300-310.) or an antibody having
increased bisecting GlcNAc structures. Such altered glycosylation
patterns have been demonstrated to increase the ADCC ability of
antibodies. Such carbohydrate modifications can be accomplished by,
for example, expressing the antibody in a host cell with altered
glycosylation machinery. Cells with altered glycosylation machinery
are provided and have been described in the art and can be used as
host cells in which to express recombinant antibodies to thereby
produce an antibody with altered glycosylation. See, for example,
Shields, R. L. et al. (2002) J. Biol. Chem. 277:26733-26740; Umana
et al., "Engineered glycoforms of an antineuroblastoma IgG1 with
optimized antibody-dependent cellular cytotoxic activity," Nat.
Biotech., 17: 176-180 (1999), as well as, European Patent No: EP
1,176,195; PCT Publication Nos. WO 03/035835 and WO 99/54342.
[0405] Protein glycosylation depends on the amino acid sequence of
the protein of interest, as well as the host cell in which the
protein is expressed. Different organisms may produce different
glycosylation enzymes (e.g., glycosyltransferases and
glycosidases), and have different substrates (nucleotide sugars)
available. Due to such factors, protein glycosylation pattern, and
composition of glycosyl residues, may differ depending on the host
system in which the particular protein is expressed. Useful
Glycosyl residues are provided and may include, but are not limited
to, glucose, galactose, mannose, fucose, n-acetylglucosamine and
sialic acid. In an embodiment, the glycosylated binding protein
comprises glycosyl residues such that the glycosylation pattern is
human.
[0406] It is known to those skilled in the art that differing
protein glycosylation may result in differing protein
characteristics. For instance, the efficacy of a therapeutic
protein produced in a microorganism host, such as yeast, and
glycosylated utilizing the yeast endogenous pathway may be reduced
compared to that of the same protein expressed in a mammalian cell,
such as a CHO cell line. Such glycoproteins may also be immunogenic
in humans and show reduced half-life in vivo after administration.
Specific receptors in humans and other animals may recognize
specific glycosyl residues and promote the rapid clearance of the
protein from the bloodstream. Other adverse effects may include
changes in protein folding, solubility, susceptibility to
proteases, trafficking, transport, compartmentalization, secretion,
recognition by other proteins or factors, antigenicity, or
allergenicity. Accordingly, a practitioner may prefer a therapeutic
protein with a specific composition and pattern of glycosylation,
for example glycosylation composition and pattern identical, or at
least similar, to that produced in human cells or in the
species-specific cells of the intended subject animal.
[0407] Expressing glycosylated proteins different from that of a
host cell may be achieved by genetically modifying the host cell to
express heterologous glycosylation enzymes. Using techniques known
in the art a practitioner may generate antibodies or
antigen-binding portions thereof exhibiting human protein
glycosylation. For example, yeast strains have been genetically
modified to express non-naturally occurring glycosylation enzymes
such that glycosylated proteins (glycoproteins) produced in these
yeast strains exhibit protein glycosylation identical to that of
animal cells, especially human cells (US patent application
publication Nos. 20040018590 and 20020137134).
[0408] In addition to the binding proteins, anti-idiotypic
(anti-Id) antibodies specific for such binding proteins are
provided. An anti-Id antibody is an antibody, which recognizes
unique determinants generally associated with the antigen-binding
region of another antibody. The anti-Id can be prepared by
immunizing an animal with the binding protein or a CDR containing
region thereof. The immunized animal will recognize, and respond to
the idiotypic determinants of the immunizing antibody and produce
an anti-Id antibody. It is readily apparent that it may be easier
to generate anti-idiotypic antibodies to the two or more parent
antibodies incorporated into a DVD-binding protein molecule; and
confirm binding studies by methods well recognized in the art
(e.g., BIAcore, ELISA) to verify that anti-idiotypic antibodies
specific for the idiotype of each parent antibody also recognize
the idiotype (e.g., antigen binding site) in the context of the
DVD-binding protein. The anti-idiotypic antibodies specific for
each of the two or more antigen binding sites of a DVD-binding
protein provide ideal reagents to measure DVD-binding protein
concentrations of a human DVD-binding protein in patient serum. For
example, DVD-binding protein concentration assays can be
established using a "sandwich assay ELISA format" with an antibody
to a first antigen binding region coated on the solid phase (e.g.,
BIAcore chip, ELISA plate, etc.), rinsed with rinsing buffer,
incubation with a serum sample, another rinsing step, and
ultimately incubation with another anti-idiotypic antibody to the
other antigen binding site, itself labeled with an enzyme for
quantitation of the binding reaction. In an embodiment, for a
DVD-binding protein with more than two different binding sites,
anti-idiotypic antibodies to the two outermost binding sites (most
distal and proximal from the constant region) will not only help in
determining the DVD-binding protein concentration in human serum
but also document the integrity of the molecule in vivo. Each
anti-Id antibody may also be used as an "immunogen" to induce an
immune response in yet another animal, producing a so-called
anti-anti-Id antibody.
[0409] Further, it will be appreciated by one skilled in the art
that a protein of interest may be expressed using a library of host
cells genetically engineered to express various glycosylation
enzymes, such that member host cells of the library produce the
protein of interest with variant glycosylation patterns. A
practitioner may then select and isolate the protein of interest
with particular novel glycosylation patterns. In an embodiment, the
protein having a particularly selected novel glycosylation pattern
exhibits improved or altered biological properties.
V. Uses of Sclerostin Binding Proteins
[0410] Given their ability to bind to human sclerostin, the
sclerostin binding proteins, or antigen binding portions thereof,
are provided and can be used to detect sclerostin (e.g., in a
biological sample, such as serum or plasma), using a conventional
immunoassay, such as an enzyme linked immunosorbent assays (ELISA),
an radioimmunoassay (RIA) or tissue immunohistochemistry. A method
for detecting sclerostin in a biological sample is provided
comprising contacting a biological sample with a provided binding
protein, or antigen binding portion, and detecting either the
binding protein (or antigen binding portion) bound to sclerostin or
unbound binding protein (or binding portion), to thereby detect
sclerostin in the biological sample. The binding protein is
directly or indirectly labeled with a detectable substance to
facilitate detection of the bound or unbound antibody. Suitable
detectable substances include various enzymes, prosthetic groups,
fluorescent materials, luminescent materials and radioactive
materials. Examples of suitable enzymes include horseradish
peroxidase, alkaline phosphatase, .beta.-galactosidase, or
acetylcholinesterase; examples of suitable prosthetic group
complexes include streptavidin/biotin and avidin/biotin; examples
of suitable fluorescent materials include umbelliferone,
fluorescein, fluorescein isothiocyanate, rhodamine,
dichlorotriazinylamine fluorescein, dansyl chloride or
phycoerythrin; an example of a luminescent material includes
luminol; and examples of suitable radioactive material include
.sup.3H, .sup.14C, .sup.35S, .sup.90Y, .sup.99Tc, .sup.111In,
.sup.125I, .sup.131I, .sup.177Lu, .sup.166Ho, or .sup.153Sm.
[0411] Alternative to labeling the binding protein, human
sclerostin can be assayed in biological fluids by a competition
immunoassay utilizing rhSOST standards labeled with a detectable
substance and an unlabeled human sclerostin binding protein. In
this assay, the biological sample, the labeled rhSOST standards,
and the human sclerostin binding protein are combined and the
amount of labeled recombinant human sclerostin standard bound to
the unlabeled antibody is determined. The amount of human
sclerostin in the biological sample is inversely proportional to
the amount of labeled rhSOST standard bound to the sclerostin
binding protein. Similarly, human sclerostin can also be assayed in
biological fluids by a competition immunoassay utilizing rhSOST
standards labeled with a detectable substance and an unlabeled
human sclerostin binding protein.
[0412] In a provided embodiment, the binding proteins and
sclerostin binding portions of are capable of neutralizing human
sclerostin activity both in vitro and in vivo. Accordingly, such
binding proteins and sclerostin binding portions thereof are
provided and can be used to inhibit hSOST activity, e.g., in a cell
culture containing hSOST, in human subjects, or in other mammalian
subjects having sclerostin with which an antibody cross-reacts. One
embodiment provides a method for inhibiting hSOST activity
comprising contacting hSOST with a sclerostin binding protein or
binding portion thereof such that hSOST activity is inhibited. For
example, in a cell culture containing, or suspected of containing
hSOST, a sclerostin binding protein or binding portion thereof can
be added to the culture medium to inhibit hSOST activity in the
culture.
[0413] Another embodiment provides a method for reducing hSOST
activity in a subject, advantageously from a subject suffering from
a disease or disorder in which sclerostin activity is detrimental.
Methods for reducing sclerostin activity in a subject suffering
from such a disease or disorder are provided, which method
comprises administering to the subject an antibody or antibody
portion such that sclerostin activity in the subject is reduced. In
an embodiment, the sclerostin is human sclerostin and the subject
is a human subject. Alternatively, the subject can be a mammal
expressing an sclerostin to which an antibody is capable of
binding. Still further, the subject can be a mammal into which
sclerostin has been introduced (e.g., by administration of
sclerostin or by expression of an SOST transgene). A sclerostin
binding protein can be administered to a human subject for
therapeutic purposes. Moreover, a binding protein can be
administered to a non-human mammal expressing an sclerostin with
which the antibody is capable of binding for veterinary purposes or
as an animal model of human disease. Regarding the latter, such
animal models may be useful for evaluating the therapeutic efficacy
of antibodies (e.g., testing of dosages and time courses of
administration).
[0414] The term "a disorder in which sclerostin activity is
detrimental" is intended to include diseases and other disorders in
which the presence of sclerostin in a subject suffering from the
disorder has been shown to be or is suspected of being either
responsible for the pathophysiology of the disorder or a factor
that contributes to a worsening of the disorder. Accordingly, a
disorder in which sclerostin activity is detrimental is a disorder
in which reduction of sclerostin activity is expected to alleviate
the symptoms and/or progression of the disorder. Such disorders may
be evidenced, for example, by an increase in the concentration of
sclerostin in a biological fluid of a subject suffering from the
disorder (e.g., an increase in the concentration of sclerostin in
serum, plasma, synovial fluid, etc. of the subject), which can be
detected, for example, using an anti-sclerostin antibody as
described above. Non-limiting examples of disorders that can be
treated with the antibodies include those disorders discussed in
the section below pertaining to pharmaceutical compositions of the
antibodies.
[0415] DVD-binding proteins capable of binding sclerostin (e.g.,
human sclerostin) alone or multiple antigens (e.g., human
sclerostin and another non-sclerostin antigen) are provided. Thus,
a DVD-binding protein may block or reduce activity of human
sclerostin and the activity of another target antigen. Such other
target antigens may include soluble targets (e.g., TNF) and cell
surface receptor targets (e.g., VEGFR, EGFR).
[0416] Such other antigens include, but are not limited to, the
targets listed in publically available databases, which databases
include those that are available on the worldwide web. These target
databases include those listing:
Therapeutic targets (hxin.cz3.nus.edu.sg/group/cjttd/ttd.asp);
Cytokines and cytokine receptors (hwww.cytokinewebfacts.com/,
hwww.copewithcytokines.de/cope.cgi, and
hcmbi.bjmu.edu.cn/cmbidata/cgf/CGF_Database/cytokine.medic.kumamoto-u.ac.-
jp/CFC/indexR.html); Chemokines
(hcytokine.medic.kumamoto-u.ac.jp/CFC/CK/Chemokine.html); Chemokine
receptors and GPCRs (hcsp.medic.kumamoto-u.ac.jp/CSP/Receptor.html,
hwww.gper.org/7tm/); Olfactory Receptors
(hsenselab.med.yale.edu/senselab/ORDB/default.asp); Receptors
(hwww.iuphar-db.org/iuphar-rd/list/index.htm); Cancer targets
(hcged.hgc.jp/cgi-bin/input.cgi); Secreted proteins as potential
antibody targets (hspd.cbi.pku.edu.cn/); Protein kinases
(hspd.cbi.pku.edu.cn/), and Human CD markers
(hcontent.labvelocity.com/tools/6/1226/CD_table_final_locked.pdf)
and (Zola H, 2005 CD molecules 2005: human cell differentiation
molecules Blood, 106:3123-6).
[0417] DVD-binding proteins are useful as therapeutic agents to
simultaneously block two or more different targets, i.e., hSOST,
and one or more other non-SOST target antigens to enhance
efficacy/safety and/or increase patient coverage. Such targets may
include soluble targets (TNF) and cell surface receptor targets
(VEGFR and EGFR).
[0418] Additionally, DVD-binding proteins that can be employed for
tissue-specific delivery (target a tissue marker and a disease
mediator for enhanced local PK thus higher efficacy and/or lower
toxicity) are provided, including intracellular delivery (targeting
an internalizing receptor and a intracellular molecule), delivering
to inside brain (targeting transferrin receptor and a CNS disease
mediator for crossing the blood-brain barrier). DVD-binding protein
can also serve as a carrier protein to deliver an antigen to a
specific location via binding to a non-neutralizing epitope of that
antigen and also to increase the half-life of the antigen.
Furthermore, DVD-binding protein can be designed to either be
physically linked to medical devices implanted into patients or
target these medical devices (see Burke, Sandra E.; Kuntz, Richard
E.; Schwartz, Lewis B., Zotarolimus eluting stents. Advanced Drug
Delivery Reviews (2006), 58(3), 437-446; Surface coatings for
biological activation and functionalization of medical devices,
Hildebrand, H. F.; Blanchemain, N.; Mayer, G.; Chai, F.; Lefebvre,
M.; Boschin, F., Surface and Coatings Technology (2006),
200(22-23), 6318-6324; Wu et al., "Drug/device combinations for
local drug therapies and infection prophylaxis," Biomaterials, 27:
2450-2467 (2006); Marques et al., "Mediation of the Cytokine
Network in the Implantation of Orthopedic Devices," Chapter 21, In
Biodegradable Systems in Tissue Engineering and Regenerative
Medicine, (Reis et al., eds.) (CRC Press LLC, Boca Raton, 2005) pp.
377-397. Briefly, directing appropriate types of cell to the site
of medical implant may promote healing and restoring normal tissue
function. Alternatively, inhibition of mediators (including but not
limited to cytokines), released upon device implantation by a
DVD-binding protein coupled to or target to a device is also
provided. For example, Stents have been used for years in
interventional cardiology to clear blocked arteries and to improve
the flow of blood to the heart muscle. However, traditional bare
metal stents have been known to cause restenosis (re-narrowing of
the artery in a treated area) in some patients and can lead to
blood clots. Recently, an anti-CD34 antibody coated stent has been
described which reduced restenosis and prevents blood clots from
occurring by capturing endothelial progenitor cells (EPC)
circulating throughout the blood. Endothelial cells are cells that
line blood vessels, allowing blood to flow smoothly. The EPCs
adhere to the hard surface of the stent forming a smooth layer that
not only promotes healing but prevents restenosis and blood clots,
complications previously associated with the use of stents (Aoji et
al. 2005 J Am Coll Cardiol. 45(10):1574-9). In addition to
improving outcomes for patients requiring stents, there are also
implications for patients requiring cardiovascular bypass surgery.
For example, a prosthetic vascular conduit (artificial artery)
coated with anti-EPC antibodies would eliminate the need to use
arteries from patients legs or arms for bypass surgery grafts. This
would reduce surgery and anesthesia times, which in turn will
reduce coronary surgery deaths. DVD-binding protein are designed in
such a way that it binds to a cell surface marker (such as CD34) as
well as a protein (or an epitope of any kind, including but not
limited to proteins, lipids and polysaccharides) that has been
coated on the implanted device to facilitate the cell recruitment.
Such approaches can also be applied to other medical implants in
general. Alternatively, DVD-binding proteins can be coated on
medical devices and upon implantation and releasing all DVDs from
the device (or any other need which may require additional fresh
DVD-binding protein, including aging and denaturation of the
already loaded DVD-binding protein) the device could be reloaded by
systemic administration of fresh DVD-binding protein to the
patient, where the DVD-binding protein is designed to binds to a
target of interest (a cytokine, a cell surface marker (such as
CD34) etc.) with one set of binding sites and to a target coated on
the device (including a protein, an epitope of any kind, including
but not limited to lipids, polysaccharides and polymers) with the
other. This technology has the advantage of extending the
usefulness of coated implants.
V.A. Use of DVD-Binding Proteins in Various Diseases
[0419] DVD-binding proteins useful as therapeutic molecules to
treat various diseases are provided. Such DVD molecules may bind
one or more targets involved in a specific disease. Examples of
such targets in various diseases are described below.
VI. Human Autoimmune and Inflammatory Response
[0420] In one aspect, a DVD-binding protein capable of binding
human sclerostin and one or more antigens that have been implicated
in general autoimmune and inflammatory responses is provided,
including C5, CCL1 (1-309), CCL11 (eotaxin), CCL13 (mcp-4), CCL15
(MIP-1d), CCL16 (HCC-4), CCL17 (TARC), CCL18 (PARC), CCL19, CCL2
(mcp-1), CCL20 (MIP-3a), CCL21 (MIP-2), CCL23 (MPIF-1), CCL24
(MPIF-2/eotaxin-2), CCL25 (TECK), CCL26, CCL3 (MIP-1a), CCL4
(MIP-1b), CCL5 (RANTES), CCL7 (mcp-3), CCL8 (mcp-2), CXCL1, CXCL10
(IP-10), CXCL11 (I-TAC/IP-9), CXCL12 (SDF1), CXCL13, CXCL14, CXCL2,
CXCL3, CXCL5 (ENA-78/LIX), CXCL6 (GCP-2), CXCL9, IL13, IL8, CCL13
(mcp-4), CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9,
CX3CR1, IL8RA, XCR1 (CCXCR1), IFNA2, IL10, IL13, IL17C, IL1A, IL1B,
IL1F10, IL1F5, IL1F6, IL1F7, IL1F8, IL1F9, IL22, IL5, IL8, IL9,
LTA, LTB, MIF, SCYE1 (endothelial Monocyte-activating cytokine),
SPP1, TNF, TNFSF5, IFNA2, IL10RA, IL10RB, IL13, IL13RA1, IL5RA,
IL9, IL9R, ABCF1, BCL6, C3, C4A, CEBPB, CRP, ICEBERG, IL1R1, IL1RN,
IL8RB, LTB4R, TOLLIP, FADD, IRAK1, IRAK2, MYD88, NCK2, TNFAIP3,
TRADD, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6, ACVR1, ACVR1B,
ACVR2, ACVR2B, ACVRL1, CD28, CD3E, CD3G, CD3Z, CD69, CD80, CD86,
CNR1, CTLA4, CYSLTR1, FCER1A, FCER2, FCGR3A, GPR44, HAVCR2, OPRD1,
P2RX7, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, BLR1,
CCL1, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL13, CCL15,
CCL16, CCL17, CCL18, CCL19, CCL20, CCL21, CCL22, CCL23, CCL24,
CCL25, CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9,
CX3CL1, CX3CR1, CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL10, CXCL11,
CXCL12, CXCL13, CXCR4, GPR2, SCYE1, SDF2, XCL1, XCL2, XCR1, AMH,
AMHR2, BMPR1A, BMPR1B, BMPR2, C19orf10 (IL27w), CER1, CSF1, CSF2,
CSF3, DKFZp451J0118, FGF2, GFI1, IFNA1, IFNB1, IFNG, IGF1, IL1A,
IL1B, IL1R1, IL1R2, IL2, IL2RA, IL2RB, IL2RG, IL3, IL4, IL4R, IL5,
IL5RA, IL6, IL6R, IL6ST, IL7, IL8, IL8RA, IL8RB, IL9, IL9R, IL10,
IL10RA, IL10RB, IL11, IL11RA, IL12A, IL12B, IL12RB1, IL12RB2, IL13,
IL13RA1, IL13RA2, IL15, IL15RA, IL16, IL17, IL17R, IL18, IL18R1,
IL19, IL20, KITLG, LEP, LTA, LTB, LTB4R, LTB4R2, LTBR, MIF, NPPB,
PDGFB, TBX21, TDGF1, TGFA, TGFB1, TGFB1I1, TGFB2, TGFB3, TGFBI,
TGFBR1, TGFBR2, TGFBR3, TH1L, TNF, TNFRSF1A, TNFRSF1B, TNFRSF7,
TNFRSF8, TNFRSF9, TNFRSF11A, TNFRSF21, TNFSF4, TNFSF5, TNFSF6,
TNFSF11, VEGF, ZFPM2, and RNF110 (ZNF144).
VI.A. Asthma
[0421] Allergic asthma is characterized by the presence of
eosinophilia, goblet cell metaplasia, epithelial cell alterations,
airway hyperreactivity (AHR), and Th2 and Th1 cytokine expression,
as well as elevated serum IgE levels. It is now widely accepted
that airway inflammation is the key factor underlying the
pathogenesis of asthma, involving a complex interplay of
inflammatory cells such as T cells, B cells, eosinophils, mast
cells and macrophages, and of their secreted mediators including
cytokines and chemokines. Corticosteroids are the most important
anti-inflammatory treatment for asthma today, however their
mechanism of action is non-specific and safety concerns exist,
especially in the juvenile patient population. The development of
more specific and targeted therapies is therefore warranted.
[0422] Animal models such as OVA-induced asthma mouse model, where
both inflammation and AHR can be assessed, are known in the art and
may be used to determine the ability of various DVD-binding
proteins to treat asthma Animal models for studying asthma are
disclosed in Coffman, et al., Journal of Experimental Medicine
(2005), 201(12), 1875-1879; Lloyd, et al., Advances in Immunology
(2001), 77, 263-295; Boyce et al., Journal of Experimental Medicine
(2005), 201(12), 1869-1873; and Snibson, et al., Journal of the
British Society for Allergy and Clinical Immunology (2005), 35(2),
146-52. In addition to routine safety assessments of these target
pairs specific tests for the degree of immunosuppression may be
warranted and helpful in selecting the best target pairs (see
Luster et al., Toxicology (1994), 92(1-3), 229-43; Descotes, et
al., Developments in biological standardization (1992), 77 99-102;
Hart et al., Journal of Allergy and Clinical Immunology (2001),
108(2), 250-257).
[0423] One aspect provides DVD-binding proteins capable of binding
SOST and one or more, for example two, of IL-4, IL-5, IL-8, IL-9,
IL-13, IL-18, IL-5R(.alpha.), TNFSF4, IL-4R(.alpha.), interferon
.alpha., eotaxin, TSLP, PAR-2, PGD2, or IgE. An embodiment includes
a dual-specific anti-sclerostin/TNF.alpha. DVD-binding protein as a
therapeutic agent beneficial for the treatment of asthma.
VI.B. Rheumatoid Arthritis (RA)
[0424] Rheumatoid arthritis (RA), a systemic disease, is
characterized by a chronic inflammatory reaction in the synovium of
joints and is associated with degeneration of cartilage and erosion
of juxta-articular bone. Many pro-inflammatory cytokines including
TNF, chemokines, and growth factors are expressed in diseased
joints. Systemic administration of anti-TNF antibody or sTNFR
fusion protein to mouse models of RA was shown to be
anti-inflammatory and joint protective. Various cytokines, included
sclerostin have been implicated in RA. Clinical investigations in
which the activity of TNF in RA patients was blocked with
intravenously administered infliximab (Harriman G, Harper L K,
Schaible T F. 1999 Summary of clinical trials in rheumatoid
arthritis using infliximab, an anti-TNFalpha treatment. Ann. Rheum.
Dis., 58 Suppl 1: I61-4), a chimeric anti-TNF mAb, has provided
evidence that TNF regulates IL-6, IL-8, MCP-1, and VEGF production,
recruitment of immune and inflammatory cells into joints,
angiogenesis, and reduction of blood levels of matrix
metalloproteinases-1 and -3. A better understanding of the
inflammatory pathway in rheumatoid arthritis has led to
identification of other therapeutic targets involved in rheumatoid
arthritis. Promising treatments such as interleukin-6 antagonists
(IL-6 receptor antibody MRA, developed by Chugai, Roche (see
Nishimoto, Norihiro et al., Arthritis & Rheumatism, (2004),
50(6): 1761-1769), CTLA4Ig (abatacept, Genovese et al. (2005)
"Abatacept for rheumatoid arthritis refractory to tumor necrosis
factor alpha inhibition," N. Engl. J. Med., 353: 1114-23.), and
anti-B cell therapy (rituximab, Okamoto H, Kamatani N. (2004)
"Rituximab for rheumatoid arthritis," N. Engl. J. Med., 351: 1909)
have already been tested in randomized controlled trials over the
past year. Sclerostin and other cytokines, such as IL-15 and IL-18,
have been identified as playing a role using RA animal models
(therapeutic antibody HuMax-IL.sub.--15, AMG 714 see Baslund, Bo et
al., Arthritis & Rheumatism (2005), 52(9): 2686-2692).
Dual-specific antibody therapy, combining anti-TNF and another
mediator, such as sclerostin, has great potential in enhancing
clinical efficacy and/or patient coverage. For example, blocking
both TNF and VEGF can potentially eradicate inflammation and
angiogenesis, both of which are involved in pathophysiology of RA.
A DVD-binding protein capable of blocking TNF-.alpha.and sclerostin
is contemplated. In addition to routine safety assessments of these
target pairs, specific tests for the degree of immunosuppression
may be warranted and helpful in selecting the best target pairs
(see Luster et al., Toxicology (1994), 92(1-3), 229-43; Descotes,
et al., Developments in biological standardization (1992), 77
99-102; Hart et al., Journal of Allergy and Clinical Immunology
(2001), 108(2), 250-257). Whether a DVD-binding protein will be
useful for the treatment of rheumatoid arthritis can be assessed
using pre-clinical animal RA models such as the collagen-induced
arthritis mouse model. Other useful models are also well known in
the art (see Brand D D., Comp. Med., (2005) 55(2):114-22). Based on
the cross-reactivity of the parental antibodies for human and mouse
orthologues (e.g., reactivity for human and mouse TNF, human and
mouse IL-15 etc.) validation studies in the mouse CIA model may be
conducted with "matched surrogate antibody" derived DVD-binding
proteins; briefly, a DVD-binding protein based on two (or more)
mouse target specific antibodies may be matched to the extent
possible to the characteristics of the parental human or humanized
antibodies used for human DVD-binding protein construction (similar
affinity, similar neutralization potency, similar half-life
etc.).
[0425] An embodiment provides a DVD-binding protein that binds
human sclerostin and another non-sclerostin target that may also be
used to treat other diseases in which SOST plays a role. Such
diseases include, but are not limited to SLE, multiple sclerosis
(MS), sepsis, various neurological diseases, and cancers (including
cervical, breast, gastric). A more extensive list of diseases and
disorders in which sclerostin plays a role is also provided
below.
[0426] An embodiment provides a DVD-binding protein capable of
binding huSOST and one or more targets of TNF.alpha., IL-12, TWEAK,
IL-23, CXCL13, CD40, CD40L, IL-18, VEGF, VLA-4, TNF.beta., CD45RB,
CD200, IFN-.gamma., GM-CSF, FGF, C5, CD52, sclerostin, or CCR2.
VI.C. SLE (Lupus)
[0427] The immunopathogenic hallmark of SLE is the polyclonal B
cell activation, which leads to hyperglobulinemia, autoantibody
production and immune complex formation. The fundamental
abnormality appears to be the failure of T cells to suppress the
forbidden B cell clones due to generalized T cell dysregulation. In
addition, B and T-cell interaction is facilitated by several
cytokines such as IL-10 as well as co-stimulatory molecules such as
CD40 and CD40L, B7 and CD28 and CTLA-4, which initiate the second
signal. These interactions together with impaired phagocytic
clearance of immune complexes and apoptotic material, perpetuate
the immune response with resultant tissue injury.
[0428] One aspect provides a DVD-binding protein capable of binding
human sclerostin and one or more of the following antigens that
have been implicated in SLE: B cell targeted therapies: CD-20,
CD-22, CD-19, CD28, CD4, CD80, HLA-DRA, IL10, IL2, IL4, TNFRSF5,
TNFRSF6, TNFSF5, TNFSF6, BLR1, HDAC4, HDAC5, HDAC7A, HDAC9, ICOSL,
IGBP1, MS4A1, RGS1, SLA2, CD81, IFNB1, IL10, TNFRSF5, TNFRSF7,
TNFSF5, AICDA, BLNK, GALNAC4S-6ST, HDAC4, HDAC5, HDAC7A, HDAC9,
IL10, IL11, IL4, INHA, INHBA, KLF6, TNFRSF7, CD28, CD38, CD69,
CD80, CD83, CD86, DPP4, FCER2, IL2RA, TNFRSF8, TNFSF7, CD24, CD37,
CD40, CD72, CD74, CD79A, CD79B, CR2, IL1R2, ITGA2, ITGA3, MS4A1,
ST6GAL1, CD1C, CHST10, HLA-A, HLA-DRA, and NT5E; co-stimulatory
signals: CTLA4 or B7.1/B7.2; inhibition of B cell survival: BlyS,
BAFF; Complement inactivation: C5; Cytokine modulation: the key
principle is that the net biologic response in any tissue is the
result of a balance between local levels of proinflammatory or
anti-inflammatory cytokines (see Sfikakis P P et al 2005 Curr Opin
Rheumatol 17:550-7). SLE is considered to be a Th-2 driven disease
with documented elevations in serum IL-4, IL-6, IL-10. DVD-binding
proteins capable of binding IL-4, IL-6, IL-10, IFN-.alpha., or
TNF-.alpha. are also contemplated. Combination of targets discussed
herein will enhance therapeutic efficacy for SLE which can be
tested in a number of lupus preclinical models (see, Peng S L
(2004) Methods Mol. Med., 102: 227-72). Based on the
cross-reactivity of the parental antibodies for human and mouse
othologues (e.g., reactivity for human and mouse CD20, human and
mouse Interferon alpha etc.) validation studies in a mouse lupus
model may be conducted with "matched surrogate antibody" derived
DVD-binding proteins; briefly, a DVD-binding protein based two (or
more) mouse target specific antibodies may be matched to the extent
possible to the characteristics of the parental human or humanized
antibodies used for human DVD-binding protein construction (similar
affinity, similar neutralization potency, similar half-life
etc.)
VI.D. Multiple Sclerosis (MS)
[0429] Multiple sclerosis (MS) is a complex human autoimmune-type
disease with a predominantly unknown etiology. Immunologic
destruction of myelin basic protein (MBP) throughout the nervous
system is the major pathology of multiple sclerosis. MS is a
disease of complex pathologies, which involves infiltration by CD4+
and CD8+ T cells and of response within the central nervous system.
Expression in the CNS of cytokines, reactive nitrogen species and
costimulator molecules have all been described in MS. Of major
consideration are immunological mechanisms that contribute to the
development of autoimmunity. In particular, antigen expression,
cytokine and leukocyte interactions, and regulatory T-cells, which
help balance/modulate other T-cells such as Th1 and Th2 cells, are
important areas for therapeutic target identification.
[0430] IL-12 is a proinflammatory cytokine that is produced by APC
and promotes differentiation of Th1 effector cells. IL-12 is
produced in the developing lesions of patients with MS as well as
in EAE-affected animals. Previously it was shown that interference
in IL-12 pathways effectively prevents EAE in rodents, and that in
vivo neutralization of IL-12p40 using a anti-IL-12 mAb has
beneficial effects in the myelin-induced EAE model in common
marmosets.
[0431] TWEAK is a member of the TNF family, constitutively
expressed in the central nervous system (CNS), with
pro-inflammatory, proliferative or apoptotic effects depending upon
cell types. Its receptor, Fn14, is expressed in CNS by endothelial
cells, reactive astrocytes and neurons. TWEAK and Fn14 mRNA
expression increased in spinal cord during experimental autoimmune
encephalomyelitis (EAE). Anti-TWEAK antibody treatment in myelin
oligodendrocyte glycoprotein (MOG) induced EAE in C57BL/6 mice
resulted in a reduction of disease severity and leukocyte
infiltration when mice were treated after the priming phase.
[0432] One aspect provides DVD-binding proteins capable of binding
SOST and one or more, for example two, targets including IL-12,
TWEAK, IL-23, CXCL13, CD40, CD40L, IL-18, VEGF, VLA-4, TNF, CD45RB,
CD200, IFNgamma, GM-CSF, FGF, C5, CD52, osteopontin, and/or CCR2.
An embodiment includes a dual-specific anti-sclerostin/TNF-.alpha.
DVD-binding protein as a therapeutic agent beneficial for the
treatment of MS.
[0433] Several animal models for assessing the usefulness of the
DVD-binding protein molecules to treat MS are known in the art (see
Steinman L, et al., (2005) Trends Immunol. 26(11):565-71; Lublin F
D., et al., (1985) Springer Semin Immunopathol. 8(3):197-208;
Genain C P, et al., (1997) J Mol Med. 75(3):187-97; Tuohy V K, et
al., (1999) J Exp Med. 189(7):1033-42; Owens T, et al., (1995)
Neurol Clin. 13(1):51-73; and 't Hart et al., J. Immunol., 175(7):
4761-4768 (2005). Based on the cross-reactivity of the parental
antibodies for human and animal species othologues (e.g.,
reactivity for human and mouse SOST, human and mouse TWEAK etc.)
validation studies in the mouse EAE model may be conducted with
"matched surrogate antibody" derived DVD-binding protein molecules;
briefly, a DVD-binding protein based on to (or more) mouse target
specific antibodies may be matched to the extent possible to the
characteristics of the parental human or humanized antibodies used
for human DVD-binding protein construction (similar affinity,
similar neutralization potency, similar half-life etc.). The same
concept applies to animal models in other non-rodent species, where
a "matched surrogate antibody" derived DVD-binding protein would be
selected for the anticipated pharmacology and possibly safety
studies. In addition to routine safety assessments of these target
pairs specific tests for the degree of immunosuppression may be
warranted and helpful in selecting the best target pairs (see
Luster et al., Toxicology (1994), 92(1-3), 229-43; Descotes, et
al., Developments in biological standardization (1992), 77 99-102;
Jones R. 2000 Rovelizumab (ICOS Corp). IDrugs. 3(4):442-6).
VI.E. Sepsis
[0434] The pathophysiology of sepsis is initiated by the outer
membrane components of both gram-negative organisms
(lipopolysaccharide [LPS], lipid A, endotoxin) and gram-positive
organisms (lipoteichoic acid, peptidoglycan). These outer membrane
components are able to bind to the CD14 receptor on the surface of
monocytes. By virtue of the recently described toll-like receptors,
a signal is then transmitted to the cell, leading to the eventual
production of the proinflammatory cytokines tumor necrosis
factor-alpha (TNF-alpha) and interleukin-1 (IL-1). Overwhelming
inflammatory and immune responses are essential features of septic
shock and play a central part in the pathogenesis of tissue damage,
multiple organ failure, and death induced by sepsis. Cytokines,
especially tumor necrosis factor (TNF) and interleukin (IL-1), have
been shown to be critical mediators of septic shock. These
cytokines have a direct toxic effect on tissues; they also activate
phospholipase A2. These and other effects lead to increased
concentrations of platelet-activating factor, promotion of nitric
oxide synthase activity, promotion of tissue infiltration by
neutrophils, and promotion of neutrophil activity.
[0435] The treatment of sepsis and septic shock remains a clinical
conundrum, and recent prospective trials with biological response
modifiers (i.e. anti-TNF, anti-MIF) aimed at the inflammatory
response have shown only modest clinical benefit. Recently,
interest has shifted toward therapies aimed at reversing the
accompanying periods of immune suppression. Studies in experimental
animals and critically ill patients have demonstrated that
increased apoptosis of lymphoid organs and some parenchymal tissues
contribute to this immune suppression, anergy, and organ system
dysfunction. During sepsis syndromes, lymphocyte apoptosis can be
triggered by the absence of IL-2 or by the release of
glucocorticoids, granzymes, or the so-called `death` cytokines:
tumor necrosis factor alpha or Fas ligand. Apoptosis proceeds via
auto-activation of cytosolic and/or mitochondrial caspases, which
can be influenced by the pro- and anti-apoptotic members of the
Bcl-2 family. In experimental animals, not only can treatment with
inhibitors of apoptosis prevent lymphoid cell apoptosis; it may
also improve outcome. Although clinical trials with anti-apoptotic
agents remain distant due in large part to technical difficulties
associated with their administration and tissue targeting,
inhibition of lymphocyte apoptosis represents an attractive
therapeutic target for the septic patient. Likewise, a
dual-specific agent targeting both inflammatory mediator and a
apoptotic mediator, may have added benefit. One aspect provides
DVD-binding proteins capable of binding sclerostin and one or more
targets involved in sepsis, including TNF, IL-1, MIF, IL-6, IL-8,
IL-18, IL-12, IL-23, FasL, LPS, Toll-like receptors, TLR-4, tissue
factor, MIP-2, ADORA2A, CASP1, CASP4, IL-10, IL-1B, NFKB1, PROC,
TNFRSF1A, CSF3, CCR3, IL1RN, MIF, NFKB1, PTAFR, TLR2, TLR4, GPR44,
HMOX1, HMG-B1, midkine, IRAK1, NFKB2, SERPINA1, SERPINE1, or TREM1.
The efficacy of such DVD-binding proteins for sepsis can be
assessed in preclinical animal models known in the art (see, Buras
J A, et al., (2005) Nat. Rev. Drug Discov., 4(10): 854-65 and
Calandra T, et al., (2000) Nat. Med., 6(2):164-70).
VI.F. Neurological Disorders and Neurodegenerative Diseases
[0436] Neurodegenerative diseases are either chronic in which case
they are usually age-dependent or acute (e.g., stroke, traumatic
brain injury, spinal cord injury, etc.). They are characterized by
progressive loss of neuronal functions (neuronal cell death,
demyelination), loss of mobility and loss of memory. Emerging
knowledge of the mechanisms underlying chronic neurodegenerative
diseases (e.g., Alzheimer's disease, AD) show a complex etiology
and a variety of factors have been recognized to contribute to
their development and progression e.g., age, glycemic status,
amyloid production and multimerization, accumulation of advanced
glycation-end products (AGE) which bind to their receptor RAGE
(receptor for AGE), increased brain oxidative stress, decreased
cerebral blood flow, neuroinflammation including release of
inflammatory cytokines and chemokines, neuronal dysfunction and
microglial activation. Thus these chronic neurodegenerative
diseases represent a complex interaction between multiple cell
types and mediators. Treatment strategies for such diseases are
limited and mostly constitute either blocking inflammatory
processes with non-specific anti-inflammatory agents (e.g.,
corticosteroids, COX inhibitors) or agents to prevent neuron loss
and/or synaptic functions. These treatments fail to stop disease
progression. Recent studies suggest that more targeted therapies
such as antibodies to soluble A.beta. peptide (including the
A.beta. oligomeric forms) can not only help stop disease
progression but may help maintain memory as well. These preliminary
observations suggest that specific therapies targeting more than
one disease mediator (e.g., A.beta. and a pro-inflammatory cytokine
such as TNF) may provide even better therapeutic efficacy for
chronic neurodegenerative diseases than observed with targeting a
single disease mechanism (e.g., soluble A.beta. alone) (see C. E.
Shepherd, et al, Neurobiol Aging. 2005 Oct. 24; Nelson R B., Curr
Pharm Des. 2005; 11:3335; William L. Klein.; Neurochem Int. 2002;
41:345; Janelsins et al., "Early correlation of microglial
activation with enhanced tumor necrosis factor-alpha and monocyte
chemoattractant protein-I expression specifically within the
entorhinal cortex of triple transgenic Alzheimer's disease mice,"
Journal of Neuroinflammation, 2(23): 1-12 (2005); Soloman B., Curr
Alzheimer Res. 2004; 1:149; Igor Klyubin, et al., Nat Med. 2005;
11:556-61; Arancio O, et al., EMBO Journal (2004) 1-10; Bornemann K
D, et al., Am J Pathol. 2001; 158:63; Deane R, et al., Nat Med.
2003; 9:907-13; and Eliezer Masliah, et al., Neuron. 2005;
46:857).
[0437] The DVD-binding proteins can bind sclerostin and one or more
targets involved in chronic neurodegenerative diseases such as
Alzheimers. Such targets include, but are not limited to, any
mediator, soluble or cell surface, implicated in AD pathogenesis,
e.g., AGE (S100 A, amphotericin), pro-inflammatory cytokines (e.g.,
IL-1), chemokines (e.g., MCP 1), molecules that inhibit nerve
regeneration (e.g., Nogo, RGM A), molecules that enhance neurite
growth (neurotrophins) and molecules that can mediate transport at
the blood brain barrier (e.g., transferrin receptor, insulin
receptor or RAGE). The efficacy of DVD-binding proteins can be
validated in pre-clinical animal models such as the transgenic mice
that over-express amyloid precursor protein or RAGE and develop
Alzheimer's disease-like symptoms. In addition, DVD-binding
proteins can be constructed and tested for efficacy in the animal
models and the best therapeutic DVD-binding protein can be selected
for testing in human patients. DVD-binding proteins can also be
employed for treatment of other neurodegenerative diseases such as
Parkinson's disease. Alpha-Synuclein is involved in Parkinson's
pathology. A DVD-binding protein capable of targeting sclerostin
and LINGO-1, alpha-synuclein, and/or inflammatory mediators such as
TNF, IL-1, MCP-1 can prove effective therapy for Parkinson's
disease and are contemplated.
VI.G. Neuronal Regeneration and Spinal Cord Injury
[0438] Despite an increase in knowledge of the pathologic
mechanisms, spinal cord injury (SCI) is still a devastating
condition and represents a medical indication characterized by a
high medical need. Most spinal cord injuries are contusion or
compression injuries and the primary injury is usually followed by
secondary injury mechanisms (inflammatory mediators e.g., cytokines
and chemokines) that worsen the initial injury and result in
significant enlargement of the lesion area, sometimes more than
10-fold. These primary and secondary mechanisms in SCI are very
similar to those in brain injury caused by other means e.g.,
stroke. No satisfying treatment exists and high dose bolus
injection of methylprednisolone (MP) is the only used therapy
within a narrow time window of 8 h post injury. This treatment,
however, is only intended to prevent secondary injury without
causing any significant functional recovery. It is heavily
criticized for the lack of unequivocal efficacy and severe adverse
effects, like immunosuppression with subsequent infections and
severe histopathological muscle alterations. No other drugs,
biologics or small molecules, stimulating the endogenous
regenerative potential are approved, but promising treatment
principles and drug candidates have shown efficacy in animal models
of SCI in recent years. To a large extent the lack of functional
recovery in human SCI is caused by factors inhibiting neurite
growth, at lesion sites, in scar tissue, in myelin as well as on
injury-associated cells. Such factors are the myelin-associated
proteins NogoA, OMgp and MAG, RGM A, the scar-associated CSPG
(Chondroitin Sulfate Proteoglycans) and inhibitory factors on
reactive astrocytes (some semaphorins and ephrins). However, at the
lesion site not only growth inhibitory molecules are found but also
neurite growth stimulating factors like neurotrophins, laminin, L1
and others. This ensemble of neurite growth inhibitory and growth
promoting molecules may explain that blocking single factors, like
NogoA or RGM A, resulted in significant functional recovery in
rodent SCI models, because a reduction of the inhibitory influences
could shift the balance from growth inhibition to growth promotion.
However, recoveries observed with blocking a single neurite
outgrowth inhibitory molecule were not complete. To achieve faster
and more pronounced recoveries either blocking two neurite
outgrowth inhibitory molecules, e.g., Nogo and RGM A, or blocking
an neurite outgrowth inhibitory molecule and enhancing functions of
a neurite outgrowth enhancing molecule, e.g., Nogo and
neurotrophins, or blocking a neurite outgrowth inhibitory molecule,
e.g., Nogo and a pro-inflammatory molecule e.g., TNF, may be
desirable (see McGee A W, et al. (2003) Trends Neurosci., 26: 193;
Marco Domeniconi, et al. (2005) J. Neurol. Sci., 233: 43; Milan
Makwanal, et al. (2005) FEBS J. 272:2628; Barry J. Dickson (2002)
Science, 298: 1959; Felicia Yu Hsuan Teng, et al. (2005) J.
Neurosci. Res. 79:273; Tara Karnezis, et al. (2004) Nature
Neuroscience, 7: 736; Gang Xu, et al. (2004) J. Neurochem., 91:
1018).
[0439] In one aspect, a DVD-binding protein that binds human
sclerostin may also bind one or both of the target pairs such as
NgR and RGM A; NogoA and RGM A; MAG and RGM A; OMGp and RGM A; RGM
A and RGM B; CSPGs and RGM A; aggrecan, midkine, neurocan,
versican, phosphacan, Te38 and TNF-.alpha.; A.beta.
globulomer-specific antibodies combined with antibodies promoting
dendrite & axon sprouting are provided. Dendrite pathology is a
very early sign of AD and it is known that NOGO A restricts
dendrite growth. One can combine such type of ab with any of the
SCI-candidate (myelin-proteins) Ab. Other DVD-binding protein
targets may include any combination of NgR-p75, NgR-Troy,
NgR-Nogo66 (Nogo), NgR-Lingo, Lingo-Troy, Lingo-p75, MAG or Omgp.
Additionally, targets may also include any mediator, soluble or
cell surface, implicated in inhibition of neurite, e.g., Nogo,
Ompg, MAG, RGM A, semaphorins, ephrins, soluble A.beta.,
pro-inflammatory cytokines (e.g., IL-1), chemokines (e.g., MIP 1a),
molecules that inhibit nerve regeneration. The efficacy of
anti-nogo/anti-RGM A or similar DVD-binding proteins can be
validated in pre-clinical animal models of spinal cord injury. In
addition, these DVD-binding proteins can be constructed and tested
for efficacy in the animal models and the best therapeutic
DVD-binding protein can be selected for testing in human patients.
In addition, DVD-binding protein can be constructed that target two
distinct ligand binding sites on a single receptor e.g., Nogo
receptor which binds three ligand Nogo, Ompg, and MAG and RAGE that
binds A.beta. and S100 A. Furthermore, neurite outgrowth inhibitors
e.g., nogo and nogo receptor, also play a role in preventing nerve
regeneration in immunological diseases like multiple sclerosis
Inhibition of nogo-nogo receptor interaction has been shown to
enhance recovery in animal models of multiple sclerosis. Therefore,
DVD-binding proteins that can block the function of one immune
mediator, e.g., a cytokine like IL-12, and a neurite outgrowth
inhibitor molecule, e.g., Nogo or RGM, may offer faster and greater
efficacy than blocking either an immune or a neurite outgrowth
inhibitor molecule alone.
In general, antibodies do not cross the blood brain barrier (BBB)
in an efficient and relevant manner. However, in certain neurologic
diseases, e.g., stroke, traumatic brain injury, multiple sclerosis,
etc., the BBB may be compromised and allows for increased
penetration of DVD-binding proteins and antibodies into the brain.
In other neurological conditions, where BBB leakage is not
occurring, one may employ the targeting of endogenous transport
systems, including carrier-mediated transporters such as glucose
and amino acid carriers and receptor-mediated
transcytosis-mediating cell structures/receptors at the vascular
endothelium of the BBB, thus enabling trans-BBB transport of the
DVD-binding protein. Structures at the BBB enabling such transport
include but are not limited to the insulin receptor, transferrin
receptor, LRP and RAGE. In addition, strategies enable the use of
DVD-binding proteins also as shuttles to transport potential drugs
into the CNS including low molecular weight drugs, nanoparticles
and nucleic acids (Coloma M J, et al. (2000) Pharm Res.
17(3):266-74; Boado R J, et al. (2007) Bioconjug. Chem.
18(2):447-55).
VI.H. Oncological Disorders
[0440] Monoclonal antibody therapy has emerged as an important
therapeutic modality for cancer (von Mehren et al., Annu. Rev.
Med., 54: 343-69 (2003)). Antibodies may exert antitumor effects by
inducing apoptosis, redirected cytotoxicity, interfering with
ligand-receptor interactions, or preventing the expression of
proteins that are critical to the neoplastic phenotype. In
addition, antibodies can target components of the tumor
microenvironment, perturbing vital structures such as the formation
of tumor-associated vasculature. Antibodies can also target
receptors whose ligands are growth factors, such as the epidermal
growth factor receptor. The antibody thus inhibits natural ligands
that stimulate cell growth from binding to targeted tumor cells.
Alternatively, antibodies may induce an anti-idiotype network,
complement-mediated cytotoxicity, or antibody-dependent cellular
cytotoxicity (ADCC). The use of dual-specific antibody that targets
two separate tumor mediators will likely give additional benefit
compared to a mono-specific therapy.
[0441] Another embodiment provides a DVD-binding protein that binds
human sclerostin may also be capable of binding another target
involved in oncological diseases including, but not limited to:
IGFR, IGF, VGFR1, PDGFRb, PDGFRa, IGF1,2, ERB3, CDCP, 1BSG2, ErbB3,
CD52, CD20, CD19, CD3, CD4, CD8, BMP6, IL12A, IL1A, IL1B, IL2,
IL24, INHA, TNF, TNFSF10, BMP6, EGF, FGF1, FGF10, FGF11, FGF12,
FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF2, FGF20, FGF21,
FGF22, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, GRP, IGF1,
IGF2, IL12A, IL1A, IL1B, IL2, INHA, TGFA, TGFB1, TGFB2, TGFB3,
VEGF, CDK2, FGF10, FGF18, FGF2, FGF4, FGF7, IGF1R, IL2, BCL2,
CD164, CDKN1A, CDKN1B, CDKN1C, CDKN2A, CDKN2B, CDKN2C, CDKN3,
GNRH1, IGFBP6, IL1A, IL1B, ODZ1, PAWR, PLG, TGFB1I1, AR, BRCA1,
CDK3, CDK4, CDK5, CDK6, CDK7, CDK9, E2F1, EGFR, ENO1, ERBB2, ESR1,
ESR2, IGFBP3, IGFBP6, IL2, INSL4, MYC, NOX5, NR6A1, PAP, PCNA,
PRKCQ, PRKD1, PRL, TP53, FGF22, FGF23, FGF9, IGFBP3, IL2, INHA,
KLK6, TP53, CHGB, GNRH1, IGF1, IGF2, INHA, INSL3, INSL4, PRL, KLK6,
SHBG, NR1D1, NR1H3, NR1I3, NR2F6, NR4A3, ESR1, ESR2, NR0B1, NR0B2,
NR1D2, NR1H2, NR1H4, NR1I2, NR2C1, NR2C2, NR2E1, NR2E3, NR2F1,
NR2F2, NR3C1, NR3C2, NR4A1, NR4A2, NR5A1, NR5A2, NR6A1, PGR, RARB,
FGF1, FGF2, FGF6, KLK3, KRT1, APOC1, BRCA1, CHGA, CHGB, CLU,
COL1A1, COL6A1, EGF, ERBB2, ERK8, FGF1, FGF10, FGF11, FGF13, FGF14,
FGF16, FGF17, FGF18, FGF2, FGF20, FGF21, FGF22, FGF23, FGF3, FGF4,
FGF5, FGF6, FGF7, FGF8, FGF9, GNRH1, IGF1, IGF2, IGFBP3, IGFBP6,
IL12A, IL1A, IL1B, IL2, IL24, INHA, INSL3, INSL4, KLK10, KLK12,
KLK13, KLK14, KLK15, KLK3, KLK4, KLK5, KLK6, KLK9, MMP2, MMP9,
MSMB, NTN4, ODZ1, PAP, PLAU, PRL, PSAP, SERPINA3, SHBG, TGFA,
TIMP3, CD44, CDH1, CDH10, CDH19, CDH20, CDH7, CDH9, CDH1, CDH10,
CDH13, CDH18, CDH19, CDH20, CDH7, CDH8, CDH9, ROBO2, CD44, ILK,
ITGA1, APC, CD164, COL6A1, MTSS1, PAP, TGFB1I1, AGR2, AIG1, AKAP1,
AKAP2, CANT1, CAV1, CDH12, CLDN3, CLN3, CYB5, CYC1, DAB2IP, DES,
DNCL1, ELAC2, ENO2, ENO3, FASN, FLJ12584, FLJ25530, GAGEB1, GAGEC1,
GGT1, GSTP1, HIP1, HUMCYT2A, IL29, K6HF, KAI1, KRT2A, MIB1, PART1,
PATE, PCA3, PIAS2, PIK3CG, PPID, PR1, PSCA, SLC2A2, SLC33A1,
SLC43A1, STEAP, STEAP2, TPM1, TPM2, TRPC6, ANGPT1, ANGPT2, ANPEP,
ECGF1, EREG, FGF1, FGF2, FIGF, FLT1, JAG1, KDR, LAMA5, NRP1, NRP2,
PGF, PLXDC1, STAB1, VEGF, VEGFC, ANGPTL3, BAI1, COL4A3, IL8, LAMA5,
NRP1, NRP2, STAB1, ANGPTL4, PECAM1, PF4, PROK2, SERPINF1, TNFAIP2,
CCL11, CCL2, CXCL1, CXCL10, CXCL3, CXCL5, CXCL6, CXCL9, IFNA1,
IFNB1, IFNG, IL1B, IL6, MDK, EDG1, EFNA1, EFNA3, EFNB2, EGF, EPHB4,
FGFR3, HGF, IGF1, ITGB3, PDGFA, TEK, TGFA, TGFB1, TGFB2, TGFBR1,
CCL2, CDH5, COL18A1, EDG1, ENG, ITGAV, ITGB3, THBS1, THBS2, BAD,
BAG1, BCL2, CCNA1, CCNA2, CCND1, CCNE1, CCNE2, CDH1 (E-cadherin),
CDKN1B (p27Kip1), CDKN2A (p16INK4a), COL6A1, CTNNB1 (b-catenin),
CTSB (cathepsin B), ERBB2 (Her-2), ESR1, ESR2, F3 (TF), FOSL1
(FRA-1), GATA3, GSN (Gelsolin), IGFBP2, IL2RA, IL6, IL6R, IL6ST
(glycoprotein 130), ITGA6 (a6 integrin), JUN, KLK5, KRT19, MAP2K7
(c-Jun), MKI67 (Ki-67), NGFB (NGF), NGFR, NME1 (NM23A), PGR, PLAU
(uPA), PTEN, SERPINB5 (maspin), SERPINE1 (PAI-1), TGFA, THBS1
(thrombospondin-1), TIE (Tie-1), TNFRSF6 (Fas), TNFSF6 (FasL),
TOP2A (topoisomerase Iia), TP53, AZGP1 (zinc-a-glycoprotein), BPAG1
(plectin), CDKN1A (p21Wap1/Cip1), CLDN7 (claudin-7), CLU
(clusterin), ERBB2 (Her-2), FGF1, FLRT1 (fibronectin), GABRP
(GABAa), GNAS1, ID2, ITGA6 (a6 integrin), ITGB4 (b 4 integrin),
KLF5 (GC Box BP), KRT19 (Keratin 19), KRTHB6 (hair-specific type II
keratin), MACMARCKS, MT3 (metallothionectin-III), MUC1 (mucin),
PTGS2 (COX-2), RAC2 (p21Rac2), S100A2, SCGB1D2 (lipophilin B),
SCGB2A1 (mammaglobin 2), SCGB2A2 (mammaglobin 1), SPRR1B (Spr1),
THBS1, THBS2, THBS4, and TNFAIP2 (B94), RON, c-Met, CD64, DLL4,
PLGF, CTLA4, phosphatidylserine, ROBO4, CD80, CD22, CD40, CD23,
CD28, CD55, CD38, CD70, CD74, CD30, CD138, CD56, CD33, CD2, CD137,
DR4, DR5, RANKL, VEGFR2, PDGFR, VEGFR1, MTSP1, MSP, EPHB2, EPHA1,
EPHA2, EpCAM, PGE2, NKG2D, LPA, SIP, APRIL, BCMA, MAPG, FLT3, PDGFR
alpha, PDGFR beta, ROR1, PSMA, PSCA, SCD1, and CD59.
VII. Pharmaceutical Composition
[0442] A pharmaceutical compositions comprising an antibody, or
antigen-binding portion thereof, of the invention and a
pharmaceutically acceptable carrier is provided. The pharmaceutical
compositions comprising antibodies are provided and are for use in,
but not limited to, diagnosing, detecting, or monitoring a
disorder, in preventing, treating, managing, or ameliorating of a
disorder or one or more symptoms thereof, and/or in research. In a
specific embodiment, a composition comprises one or more antibodies
is provided. In another provided embodiment, the pharmaceutical
composition comprises one or more antibodies and one or more
prophylactic or therapeutic agents other than antibodies treating a
disorder in which SOST activity is detrimental. In an embodiment,
the prophylactic or therapeutic agents are known to be useful for
or having been or currently being used in the prevention,
treatment, management, or amelioration of a disorder or one or more
symptoms thereof. In accordance with these embodiments, the
composition may further comprise of a carrier, diluent or
excipient.
[0443] The antibodies and antibody portions can be incorporated
into pharmaceutical compositions suitable for administration to a
subject. Typically, the pharmaceutical composition comprises an
antibody or antibody portion and a pharmaceutically acceptable
carrier. The term "pharmaceutically acceptable carrier" includes
any and all solvents, dispersion media, coatings, antibacterial and
antifungal agents, isotonic and absorption delaying agents, and the
like that are physiologically compatible. Examples of
pharmaceutically acceptable carriers include one or more of water,
saline, phosphate buffered saline, dextrose, glycerol, ethanol and
the like, as well as combinations thereof. In many cases, it will
be preferable to include isotonic agents, for example, sugars,
polyalcohols such as mannitol, sorbitol, or sodium chloride in the
composition. Pharmaceutically acceptable carriers may further
comprise minor amounts of auxiliary substances such as wetting or
emulsifying agents, preservatives or buffers, which enhance the
shelf life or effectiveness of the antibody or antibody
portion.
[0444] Various delivery systems are known and can be used to
administer one or more antibodies or the combination of one or more
antibodies and a prophylactic agent or therapeutic agent useful for
preventing, managing, treating, or ameliorating a disorder or one
or more symptoms thereof, e.g., encapsulation in liposomes,
microparticles, microcapsules, recombinant cells capable of
expressing the antibody or antibody fragment, receptor-mediated
endocytosis (see, e. g., Wu and Wu, J. Biol. Chem., 262: 4429-4432
(1987)), construction of a nucleic acid as part of a retroviral or
other vector. Methods of administering a prophylactic or
therapeutic agent are provided and include, but are not limited to,
parenteral administration (e.g., intradermal, intramuscular,
intraperitoneal, intravenous and subcutaneous), epidural
administration, intratumoral administration, and mucosal
administration (e.g., intranasal and oral routes). In addition,
pulmonary administration can be employed, e.g., by use of an
inhaler or nebulizer, and formulation with an aerosolizing agent.
See, e.g., U.S. Pat. Nos. 6,019,968; 5,985,320; 5,985,309; 5,934,
272; 5,874,064; 5,855,913; 5,290,540; and 4,880,078; and PCT
Publication Nos. WO 92/19244, WO 97/32572, WO 97/44013, WO
98/31346, and WO 99/66903. One embodiment provides an antibody or
antibody portion, combination therapy, or a composition
administered using Alkermes AIR.RTM. pulmonary drug delivery
technology (Alkermes, Inc., Cambridge, Mass., US). In a provided
specific embodiment, prophylactic or therapeutic agents are
administered intramuscularly, intravenously, intratumorally,
orally, intranasally, pulmonary, or subcutaneously. The
prophylactic or therapeutic agents may be administered by any
convenient route, for example by infusion or bolus injection, by
absorption through epithelial or mucocutaneous linings (e.g., oral
mucosa, rectal, and intestinal mucosa, etc.) and may be
administered together with other biologically active agents
Administration can be systemic or local.
[0445] In an embodiment, specific binding of antibody-coupled
carbon nanotubes (CNTs) to tumor cells in vitro, followed by their
highly specific ablation with near-infrared (NIR) light can be used
to target tumor cells. For example, biotinylated polar lipids can
be used to prepare stable, biocompatible, noncytotoxic CNT
dispersions that are then attached to one or two different
neutralite avidin-derivatized DVD-binding proteins directed against
one or more tumor antigens (e.g., CD22) (Chakravarty, P. et al.
(2008) Proc. Natl. Acad. Sci. USA, 105:8697-8702).
[0446] A specific embodiment provides it may be desirable to
administer the prophylactic or therapeutic agents locally to the
area in need of treatment; this may be achieved by, for example,
and not by way of limitation, local infusion, by injection, or by
means of an implant, said implant being of a porous or non-porous
material, including membranes and matrices, such as sialastic
membranes, polymers, fibrous matrices (e.g., Tissuel.RTM.), or
collagen matrices. One embodiment provides an effective amount of
one or more antibody antagonists is administered locally to the
affected area to a subject to prevent, treat, manage, and/or
ameliorate a disorder or a symptom thereof. Another embodiment
provides an effective amount of one or more antibodies administered
locally to the affected area of a subject in combination with an
effective amount of one or more therapies (e.g., one or more
prophylactic or therapeutic agents) other than an antibody to
prevent, treat, manage, and/or ameliorate a disorder or one or more
symptoms thereof.
[0447] In another embodiment, the prophylactic or therapeutic agent
can be delivered in a controlled release or sustained release
system. In one embodiment, a pump may be used to achieve controlled
or sustained release (see Langer, supra; Sefton, 1987, CRC Crit.
Ref. Biomed. Eng., 14: 20; Buchwald et al., 1980, Surgery, 88: 507;
Saudek et al., 1989, N. Engl. J. Med., 321: 574). Another
embodiment provides polymeric materials can be used to achieve
controlled or sustained release of the therapies (see, e.g.,
Goodson, J. M., Chapter 6, In Medical Applications of Controlled
Release, Vol. II, Applications and Evaluation, (Langer and Wise,
eds.) (CRC Press, Inc., Boca Raton, 1984), pp. 115-138; Ranger and
Peppas, 1983, J. Macromol. Sci. Rev. Macromol. Chem. 23:61; see
also Levy et al., 1985, Science 228:190; During et al., 1989, Ann.
Neurol. 25:351; Howard et al., 1989, J. Neurosurg. 7 1:105); U.S.
Pat. No. 5,679,377; U.S. Pat. No. 5,916,597; U.S. Pat. No.
5,912,015; U.S. Pat. No. 5,989,463; U.S. Pat. No. 5,128,326; PCT
Publication No. WO 99/15154; and PCT Publication No. WO 99/20253.
Examples of polymers used in sustained release formulations
include, but are not limited to, poly(2-hydroxy ethyl
methacrylate), poly(methyl methacrylate), poly(acrylic acid),
poly(ethylene-co-vinyl acetate), poly(methacrylic acid),
polyglycolides (PLG), polyanhydrides, poly(N-vinyl pyrrolidone),
poly(vinyl alcohol), polyacrylamide, poly(ethylene glycol),
polylactides (PLA), poly(lactide-co-glycolides) (PLGA), and
polyorthoesters. In an embodiment, the polymer used in a sustained
release formulation is inert, free of leachable impurities, stable
on storage, sterile, and biodegradable. In yet another embodiment,
a controlled or sustained release system can be placed in proximity
of the prophylactic or therapeutic target, thus requiring only a
fraction of the systemic dose (see, e.g., Goodson, in Medical
Applications of Controlled Release, supra, vol. 2, pp. 115-138
(1984)).
[0448] Controlled release systems are discussed in the review by
Langer (1990, Science 249:1527-1533). Any technique known to one of
skill in the art are provided and can be used to produce sustained
release formulations comprising one or more therapeutic agents.
See, e.g., U.S. Pat. No. 4,526,938, PCT publication WO 91/05548,
PCT publication WO 96/20698, Ning et al., 1996, "Intratumoral
Radioimmunotherapy of a Human Colon Cancer Xenograft Using a
Sustained-Release Gel," Radiotherapy & Oncology, 39: 179-189,
Song et al., 1995, "Antibody Mediated Lung Targeting of
Long-Circulating Emulsions," PDA Journal of Pharmaceutical Science
& Technology, 50: 372-397, Cleek et al., 1997, "Biodegradable
Polymeric Carriers for a bFGF Antibody for Cardiovascular
Application," Pro. Int'l. Symp. Control. Rd. Bioact. Mater. 24:
853-854, and Lam et al., 1997, "Microencapsulation of Recombinant
Humanized Monoclonal Antibody for Local Delivery," Proc. Int'l.
Symp. Control Rd. Bioact. Mater. 24:759-760.
[0449] A specific embodiment provides where the composition is a
nucleic acid encoding a prophylactic or therapeutic agent, the
nucleic acid can be administered in vivo to promote expression of
its encoded prophylactic or therapeutic agent, by constructing it
as part of an appropriate nucleic acid expression vector and
administering it so that it becomes intracellular, e.g., by use of
a retroviral vector (see U.S. Pat. No. 4,980,286), or by direct
injection, or by use of microparticle bombardment (e.g., a gene
gun; Biolistic, Dupont), or coating with lipids or cell-surface
receptors or transfecting agents, or by administering it in linkage
to a homeobox-like peptide which is known to enter the nucleus
(see, e.g., Joliot et al., 1991, Proc. Natl. Acad. Sci. USA, 88:
1864-1868). Alternatively, a nucleic acid can be introduced
intracellularly and incorporated within host cell DNA for
expression by homologous recombination.
[0450] A pharmaceutical composition formulated to be compatible
with its intended route of administration is provided. Examples of
routes of administration include, but are not limited to,
parenteral, e.g., intravenous, intradermal, subcutaneous, oral,
intranasal (e.g., inhalation), transdermal (e.g., topical),
transmucosal, and rectal administration. In a specific embodiment,
the composition is formulated in accordance with routine procedures
as a pharmaceutical composition adapted for intravenous,
subcutaneous, intramuscular, oral, intranasal, or topical
administration to human beings. Typically, compositions for
intravenous administration are solutions in sterile isotonic
aqueous buffer. Where necessary, the composition may also include a
solubilizing agent and a local anesthetic, such as lignocamne, to
ease pain at the site of the injection.
[0451] If the compositions are to be administered topically, the
compositions can be formulated in the form of an ointment, cream,
transdermal patch, lotion, gel, shampoo, spray, aerosol, solution,
emulsion, or other form well-known to one of skill in the art. See,
e.g., Remington's Pharmaceutical Sciences and Introduction to
Pharmaceutical Dosage Forms, 19th ed., Mack Pub. Co., Easton, Pa.
(1995). For non-sprayable topical dosage forms, viscous to
semi-solid or solid forms comprising a carrier or one or more
excipients compatible with topical application and having a dynamic
viscosity, e.g., greater than water are typically employed.
Suitable formulations include, without limitation, solutions,
suspensions, emulsions, creams, ointments, powders, liniments,
salves, and the like, which are, if desired, sterilized or mixed
with auxiliary agents (e.g., preservatives, stabilizers, wetting
agents, buffers, or salts) for influencing various properties, such
as, for example, osmotic pressure. Other suitable topical dosage
forms include sprayable aerosol preparations wherein the active
ingredient, e.g., in combination with a solid or liquid inert
carrier, is packaged in a mixture with a pressurized volatile
(e.g., a gaseous propellant, such as FREON.RTM.) or in a squeeze
bottle. Moisturizers or humectants can also be added to
pharmaceutical compositions and dosage forms if desired. Examples
of such additional ingredients are well known in the art.
[0452] If the method comprises intranasal administration of a
composition, the composition can be formulated in an aerosol form,
spray, mist or in the form of drops. In particular, prophylactic or
therapeutic agents are provided and can be conveniently delivered
in the form of an aerosol spray presentation from pressurized packs
or a nebulizer, with the use of a suitable propellant (e.g.,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas).
In the case of a pressurized aerosol the dosage unit may be
determined by providing a valve to deliver a metered amount.
Capsules and cartridges (composed of, e.g., gelatin) for use in an
inhaler or insufflator may be formulated containing a powder mix of
the compound and a suitable powder base such as lactose or
starch.
[0453] If the method comprises oral administration, compositions
can be formulated orally in the form of tablets, capsules, cachets,
gelcaps, solutions, suspensions, and the like. Tablets or capsules
can be prepared by conventional means with pharmaceutically
acceptable excipients such as binding agents (e.g., pregelatinised
maize starch, polyvinylpyrrolidone, or hydroxypropyl
methylcellulose); fillers (e.g., lactose, microcrystalline
cellulose, or calcium hydrogen phosphate); lubricants (e.g.,
magnesium stearate, talc, or silica); disintegrants (e.g., potato
starch or sodium starch glycolate); or wetting agents (e.g., sodium
lauryl sulphate). The tablets may be coated by methods well-known
in the art. Liquid preparations for oral administration may take
the form of, but not limited to, solutions, syrups or suspensions,
or they may be presented as a dry product for constitution with
water or other suitable vehicle before use. Such liquid
preparations may be prepared by conventional means with
pharmaceutically acceptable additives such as suspending agents
(e.g., sorbitol syrup, cellulose derivatives, or hydrogenated
edible fats); emulsifying agents (e.g., lecithin or acacia);
non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol,
or fractionated vegetable oils); and preservatives (e.g., methyl or
propyl-p-hydroxybenzoates or sorbic acid). The preparations may
also contain buffer salts, flavoring, coloring, and sweetening
agents as appropriate. Preparations for oral administration may be
suitably formulated for slow release, controlled release, or
sustained release of a prophylactic or therapeutic agent(s).
[0454] The provided method may comprise pulmonary administration,
e.g., by use of an inhaler or nebulizer, of a composition
formulated with an aerosolizing agent. See, e.g., U.S. Pat. Nos.
6,019,968; 5,985,320; 5,985,309; 5,934,272; 5,874,064; 5,855,913;
5,290,540; and 4,880,078; and PCT Publication Nos. WO 92/19244, WO
97/32572, WO 97/44013, WO 98/31346, and WO 99/66903. A specific
embodiment provides an antibody, combination therapy, and/or
composition administered using Alkermes AIR.RTM. pulmonary drug
delivery technology (Alkermes, Inc., Cambridge, Mass.).
[0455] The provided method may comprise administration of a
composition formulated for parenteral administration by injection
(e. g., by bolus injection or continuous infusion). Formulations
for injection may be presented in unit dosage form (e.g., in
ampoules or in multi-dose containers) with an added preservative.
The compositions may take such forms as suspensions, solutions or
emulsions in oily or aqueous vehicles, and may contain formulatory
agents such as suspending, stabilizing and/or dispersing agents.
Alternatively, the active ingredient may be in powder form for
constitution with a suitable vehicle (e.g., sterile pyrogen-free
water) before use.
[0456] The provided methods may additionally comprise of
administration of compositions formulated as depot preparations.
Such long acting formulations may be administered by implantation
(e.g., subcutaneously or intramuscularly) or by intramuscular
injection. Thus, for example, the compositions may be formulated
with suitable polymeric or hydrophobic materials (e.g., as an
emulsion in an acceptable oil) or ion exchange resins, or as
sparingly soluble derivatives (e.g., as a sparingly soluble
salt).
[0457] The provided methods encompass administration of
compositions formulated as neutral or salt forms. Pharmaceutically
acceptable salts include those formed with anions such as those
derived from hydrochloric, phosphoric, acetic, oxalic, tartaric
acids, etc., and those formed with cations such as those derived
from sodium, potassium, ammonium, calcium, ferric hydroxides,
isopropylamine, triethylamine, 2-ethylamino ethanol, histidine,
procaine, etc.
[0458] Generally, the ingredients of compositions are supplied
either separately or mixed together in unit dosage form, for
example, as a dry lyophilized powder or water free concentrate in a
hermetically sealed container such as an ampoule or sachet
indicating the quantity of active agent. Where the mode of
administration is infusion, composition can be dispensed with an
infusion bottle containing sterile pharmaceutical grade water or
saline. Where the mode of administration is by injection, an
ampoule of sterile water for injection or saline can be provided so
that the ingredients may be mixed prior to administration.
[0459] In particular, it is also provided that one or more of the
prophylactic or therapeutic agents, or pharmaceutical compositions
is packaged in a hermetically sealed container such as an ampoule
or sachette indicating the quantity of the agent. One embodiment
provides one or more of the prophylactic or therapeutic agents, or
pharmaceutical compositions is supplied as a dry sterilized
lyophilized powder or water free concentrate in a hermetically
sealed container and can be reconstituted (e.g., with water or
saline) to the appropriate concentration for administration to a
subject. An embodiment provides one or more of the prophylactic or
therapeutic agents or pharmaceutical compositions is supplied as a
dry sterile lyophilized powder in a hermetically sealed container
at a unit dosage of at least 5 mg, e.g., at least 10 mg, at least
15 mg, at least 25 mg, at least 35 mg, at least 45 mg, at least 50
mg, at least 75 mg, or at least 100 mg. The lyophilized
prophylactic or therapeutic agents or pharmaceutical compositions
should be stored at between 2.degree. C. and 8.degree. C. in its
original container and the prophylactic or therapeutic agents, or
pharmaceutical compositions should be administered within 1 week,
e.g., within 5 days, within 72 hours, within 48 hours, within 24
hours, within 12 hours, within 6 hours, within 5 hours, within 3
hours, or within 1 hour after being reconstituted. An alternative
embodiment provides one or more of the prophylactic or therapeutic
agents or pharmaceutical compositions is supplied in liquid form in
a hermetically sealed container indicating the quantity and
concentration of the agent. In an embodiment, the liquid form of
the administered composition is supplied in a hermetically sealed
container at least 0.25 mg/ml, e.g., at least 0.5 mg/ml, at least 1
mg/ml, at least 2.5 mg/ml, at least 5 mg/ml, at least 8 mg/ml, at
least 10 mg/ml, at least 15 mg/kg, at least 25 mg/ml, at least 50
mg/ml, at least 75 mg/ml or at least 100 mg/ml. The liquid form
should be stored at between 2.degree. C. and 8.degree. C. in its
original container.
[0460] Antibodies and antibody portions that can be incorporated
into a pharmaceutical composition suitable for parenteral
administration are provided. In an embodiment, the antibody or
antibody-portions will be prepared as an injectable solution
containing 0.1-250 mg/ml antibody. The injectable solution can be
composed of either a liquid or lyophilized dosage form in a flint
or amber vial, ampoule or pre-filled syringe. The buffer can be
L-histidine (1-50 mM), optimally 5-10 mM, at pH 5.0 to 7.0
(optimally pH 6.0). Other suitable buffers include but are not
limited to, sodium succinate, sodium citrate, sodium phosphate or
potassium phosphate. Sodium chloride can be used to modify the
toxicity of the solution at a concentration of 0-300 mM (optimally
150 mM for a liquid dosage form). Cryoprotectants can be included
for a lyophilized dosage form, principally 0-10% sucrose (optimally
0.5-1.0%). Other suitable cryoprotectants include trehalose and
lactose. Bulking agents can be included for a lyophilized dosage
form, principally 1-10% mannitol (optimally 2-4%). Stabilizers can
be used in both liquid and lyophilized dosage forms, principally
1-50 mM L-Methionine (optimally 5-10 mM). Other suitable bulking
agents include glycine, arginine, can be included as 0-0.05%
polysorbate-80 (optimally 0.005-0.01%). Additional surfactants
include but are not limited to polysorbate 20 and BRIJ surfactants.
The pharmaceutical composition comprising an antibody or antibody
portion prepared as an injectable solution for parenteral
administration is provided, and can further comprise an agent
useful as an adjuvant, such as those used to increase the
absorption, or dispersion of a therapeutic protein (e.g.,
antibody). A particularly useful adjuvant is hyaluronidase (such as
Hylenex.RTM. recombinant human hyaluronidase). Addition of
hyaluronidase in the injectable solution improves human
bioavailability following parenteral administration, particularly
subcutaneous administration. It also allows for greater injection
site volumes (i.e. greater than 1 ml) with less pain and
discomfort, and minimum incidence of injection site reactions (see,
WO 2004/078140 and US patent application publication No.
2006104968).
[0461] The compositions provided may be in a variety of forms.
These include, for example, liquid, semi-solid and solid dosage
forms, such as liquid solutions (e.g., injectable and infusible
solutions), dispersions or suspensions, tablets, pills, powders,
liposomes and suppositories. An exemplary form depends on the
intended mode of administration and therapeutic application.
Typical compositions are in the form of injectable or infusible
solutions, such as compositions similar to those used for passive
immunization of humans with other antibodies. An exemplary mode of
administration is parenteral (e.g., intravenous, subcutaneous,
intraperitoneal, intramuscular). In an embodiment, the antibody is
administered by intravenous infusion or injection. In another
embodiment, the antibody is administered by intramuscular or
subcutaneous injection.
[0462] Therapeutic compositions typically must be sterile and
stable under the conditions of manufacture and storage. The
composition can be formulated as a solution, microemulsion,
dispersion, liposome, or other ordered structure suitable to high
drug concentration. Sterile injectable solutions can be prepared by
incorporating the active compound (i.e., antibody or antibody
portion) in the required amount in an appropriate solvent with one
or a combination of ingredients enumerated above, as required,
followed by filtered sterilization. Generally, dispersions are
prepared by incorporating the active compound into a sterile
vehicle that contains a basic dispersion medium and the required
other ingredients from those enumerated above. In the case of
sterile, lyophilized powders for the preparation of sterile
injectable solutions, exemplary methods of preparation are vacuum
drying and spray-drying that yields a powder of the active
ingredient plus any additional desired ingredient from a previously
sterile-filtered solution thereof. The proper fluidity of a
solution can be maintained, for example, by the use of a coating
such as lecithin, by the maintenance of the required particle size
in the case of dispersion and by the use of surfactants. Prolonged
absorption of injectable compositions can be brought about by
including, in the composition, an agent that delays absorption, for
example, monostearate salts and gelatin.
[0463] Antibodies and antibody-portions administered by a variety
of methods known in the art are provided, although for many
therapeutic applications, an exemplary route/mode of administration
is subcutaneous injection, intravenous injection or infusion. As
will be appreciated by the skilled artisan, the route and/or mode
of administration will vary depending upon the desired results. In
certain embodiments, the active compound may be prepared with a
carrier that will protect the compound against rapid release, such
as a controlled release formulation, including implants,
transdermal patches, and microencapsulated delivery systems.
Biodegradable, biocompatible polymers can be used, such as ethylene
vinyl acetate, polyanhydrides, polyglycolic acid, collagen,
polyorthoesters, and polylactic acid. Many methods for the
preparation of such formulations are patented or generally known to
those skilled in the art. See, e.g., Sustained and Controlled
Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker,
Inc., New York, 1978.
[0464] In certain provided embodiments, an antibody or antibody
portion may be orally administered, for example, with an inert
diluent or an assimilable edible carrier. The compound (and other
ingredients, if desired) may also be enclosed in a hard or soft
shell gelatin capsule, compressed into tablets, or incorporated
directly into the subject's diet. For oral therapeutic
administration, the compounds may be incorporated with excipients
and used in the form of ingestible tablets, buccal tablets,
troches, capsules, elixirs, suspensions, syrups, wafers, and the
like. To administer a compound by other than parenteral
administration, it may be necessary to coat the compound with, or
co-administer the compound with, a material to prevent its
inactivation.
[0465] Supplementary active compounds can also be incorporated into
the compositions. Embodiments provide an antibody or antibody
portion coformulated with and/or coadministered with one or more
additional therapeutic agents that are useful for treating
disorders in which SOST activity is detrimental. For example, an
anti-hSOST antibody or antibody portion may be coformulated and/or
coadministered with one or more additional antibodies that bind
other targets (e.g., antibodies that bind other cytokines or that
bind cell surface molecules). Furthermore, one or more antibodies
may be used in combination with two or more of the foregoing
therapeutic agents. Such combination therapies may advantageously
utilize lower dosages of the administered therapeutic agents, thus
avoiding possible toxicities or complications associated with the
various monotherapies.
[0466] In certain embodiments, an antibody to SOST or fragment
thereof is linked to a half-life extending vehicle known in the
art. Such vehicles include, but are not limited to, the Fc domain,
polyethylene glycol, and dextran. Such vehicles are described,
e.g., in U.S. Ser. No. 09/428,082 and published PCT Publication No.
WO 99/25044.
[0467] In a specific embodiment, nucleic acid sequences comprising
nucleotide sequences encoding an antibody of the invention or
another prophylactic or therapeutic agent of the invention are
administered to treat, prevent, manage, or ameliorate a disorder or
one or more symptoms thereof by way of gene therapy are provided.
Gene therapy refers to therapy performed by the administration to a
subject of an expressed or expressible nucleic acid. In this
provided embodiment, the nucleic acids produce their encoded
antibody or prophylactic or therapeutic agent that mediates a
prophylactic or therapeutic effect.
[0468] Any of the methods for gene therapy available in the art are
provided. For general reviews of the methods of gene therapy, see
Goldspiel et al., 1993, Clinical Pharmacy, 12: 488-505; Wu et al.,
"Delivery systems for gene therapy," Biotherapy, 3: 87-95 (1991);
Tolstoshev, 1993, Ann. Rev. Pharmacol. Toxicol., 32: 573-596;
Mulligan, Science, 260: 926-932 (1993); and Morgan and Anderson,
"Human Gene Therapy," Ann. Rev. Biochem., 62:191-217 (1993);
Robinson, C., Trends Biotechnol., 11:155 (1993). Methods commonly
known in the art of recombinant DNA technology which can be used
are described in Ausubel et al. (eds.), Current Protocols in
Molecular Biology, John Wiley &Sons, NY (1993); and Kriegler,
Gene Transfer and Expression, A Laboratory Manual, Stockton Press,
NY (1990). Detailed description of various methods of gene therapy
are disclosed in US application publication No. US
2005/0042664.
[0469] Sclerostin plays a critical role in the pathology associated
with a variety of diseases involving immune and inflammatory
elements. These diseases include, but are not limited to, Acquired
Immunodeficiency Disease Syndrome; Acquired Immunodeficiency
Related Diseases; acquired pernicious anaemia; Acute coronary
syndromes; acute and chronic pain (different forms of pain); Acute
Idiopathic Polyneuritis; acute immune disease associated with organ
transplantation; acute or chronic immune disease associated with
organ transplantation; Acute Inflammatory Demyelinating
Polyradiculoneuropathy; Acute ischemia; acute liver disease; acute
rheumatic fever; acute transverse myelitis; Addison's disease;
adult (acute) respiratory distress syndrome; Adult Still's Disease;
alcoholic cirrhosis; alcohol-induced liver injury; allergic
diseases; allergy; alopecia; Alopecia areata; Alzheimer's disease;
Anaphylaxis; ankylosing spondylitis; ankylosing spondylitis
associated lung disease; Anti-Phospholipid Antibody Syndrome;
Aplastic anemia; Arteriosclerosis; arthropathy; asthma;
atheromatous disease/arteriosclerosis; atherosclerosis; atopic
allergy; Atopic eczema; Atopic dermatitis; atrophic autoimmune
hypothyroidism; autoimmune bullous disease; Autoimmune dermatitis;
autoimmune diabetes; Autoimmune disorder associated with
Streptococcus infection; Autoimmune Enteropathy; autoimmune
haemolytic anaemia; autoimmune hepatitis; Autoimmune hearing loss;
Autoimmune Lymphoproliferative Syndrome (ALPS); autoimmune mediated
hypoglycaemia; Autoimmune myocarditis; autoimmune neutropenia;
Autoimmune premature ovarian failure; autoimmune thrombocytopenia
(AITP); autoimmune thyroid disease; autoimmune uveitis;
bronchiolitis obliterans; Behcet's disease; Blepharitis;
Bronchiectasis; Bullous pemphigoid; cachexia; Cardiovascular
Disease; Catastrophic Antiphospholipid Syndrome; Celiac Disease;
Cervical Spondylosis; chlamydia; choleosatatis; chronic active
hepatitis; chronic eosinophilic pneumonia; chronic fatigue
syndrome; chronic immune disease associated with organ
transplantation; Chronic ischemia; chronic liver diseases; chronic
mucocutaneous candidiasis; Cicatricial pemphigoid; Clinically
isolated Syndrome (CIS) with Risk for Multiple Sclerosis; common
varied immunodeficiency (common variable hypogammaglobulinaemia);
connective tissue disease associated interstitial lung disease;
Conjunctivitis; Coombs positive haemolytic anaemia; Childhood Onset
Psychiatric Disorder; Chronic obstructive pulmonary disease (COPD);
Crohn's disease; cryptogenic autoimmune hepatitis; cryptogenic
fibrosing alveolitis; Dacryocystitis; depression; dermatitis
scleroderma; dermatomyositis; dermatomyositis/polymyositis
associated lung disease; Diabetic retinopathy; Diabetes mellitus;
dilated cardiomyopathy; discoid lupus erythematosus; Disk
herniation; Disk prolapse; disseminated intravascular coagulation;
Drug-Induced hepatitis; drug-induced interstitial lung disease;
Drug induced immune hemolytic anemia; Endocarditis; Endometriosis;
endophthalmitis; enteropathic synovitis; Episcleritis; Erythema
multiforme; erythema multiforme major; female infertility;
fibrosis; fibrotic lung disease; Gestational pemphigoid; giant cell
arteritis (GCA); glomerulonephritides; goitrous autoimmune
hypothyroidism (Hashimoto's disease); Goodpasture's syndrome; gouty
arthritis; graft versus host disease (GVHD); Grave's disease; group
B streptococci (GBS) infection; Guillain-Barre Syndrome (GBS);
haemosiderosis associated lung disease; Hay Fever; heart failure;
hemolytic anemia; Henoch-Schoenlein purpurea; Hepatitis B;
Hepatitis C; Hughes Syndrome; Huntington's chorea; hyperthyroidism;
hypoparathyroidism; idiopathic leucopaenia; idiopathic
thrombocytopaenia; Idiopathic Parkinson's Disease; idiopathic
interstitial pneumonia; idiosyncratic liver disease; IgE-mediated
Allergy; Immune hemolytic anemiae; Inclusion Body Myositis;
infectious diseases; Infectious ocular inflammatory disease;
inflammatory bowel disease; Inflammatory demyelinating disease;
Inflammatory heart disease; Inflammatory kidney disease; insulin
dependent diabetes mellitus; interstitial pneumonitis; IPF/UIP;
Iritis; juvenile chronic arthritis; juvenile pernicious anaemia;
Juvenile rheumatoid arthritis; Kawasaki's diseasee; Keratitis;
Keratojunctivitis sicca; Kussmaul disease or Kussmaul-Meier
Disease; Landry's Paralysis; Langerhan's Cell Histiocytosis; linear
IgA disease; Livedo reticularis; Lyme arthritis; lymphocytic
infiltrative lung disease; Macular Degeneration; male infertility
idiopathic or NOS; malignancies; microscopic vasculitis of the
kidneys; Microscopic Polyangiitis; mixed connective tissue disease
associated lung disease; Morbus Bechterev; Motor Neuron Disorders;
Mucous membrane pemphigoid; multiple sclerosis (all subtypes:
primary progressive, secondary progressive, relapsing remitting
etc.); Multiple Organ failure; myalgic encephalitis/Royal Free
Disease; Myasthenia Gravis; Myelodysplastic Syndrome; myocardial
infarction; Myocarditis; nephrotic syndrome; Nerve Root Disorders;
Neuropathy; Non-alcoholic Steatohepatitis; Non-A Non-B Hepatitis;
Optic Neuritis; organ transplant rejection; osteoarthritis;
Osteolysis; Ovarian cancer; ovarian failure; Pancreatitis;
Parasitic diseases; Parkinson's disease; Pauciarticular JRA;
pemphigoid; pemphigus foliaceus; pemphigus vulgaris; peripheral
artery occlusive disease (PAOD); peripheral vascular disease (PVD);
peripheral artery disease (PAD); phacogenic uveitis; Phlebitis;
Polyarteritis nodosa (or periarteritis nodosa); Polychondritis;
Polymyalgia Rheumatica; Poliosis; Polyarticular JRA; Polyendocrine
Deficiency Syndrome; Polymyositis; polyglandular deficiency type I
and polyglandular deficiency type II; polymyalgia rheumatica (PMR);
postinfectious interstitial lung disease; post-inflammatory
interstitial lung disease; Post-Pump Syndrome; premature ovarian
failure; primary biliary cirrhosis; primary myxoedema; primary
parkinsonism; primary sclerosing cholangitis; primary sclerosing
hepatitis; primary vasculitis; prostate and rectal cancer and
hematopoietic malignancies (leukemia and lymphoma); Prostatitis;
psoriasis; psoriasis type 1; psoriasis type 2; psoriatic arthritis;
psoriatic arthropathy; pulmonary hypertension secondary to
connective tissue disease; pulmonary manifestation of polyarteritis
nodosa; Pure red cell aplasia; Primary Adrenal Insufficiency;
radiation fibrosis; reactive arthritis; Reiter's disease; Recurrent
Neuromyelitis Optica; renal disease NOS; Restenosis; rheumatoid
arthritis; rheumatoid arthritis associated interstitial lung
disease; Rheumatic heart disease; SAPHO (synovitis, acne,
pustulosis, hyperostosis, and osteitis); sarcoidosis;
Schizophreniae; Schmidt's syndrome; Scleroderma; Secondary
Amyloidosis; Shock lung; Scleritis; Sciatica; Secondary Adrenal
Insufficiency; sepsis syndrome; septic arthritis; septic shock;
seronegative arthropathy; Silicone associated connective tissue
disease; Sjogren's disease associated lung disease; Sjorgren's
syndrome; Sneddon-Wilkinson Dermatosis; sperm autoimmunity;
spondyloarthropathy; spondylitis ankylosans; Stevens-Johnson
Syndrome (SJS); Still's disease; stroke; sympathetic ophthalmia;
Systemic inflammatory response syndrome; systemic lupus
erythematosus; systemic lupus erythematosus associated lung
disease; systemic sclerosis; systemic sclerosis associated
interstitial lung disease; Takayasu's disease/arteritis; Temporal
arteritis; Th2 Type and Th1 Type mediated diseases; thyroiditis;
toxic shock syndrome; toxoplasmic retinitis; toxic epidermal
necrolysis; Transverse myelitis; TRAPS (Tumor-necrosis factor
receptor type 1 (TNFR)-Associated Periodic Syndrome); type B
insulin resistance with acanthosis nigricans; Type 1 allergic
reaction; type-1 autoimmune hepatitis (classical autoimmune or
lupoid hepatitis); type-2 autoimmune hepatitis (anti-LKM antibody
hepatitis)e; Type II Diabetes; ulcerative colitic arthropathy;
ulcerative colitis; Urticaria; Usual interstitial pneumonia (UIP);
uveitis; vasculitic diffuse lung disease; Vasculitis; Vernal
conjunctivitis; viral retinitis; vitiligo; Vogt-Koyanagi-Harada
syndrome (VKH syndrome); Wegener's granulomatosis; Wet macular
degeneration; Wound healing; or yersinia and salmonella associated
arthropathy.
[0470] The antibodies and antibody portions can be used to treat
humans suffering from autoimmune diseases, in particular those
associated with inflammation, rheumatoid arthritis (RA),
osteoarthritis, psoriasis, multiple sclerosis (MS), and other
autoimmune diseases.
[0471] An antibody or antibody portion also can be administered
with one or more additional therapeutic agents useful in the
treatment of autoimmune and inflammatory diseases.
[0472] In a provided embodiment, diseases that can be treated or
diagnosed with the compositions and methods include, but are not
limited to, primary and metastatic cancers, including carcinomas of
breast, colon, rectum, lung, oropharynx, hypopharynx, esophagus,
stomach, pancreas, liver, gallbladder and bile ducts, small
intestine, urinary tract (including kidney, bladder and
urothelium), female genital tract (including cervix, uterus, and
ovaries as well as choriocarcinoma and gestational trophoblastic
disease), male genital tract (including prostate, seminal vesicles,
testes and germ cell tumors), endocrine glands (including the
thyroid, adrenal, and pituitary glands), and skin, as well as
hemangiomas, melanomas, sarcomas (including those arising from bone
and soft tissues as well as Kaposi's sarcoma), tumors of the brain,
nerves, eyes, and meninges (including astrocytomas, gliomas,
glioblastomas, retinoblastomas, neuromas, neuroblastomas,
Schwannomas, and meningiomas), solid tumors arising from
hematopoietic malignancies such as leukemias, and lymphomas (both
Hodgkin's and non-Hodgkin's lymphomas).
[0473] In another embodiment, an antibody or antigen binding
portion thereof used to treat cancer or in the prevention of
metastases from a tumor is provided. Such treatment may involve
administration of the antibody or antigen binding portion thereof
alone or in combination with another therapeutic agent or
treatment, such as radiotherapy and/or a chemotherapeutic
agent.
[0474] Antibodies or antigen binding portions thereof are provided
that may be combined with agents that include but are not limited
to, antineoplastic agents, radiotherapy, chemotherapy such as DNA
alkylating agents, cisplatin, carboplatin, anti-tubulin agents,
paclitaxel, docetaxel, taxol, doxorubicin, gemcitabine, gemzar,
anthracyclines, adriamycin, topoisomerase I inhibitors,
topoisomerase II inhibitors, 5-fluorouracil (5-FU), leucovorin,
irinotecan, receptor tyrosine kinase inhibitors (e.g., erlotinib,
gefitinib), COX-2 inhibitors (e.g., celecoxib), kinase inhibitors,
and siRNAs.
[0475] A binding protein administered with one or more additional
therapeutic agents useful in the treatment of various diseases is
also provided.
[0476] Antibodies or antigen binding portions thereof that can be
used alone or in combination to treat such diseases are provided.
It should be understood that the antibodies or antigen binding
portion thereof can be used alone or in combination with an
additional agent, e.g., a therapeutic agent, said additional agent
being selected by the skilled artisan for its intended purpose. For
example, the additional agent can be a therapeutic agent
art-recognized as being useful to treat the disease or condition
being treated by the antibody. The additional agent also can be an
agent that imparts a beneficial attribute to the therapeutic
composition, e.g., an agent that affects the viscosity of the
composition.
[0477] It should further be understood that the combinations which
are to be included are those combinations useful for their intended
purpose. The agents set forth below are illustrative for purposes
and not intended to be limited. The combinations can be the
antibodies and at least one additional agent selected from the
lists below. The combination can also include more than one
additional agent, e.g., two or three additional agents if the
combination is such that the formed composition can perform its
intended function.
[0478] Exemplary combinations are non-steroidal anti-inflammatory
drug(s) also referred to as NSAIDS which include drugs like
ibuprofen. Other exemplary combinations are corticosteroids
including prednisolone; the well known side-effects of steroid use
can be reduced or even eliminated by tapering the steroid dose
required when treating patients in combination with the
anti-sclerostin antibodies. Non-limiting examples of therapeutic
agents for rheumatoid arthritis with which an antibody or antibody
portion can be combined are provided and include, but are not
limited to, the following: cytokine suppressive anti-inflammatory
drug(s) (CSAIDs); antibodies to or antagonists of other human
cytokines or growth factors, for example, TNF, LT, IL-1, IL-2,
IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-15, IL-16, IL-18, IL-21,
interferons, EMAP-II, GM-CSF, FGF, and PDGF. Antibodies or antigen
binding portions thereof, can be combined with antibodies to cell
surface molecules such as CD2, CD3, CD4, CD8, CD25, CD28, CD30,
CD40, CD45, CD69, CD80 (B7.1), CD86 (B7.2), CD90, CTLA or their
ligands including CD154 (39 or CD40L).
[0479] Exemplary combinations of therapeutic agents may interfere
at different points in the autoimmune and subsequent inflammatory
cascade; exemplary examples include TNF antagonists like chimeric,
humanized or human TNF antibodies, D2E7, (PCT Publication No. WO
97/29131), CA2 (Remicade.TM.), CDP 571, and soluble p55 or p75 TNF
receptors, derivatives, thereof, (p75TNFR1gG (Enbrel.TM.) or
p55TNFR1gG (Lenercept), and also TNF.alpha. converting enzyme
(TACE) inhibitors; similarly IL-1 inhibitors
(Interleukin-1-converting enzyme inhibitors, IL-1RA etc.) may be
effective for the same reason. Other exemplary combinations include
Interleukin 11. Yet another exemplary combination are other key
players of the autoimmune response which may act parallel to,
dependent on or in concert with SOST function. Yet another
exemplary combination are non-depleting anti-CD4 inhibitors. Yet
other exemplary combinations include antagonists of the
co-stimulatory pathway CD80 (B7.1) or CD86 (B7.2) including
antibodies, soluble receptors or antagonistic ligands.
[0480] The antibodies or antigen binding portions thereof, may also
be combined with agents, such as methotrexate, 6-MP, azathioprine
sulphasalazine, mesalazine, olsalazine
chloroquinine/hydroxychloroquine, pencillamine, aurothiomalate
(intramuscular and oral), azathioprine, colchicine, corticosteroids
(oral, inhaled and local injection), beta-2 adrenoreceptor agonists
(salbutamol, terbutaline, salmeteral), xanthines (theophylline,
aminophylline), cromoglycate, nedocromil, ketotifen, ipratropium
and oxitropium, cyclosporin, FK506, rapamycin, mycophenolate
mofetil, leflunomide, NSAIDs, for example, ibuprofen,
corticosteroids such as prednisolone, phosphodiesterase inhibitors,
adensosine agonists, antithrombotic agents, complement inhibitors,
adrenergic agents, agents which interfere with signaling by
proinflammatory cytokines such as TNF-.alpha. or IL-1 (e.g., IRAK,
NIK, IKK, p38, or MAP kinase inhibitors), IL-1.beta. converting
enzyme inhibitors, TNF.alpha. converting enzyme (TACE) inhibitors,
T-cell signaling inhibitors such as kinase inhibitors,
metalloproteinase inhibitors, sulfasalazine, azathioprine,
6-mercaptopurines, angiotensin converting enzyme inhibitors,
soluble cytokine receptors and derivatives thereof (e.g. soluble
p55 or p75 TNF receptors and the derivatives p75TNFRIgG (Enbrel.TM.
and p55TNFRIgG (Lenercept)), sIL-1RI, sIL-1RII, sIL-6R),
antiinflammatory cytokines (e.g. IL-4, IL-10, IL-11, IL-13 and
TGF.beta.), celecoxib, folic acid, hydroxychloroquine sulfate,
rofecoxib, etanercept, infliximab, naproxen, valdecoxib,
sulfasalazine, methylprednisolone, meloxicam, methylprednisolone
acetate, gold sodium thiomalate, aspirin, triamcinolone acetonide,
propoxyphene napsylate/apap, folate, nabumetone, diclofenac,
piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone hcl,
hydrocodone bitartrate/apap, diclofenac sodium/misoprostol,
fentanyl, anakinra, human recombinant, tramadol hcl, salsalate,
sulindac, cyanocobalamin/fa/pyridoxine, acetaminophen, alendronate
sodium, prednisolone, morphine sulfate, lidocaine hydrochloride,
indomethacin, glucosamine sulf/chondroitin, amitriptyline hcl,
sulfadiazine, oxycodone hcl/acetaminophen, olopatadine hcl,
misoprostol, naproxen sodium, omeprazole, cyclophosphamide,
rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18 BP, anti-IL-18,
Anti-IL15, BIRB-796, SCIO-469, VX-702, AMG-548, VX-740,
Roflumilast, IC-485, CDC-801, and Mesopram. Exemplary combinations
include methotrexate or leflunomide and in moderate or severe
rheumatoid arthritis cases, cyclosporine.
[0481] Non-limiting additional agents which can also be used in
combination with a binding protein to treat rheumatoid arthritis
(RA) include, but are not limited to, the following: non-steroidal
anti-inflammatory drug(s) (NSAIDs); cytokine suppressive
anti-inflammatory drug(s) (CSAIDs); CDP-571/BAY-10-3356 (humanized
anti-TNF.alpha. antibody; Celltech/Bayer); cA2/infliximab (chimeric
anti-TNF.alpha. antibody; Centocor); 75 kdTNFR-IgG/etanercept (75
kD TNF receptor-IgG fusion protein; Immunex; see e.g., Arthritis
& Rheumatism (1994) Vol. 37, S295; J. Invest. Med. (1996) Vol.
44, 235A); 55 kdTNF-IgG (55 kD TNF receptor-IgG fusion protein;
Hoffmann-LaRoche); IDEC-CE9.1/SB 210396 (non-depleting primatized
anti-CD4 antibody; IDEC/SmithKline; see e.g., Arthritis &
Rheumatism (1995) Vol. 38, S185); DAB 486-IL-2 and/or DAB 389-IL-2
(IL-2 fusion proteins; Seragen; see e.g., Arthritis &
Rheumatism (1993) Vol. 36, 1223); Anti-Tac (humanized
anti-IL-2R.alpha.; Protein Design Labs/Roche); IL-4
(anti-inflammatory cytokine; DNAX/Schering); IL-10 (SCH 52000;
recombinant IL-10, anti-inflammatory cytokine; DNAX/Schering);
IL-4; IL-10 and/or IL-4 agonists (e.g., agonist antibodies); IL-1RA
(IL-1 receptor antagonist; Synergen/Amgen); anakinra
(Kineret.RTM./Amgen); TNF-bp/s-TNF (soluble TNF binding protein;
see e.g., Arthritis & Rheumatism (1996) Vol. 39, No. 9
(supplement), S284; Amer. J. Physiol.--Heart and Circulatory
Physiology (1995) Vol. 268, pp. 37-42); R973401 (phosphodiesterase
Type IV inhibitor; see e.g., Arthritis & Rheumatism (1996) Vol.
39, No. 9 (supplement), S282); MK-966 (COX-2 Inhibitor; see e.g.,
Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement),
S81); Iloprost (see e.g., Arthritis & Rheumatism (1996) Vol.
39, No. 9 (supplement), S82); methotrexate; thalidomide (see e.g.,
Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement),
S282) and thalidomide-related drugs (e.g., Celgen); leflunomide
(anti-inflammatory and cytokine inhibitor; see e.g., Arthritis
& Rheumatism (1996) Vol. 39, No. 9 (supplement), S131;
Inflammation Research (1996) Vol. 45, pp. 103-107); tranexamic acid
(inhibitor of plasminogen activation; see e.g., Arthritis &
Rheumatism (1996) Vol. 39, No. 9 (supplement), S284); T-614
(cytokine inhibitor; see e.g., Arthritis & Rheumatism (1996)
Vol. 39, No. 9 (supplement), S282); prostaglandin E1 (see e.g.,
Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement),
S282); Tenidap (non-steroidal anti-inflammatory drug; see e.g.,
Arthritis & Rheumatism (1996) Vol. 39, No. 9 (supplement),
S280); Naproxen (non-steroidal anti-inflammatory drug; see e.g.,
Neuro Report (1996) Vol. 7, pp. 1209-1213); Meloxicam
(non-steroidal anti-inflammatory drug); Ibuprofen (non-steroidal
anti-inflammatory drug); Piroxicam (non-steroidal anti-inflammatory
drug); Diclofenac (non-steroidal anti-inflammatory drug);
Indomethacin (non-steroidal anti-inflammatory drug); Sulfasalazine
(see e.g., Arthritis & Rheumatism (1996) Vol. 39, No. 9
(supplement), S281); Azathioprine (see e.g., Arthritis &
Rheumatism (1996) Vol. 39, No. 9 (supplement), S281); ICE inhibitor
(inhibitor of the enzyme interleukin-1.beta. converting enzyme);
zap-70 and/or lck inhibitor (inhibitor of the tyrosine kinase
zap-70 or lck); VEGF inhibitor and/or VEGF-R inhibitor (inhibitors
of vascular endothelial cell growth factor or vascular endothelial
cell growth factor receptor; inhibitors of angiogenesis);
corticosteroid anti-inflammatory drugs (e.g., SB203580);
TNF-convertase inhibitors; anti-IL-12 antibodies; anti-IL-18
antibodies; interleukin-11 (see e.g., Arthritis & Rheumatism
(1996) Vol. 39, No. 9 (supplement), S296); interleukin-13 (see
e.g., Arthritis & Rheumatism (1996) Vol. 39, No. 9
(supplement), S308); interleukin-17 inhibitors (see e.g., Arthritis
& Rheumatism (1996) Vol. 39, No. 9 (supplement), S120); gold;
penicillamine; chloroquine; chlorambucil; hydroxychloroquine;
cyclosporine; cyclophosphamide; total lymphoid irradiation;
anti-thymocyte globulin; anti-CD4 antibodies; CD5-toxins;
orally-administered peptides and collagen; lobenzarit disodium;
Cytokine Regulating Agents (CRAs) HP228 and HP466 (Houghten
Pharmaceuticals, Inc.); ICAM-1 antisense phosphorothioate
oligo-deoxynucleotides (ISIS 2302; Isis Pharmaceuticals, Inc.);
soluble complement receptor 1 (TP10; T Cell Sciences, Inc.);
prednisone; orgotein; glycosaminoglycan polysulphate; minocycline;
anti-IL2R antibodies; marine and botanical lipids (fish and plant
seed fatty acids; see e.g., DeLuca et al. (1995) Rheum. Dis. Clin.
North Am., 21: 759-777); auranofin; phenylbutazone; meclofenamic
acid; flufenamic acid; intravenous immune globulin; zileuton;
azaribine; mycophenolic acid (RS-61443); tacrolimus (FK-506);
sirolimus (rapamycin); amiprilose (therafectin); cladribine
(2-chlorodeoxyadenosine); methotrexate; bcl-2 inhibitors (see
Bruncko, Milan et al., Journal of Medicinal Chemistry (2007),
50(4), 641-662); antivirals and immune modulating agents.
[0482] In one embodiment, the binding protein or antigen-binding
portion thereof, is administered in combination with one of the
following agents for the treatment of rheumatoid arthritis (RA):
small molecule inhibitor of KDR, small molecule inhibitor of Tie-2;
methotrexate; prednisone; celecoxib; folic acid; hydroxychloroquine
sulfate; rofecoxib; etanercept; infliximab; leflunomide; naproxen;
valdecoxib; sulfasalazine; methylprednisolone; ibuprofen;
meloxicam; methylprednisolone acetate; gold sodium thiomalate;
aspirin; azathioprine; triamcinolone acetonide; propoxyphene
napsylate/apap; folate; nabumetone; diclofenac; piroxicam;
etodolac; diclofenac sodium; oxaprozin; oxycodone hcl; hydrocodone
bitartrate/apap; diclofenac sodium/misoprostol; fentanyl; anakinra,
human recombinant; tramadol hcl; salsalate; sulindac;
cyanocobalamin/fa/pyridoxine; acetaminophen; alendronate sodium;
prednisolone; morphine sulfate; lidocaine hydrochloride;
indomethacin; glucosamine sulfate/chondroitin; cyclosporine;
amitriptyline hcl; sulfadiazine; oxycodone hcl/acetaminophen;
olopatadine hcl; misoprostol; naproxen sodium; omeprazole;
mycophenolate mofetil; cyclophosphamide; rituximab; IL-1 TRAP; MRA;
CTLA4-IG; IL-18 BP; IL-12/23; anti-IL 18; anti-IL 15; BIRB-796;
SCIO-469; VX-702; AMG-548; VX-740; Roflumilast; IC-485; CDC-801;
and mesopram.
[0483] Non-limiting examples of therapeutic agents for inflammatory
bowel disease with which a binding protein can be combined include
the following: budenoside; epidermal growth factor;
corticosteroids; cyclosporin, sulfasalazine; aminosalicylates;
6-mercaptopurine; azathioprine; metronidazole; lipoxygenase
inhibitors; mesalamine; olsalazine; balsalazide; antioxidants;
thromboxane inhibitors; IL-1 receptor antagonists; anti-IL-1.beta.
mAbs; anti-IL-6 mAbs; growth factors; elastase inhibitors;
pyridinyl-imidazole compounds; antibodies to or antagonists of
other human cytokines or growth factors, for example, TNF, LT,
IL-1, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-17, IL-18, EMAP-II,
GM-CSF, FGF, and PDGF. Antibodies or antigen binding portions
thereof, can be combined with antibodies to cell surface molecules
such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69,
CD90 or their ligands. The antibodies or antigen binding portions
thereof, may also be combined with agents, such as methotrexate,
cyclosporin, FK506, rapamycin, mycophenolate mofetil, leflunomide,
NSAIDs, for example, ibuprofen, corticosteroids such as
prednisolone, phosphodiesterase inhibitors, adenosine agonists,
antithrombotic agents, complement inhibitors, adrenergic agents,
agents which interfere with signaling by proinflammatory cytokines
such as TNF.alpha. or IL-1 (e.g., IRAK, NIK, IKK, p38 or MAP kinase
inhibitors), IL-1.beta. converting enzyme inhibitors, TNF.alpha.
converting enzyme inhibitors, T-cell signaling inhibitors such as
kinase inhibitors, metalloproteinase inhibitors, sulfasalazine,
azathioprine, 6-mercaptopurines, angiotensin converting enzyme
inhibitors, soluble cytokine receptors and derivatives thereof
(e.g., soluble p55 or p75 TNF receptors, sIL-1RI, sIL-1RII, sIL-6R)
and antiinflammatory cytokines (e.g., IL-4, IL-10, IL-11, IL-13 and
TGF.beta.) and bcl-2 inhibitors.
[0484] Examples of therapeutic agents for Crohn's disease in which
a binding protein can be combined include the following: TNF
antagonists, for example, anti-TNF antibodies, Adalimumab (PCT
Publication No. WO 97/29131; HUMIRA.RTM.), CA2 (REMICADE), CDP 571,
TNFR-Ig constructs, (p75TNFRIgG (ENBREL.RTM.) and p55TNFRIgG
(LENERCEPT.TM.)) inhibitors and PDE4 inhibitors. Antibodies or
antigen binding portions thereof, can be combined with
corticosteroids, for example, budenoside and dexamethasone. Binding
proteins or antigen binding portions thereof, may also be combined
with agents such as sulfasalazine, 5-aminosalicylic acid and
olsalazine, and agents which interfere with synthesis or action of
proinflammatory cytokines such as IL-1, for example, IL-1.beta.
converting enzyme inhibitors and IL-1ra. Antibodies or antigen
binding portion thereof may also be used with T cell signaling
inhibitors, for example, tyrosine kinase inhibitors
6-mercaptopurines. Binding proteins or antigen binding portions
thereof, can be combined with IL-11. Binding proteins or antigen
binding portions thereof, can be combined with mesalamine,
prednisone, azathioprine, mercaptopurine, infliximab,
methylprednisolone sodium succinate, diphenoxylate/atrop sulfate,
loperamide hydrochloride, methotrexate, omeprazole, folate,
ciprofloxacin/dextrose-water, hydrocodone bitartrate/apap,
tetracycline hydrochloride, fluocinonide, metronidazole,
thimerosal/boric acid, cholestyramine/sucrose, ciprofloxacin
hydrochloride, hyoscyamine sulfate, meperidine hydrochloride,
midazolam hydrochloride, oxycodone hcl/acetaminophen, promethazine
hydrochloride, sodium phosphate, sulfamethoxazole/trimethoprim,
celecoxib, polycarbophil, propoxyphene napsylate, hydrocortisone,
multivitamins, balsalazide disodium, codeine phosphate/apap,
colesevelam hcl, cyanocobalamin, folic acid, levofloxacin,
methylprednisolone, natalizumab and interferon-gamma
[0485] Non-limiting examples of therapeutic agents for multiple
sclerosis (MS) with which binding proteins can be combined include
the following: corticosteroids; prednisolone; methylprednisolone;
azathioprine; cyclophosphamide; cyclosporine; methotrexate;
4-aminopyridine; tizanidine; interferon-.beta.1a (AVONEX; Biogen);
interferon-.beta.1b (BETASERON; Chiron/Berlex); interferon
.alpha.-n3) (Interferon Sciences/Fujimoto), interferon-.alpha.
(Alfa Wassermann/J&J), interferon .beta.1A-IF (Serono/Inhale
Therapeutics), Peginterferon .alpha. 2b (Enzon/Schering-Plough),
Copolymer 1 (Cop-1; COPAXONE; Teva Pharmaceutical Industries,
Inc.); hyperbaric oxygen; intravenous immunoglobulin; clabribine;
antibodies to or antagonists of other human cytokines or growth
factors and their receptors, for example, TNF, LT, IL-1, IL-2,
IL-6, IL-7, IL-8, IL-23, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF,
and PDGF. Binding proteins can be combined with antibodies to cell
surface molecules such as CD2, CD3, CD4, CD8, CD19, CD20, CD25,
CD28, CD30, CD40, CD45, CD69, CD80, CD86, CD90 or their ligands.
Binding proteins may also be combined with agents, such as
methotrexate, cyclosporine, FK506, rapamycin, mycophenolate
mofetil, leflunomide, NSAIDs, for example, ibuprofen,
corticosteroids such as prednisolone, phosphodiesterase inhibitors,
adensosine agonists, antithrombotic agents, complement inhibitors,
adrenergic agents, agents which interfere with signaling by
proinflammatory cytokines such as TNF.alpha. or IL-1 (e.g., IRAK,
NIK, IKK, p38 or MAP kinase inhibitors), IL-1.beta. converting
enzyme inhibitors, TACE inhibitors, T-cell signaling inhibitors
such as kinase inhibitors, metalloproteinase inhibitors,
sulfasalazine, azathioprine, 6-mercaptopurines, angiotensin
converting enzyme inhibitors, soluble cytokine receptors and
derivatives thereof (e.g., soluble p55 or p75 TNF receptors,
sIL-1RI, sIL-1RII, sIL-6R), antiinflammatory cytokines (e.g., IL-4,
IL-10, IL-13 and TGF.beta.) and bcl-2 inhibitors.
[0486] Examples of therapeutic agents for multiple sclerosis in
which binding proteins can be combined include interferon-.beta.,
for example, IFN.beta.1a and IFN.beta.1b; copaxone,
corticosteroids, caspase inhibitors, for example inhibitors of
caspase-1, IL-1 inhibitors, TNF inhibitors, and antibodies to CD40
ligand and CD80.
[0487] The binding proteins may also be combined with agents, such
as alemtuzumab, dronabinol, Unimed, daclizumab, mitoxantrone,
xaliproden hydrochloride, fampridine, glatiramer acetate,
natalizumab, sinnabidol, a-immunokine NNSO3, ABR-215062,
AnergiX.MS, chemokine receptor antagonists, BBR-2778, calagualine,
CPI-1189, LEM (liposome encapsulated mitoxantrone), THC.CBD
(cannabinoid agonist) MBP-8298, mesopram (PDE4 inhibitor), MNA-715,
anti-IL-6 receptor antibody, neurovax, pirfenidone allotrap 1258
(RDP-1258), sTNF-R1, talampanel, teriflunomide, TGF-beta2,
tiplimotide, VLA-4 antagonists (for example, TR-14035, VLA4
Ultrahaler, Antegran-ELAN/Biogen), interferon gamma antagonists,
IL-4 agonists.
[0488] Non-limiting examples of therapeutic agents for Angina with
which binding proteins can be combined are provided and include the
following: aspirin, nitroglycerin, isosorbide mononitrate,
metoprolol succinate, atenolol, metoprolol tartrate, amlodipine
besylate, diltiazem hydrochloride, isosorbide dinitrate,
clopidogrel bisulfate, nifedipine, atorvastatin calcium, potassium
chloride, furosemide, simvastatin, verapamil hcl, digoxin,
propranolol hydrochloride, carvedilol, lisinopril, spironolactone,
hydrochlorothiazide, enalapril maleate, nadolol, ramipril,
enoxaparin sodium, heparin sodium, valsartan, sotalol
hydrochloride, fenofibrate, ezetimibe, bumetanide, losartan
potassium, lisinopril/hydrochlorothiazide, felodipine, captopril,
bisoprolol fumarate.
[0489] Non-limiting examples of therapeutic agents for Ankylosing
Spondylitis with which binding proteins can be combined are
provided and include the following: ibuprofen, diclofenac and
misoprostol, naproxen, meloxicam, indomethacin, diclofenac,
celecoxib, rofecoxib, Sulfasalazine, Methotrexate, azathioprine,
minocyclin, prednisone, etanercept, infliximab.
[0490] Non-limiting examples of therapeutic agents for Asthma with
which binding proteins can be combined are provided and include the
following: albuterol, salmeterol/fluticasone, montelukast sodium,
fluticasone propionate, budesonide, prednisone, salmeterol
xinafoate, levalbuterol hcl, albuterol sulfate/ipratropium,
prednisolone sodium phosphate, triamcinolone acetonide,
beclomethasone dipropionate, ipratropium bromide, azithromycin,
pirbuterol acetate, prednisolone, theophylline anhydrous,
methylprednisolone sodium succinate, clarithromycin, zafirlukast,
formoterol fumarate, influenza virus vaccine, methylprednisolone,
amoxicillin trihydrate, flunisolide, allergy injection, cromolyn
sodium, fexofenadine hydrochloride, flunisolide/menthol,
amoxicillin/clavulanate, levofloxacin, inhaler assist device,
guaifenesin, dexamethasone sodium phosphate, moxifloxacin hcl,
doxycycline hyclate, guaifenesin/d-methorphan,
p-ephedrine/cod/chlorphenir, gatifloxacin, cetirizine
hydrochloride, mometasone furoate, salmeterol xinafoate,
benzonatate, cephalexin, pe/hydrocodone/chlorphenir, cetirizine
hcl/pseudoephed, phenylephrine/cod/promethazine,
codeine/promethazine, cefprozil, dexamethasone,
guaifenesin/pseudoephedrine, chlorpheniramine/hydrocodone,
nedocromil sodium, terbutaline sulfate, epinephrine,
methylprednisolone, metaproterenol sulfate.
[0491] Non-limiting examples of therapeutic agents for COPD with
which binding proteins can be combined are provided and include the
following: albuterol sulfate/ipratropium, ipratropium bromide,
salmeterol/fluticasone, albuterol, salmeterol xinafoate,
fluticasone propionate, prednisone, theophylline anhydrous,
methylprednisolone sodium succinate, montelukast sodium,
budesonide, formoterol fumarate, triamcinolone acetonide,
levofloxacin, guaifenesin, azithromycin, beclomethasone
dipropionate, levalbuterol hcl, flunisolide, ceftriaxone sodium,
amoxicillin trihydrate, gatifloxacin, zafirlukast,
amoxicillin/clavulanate, flunisolide/menthol,
chlorpheniramine/hydrocodone, metaproterenol sulfate,
methylprednisolone, mometasone furoate,
p-ephedrine/cod/chlorphenir, pirbuterol acetate,
p-ephedrine/loratadine, terbutaline sulfate, tiotropium bromide,
(R,R)-formoterol, TgAAT, Cilomilast, Roflumilast.
[0492] Non-limiting examples of therapeutic agents for HCV with
which binding proteins can be combined are provided and include the
following: Interferon-alpha-2a, Interferon-alpha-2b,
Interferon-alpha coni, Interferon-alpha-nl, Pegylated
interferon-alpha-2a, Pegylated interferon-alpha-2b, ribavirin,
Peginterferon alfa-2b+ribavirin, Ursodeoxycholic Acid, Glycyrrhizic
Acid, Thymalfasin, Maxamine, VX-497 and any compounds that are used
to treat HCV through intervention with the following targets: HCV
polymerase, HCV protease, HCV helicase, HCV IRES (internal ribosome
entry site).
[0493] Non-limiting examples of therapeutic agents for Idiopathic
Pulmonary Fibrosis with which binding proteins can be combined are
provided and include the following: prednisone, azathioprine,
albuterol, colchicine, albuterol sulfate, digoxin, gamma
interferon, methylprednisolone sod succ, lorazepam, furosemide,
lisinopril, nitroglycerin, spironolactone, cyclophosphamide,
ipratropium bromide, actinomycin d, alteplase, fluticasone
propionate, levofloxacin, metaproterenol sulfate, morphine sulfate,
oxycodone hcl, potassium chloride, triamcinolone acetonide,
tacrolimus anhydrous, calcium, interferon-alpha, methotrexate,
mycophenolate mofetil, Interferon-gamma-1.beta..
[0494] Non-limiting examples of therapeutic agents for Myocardial
Infarction with which binding proteins can be combined are provided
and include the following: aspirin, nitroglycerin, metoprolol
tartrate, enoxaparin sodium, heparin sodium, clopidogrel bisulfate,
carvedilol, atenolol, morphine sulfate, metoprolol succinate,
warfarin sodium, lisinopril, isosorbide mononitrate, digoxin,
furosemide, simvastatin, ramipril, tenecteplase, enalapril maleate,
torsemide, retavase, losartan potassium, quinapril hcl/mag carb,
bumetanide, alteplase, enalaprilat, amiodarone hydrochloride,
tirofiban hcl m-hydrate, diltiazem hydrochloride, captopril,
irbesartan, valsartan, propranolol hydrochloride, fosinopril
sodium, lidocaine hydrochloride, eptifibatide, cefazolin sodium,
atropine sulfate, aminocaproic acid, spironolactone, interferon,
sotalol hydrochloride, potassium chloride, docusate sodium,
dobutamine hcl, alprazolam, pravastatin sodium, atorvastatin
calcium, midazolam hydrochloride, meperidine hydrochloride,
isosorbide dinitrate, epinephrine, dopamine hydrochloride,
bivalirudin, rosuvastatin, ezetimibe/simvastatin, avasimibe,
cariporide.
[0495] Non-limiting examples of therapeutic agents for Psoriasis
with which binding proteins can be combined are provided and
include the following: small molecule inhibitor of KDR, small
molecule inhibitor of Tie-2, calcipotriene, clobetasol propionate,
triamcinolone acetonide, halobetasol propionate, tazarotene,
methotrexate, fluocinonide, betamethasone diprop augmented,
fluocinolone acetonide, acitretin, tar shampoo, betamethasone
valerate, mometasone furoate, ketoconazole, pramoxine/fluocinolone,
hydrocortisone valerate, flurandrenolide, urea, betamethasone,
clobetasol propionate/emoll, fluticasone propionate, azithromycin,
hydrocortisone, moisturizing formula, folic acid, desonide,
pimecrolimus, coal tar, diflorasone diacetate, etanercept folate,
lactic acid, methoxsalen, hc/bismuth subgal/znox/resor,
methylprednisolone acetate, prednisone, sunscreen, halcinonide,
salicylic acid, anthralin, clocortolone pivalate, coal extract,
coal tar/salicylic acid, coal tar/salicylic acid/sulfur,
desoximetasone, diazepam, emollient, fluocinonide/emollient,
mineral oil/castor oillna lact, mineral oil/peanut oil,
petroleum/isopropyl myristate, psoralen, salicylic acid,
soap/tribromsalan, thimerosal/boric acid, celecoxib, infliximab,
cyclosporine, alefacept, efalizumab, tacrolimus, pimecrolimus,
PUVA, UVB, sulfasalazine.
[0496] Non-limiting examples of therapeutic agents for Psoriatic
Arthritis with which binding proteins can be combined are provided
and include the following: methotrexate, etanercept, rofecoxib,
celecoxib, folic acid, sulfasalazine, naproxen, leflunomide,
methylprednisolone acetate, indomethacin, hydroxychloroquine
sulfate, prednisone, sulindac, betamethasone diprop augmented,
infliximab, methotrexate, folate, triamcinolone acetonide,
diclofenac, dimethylsulfoxide, piroxicam, diclofenac sodium,
ketoprofen, meloxicam, methylprednisolone, nabumetone, tolmetin
sodium, calcipotriene, cyclosporine, diclofenac sodium/misoprostol,
fluocinonide, glucosamine sulfate, gold sodium thiomalate,
hydrocodone bitartrate/apap, ibuprofen, risedronate sodium,
sulfadiazine, thioguanine, valdecoxib, alefacept, efalizumab and
bcl-2 inhibitors.
[0497] Non-limiting examples of therapeutic agents for Restenosis
with which binding proteins can be combined are provided and
include the following: sirolimus, paclitaxel, everolimus,
tacrolimus, Zotarolimus, acetaminophen.
[0498] Non-limiting examples of therapeutic agents for Sciatica
with which binding proteins can be combined are provided and
include the following: hydrocodone bitartrate/apap, rofecoxib,
cyclobenzaprine hcl, methylprednisolone, naproxen, ibuprofen,
oxycodone hcl/acetaminophen, celecoxib, valdecoxib,
methylprednisolone acetate, prednisone, codeine phosphate/apap,
tramadol hcl/acetaminophen, metaxalone, meloxicam, methocarbamol,
lidocaine hydrochloride, diclofenac sodium, gabapentin,
dexamethasone, carisoprodol, ketorolac tromethamine, indomethacin,
acetaminophen, diazepam, nabumetone, oxycodone hcl, tizanidine hcl,
diclofenac sodium/misoprostol, propoxyphene napsylate/apap,
asa/oxycod/oxycodone ter, ibuprofen/hydrocodone bit, tramadol hcl,
etodolac, propoxyphene hcl, amitriptyline hcl, carisoprodol/codeine
phos/asa, morphine sulfate, multivitamins, naproxen sodium,
orphenadrine citrate, temazepam.
[0499] Examples of therapeutic agents for SLE (Lupus) in which
binding proteins can be combined are provided and include the
following: NSAIDS, for example, diclofenac, naproxen, ibuprofen,
piroxicam, indomethacin; COX2 inhibitors, for example, Celecoxib,
rofecoxib, valdecoxib; anti-malarials, for example,
hydroxychloroquine; Steroids, for example, prednisone,
prednisolone, budenoside, dexamethasone; Cytotoxics, for example,
azathioprine, cyclophosphamide, mycophenolate mofetil,
methotrexate; inhibitors of PDE4 or purine synthesis inhibitor, for
example Cellcept. Binding proteins may also be combined with agents
such as sulfasalazine, 5-aminosalicylic acid, olsalazine, Imuran
and agents which interfere with synthesis, production or action of
proinflammatory cytokines such as IL-1, for example, caspase
inhibitors like IL-1.beta. converting enzyme inhibitors and IL-1ra.
Binding proteins may also be used with T cell signaling inhibitors,
for example, tyrosine kinase inhibitors; or molecules that target T
cell activation molecules, for example, CTLA-4-IgG or anti-B7
family antibodies, anti-PD-1 family antibodies. Binding proteins
can be combined with IL-11 or anti-cytokine antibodies, for
example, fonotolizumab (anti-IFNg antibody), or anti-receptor
receptor antibodies, for example, anti-IL-6 receptor antibody and
antibodies to B-cell surface molecules. Antibodies or antigen
binding portion thereof may also be used with LJP 394 (abetimus),
agents that deplete or inactivate B-cells, for example, Rituximab
(anti-CD20 antibody), lymphostat-B (anti-BlyS antibody), TNF
antagonists, for example, anti-TNF antibodies, Adalimumab (PCT
Publication No. WO 97/29131; HUMIRA.RTM.), CA2 (REMICADE.RTM.), CDP
571, TNFR-Ig constructs, (p75TNFRIgG (ENBREL.RTM.) and p55TNFRIgG
(LENERCEPT.RTM.)) and bcl-2 inhibitors, because bcl-2
overexpression in transgenic mice has been demonstrated to cause a
lupus like phenotype (see Marquina, Regina et al., Journal of
Immunology (2004), 172(11), 7177-7185), therefore inhibition is
expected to have therapeutic effects.
[0500] The pharmaceutical compositions may include a
"therapeutically effective amount" or a "prophylactically effective
amount" of an antibody or antibody portion thereof. A
"therapeutically effective amount" refers to an amount effective,
at dosages and for periods of time necessary, to achieve the
desired therapeutic result. A therapeutically effective amount of
the antibody or antibody portion may be determined by a person
skilled in the art and may vary according to factors such as the
disease state, age, sex, and weight of the individual, and the
ability of the antibody or antibody portion to elicit a desired
response in the individual. A therapeutically effective amount is
also one in which any toxic or detrimental effects of the antibody,
or antibody portion, are outweighed by the therapeutically
beneficial effects. A "prophylactically effective amount" refers to
an amount effective, at dosages and for periods of time necessary,
to achieve the desired prophylactic result. Typically, since a
prophylactic dose is used in subjects prior to or at an earlier
stage of disease, the prophylactically effective amount will be
less than the therapeutically effective amount.
[0501] Dosage regimens may be adjusted to provide the optimum
desired response (e.g., a therapeutic or prophylactic response).
For example, a single bolus may be administered, several divided
doses may be administered over time or the dose may be
proportionally reduced or increased as indicated by the exigencies
of the therapeutic situation. It is especially advantageous to
formulate parenteral compositions in dosage unit form for ease of
administration and uniformity of dosage. The term "dosage unit
form" refers to physically discrete units suited as unitary dosages
for the mammalian subjects to be treated; each unit containing a
predetermined quantity of active compound calculated to produce the
desired therapeutic effect in association with the required
pharmaceutical carrier. The specification for the dosage unit forms
are dictated by and directly dependent on (a) the unique
characteristics of the active compound and the particular
therapeutic or prophylactic effect to be achieved, and (b) the
limitations inherent in the art of compounding such an active
compound for the treatment of sensitivity in individuals.
[0502] It is to be noted that dosage values may vary with the type
and severity of the condition to be alleviated. It is to be further
understood that for any particular subject, specific dosage
regimens should be adjusted over time according to the individual
need and the professional judgment of the person administering or
supervising the administration of the compositions, and that dosage
ranges set forth herein are exemplary only and are not intended to
limit the scope or practice of the claimed composition.
VIII. Diagnostics
[0503] The disclosure herein also provides diagnostic applications.
This is further elucidated below. Antibodies that bind sclerostin
are provided and may be employed in any of a variety of formats to
detect sclerostin in vivo, in vitro, or ex vivo (i.e., in cells or
tissues that have been obtained from a living individual, subjected
to a procedure, then returned to the individual). DVD-binding
proteins offer the further advantage of being capable of binding to
an epitope of sclerostin as well as other antigens or epitopes in
various diagnostic and detection assay formats.
I. Method of Assay
[0504] The present disclosure also provides a method for
determining the presence, amount or concentration of a sclerostin,
or a fragment thereof, ("analyte") in a test sample using at least
one anti-sclerostin binding protein or antigen binding portion
thereof, including a DVD-binding protein, as described herein. Any
suitable assay as is known in the art can be used in the method.
Examples include, but are not limited to, immunoassay, such as
sandwich immunoassay (e.g., monoclonal, polyclonal and/or
DVD-binding protein sandwich immunoassays or any variation thereof
(e.g., monoclonal/DVD-binding protein, DVD-binding
protein/polyclonal, etc.), including radioisotope detection
(radioimmunoassay (RIA)) and enzyme detection (enzyme immunoassay
(EIA) or enzyme-linked immunosorbent assay (ELISA) (e.g.,
Quantikine ELISA assays, R&D Systems, Minneapolis, Minn.))),
competitive inhibition immunoassay (e.g., forward and reverse),
fluorescence polarization immunoassay (FPIA), enzyme multiplied
immunoassay technique (EMIT), bioluminescence resonance energy
transfer (BRET), and homogeneous chemiluminescent assay, etc. In a
SELDI-based immunoassay, a capture reagent that specifically binds
an analyte (or a fragment thereof) of interest is attached to the
surface of a mass spectrometry probe, such as a pre-activated
protein chip array. The analyte (or a fragment thereof) is then
specifically captured on the biochip, and the captured analyte (or
a fragment thereof) is detected by mass spectrometry.
Alternatively, the analyte (or a fragment thereof) can be eluted
from the capture reagent and detected by traditional MALDI
(matrix-assisted laser desorption/ionization) or by SELDI. A
chemiluminescent microparticle immunoassay, in particular one
employing the ARCHITECT.RTM. automated analyzer (Abbott
Laboratories, Abbott Park, Ill.), is an example of an
immunoassay.
[0505] Methods well-known in the art for collecting, handling and
processing urine, blood, serum and plasma, and other body fluids,
are used in the practice of the present disclosure, for instance,
when anti-sclerostin binding protein as described herein is
employed as an immunodiagnostic reagent and/or in an analyte
immunoassay kit. The test sample can comprise further moieties in
addition to the analyte of interest, such as antibodies, antigens,
haptens, hormones, drugs, enzymes, receptors, proteins, peptides,
polypeptides, oligonucleotides and/or polynucleotides. For example,
the sample can be a whole blood sample obtained from a subject. It
can be necessary or desired that a test sample, particularly whole
blood, be treated prior to immunoassay as described herein, e.g.,
with a pretreatment reagent. Even in cases where pretreatment is
not necessary (e.g., most urine samples), pretreatment optionally
can be done (e.g., as part of a regimen on a commercial
platform).
[0506] The pretreatment reagent can be any reagent appropriate for
use with the immunoassay and kits. The pretreatment optionally
comprises: (a) one or more solvents (e.g., methanol and ethylene
glycol) and optionally, salt, (b) one or more solvents and salt,
and optionally, detergent, (c) detergent, or (d) detergent and
salt. Pretreatment reagents are known in the art, and such
pretreatment can be employed, e.g., as used for assays on Abbott
TDx, AxSYM.RTM., and ARCHITECT.RTM. analyzers (Abbott Laboratories,
Abbott Park, Ill.), as described in the literature (see, e.g.,
Yatscoff et al., Abbott TDx Monoclonal Antibody Assay Evaluated for
Measuring Cyclosporine in Whole Blood, Clin. Chem. 36: 1969-1973
(1990), and Wallemacq et al., Evaluation of the New AxSYM
Cyclosporine Assay: Comparison with TDx Monoclonal Whole Blood and
EMIT Cyclosporine Assays, Clin. Chem. 45: 432-435 (1999)), and/or
as commercially available. Additionally, pretreatment can be done
as described in Abbott's U.S. Pat. No. 5,135,875; European Patent
Publication No. 0 471 293; PCT Publication No. WO 2008/082984; and
US Patent Application Publication No. 2008/0020401. The
pretreatment reagent can be a heterogeneous agent or a homogeneous
agent.
[0507] With use of a heterogeneous pretreatment reagent, the
pretreatment reagent precipitates analyte binding protein (e.g.,
protein that can bind to an analyte or a fragment thereof) present
in the sample. Such a pretreatment step comprises removing any
analyte binding protein by separating from the precipitated analyte
binding protein the supernatant of the mixture formed by addition
of the pretreatment agent to sample. In such an assay, the
supernatant of the mixture absent any binding protein is used in
the assay, proceeding directly to the antibody capture step.
[0508] With use of a homogeneous pretreatment reagent there is no
such separation step. The entire mixture of test sample and
pretreatment reagent are contacted with a labeled specific binding
partner for analyte (or a fragment thereof), such as a labeled
anti-analyte antibody (or an antigenically reactive fragment
thereof). The pretreatment reagent employed for such an assay
typically is diluted in the pretreated test sample mixture, either
before or during capture by the first specific binding partner.
Despite such dilution, a certain amount of the pretreatment reagent
is still present (or remains) in the test sample mixture during
capture. An exemplary labeled specific binding partner can be a
DVD-binding protein (or a fragment, a variant, or a fragment of a
variant thereof).
[0509] In a heterogeneous format, after the test sample is obtained
from a subject, a first mixture is prepared. The mixture contains
the test sample being assessed for an analyte (or a fragment
thereof) and a first specific binding partner, wherein the first
specific binding partner and any analyte contained in the test
sample form a first specific binding partner-analyte complex. In an
embodiment, the first specific binding partner is an anti-analyte
antibody or a fragment thereof. The first specific binding partner
can be a DVD-binding protein (or a fragment, a variant, or a
fragment of a variant thereof) as described herein. The order in
which the test sample and the first specific binding partner are
added to form the mixture is not critical. In an embodiment, the
first specific binding partner is immobilized on a solid phase. The
solid phase used in the immunoassay (for the first specific binding
partner and, optionally, the second specific binding partner) can
be any solid phase known in the art, such as, but not limited to, a
magnetic particle, a bead, a test tube, a microtiter plate, a
cuvette, a membrane, a scaffolding molecule, a film, a filter
paper, a disc and a chip.
[0510] After the mixture containing the first specific binding
partner-analyte complex is formed, any unbound analyte is removed
from the complex using any technique known in the art. For example,
the unbound analyte can be removed by washing. Desirably, however,
the first specific binding partner is present in excess of any
analyte present in the test sample, such that all analyte that is
present in the test sample is bound by the first specific binding
partner.
[0511] After any unbound analyte is removed, a second specific
binding partner is added to the mixture to form a first specific
binding partner-analyte-second specific binding partner complex.
The second specific binding partner is, e.g., an anti-analyte
antibody that binds to an epitope on analyte that differs from the
epitope on analyte bound by the first specific binding partner.
Moreover, in an embodiment, the second specific binding partner is
labeled with or contains a detectable label as described above. The
second specific binding partner can be a DVD-binding protein (or a
fragment, a variant, or a fragment of a variant thereof) as
described herein.
[0512] Any suitable detectable label as is known in the art can be
used. For example, the detectable label can be a radioactive label
(such as .sup.3H, .sup.125I, .sup.35S, .sup.14C, .sup.32P, and
33P), an enzymatic label (such as horseradish peroxidase, alkaline
peroxidase, glucose 6-phosphate dehydrogenase, and the like), a
chemiluminescent label (such as acridinium esters, thioesters, or
sulfonamides; luminol, isoluminol, phenanthridinium esters, and the
like), a fluorescent label (such as fluorescein (e.g.,
5-fluorescein, 6-carboxyfluorescein, 3'6-carboxyfluorescein,
5(6)-carboxyfluorescein, 6-hexachloro-fluorescein,
6-tetrachlorofluorescein, fluorescein isothiocyanate, and the
like)), rhodamine, phycobiliproteins, R-phycoerythrin, quantum dots
(e.g., zinc sulfide-capped cadmium selenide), a thermometric label,
or an immuno-polymerase chain reaction label. An introduction to
labels, labeling procedures and detection of labels is found in
Polak and Van Noorden, Introduction to Immunocytochemistry, 2nd
ed., Springer Verlag, N.Y. (1997), and in Haugland, Handbook of
Fluorescent Probes and Research Chemicals (1996), which is a
combined handbook and catalogue published by Molecular Probes,
Inc., Eugene, Oreg. A fluorescent label can be used in FPIA (see,
e.g., U.S. Pat. Nos. 5,593,896, 5,573,904, 5,496,925, 5,359,093,
and 5,352,803). An acridinium compound can be used as a detectable
label in a homogeneous or heterogeneous chemiluminescent assay
(see, e.g., Adamczyk et al., Bioorg. Med. Chem. Lett. 16: 1324-1328
(2006); Adamczyk et al., Bioorg. Med. Chem. Lett. 4: 2313-2317
(2004); Adamczyk et al., Biorg. Med. Chem. Lett. 14: 3917-3921
(2004); and Adamczyk et al., Org. Lett. 5: 3779-3782 (2003)).
[0513] An exemplary acridinium compound is an
acridinium-9-carboxamide. Methods for preparing acridinium
9-carboxamides are described in Mattingly, J. Biolumin. Chemilumin.
6: 107-114 (1991); Adamczyk et al., J. Org. Chem., 63: 5636-5639
(1998); Adamczyk et al., Tetrahedron, 55: 10899-10914 (1999);
Adamczyk et al., Org. Lett., 1: 779-781 (1999); Adamczyk et al.,
Bioconjugate Chem., 11: 714-724 (2000); Mattingly et al., In
Luminescence Biotechnology: Instruments and Applications; Dyke, K.
V. Ed.; CRC Press: Boca Raton, pp. 77-105 (2002); Adamczyk et al.,
Org. Lett., 5: 3779-3782 (2003); and U.S. Pat. Nos. 5,468,646,
5,543,524 and 5,783,699. Another exemplary acridinium compound is
an acridinium-9-carboxylate aryl ester. An example of an
acridinium-9-carboxylate aryl ester is
10-methyl-9-(phenoxycarbonyfiacridinium fluorosulfonate (available
from Cayman Chemical, Ann Arbor, Mich.). Methods for preparing
acridinium 9-carboxylate aryl esters are described in McCapra et
al., Photochem. Photobiol., 4: 1111-21 (1965); Razavi et al.,
Luminescence, 15: 245-249 (2000); Razavi et al., Luminescence, 15:
239-244 (2000); and U.S. Pat. No. 5,241,070. Further details
regarding acridinium-9-carboxylate aryl ester and its use are set
forth in US 2008-0248493.
[0514] Chemiluminescent assays (e.g., using acridinium as described
above or other chemiluminescent agents) can be performed in
accordance with the methods described in Adamczyk et al., Anal.
Chim. Acta, 579(1): 61-67 (2006). While any suitable assay format
can be used, a microplate chemiluminometer (Mithras LB-940,
Berthold Technologies U.S.A., LLC, Oak Ridge, Tenn.) enables the
assay of multiple samples of small volumes rapidly.
[0515] The order in which the test sample and the specific binding
partner(s) are added to form the mixture for chemiluminescent assay
is not critical. If the first specific binding partner is
detectably labeled with a chemiluminescent agent such as an
acridinium compound, detectably labeled first specific binding
partner-analyte complexes form. Alternatively, if a second specific
binding partner is used and the second specific binding partner is
detectably labeled with a chemiluminescent agent such as an
acridinium compound, detectably labeled first specific binding
partner-analyte-second specific binding partner complexes form. Any
unbound specific binding partner, whether labeled or unlabeled, can
be removed from the mixture using any technique known in the art,
such as washing.
[0516] Hydrogen peroxide can be generated in situ in the mixture or
provided or supplied to the mixture (e.g., the source of the
hydrogen peroxide being one or more buffers or other solutions that
are known to contain hydrogen peroxide) before, simultaneously
with, or after the addition of an above-described acridinium
compound. Hydrogen peroxide can be generated in situ in a number of
ways such as would be apparent to one skilled in the art.
[0517] Upon the simultaneous or subsequent addition of at least one
basic solution to the sample, a detectable signal, namely, a
chemiluminescent signal, indicative of the presence of analyte is
generated. The basic solution contains at least one base and has a
pH greater than or equal to 10, e.g., greater than or equal to 12.
Examples of basic solutions include, but are not limited to, sodium
hydroxide, potassium hydroxide, calcium hydroxide, ammonium
hydroxide, magnesium hydroxide, sodium carbonate, sodium
bicarbonate, calcium hydroxide, calcium carbonate, and calcium
bicarbonate. The amount of basic solution added to the sample
depends on the concentration of the basic solution. Based on the
concentration of the basic solution used, one skilled in the art
can easily determine the amount of basic solution to add to the
sample.
[0518] The chemiluminescent signal that is generated can be
detected using routine techniques known to those skilled in the
art. Based on the intensity of the signal generated, the amount of
analyte in the sample can be quantified. Specifically, the amount
of analyte in the sample is proportional to the intensity of the
signal generated. The amount of analyte present can be quantified
by comparing the amount of light generated to a standard curve for
analyte or by comparison to a reference standard. The standard
curve can be generated using serial dilutions or solutions of known
concentrations of analyte by mass spectroscopy, gravimetric
methods, and other techniques known in the art. While the above is
described with emphasis on use of an acridinium compound as the
chemiluminescent agent, one of ordinary skill in the art can
readily adapt this description for use of other chemiluminescent
agents.
[0519] Analyte immunoassays generally can be conducted using any
format known in the art, such as, but not limited to, a sandwich
format. Specifically, in one immunoassay format, at least two
antibodies are employed to separate and quantify analyte, such as
human analyte, or a fragment thereof in a sample. More
specifically, the at least two antibodies bind to different
epitopes on an analyte (or a fragment thereof) forming an immune
complex, which is referred to as a "sandwich." Generally, in the
immunoassays one or more antibodies can be used to capture the
analyte (or a fragment thereof) in the test sample (these
antibodies are frequently referred to as a "capture" antibody or
"capture" antibodies) and one or more antibodies can be used to
bind a detectable (namely, quantifiable) label to the sandwich
(these antibodies are frequently referred to as the "detection
antibody," the "detection antibodies," the "conjugate," or the
"conjugates"). Thus, in the context of a sandwich immunoassay
format, a DVD-binding protein (or a fragment, a variant, or a
fragment of a variant thereof) as described herein can be used as a
capture antibody, a detection antibody, or both. For example, one
DVD-binding protein having a domain that can bind a first epitope
on an analyte (or a fragment thereof) can be used as a capture
antibody and/or another DVD-binding protein having a domain that
can bind a second epitope on an analyte (or a fragment thereof) can
be used as a detection antibody. In this regard, a DVD-binding
protein having a first domain that can bind a first epitope on an
analyte (or a fragment thereof) and a second domain that can bind a
second epitope on an analyte (or a fragment thereof) can be used as
a capture antibody and/or a detection antibody. Alternatively, one
DVD-binding protein having a first domain that can bind an epitope
on a first analyte (or a fragment thereof) and a second domain that
can bind an epitope on a second analyte (or a fragment thereof) can
be used as a capture antibody and/or a detection antibody to
detect, and optionally quantify, two or more analytes. In the event
that an analyte can be present in a sample in more than one form,
such as a monomeric form and a dimeric/multimeric form, which can
be homomeric or heteromeric, one DVD-binding protein having a
domain that can bind an epitope that is only exposed on the
monomeric form and another DVD-binding protein having a domain that
can bind an epitope on a different part of a dimeric/multimeric
form can be used as capture antibodies and/or detection antibodies,
thereby enabling the detection, and optional quantification, of
different forms of a given analyte. Furthermore, employing
DVD-binding proteins with differential affinities within a single
DVD-binding protein and/or between DVD-binding proteins can provide
an avidity advantage. In the context of immunoassays as described
herein, it generally may be helpful or desired to incorporate one
or more linkers within the structure of a DVD-binding protein. When
present, optimally the linker should be of sufficient length and
structural flexibility to enable binding of an epitope by the inner
domains as well as binding of another epitope by the outer domains.
In this regard, if a DVD-binding protein can bind two different
analytes and one analyte is larger than the other, desirably the
larger analyte is bound by the outer domains.
[0520] Generally speaking, a sample being tested for (for example,
suspected of containing) an SOST protein (or a fragment thereof)
can be contacted with at least one capture antibody (or antibodies)
and at least one detection antibody (which can be a second
detection antibody or a third detection antibody or even a
successively numbered antibody, e.g., as where the capture and/or
detection antibody comprise multiple antibodies) either
simultaneously or sequentially and in any order. For example, the
test sample can be first contacted with at least one capture
antibody and then (sequentially) with at least one detection
antibody. Alternatively, the test sample can be first contacted
with at least one detection antibody and then (sequentially) with
at least one capture antibody. In yet another alternative, the test
sample can be contacted simultaneously with a capture antibody and
a detection antibody.
[0521] In the sandwich assay format, a sample suspected of
containing SOST (or a fragment thereof) is first brought into
contact with at least one first capture binding protein (e.g., SOST
antibody) under conditions that allow the formation of a first
binding protein/SOST complex. If more than one capture binding
protein is used, a first capture binding protein/SOST complex
comprising two or more capture binding proteins forms. In a
sandwich assay, the binding proteins, i.e., e.g., the at least one
capture binding protein, are used in molar excess amounts of the
maximum amount of SOST analyte (or a fragment thereof) expected in
the test sample. For example, from about 5 .mu.g to about 1 mg of
antibody per mL of buffer (e.g., microparticle coating buffer) can
be used.
[0522] Competitive inhibition immunoassays, which are often used to
measure small analytes because binding by only one antibody is
required, comprise sequential and classic formats. In a sequential
competitive inhibition immunoassay a capture binding protein to
sclerostin is coated onto a well of a microtiter plate or other
solid support. When the sample containing the sclerostin is added
to the well, the sclerostin binds to the capture binding protein.
After washing, a known amount of labeled (e.g., biotin or
horseradish peroxidase (HRP)) sclerostin is added to the well. A
substrate for an enzymatic label is necessary to generate a signal.
An example of a suitable substrate for HRP is
3,3',5,5'-tetramethylbenzidine (TMB). After washing, the signal
generated by the labeled analyte is measured and is inversely
proportional to the amount of sclerostin in the sample. In a
classic competitive inhibition immunoassay, a binding protein to
sclerostin is coated onto a solid support (e.g., a well of a
microtiter plate). However, unlike the sequential competitive
inhibition immunoassay, the sample and the labeled sclerostin are
added to the well at the same time. Any sclerostin in the sample
competes with labeled sclerostin for binding to the capture binding
protein. After washing, the signal generated by the labeled
sclerostin is measured and is inversely proportional to the amount
of sclerostin in the sample.
[0523] Optionally, prior to contacting the test sample with the at
least one capture binding protein (for example, the first capture
antibody), the at least one capture binding protein can be bound to
a solid support, which facilitates the separation of the first
binding protein/sclerostin (or a fragment thereof) complex from the
test sample. The substrate to which the capture binding protein is
bound can be any suitable solid support or solid phase that
facilitates separation of the capture antibody-analyte complex from
the sample.
[0524] Examples include a well of a plate, such as a microtiter
plate, a test tube, a porous gel (e.g., silica gel, agarose,
dextran, or gelatin), a polymeric film (e.g., polyacrylamide),
beads (e.g., polystyrene beads or magnetic beads), a strip of a
filter/membrane (e.g., nitrocellulose or nylon), microparticles
(e.g., latex particles, magnetizable microparticles (e.g.,
microparticles having ferric oxide or chromium oxide cores and
homo- or hetero-polymeric coats and radii of about 1-10 microns).
The substrate can comprise a suitable porous material with a
suitable surface affinity to bind antigens and sufficient porosity
to allow access by detection antibodies. A microporous material is
generally preferred, although a gelatinous material in a hydrated
state can be used. Such porous substrates are, e.g., in the form of
sheets having a thickness of about 0.01 to about 0.5 mm, e.g.,
about 0.1 mm. While the pore size may vary quite a bit, e.g., the
pore size is from about 0.025 to about 15 microns, e.g., from about
0.15 to about 15 microns. The surface of such substrates can be
activated by chemical processes that cause covalent linkage of an
antibody to the substrate. Irreversible binding, generally by
adsorption through hydrophobic forces, of the antigen or the
antibody to the substrate results; alternatively, a chemical
coupling agent or other means can be used to bind covalently the
antibody to the substrate, provided that such binding does not
interfere with the ability of the antibody to bind to analyte.
Alternatively, the antibody can be bound with microparticles, which
have been previously coated with streptavidin (e.g., DYNAL.RTM.
Magnetic Beads, Invitrogen, Carlsbad, Calif.) or biotin (e.g.,
using Power-Bind.TM.-SA-MP streptavidin-coated microparticles
(Seradyn, Indianapolis, Ind.)) or anti-species-specific monoclonal
antibodies. If necessary, the substrate can be derivatized to allow
reactivity with various functional groups on the antibody. Such
derivatization requires the use of certain coupling agents,
examples of which include, but are not limited to, maleic
anhydride, N-hydroxysuccinimide, and
1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. If desired, one or
more capture reagents, such as antibodies (or fragments thereof),
each of which is specific for analyte(s) can be attached to solid
phases in different physical or addressable locations (e.g., such
as in a biochip configuration (see, e.g., U.S. Pat. No. 6,225,047;
PCT Publication No. WO 99/51773; U.S. Pat. No. 6,329,209; PCT
Publication No. WO 00/56934; and U.S. Pat. No. 5,242,828). If the
capture reagent is attached to a mass spectrometry probe as the
solid support, the amount of analyte bound to the probe can be
detected by laser desorption ionization mass spectrometry.
Alternatively, a single column can be packed with different beads,
which are derivatized with the one or more capture reagents,
thereby capturing the analyte in a single place (see,
antibody-derivatized, bead-based technologies, e.g., the xMAP
technology of Luminex (Austin, Tex.)).
[0525] After the test sample being assayed for analyte (or a
fragment thereof) is brought into contact with the at least one
capture antibody (for example, the first capture antibody), the
mixture is incubated in order to allow for the formation of a first
antibody (or multiple antibody)-analyte (or a fragment thereof)
complex. The incubation can be carried out at a pH of from about
4.5 to about 10.0, at a temperature of from about 2.degree. C. to
about 45.degree. C., and for a period from at least about one (1)
minute to about eighteen (18) hours, e.g., from about 1 to about 24
minutes, e.g., for about 4 to about 18 minutes. The immunoassay
described herein can be conducted in one step (meaning the test
sample, at least one capture antibody and at least one detection
antibody are all added sequentially or simultaneously to a reaction
vessel) or in more than one step, such as two steps, three steps,
etc.
[0526] After formation of the (first or multiple) capture
antibody/analyte (or a fragment thereof) complex, the complex is
then contacted with at least one detection antibody under
conditions which allow for the formation of a (first or multiple)
capture antibody/analyte (or a fragment thereof)/second detection
antibody complex). While captioned for clarity as the "second"
antibody (e.g., second detection antibody), in fact, where multiple
antibodies are used for capture and/or detection, the at least one
detection antibody can be the second, third, fourth, etc.
antibodies used in the immunoassay. If the capture antibody/analyte
(or a fragment thereof) complex is contacted with more than one
detection antibody, then a (first or multiple) capture
antibody/analyte (or a fragment thereof)/(multiple) detection
antibody complex is formed. As with the capture antibody (e.g., the
first capture antibody), when the at least one (e.g., second and
any subsequent) detection antibody is brought into contact with the
capture antibody/analyte (or a fragment thereof) complex, a period
of incubation under conditions similar to those described above is
required for the formation of the (first or multiple) capture
antibody/analyte (or a fragment thereof)/(second or multiple)
detection antibody complex. In an embodiment, at least one
detection antibody contains a detectable label. The detectable
label can be bound to the at least one detection antibody (e.g.,
the second detection antibody) prior to, simultaneously with, or
after the formation of the (first or multiple) capture
antibody/analyte (or a fragment thereof)/(second or multiple)
detection antibody complex. Any detectable label known in the art
can be used (see discussion above, including of the Polak and Van
Noorden (1997) and Haugland (1996) references).
[0527] The detectable label can be bound to the antibodies either
directly or through a coupling agent. An example of a coupling
agent that can be used is EDAC (1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide, hydrochloride), which is commercially available from
Sigma-Aldrich, St. Louis, Mo. Other coupling agents that can be
used are known in the art. Methods for binding a detectable label
to an antibody are known in the art. Additionally, many detectable
labels can be purchased or synthesized that already contain end
groups that facilitate the coupling of the detectable label to the
antibody, such as CPSP-Acridinium Ester (i.e.,
9-[N-tosyl-N-(3-carboxypropyl)]-10-(3-sulfopropyl)acridinium
carboxamide) or SPSP-Acridinium Ester (i.e.,
N10-(3-sulfopropyl)-N-(3-sulfopropyl)-acridinium-9-carboxamide).
[0528] The (first or multiple) capture antibody/analyte/(second or
multiple) detection antibody complex can be, but does not have to
be, separated from the remainder of the test sample prior to
quantification of the label. For example, if the at least one
capture antibody (e.g., the first capture antibody) is bound to a
solid support, such as a well or a bead, separation can be
accomplished by removing the fluid (of the test sample) from
contact with the solid support. Alternatively, if the at least
first capture antibody is bound to a solid support, it can be
simultaneously contacted with the analyte-containing sample and the
at least one second detection antibody to form a first (multiple)
antibody/analyte/second (multiple) antibody complex, followed by
removal of the fluid (test sample) from contact with the solid
support. If the at least one first capture antibody is not bound to
a solid support, then the (first or multiple) capture
antibody/analyte/(second or multiple) detection antibody complex
does not have to be removed from the test sample for quantification
of the amount of the label.
[0529] After formation of the labeled capture
antibody/analyte/detection antibody complex (e.g., the first
capture antibody/analyte/second detection antibody complex), the
amount of label in the complex is quantified using techniques known
in the art. For example, if an enzymatic label is used, the labeled
complex is reacted with a substrate for the label that gives a
quantifiable reaction such as the development of color. If the
label is a radioactive label, the label is quantified using
appropriate means, such as a scintillation counter. If the label is
a fluorescent label, the label is quantified by stimulating the
label with a light of one color (which is known as the "excitation
wavelength") and detecting another color (which is known as the
"emission wavelength") that is emitted by the label in response to
the stimulation. If the label is a chemiluminescent label, the
label is quantified by detecting the light emitted either visually
or by using luminometers, x-ray film, high speed photographic film,
a CCD camera, etc. Once the amount of the label in the complex has
been quantified, the concentration of analyte or a fragment thereof
in the test sample is determined by appropriate means, such as by
use of a standard curve that has been generated using serial
dilutions of analyte or a fragment thereof of known concentration.
Other than using serial dilutions of analyte or a fragment thereof,
the standard curve can be generated gravimetrically, by mass
spectroscopy and by other techniques known in the art.
[0530] In a chemiluminescent microparticle assay employing the
ARCHITECT.RTM. analyzer, the conjugate diluent pH should be about
6.0+/-0.2, the microparticle coating buffer should be maintained at
about room temperature (i.e., at from about 17 to about 27 {hacek
over (.differential.)}C), the microparticle coating buffer pH
should be about 6.5+/-0.2, and the microparticle diluent pH should
be about 7.8+/-0.2. In an embodiment, solids are less than about
0.2%, such as less than about 0.15%, less than about 0.14%, less
than about 0.13%, less than about 0.12%, or less than about 0.11%,
such as about 0.10%.
[0531] FPIAs are based on competitive binding immunoassay
principles. A fluorescently labeled compound, when excited by a
linearly polarized light, will emit fluorescence having a degree of
polarization inversely proportional to its rate of rotation. When a
fluorescently labeled tracer-antibody complex is excited by a
linearly polarized light, the emitted light remains highly
polarized because the fluorophore is constrained from rotating
between the time light is absorbed and the time light is emitted.
When a "free" tracer compound (i.e., a compound that is not bound
to an antibody) is excited by linearly polarized light, its
rotation is much faster than the corresponding tracer-antibody
conjugate produced in a competitive binding immunoassay. FPIAs are
advantageous over RIAs inasmuch as there are no radioactive
substances requiring special handling and disposal. In addition,
FPIAs are homogeneous assays that can be easily and rapidly
performed.
[0532] In view of the above, a method of determining the presence,
amount, or concentration of analyte (or a fragment thereof) in a
test sample is provided. The method comprises assaying the test
sample for an analyte (or a fragment thereof) by an assay (i)
employing (i') at least one of an antibody, a fragment of an
antibody that can bind to an analyte, a variant of an antibody that
can bind to an analyte, a fragment of a variant of an antibody that
can bind to an analyte, and a DVD-binding protein (or a fragment, a
variant, or a fragment of a variant thereof) that can bind to an
analyte, and (ii') at least one detectable label and (ii)
comprising comparing a signal generated by the detectable label as
a direct or indirect indication of the presence, amount or
concentration of analyte (or a fragment thereof) in the test sample
to a signal generated as a direct or indirect indication of the
presence, amount or concentration of analyte (or a fragment
thereof) in a control or calibrator. The calibrator is optionally
part of a series of calibrators, in which each of the calibrators
differs from the other calibrators by the concentration of
analyte.
[0533] The method can comprise (i) contacting the test sample with
at least one first specific binding partner for analyte (or a
fragment thereof) of an antibody, a fragment of an antibody that
can bind to an analyte, a variant of an antibody that can bind to
an analyte, a fragment of a variant of an antibody that can bind to
an analyte, or a DVD-binding protein (or a fragment, a variant, or
a fragment of a variant thereof) that can bind to an analyte so as
to form a first specific binding partner/analyte (or fragment
thereof) complex, (ii) contacting the first specific binding
partner/analyte (or fragment thereof) complex with at least one
second specific binding partner for analyte (or fragment thereof)
of a detectably labeled anti-analyte antibody, a detectably labeled
fragment of an anti-analyte antibody that can bind to analyte, a
detectably labeled variant of an anti-analyte antibody that can
bind to analyte, a detectably labeled fragment of a variant of an
anti-analyte antibody that can bind to analyte, or a detectably
labeled DVD-binding protein (or a fragment, a variant, or a
fragment of a variant thereof) so as to form a first specific
binding partner/analyte (or fragment thereof)/second specific
binding partner complex, and (iii) determining the presence, amount
or concentration of analyte in the test sample by detecting or
measuring the signal generated by the detectable label in the first
specific binding partner/analyte (or fragment thereof)/second
specific binding partner complex formed in (ii). A method in which
at least one first specific binding partner for analyte (or a
fragment thereof) and/or at least one second specific binding
partner for analyte (or a fragment thereof) is a DVD-binding
protein (or a fragment, a variant, or a fragment of a variant
thereof) as described herein can be preferred.
[0534] Alternatively, the method can comprise contacting the test
sample with at least one first specific binding partner for an SOST
analyte (or a fragment thereof) of an antibody, a fragment of an
antibody that can bind to an analyte, a variant of an antibody that
can bind to an analyte, a fragment of a variant of an antibody that
can bind to an analyte, or a DVD-binding protein (or a fragment, a
variant, or a fragment of a variant thereof) and simultaneously or
sequentially, in either order, contacting the test sample with at
least one second specific binding partner, which can compete with
analyte (or a fragment thereof) for binding to the at least one
first specific binding partner and which is a detectably labeled
analyte, a detectably labeled fragment of analyte that can bind to
the first specific binding partner, a detectably labeled variant of
analyte that can bind to the first specific binding partner, or a
detectably labeled fragment of a variant of analyte that can bind
to the first specific binding partner. Any SOST (or a fragment
thereof) present in the test sample and the at least one second
specific binding partner compete with each other to form a first
specific binding partner/analyte (or fragment thereof) complex and
a first specific binding partner/second specific binding partner
complex, respectively. The method further comprises determining the
presence, amount or concentration of analyte in the test sample by
detecting or measuring the signal generated by the detectable label
in the first specific binding partner/second specific binding
partner complex formed in (ii), wherein the signal generated by the
detectable label in the first specific binding partner/second
specific binding partner complex is inversely proportional to the
amount or concentration of analyte in the test sample.
[0535] The above methods can further comprise diagnosing,
prognosticating, or assessing the efficacy of a
therapeutic/prophylactic treatment of a patient from whom the test
sample was obtained. If the method further comprises assessing the
efficacy of a therapeutic/prophylactic treatment of the patient
from whom the test sample was obtained, the method optionally
further comprises modifying the therapeutic/prophylactic treatment
of the patient as needed to improve efficacy. The method can be
adapted for use in an automated system or a semi-automated
system.
[0536] With regard to the methods of assay (and kit therefor), it
may be possible to employ commercially available anti-analyte
antibodies or methods for production of anti-analyte as described
in the literature. Commercial supplies of various antibodies
include, but are not limited to, Santa Cruz Biotechnology Inc.
(Santa Cruz, Calif.), GenWay Biotech, Inc. (San Diego, Calif.), and
R&D Systems (RDS; Minneapolis, Minn.).
[0537] Generally, a predetermined level can be employed as a
benchmark against which to assess results obtained upon assaying a
test sample for analyte or a fragment thereof, e.g., for detecting
disease or risk of disease. Generally, in making such a comparison,
the predetermined level is obtained by running a particular assay a
sufficient number of times and under appropriate conditions such
that a linkage or association of analyte presence, amount or
concentration with a particular stage or endpoint of a disease,
disorder or condition or with particular clinical indicia can be
made. Typically, the predetermined level is obtained with assays of
reference subjects (or populations of subjects). The analyte
measured can include fragments thereof, degradation products
thereof, and/or enzymatic cleavage products thereof.
[0538] In particular, with respect to a predetermined level as
employed for monitoring disease progression and/or treatment, the
amount or concentration of analyte or a fragment thereof may be
"unchanged," "favorable" (or "favorably altered"), or "unfavorable"
(or "unfavorably altered"). "Elevated" or "increased" refers to an
amount or a concentration in a test sample that is higher than a
typical or normal level or range (e.g., predetermined level), or is
higher than another reference level or range (e.g., earlier or
baseline sample). The term "lowered" or "reduced" refers to an
amount or a concentration in a test sample that is lower than a
typical or normal level or range (e.g., predetermined level), or is
lower than another reference level or range (e.g., earlier or
baseline sample). The term "altered" refers to an amount or a
concentration in a sample that is altered (increased or decreased)
over a typical or normal level or range (e.g., predetermined
level), or over another reference level or range (e.g., earlier or
baseline sample).
[0539] The typical or normal level or range for analyte is defined
in accordance with standard practice. Because the levels of analyte
in some instances will be very low, a so-called altered level or
alteration can be considered to have occurred when there is any net
change as compared to the typical or normal level or range, or
reference level or range, that cannot be explained by experimental
error or sample variation. Thus, the level measured in a particular
sample will be compared with the level or range of levels
determined in similar samples from a so-called normal subject. In
this context, a "normal subject" is an individual with no
detectable disease, for example, and a "normal" (sometimes termed
"control") patient or population is/are one(s) that exhibit(s) no
detectable disease, respectively, for example. Furthermore, given
that analyte is not routinely found at a high level in the majority
of the human population, a "normal subject" can be considered an
individual with no substantial detectable increased or elevated
amount or concentration of analyte, and a "normal" (sometimes
termed "control") patient or population is/are one(s) that
exhibit(s) no substantial detectable increased or elevated amount
or concentration of analyte. An "apparently normal subject" is one
in which analyte has not yet been or currently is being assessed.
The level of an analyte is said to be "elevated" when the analyte
is normally undetectable (e.g., the normal level is zero, or within
a range of from about 25 to about 75 percentiles of normal
populations), but is detected in a test sample, as well as when the
analyte is present in the test sample at a higher than normal
level. Thus, inter alia, the disclosure provides a method of
screening for a subject having, or at risk of having, a particular
disease, disorder, or condition. The method of assay can also
involve the assay of other markers and the like.
[0540] Accordingly, the methods described herein also can be used
to determine whether or not a subject has or is at risk of
developing a given disease, disorder or condition. Specifically,
such a method can comprise the steps of:
[0541] (a) determining the concentration or amount in a test sample
from a subject of sclerostin (or a fragment thereof) (e.g., using
the methods described herein, or methods known in the art); and
[0542] (b) comparing the concentration or amount of sclerostin (or
a fragment thereof) determined in step (a) with a predetermined
level, wherein, if the concentration or amount of analyte
determined in step (a) is favorable with respect to a predetermined
level, then the subject is determined not to have or be at risk for
a given disease, disorder or condition. However, if the
concentration or amount of sclerostin determined in step (a) is
unfavorable with respect to the predetermined level, then the
subject is determined to have or be at risk for a given disease,
disorder or condition.
[0543] Additionally, provided herein is method of monitoring the
progression of disease in a subject. Optimally the method
comprising the steps of:
[0544] (a) determining the concentration or amount in a test sample
from a subject of sclerostin;
[0545] (b) determining the concentration or amount in a later test
sample from the subject of SOST; and
[0546] (c) comparing the concentration or amount of analyte as
determined in step (b) with the concentration or amount of
sclerostin determined in step (a), wherein if the concentration or
amount determined in step (b) is unchanged or is unfavorable when
compared to the concentration or amount of sclerostin determined in
step (a), then the disease in the subject is determined to have
continued, progressed or worsened. By comparison, if the
concentration or amount of sclerostin as determined in step (b) is
favorable when compared to the concentration or amount of
sclerostin as determined in step (a), then the disease in the
subject is determined to have discontinued, regressed or
improved.
[0547] Optionally, the method further comprises comparing the
concentration or amount of sclerostin analyte as determined in step
(b), for example, with a predetermined level. Further, optionally
the method comprises treating the subject with one or more
pharmaceutical compositions for a period of time if the comparison
shows that the concentration or amount of analyte as determined in
step (b), for example, is unfavorably altered with respect to the
predetermined level.
[0548] Still further, the methods can be used to monitor treatment
in a subject receiving treatment with one or more pharmaceutical
compositions. Specifically, such methods involve providing a first
test sample from a subject before the subject has been administered
one or more pharmaceutical compositions. Next, the concentration or
amount in a first test sample from a subject of sclerostin is
determined (e.g., using the methods described herein or as known in
the art). After the concentration or amount of sclerostin is
determined, optionally the concentration or amount of sclerostin is
then compared with a predetermined level. If the concentration or
amount of sclerostin as determined in the first test sample is
lower than the predetermined level, then the subject is not treated
with one or more pharmaceutical compositions. However, if the
concentration or amount of sclerostin as determined in the first
test sample is higher than the predetermined level, then the
subject is treated with one or more pharmaceutical compositions for
a period of time. The period of time that the subject is treated
with the one or more pharmaceutical compositions can be determined
by one skilled in the art (for example, the period of time can be
from about seven (7) days to about two years, e.g., from about
fourteen (14) days to about one (1) year).
[0549] During the course of treatment with the one or more
pharmaceutical compositions, second and subsequent test samples are
then obtained from the subject. The number of test samples and the
time in which said test samples are obtained from the subject are
not critical. For example, a second test sample could be obtained
seven (7) days after the subject is first administered the one or
more pharmaceutical compositions, a third test sample could be
obtained two (2) weeks after the subject is first administered the
one or more pharmaceutical compositions, a fourth test sample could
be obtained three (3) weeks after the subject is first administered
the one or more pharmaceutical compositions, a fifth test sample
could be obtained four (4) weeks after the subject is first
administered the one or more pharmaceutical compositions, etc.
[0550] After each second or subsequent test sample is obtained from
the subject, the concentration or amount of sclerostin analyte is
determined in the second or subsequent test sample is determined
(e.g., using the methods described herein or as known in the art).
The concentration or amount of sclerostin as determined in each of
the second and subsequent test samples is then compared with the
concentration or amount of analyte as determined in the first test
sample (e.g., the test sample that was originally optionally
compared to the predetermined level). If the concentration or
amount of sclerostin as determined in step (c) is favorable when
compared to the concentration or amount of analyte as determined in
step (a), then the disease in the subject is determined to have
discontinued, regressed or improved, and the subject should
continue to be administered the one or pharmaceutical compositions
of step (b). However, if the concentration or amount determined in
step (c) is unchanged or is unfavorable when compared to the
concentration or amount of analyte as determined in step (a), then
the disease in the subject is determined to have continued,
progressed or worsened, and the subject should be treated with a
higher concentration of the one or more pharmaceutical compositions
administered to the subject in step (b) or the subject should be
treated with one or more pharmaceutical compositions that are
different from the one or more pharmaceutical compositions
administered to the subject in step (b). Specifically, the subject
can be treated with one or more pharmaceutical compositions that
are different from the one or more pharmaceutical compositions that
the subject had previously received to decrease or lower said
subject's analyte level.
[0551] Generally, for assays in which repeat testing may be done
(e.g., monitoring disease progression and/or response to
treatment), a second or subsequent test sample is obtained at a
period in time after the first test sample has been obtained from
the subject. Specifically, a second test sample from the subject
can be obtained minutes, hours, days, weeks or years after the
first test sample has been obtained from the subject. For example,
the second test sample can be obtained from the subject at a time
period of about 1 minute, about 5 minutes, about 10 minutes, about
15 minutes, about 30 minutes, about 45 minutes, about 60 minutes,
about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6
hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours,
about 11 hours, about 12 hours, about 13 hours, about 14 hours,
about 15 hours, about 16 hours, about 17 hours, about 18 hours,
about 19 hours, about 20 hours, about 21 hours, about 22 hours,
about 23 hours, about 24 hours, about 2 days, about 3 days, about 4
days, about 5 days, about 6 days, about 7 days, about 2 weeks,
about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7
weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11
weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15
weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19
weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23
weeks, about 24 weeks, about 25 weeks, about 26 weeks, about 27
weeks, about 28 weeks, about 29 weeks, about 30 weeks, about 31
weeks, about 32 weeks, about 33 weeks, about 34 weeks, about 35
weeks, about 36 weeks, about 37 weeks, about 38 weeks, about 39
weeks, about 40 weeks, about 41 weeks, about 42 weeks, about 43
weeks, about 44 weeks, about 45 weeks, about 46 weeks, about 47
weeks, about 48 weeks, about 49 weeks, about 50 weeks, about 51
weeks, about 52 weeks, about 1.5 years, about 2 years, about 2.5
years, about 3.0 years, about 3.5 years, about 4.0 years, about 4.5
years, about 5.0 years, about 5.5. years, about 6.0 years, about
6.5 years, about 7.0 years, about 7.5 years, about 8.0 years, about
8.5 years, about 9.0 years, about 9.5 years or about 10.0 years
after the first test sample from the subject is obtained.
[0552] When used to monitor disease progression, the above assay
can be used to monitor the progression of disease in subjects
suffering from acute conditions. Acute conditions, also known as
critical care conditions, refer to acute, life-threatening diseases
or other critical medical conditions involving, for example, the
cardiovascular system or excretory system. Typically, critical care
conditions refer to those conditions requiring acute medical
intervention in a hospital-based setting (including, but not
limited to, the emergency room, intensive care unit, trauma center,
or other emergent care setting) or administration by a paramedic or
other field-based medical personnel. For critical care conditions,
repeat monitoring is generally done within a shorter time frame,
namely, minutes, hours or days (e.g., about 1 minute, about 5
minutes, about 10 minutes, about 15 minutes, about 30 minutes,
about 45 minutes, about 60 minutes, about 2 hours, about 3 hours,
about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8
hours, about 9 hours, about 10 hours, about 11 hours, about 12
hours, about 13 hours, about 14 hours, about 15 hours, about 16
hours, about 17 hours, about 18 hours, about 19 hours, about 20
hours, about 21 hours, about 22 hours, about 23 hours, about 24
hours, about 2 days, about 3 days, about 4 days, about 5 days,
about 6 days or about 7 days), and the initial assay likewise is
generally done within a shorter timeframe, e.g., about minutes,
hours or days of the onset of the disease or condition.
[0553] The assays also can be used to monitor the progression of
disease in subjects suffering from chronic or non-acute conditions.
Non-critical care or, non-acute conditions, refers to conditions
other than acute, life-threatening disease or other critical
medical conditions involving, for example, the cardiovascular
system and/or excretory system. Typically, non-acute conditions
include those of longer-term or chronic duration. For non-acute
conditions, repeat monitoring generally is done with a longer
timeframe, e.g., hours, days, weeks, months or years (e.g., about 1
hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours,
about 6 hours, about 7 hours, about 8 hours, about 9 hours, about
10 hours, about 11 hours, about 12 hours, about 13 hours, about 14
hours, about 15 hours, about 16 hours, about 17 hours, about 18
hours, about 19 hours, about 20 hours, about 21 hours, about 22
hours, about 23 hours, about 24 hours, about 2 days, about 3 days,
about 4 days, about 5 days, about 6 days, about 7 days, about 2
weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks,
about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about
11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15
weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19
weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23
weeks, about 24 weeks, about 25 weeks, about 26 weeks, about 27
weeks, about 28 weeks, about 29 weeks, about 30 weeks, about 31
weeks, about 32 weeks, about 33 weeks, about 34 weeks, about 35
weeks, about 36 weeks, about 37 weeks, about 38 weeks, about 39
weeks, about 40 weeks, about 41 weeks, about 42 weeks, about 43
weeks, about 44 weeks, about 45 weeks, about 46 weeks, about 47
weeks, about 48 weeks, about 49 weeks, about 50 weeks, about 51
weeks, about 52 weeks, about 1.5 years, about 2 years, about 2.5
years, about 3.0 years, about 3.5 years, about 4.0 years, about 4.5
years, about 5.0 years, about 5.5. years, about 6.0 years, about
6.5 years, about 7.0 years, about 7.5 years, about 8.0 years, about
8.5 years, about 9.0 years, about 9.5 years or about 10.0 years),
and the initial assay likewise generally is done within a longer
time frame, e.g., about hours, days, months or years of the onset
of the disease or condition.
[0554] Furthermore, the above assays can be performed using a first
test sample obtained from a subject where the first test sample is
obtained from one source, such as urine, serum or plasma.
Optionally, the above assays can then be repeated using a second
test sample obtained from the subject where the second test sample
is obtained from another source. For example, if the first test
sample was obtained from urine, the second test sample can be
obtained from serum or plasma. The results obtained from the assays
using the first test sample and the second test sample can be
compared. The comparison can be used to assess the status of a
disease or condition in the subject.
[0555] Moreover, the present disclosure also relates to methods of
determining whether a subject predisposed to or suffering from a
given disease, disorder or condition will benefit from treatment.
In particular, the disclosure relates to analyte companion
diagnostic methods and products. Thus, the method of "monitoring
the treatment of disease in a subject" as described herein further
optimally also can encompass selecting or identifying candidates
for therapy.
[0556] Thus, in particular embodiments, the disclosure also
provides a method of determining whether a subject having, or at
risk for, a given disease, disorder or condition is a candidate for
therapy. Generally, the subject is one who has experienced some
symptom of a given disease, disorder or condition or who has
actually been diagnosed as having, or being at risk for, a given
disease, disorder or condition, and/or who demonstrates an
unfavorable concentration or amount of analyte or a fragment
thereof, as described herein.
[0557] The method optionally comprises an assay as described
herein, where SOST is assessed before and following treatment of a
subject with one or more pharmaceutical compositions (e.g.,
particularly with a pharmaceutical related to a mechanism of action
involving analyte), with immunosuppressive therapy, or by
immunoabsorption therapy, or where analyte is assessed following
such treatment and the concentration or the amount of analyte is
compared against a predetermined level. An unfavorable
concentration of amount of SOST observed following treatment
confirms that the subject will not benefit from receiving further
or continued treatment, whereas a favorable concentration or amount
of analyte observed following treatment confirms that the subject
will benefit from receiving further or continued treatment. This
confirmation assists with management of clinical studies, and
provision of improved patient care.
[0558] It goes without saying that, while certain embodiments
herein are advantageous when employed to assess a given disease,
disorder or condition as discussed herein, the assays and kits can
be employed to assess analyte in other diseases, disorders and
conditions. The method of assay can also involve the assay of other
markers and the like.
[0559] The method of assay also can be used to identify a compound
that ameliorates a given disease, disorder or condition. For
example, a cell that expresses analyte can be contacted with a
candidate compound. The level of expression of analyte in the cell
contacted with the compound can be compared to that in a control
cell using the method of assay described herein.
II. Kits
[0560] A kit for assaying a test sample for the presence, amount or
concentration of an analyte (or a fragment thereof) in a test
sample is also provided. The kit comprises at least one component
for assaying the test sample for sclerostin (or a fragment thereof)
and instructions for assaying the test sample for the analyte (or a
fragment thereof). The at least one component for assaying the test
sample for the analyte (or a fragment thereof) can include a
composition comprising an anti-sclerostin binding protein, such as
a monoclonal antibody or DVD-binding protein (or a fragment, a
variant, or a fragment of a variant thereof), as described herein
and which is optionally immobilized on a solid phase.
[0561] The kit can comprise at least one component for assaying the
test sample for an SOST analyte by immunoassay, e.g.,
chemiluminescent microparticle immunoassay, and instructions for
assaying the test sample for an SOST analyte by immunoassay, e.g.,
chemiluminescent microparticle immunoassay. For example, the kit
can comprise at least one specific binding partner for SOST, such
as an anti-sclerostin monoclonal/polyclonal antibody (or a fragment
thereof that can bind to the SOST analyte, a variant thereof that
can bind to the analyte, or a fragment of a variant that can bind
to the analyte) or an anti-sclerostin DVD-binding protein (or a
fragment, a variant, or a fragment of a variant thereof), either of
which can be detectably labeled. Alternatively or additionally, the
kit can comprise detectably labeled SOST analyte (or a fragment
thereof that can bind to an anti-analyte, monoclonal/polyclonal
antibody or an anti-analyte DVD-binding protein (or a fragment, a
variant, or a fragment of a variant thereof)), which can compete
with any analyte in a test sample for binding to an anti-analyte
monoclonal/polyclonal antibody (or a fragment thereof that can bind
to the analyte, a variant thereof that can bind to the analyte, or
a fragment of a variant that can bind to the analyte) or an
anti-analyte DVD-binding protein (or a fragment, a variant, or a
fragment of a variant thereof), either of which can be immobilized
on a solid support. The kit can comprise a calibrator or control,
e.g., isolated or purified analyte. The kit can comprise at least
one container (e.g., tube, microtiter plates or strips, which can
be already coated with a first specific binding partner, for
example) for conducting the assay, and/or a buffer, such as an
assay buffer or a wash buffer, either one of which can be provided
as a concentrated solution, a substrate solution for the detectable
label (e.g., an enzymatic label), or a stop solution. In an
embodiment, the kit comprises all components, i.e., reagents,
standards, buffers, diluents, etc., which are necessary to perform
the assay. The instructions can be in paper form or
computer-readable form, such as a disk, CD, DVD, or the like.
[0562] Any binding protein, such as an anti-sclerostin binding
protein or an anti-analyte DVD-binding protein, or tracer can
incorporate a detectable label as described herein, such as a
fluorophore, a radioactive moiety, an enzyme, a biotin/avidin
label, a chromophore, a chemiluminescent label, or the like, or the
kit can include reagents for carrying out detectable labeling. The
antibodies, calibrators and/or controls can be provided in separate
containers or pre-dispensed into an appropriate assay format, for
example, into microtiter plates.
[0563] Optionally, the kit includes quality control components (for
example, sensitivity panels, calibrators, and positive controls).
Preparation of quality control reagents is well-known in the art
and is described on insert sheets for a variety of immunodiagnostic
products. Sensitivity panel members optionally are used to
establish assay performance characteristics, and further optionally
are useful indicators of the integrity of the immunoassay kit
reagents, and the standardization of assays.
[0564] The kit can also optionally include other reagents required
to conduct a diagnostic assay or facilitate quality control
evaluations, such as buffers, salts, enzymes, enzyme co-factors,
enzyme substrates, detection reagents, and the like. Other
components, such as buffers and solutions for the isolation and/or
treatment of a test sample (e.g., pretreatment reagents), also can
be included in the kit. The kit can additionally include one or
more other controls. One or more of the components of the kit can
be lyophilized, in which case the kit can further comprise reagents
suitable for the reconstitution of the lyophilized components.
[0565] The various components of the kit optionally are provided in
suitable containers as necessary, e.g., a microtiter plate. The kit
can further include containers for holding or storing a sample
(e.g., a container or cartridge for a urine sample). Where
appropriate, the kit optionally also can contain reaction vessels,
mixing vessels, and other components that facilitate the
preparation of reagents or the test sample. The kit can also
include one or more instruments for assisting with obtaining a test
sample, such as a syringe, pipette, forceps, measured spoon, or the
like.
[0566] If the detectable label is at least one acridinium compound,
the kit can comprise at least one acridinium-9-carboxamide, at
least one acridinium-9-carboxylate aryl ester, or any combination
thereof. If the detectable label is at least one acridinium
compound, the kit also can comprise a source of hydrogen peroxide,
such as a buffer, a solution, and/or at least one basic solution.
If desired, the kit can contain a solid phase, such as a magnetic
particle, bead, test tube, microtiter plate, cuvette, membrane,
scaffolding molecule, film, filter paper, disc or chip.
III. Adaptation of Kit and Method
[0567] The kit (or components thereof), as well as the method of
determining the presence, amount or concentration of an analyte in
a test sample by an assay, such as an immunoassay as described
herein, can be adapted for use in a variety of automated and
semi-automated systems (including those wherein the solid phase
comprises a microparticle), as described, e.g., in U.S. Pat. Nos.
5,089,424 and 5,006,309, and as commercially marketed, e.g., by
Abbott Laboratories (Abbott Park, Ill.) as ARCHITECT.RTM..
[0568] Some of the differences between an automated or
semi-automated system as compared to a non-automated system (e.g.,
ELISA) include the substrate to which the first specific binding
partner (e.g., an anti-analyte, monoclonal/polyclonal antibody (or
a fragment thereof, a variant thereof, or a fragment of a variant
thereof) or an anti-analyte DVD-binding protein (or a fragment
thereof, a variant thereof, or a fragment of a variant thereof) is
attached; either way, sandwich formation and analyte reactivity can
be impacted), and the length and timing of the capture, detection
and/or any optional wash steps. Whereas a non-automated format,
such as an ELISA, may require a relatively longer incubation time
with sample and capture reagent (e.g., about 2 hours), an automated
or semi-automated format (e.g., ARCHITECT.RTM., Abbott
Laboratories) may have a relatively shorter incubation time (e.g.,
approximately 18 minutes for ARCHITECT.RTM.). Similarly, whereas a
non-automated format, such as an ELISA, may incubate a detection
antibody, such as the conjugate reagent, for a relatively longer
incubation time (e.g., about 2 hours), an automated or
semi-automated format (e.g., ARCHITECT.RTM.) may have a relatively
shorter incubation time (e.g., approximately 4 minutes for the
ARCHITECT.RTM.).
[0569] Other platforms available from Abbott Laboratories include,
but are not limited to, AxSYM.RTM., IMx.RTM. (see, e.g., U.S. Pat.
No. 5,294,404), PRISM.RTM., EIA (bead), and Quantum.TM. II, as well
as other platforms. Additionally, the assays, kits and kit
components can be employed in other formats, for example, on
electrochemical or other hand-held or point-of-care assay systems.
The present disclosure is, for example, applicable to the
commercial Abbott Point of Care (i-STAT.RTM., Abbott Laboratories)
electrochemical immunoassay system that performs sandwich
immunoassays Immunosensors and their methods of manufacture and
operation in single-use test devices are described, for example in,
U.S. Pat. No. 5,063,081, US Patent Application Publication No.
2003/0170881, US Patent Application Publication No. 2004/0018577,
US Patent Application Publication No. 2005/0054078, and US Patent
Application Publication No. 2006/0160164.
[0570] In particular, with regard to the adaptation of an analyte
assay to the I-STAT.RTM. system, the following configuration is
exemplary. A microfabricated silicon chip is manufactured with a
pair of gold amperometric working electrodes and a silver-silver
chloride reference electrode. On one of the working electrodes,
polystyrene beads (0.2 mm diameter) with immobilized anti-analyte,
monoclonal/polyclonal antibody (or a fragment thereof, a variant
thereof, or a fragment of a variant thereof) or anti-analyte
DVD-binding protein (or a fragment thereof, a variant thereof, or a
fragment of a variant thereof), are adhered to a polymer coating of
patterned polyvinyl alcohol over the electrode. This chip is
assembled into an I-STAT.RTM. cartridge with a fluidics format
suitable for immunoassay. On a portion of the wall of the
sample-holding chamber of the cartridge there is a layer comprising
a specific binding partner for an analyte, such as an anti-analyte,
monoclonal/polyclonal antibody (or a fragment thereof, a variant
thereof, or a fragment of a variant thereof that can bind the
analyte) or an anti-analyte DVD-binding protein (or a fragment
thereof, a variant thereof, or a fragment of a variant thereof that
can bind the analyte), either of which can be detectably labeled.
Within the fluid pouch of the cartridge is an aqueous reagent that
includes p-aminophenol phosphate.
[0571] In operation, a sample suspected of containing an analyte is
added to the holding chamber of the test cartridge, and the
cartridge is inserted into the I-STAT.RTM. reader. After the
specific binding partner for an analyte has dissolved into the
sample, a pump element within the cartridge forces the sample into
a conduit containing the chip. Here it is oscillated to promote
formation of the sandwich. In the penultimate step of the assay,
fluid is forced out of the pouch and into the conduit to wash the
sample off the chip and into a waste chamber. In the final step of
the assay, the alkaline phosphatase label reacts with p-aminophenol
phosphate to cleave the phosphate group and permit the liberated
p-aminophenol to be electrochemically oxidized at the working
electrode. Based on the measured current, the reader is able to
calculate the amount of analyte in the sample by means of an
embedded algorithm and factory-determined calibration curve.
[0572] It further goes without saying that the methods and kits as
described herein necessarily encompass other reagents and methods
for carrying out the immunoassay. For instance, encompassed are
various buffers such as are known in the art and/or which can be
readily prepared or optimized to be employed, e.g., for washing, as
a conjugate diluent, microparticle diluent, and/or as a calibrator
diluent. An exemplary conjugate diluent is ARCHITECT.RTM. conjugate
diluent employed in certain kits (Abbott Laboratories, Abbott Park,
Ill.) and containing 2-(N-morpholino)ethanesulfonic acid (MES), a
salt, a protein blocker, an antimicrobial agent, and a detergent.
An exemplary calibrator diluent is ARCHITECT.RTM. human calibrator
diluent employed in certain kits (Abbott Laboratories, Abbott Park,
Ill.), which comprises a buffer containing MES, other salt, a
protein blocker, and an antimicrobial agent. Additionally, as
described in U.S. patent application Ser. No. 12/650,241 (see, also
PCT/US2009/069846), improved signal generation may be obtained,
e.g., in an I-Stat cartridge format, using a nucleic acid sequence
linked to the signal antibody as a signal amplifier.
[0573] It will be readily apparent to those skilled in the art that
other suitable modifications and adaptations of the methods
described herein are obvious and may be made using suitable
equivalents without departing from the scope or the embodiments
disclosed herein.
[0574] Having now described that which is provided in detail, the
same will be more clearly understood by reference to the following
examples, which are included for purposes of illustration only and
are not intended to be limiting.
EXAMPLES
Example 1
Anti-Human SOST Antibodies
Example 1.1
Identification of Fully Human Binding Proteins to Sclerostin by In
Vitro Display Systems
Example 1.1.1
Antibody Selections
[0575] Fully human anti-human sclerostin monoclonal antibodies were
isolated by in vitro display technologies from human antibody
libraries by their ability to bind recombinant human sclerostin
proteins. The amino acid sequences of the variable heavy (VH) and
variable light (VL) chains were determined from DNA sequencing.
Example 1.1.2
Affinity Maturation of the Fully Human Anti-Human Sclerostin
Binding Protein AE10-6
[0576] The AE10-6 human binding protein to human sclerostin was
affinity matured by in vitro display technology. Sequence alignment
shows that the Sclerostin antibody AE10-6 shares the highest
identity to human germlines VH1-24/JH1 and IGKV7-46/JL2. To improve
the affinity of AE10-6 to Sclerostin, hypermutated CDR residues
were identified from other human antibody sequences in the IgBLAST
database that also shared high identity to germlines VH1-24 and
IGKV7-46. The corresponding AE10-6 CDR residues were then subjected
to limited mutagenesis by PCR with primers having low degeneracy at
these positions to create two antibody libraries in the scFv format
suitable for surface display. The first library contained mutations
at residues 34, 51, 54, 57 and 95 to 100c in the VH CDR1, 2 and 3
(Kabat numbering); the second library at residues 27b, 29, 30, 52,
53, 55 and 91 to 96 in the three VL CDRs. To further increase the
identity of AE10-6 to the human germline framework sequences, a
binary degeneracy at VH positions 30 (T/S), 50 (G/R) and 52 (D/N)
were introduced into the first library. Also, VH position 105 was
germlined (P/Q) in the first library. Binary degeneracy at VL
positions 24 (K/R), 33 (V/L), 54 (R/L), 55 (H/Q), 56 (T/S), 91
(H/S) and 96 (F/Y) were introduced into the second library. (see
table 1). Also, VL position 2 was germlined (T/A) in the second
library.
[0577] The table below (Table 5) provides a list of amino acid
sequences of VH and VL of the fully human AE10-6 binding protein
which were subjected to the affinity maturation selection protocol
Amino acid residues of individual CDRs of each VH and VL sequence
are indicated in bold.
TABLE-US-00006 TABLE 5 Amino acid residues found during the
affinity maturation of anti-Sclerostin antibody AE10-6. AE10-6
Heavy chain variable region (SEQ ID NO: 1) SOST 1 2 3 4 5 6 AE10-6
1234567890123456789012345678901234567890123456789012a345678901
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQ SG V
SN E I L V L Y M N N G A R Q H F I 1 1 7 8 9 0 1
234567890123456789012abc345678901234567890abcd1234567890123
KFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCATDSEGYWEKYFQHWGPGTLVTVSS F I M
ETDSF YQF Q AV R FI A N V L V Y S W P Q I N M G AE10-6 Light chain
variable region (SEQ ID NO: 2) SOST 1 2 3 4 5 AE10-6
1234567891234567abc89012345678901234567890123456789012345678
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGHYPYWFQQKPGQAPRTLISDTNDKHSWT D IE T
FVY DE Q P VD I N G TW R D S GY V N MN T E T H 1 6 7 8 9 0
901234567890123456789012345678901234567890123456a
PARFSGSLLGGKAALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTVL DDRSSL N F NM N
RF
[0578] These AE10-6 libraries were transformed and displayed on
cell surfaces to be selected against a low concentration of
biotinylated Sclerostin by magnetic then fluorescence activated
cell sorting. Selections to improve on-rate, off-rate, or both were
carried out and antibody protein sequences of affinity-modulated
AE10-6 clones were recovered from cells and converted back to IgG
format for further characterization.
[0579] The tables below provides a list of amino acid sequences of
VH (Table 6) and VL (Table 7) regions of affinity matured fully
human Sclerostin antibodies derived from AE10-6. Amino acid
residues of individual CDRs of each VH sequence are indicated in
bold
TABLE-US-00007 TABLE 6 VH sequences of affinity matured AE10-6
variants SEQ ID Clone NO: VH HC-38 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1719
GFDPEVGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DAGGFWYKFFQHWGPGTLVTVSS HC-41 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1720
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
NSEGYWWKDFQHWGPGTLVTVSS HC-42 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSIHWVRQAPGKGLEWMG NO: 1721
GFDPEEGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGPGTLVTVSS HC-47 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1722
GFDPEAGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DGELYWYKFFQHWGPGTLVTVSS HC-48 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1723
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DAGGYWYKFFQHWGPGTLVTVSS HC-5 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1724
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DAAGYWYKFFQHWGPGTLVTVSS HC-77 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1725
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGFWYKFFQHWGPGTLVTVSS HC-S1 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1726
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGPGTLVTVSS HC-S10 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1727
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGRWEKYFQHWGPGTLVTVSS HC-S11 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG NO: 1728
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTMGYWEKYFQHWGPGTLVTVSS HC-S12 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1729
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S14 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1730
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSYGYWEKFFQHWGPGTLVTVSS HC-S15 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1731
GFDPENGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTAGYWEKFFQHWGPGTLVTVSS HC-S16 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1732
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGVWEKYFQHWGPGTLVTVSS HC-S17 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1733
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGFWEKFFQHWGPGTLVTVSS HC-S18 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1734
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVGYWEKFFQHWGPGTLVTVSS HC-S19 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1735
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGYWEKFFQHWGPGTLVTVSS HC-S2 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1736
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVGYWEKFFQHWGPGTLVTVSS HC-S21 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1737
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGNWEKFFQHWGPGTLVTVSS HC-S23 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1738
GFDPEDGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKYFQHWGPGTLVTVSS HC-S24 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1739
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKYFQHWGPGTLVTVSS HC-S25 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1740
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDSYWEKFFQHWGPGTLVTVSS HC-S26 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1741
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S27 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSGLSMHWVRQAPGKGLEWMG NO: 1742
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSWGYWEKFFQHWGPGTLVTVSS HC-S29 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG NO: 1743
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGSWYKFFQHWGPGTLVTVSS HC-S30 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG NO: 1744
GFDPENGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGFWEKFFQHWGPGTLVTVSS HC-S31 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1745
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSQGYWYKFFQHWGPGTLVTVSS HC-S32 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1746
GFDPEQGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DADGYWEKFFQHWGPGTLVTVSS HC-S34 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1747
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVGRWYKFFQHWGPGTLVTVSS HC-S36 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1748
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTEGYWFKYFQHWGPGTLVTVSS HC-S38 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1749
GFDPEEGEMIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
ESEGFWFKYFQHWGPGTLVTVSS HC-S39 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1750
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVSYWEKYFQHWGPGTLVTVSS HC-S40 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1751
GFDPENGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S41 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1752
GFDPEDGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTEGYWEKYFQHWGPGTLVTVSS HC-S42 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSLHWVRQAPGKGLEWMG NO: 1753
GFNPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWVKYFQHWGPGTLVTVSS HC-S44 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1754
GFDPENGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGPGTLVTVSS HC-S45 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1755
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S46 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1756
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTLGYWEKFFQHWGPGTLVTVSS HC-S48 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1757
GFDPEDGENIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWFKYFQHWGPGTLVTVSS HC-S5 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1758
GFDPEEGEMIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S50 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1759
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSNGYWEKFFQHWGPGTLVTVSS HC-S51 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSGLSMHWVRQAPGKGLEWMG NO: 1760
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGPGTLVTVSS HC-S52 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1761
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYQFFQHWGPGTLVTVSS HC-S53 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1762
GFDPEVGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGFWFKYFQHWGPGTLVTVSS HC-S54 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1763
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGPGTLVTVSS HC-S55 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1764
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGPGTLVTVSS HC-S56 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSLHWVRQAPGKGLEWMG NO: 1765
GFDPEAGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGFWYKYFQHWGPGTLVTVSS HC-S57 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1766
GFDPEAGENIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWVKFFQHWGPGTLVTVSS HC-S58 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1767
GFDPEVGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKYFQHWGPGTLVTVSS HC-S59 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1768
GFDPEAGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWYKFFQHWGPGTLVTVSS HC-S6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1769
GFDPEGGENIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S63 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1770
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWYKFFQHWGPGTLVTVSS HC-S64 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG NO: 1771
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S65 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1772
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGNWEKFFQHWGPGTLVTVSS HC-S66 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1773
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGPGTLVTVSS HC-S67 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1774
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S69 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSLHWVRQAPGKGLEWMG NO: 1775
GFDPEHGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSLGYWYKFFQHWGPGTLVTVSS HC-S7 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1776
GFDPEYGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWEKYFQHWGPGTLVTVSS HC-S71 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1777
GFDPEDGEMIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKYFQHWGPGTLVTVSS HC-S72 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG NO: 1778
GFDPEAGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKYFQHWGPGTLVTVSS HC-S74 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1779
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGPGTLVTVSS
HC-S75 SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSLHWVRQAPGKGLEWMG NO:
1780 GSDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWFKYFQHWGPGTLVTVSS HC-S76 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1781
GFDPEVGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKYFQHWGPGTLVTVSS HC-S78 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1782
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGPWYKFFQHWGPGTLVTVSS HC-S79 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1783
GFDPERGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGPGTLVTVSS HC-S8 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1784
GFDPEDGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGFWFKYFQHWGPGTLVTVSS HC-S80 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1785
GFDPEDGEMIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S81 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1786
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGPGTLVTVSS HC-S83 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG NO: 1787
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S87 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1788
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGYWEKFFQHWGPGTLVTVSS HC-S89 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1789
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S9 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1790
GFDPEFGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGPGTLVTVSS HC-S90 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1791
GFDPEYGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKYFQHWGPGTLVTVSS HC-S92 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1792
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGIWEKFFQHWGPGTLVTVSS HC-S94 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1793
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGPGTLVTVSS HC-S95 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1794
GFDPEAGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKYFQHWGPGTLVTVSS rHC + LC-1 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG NO: 1795
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 100 NO: 1796
GFDPEEGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 109 NO: 1797
GFDPEYGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKYFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 113 NO: 1798
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DAGGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 143 NO: 1799
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG 149 NO: 1800
GFDPEAGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DGELYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 21 NO: 1801
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 26 NO: 1802
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG 43 NO: 1803
GFDPEHGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVSFWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSTHWVRQAPGKGLEWMG 52 NO: 1804
GFDPEYGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGNWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 60 NO: 1805
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTQGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG 66 NO: 1806
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSIHWVRQAPGKGLEWMG 69 NO: 1807
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 82 NO: 1808
GFDPEVGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 83 NO: 1809
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTAGNWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG 87 NO: 1810
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 93 NO: 1811
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG 94 NO: 1812
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSLHWVRQAPGKGLEWMG 99 NO: 1813
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSLGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG A1 NO: 1814
GFDPEEGETIYAQKFQGRVNMTEDTSTDTAYMELSSLRSEDTAVYYCAT
NSEGYWWKDFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG A10 NO: 1815
GFDPEGGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
NSDGYWWKDFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG A2 NO: 1816
GFDPENGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG A7 NO: 1817
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSGGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG A8 NO: 1818
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG B12 NO: 1819
GFDPEGGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG B2 NO: 1820
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYSLSELSMHWVRQAPGKGLEWMG B6 NO: 1821
GFDPEVGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKYFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG B9 NO: 1822
GFDPENGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG C12 NO: 1823
GFDPEEGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKHGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG C4 NO: 1824
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG C6 NO: 1825
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAMYYCAT
DSDGYWEKFFQHWGPGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG C7 NO: 1826
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG C9 NO: 1827
GFDPEDGEVIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D1 NO: 1828
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG D10 NO: 1829
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D11 NO: 1830
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
NSDGYWEKYFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D12 NO: 1831
GFDPEAGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D2 NO: 1832
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSIHWVRQAPGKGLEWMG D4 NO: 1833
GFDPEEGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D5 NO: 1834
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D6 NO: 1835
GFDPEDGETIYAQKFEGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG D7 NO: 1836
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG D8 NO: 1837
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG E1 NO: 1838
GFDPEAGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG E10 NO: 1839
GFDPEDGELIYAQKFQGRVTMTEDTSTDTAYMELSSLGSEDTAVYYCAT
NSAGYWWKDFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG E11 NO: 1840
GFDPENGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGFWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG E4 NO: 1841
GFDPEEGELIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVMKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG E6 NO: 1842
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGYWEKFFQHWARGTLVTVSS
rHC + LC- SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG
E9 NO: 1843 GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG F1 NO: 1844
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTISELSMHWVRQAPGKGLEWMG F12 NO: 1845
GFDPEVGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG F3 NO: 1846
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGFWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG F4 NO: 1847
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG F5 NO: 1848
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGFWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG F6 NO: 1849
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSIHWVRQAPGKGLEWMG F7 NO: 1850
GFDPENGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG G1 NO: 1851
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVGRWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG G11 NO: 1852
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTVGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG G12 NO: 1853
GFDPEVGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG G2 NO: 1854
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG G4 NO: 1855
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSVGRWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG G6 NO: 1856
GFDPEYGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG G7 NO: 1857
GLDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
ESVGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMG G8 NO: 1858
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
NSVGYWWKDFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG G9 NO: 1859
GFDPEYGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKYFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG H11 NO: 1860
GFDPEEGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSEGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG H12 NO: 1861
GFDPEAGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSAGYWYKYFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG H2 NO: 1862
GFDPEDGEIIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGFWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG H3 NO: 1863
GFDPEVGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DSDGFWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG H6 NO: 1864
GFDPEGGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DAGGYWYKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSVHWVRQAPGKGLEWMG H7 NO: 1865
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DTDGYWEKFFQHWGQGTLVTVSS rHC + LC- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYTLSELSMHWVRQAPGKGLEWMG H8 NO: 1866
GFDPEDGETIYAQKFQGRVTMTEDTSTDTAYMELSSLRSEDTAVYYCAT
DAAGYWYKFFQHWGQGTLVTVSS
TABLE-US-00008 TABLE 7 VL sequences of affinity matured AE10-6
variants Clone SEQ ID NO: VL LC-10 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1867
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFDGGTLVFGGGTKLTV L LC-31 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGLSTGNVTIWHYPY NO: 1868
WFQQKPGQAPRTLIFDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-45 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTIWHYPY NO: 1869
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-50 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1870
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-67 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1871
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-69 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1872
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-88 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIDHYPY NO: 1873
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S10 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTSGHYPY NO: 1874
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTSEHYPY NO: 1875
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S13 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTVNHYPY NO: 1876
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S15 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTIEHYPY NO: 1877
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTMVFGGGTKLTV L LC-S17 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1878
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFNGGTLVFGGGTKLTV L LC-S18 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1879
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S2 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTSDHYPY NO: 1880
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S20 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTIDHYPY NO: 1881
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGSVVFGGGTKLTV L LC-S21 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTIDHYPY NO: 1882
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S22 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTSDHYPY NO: 1883
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S23 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIGHYPY NO: 1884
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S24 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSEHYPY NO: 1885
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGNLVFGGGTKLTV L LC-S26 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1886
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S27 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSDHYPY NO: 1887
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S28 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIYHYPY NO: 1888
WFQQKPGQAPRTFISDTRDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S29 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSEHYPY NO: 1889
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S30 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSEHYPY NO: 1890
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S31 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGHYPY NO: 1891
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L LC-S32 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTIDHYPY NO: 1892
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S33 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1893
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGNVVFGGGTKLTV L LC-S35 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIDHYPY NO: 1894
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S36 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTIDHYPY NO: 1895
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S37 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1896
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S38 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIEHYPY NO: 1897
WFQQKPGQAPRTLVSDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFFGGTMVFGGGTKLTV L LC-S39 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1898
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSVVFGGGTKLTV L LC-S40 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1899
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S41 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTSGHYPY NO: 1900
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S42 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTSGHYTY NO: 1901
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGRLVFGGGTKLTV L LC-S44 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTSEHYPY NO: 1902
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTNLTV L LC-S45 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTEHYPY NO: 1903
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S46 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTIGHYPY NO: 1904
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S49 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTSEHYPY NO: 1905
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S50 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSDHYPY NO: 1906
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S51 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSEHYPY NO: 1907
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGNLVFGGGTKLTV L LC-S52 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTGDHYPY NO: 1908
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S54 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSDHYPY NO: 1909
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S57 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTIDHYPY NO: 1910
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGSLVFGGGTKLTV L LC-S58 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTSEHYPY NO: 1911
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S6 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTSEHYPY NO: 1912
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTMVFGGGTKLTV L LC-S60 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTSEHYPY NO: 1913
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S61 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTSEHYPY NO: 1914
WFQQKPGQAPRTLIYDTIEKDSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S64 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGHYPY NO: 1915
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L
LC-S66 SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSEHYPY NO: 1916
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S7 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1917
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S70 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1918
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGNVVFGGGTKLTV L LC-S74 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1919
WFQQKPGQAPRTLISDTDDKQSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S75 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIDHYPY NO: 1920
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S76 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1921
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFFGGTVVFGGGTKLTV L LC-S8 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIWHYPY NO: 1922
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDDGSTVVFGGGTKLTV L LC-S80 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGHYPY NO: 1923
WFQQKPGQAPRTLIYDTDDKNSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L LC-S82 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIWHYPY NO: 1924
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFDGGTVVFGGGTKLTV L LC-S83 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTSGHYPY NO: 1925
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S85 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTSEHYPY NO: 1926
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S86 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTSEHYPY NO: 1927
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S88 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIGHYPY NO: 1928
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTMVFGGGTKLTV L LC-S89 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTSEHYPY NO: 1929
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFDGGTLVFGGGTKLTV L LC-S9 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1930
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S90 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIEHYPY NO: 1931
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S91 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTSEHYPY NO: 1932
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L LC-S92 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1933
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGSMVFGGGTKLTV L LC-S93 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1934
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFDRGSMVFGGGTKLTV L LC-S94 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGTVTSEHYPY NO: 1935
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L LC-S95 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1936
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-1 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1937
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-100 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1938
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYEGTFVFGGGTKLTV L rHC + LC-109 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGGVTIDHYPY NO: 1939
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSFVFGGGTKLTV L rHC + LC-113 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIEHYPY NO: 1940
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-143 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1941
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-149 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIWHYPY NO: 1942
WFQQKPGQAPRTLISDTXDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSFVFGGGTKLTV L rHC + LC-175 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTIEHYPY NO: 1943
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYRGTFVFGGGTKLTV L rHC + LC-26 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1944
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYRGTFVFGGGTKLTV L rHC + LC-43 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTIDHYPY NO: 1945
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSFVFGGGTKLTV L rHC + LC-5 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIDHYPY NO: 1946
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSFVFGGGTKLTV L rHC + LC-60 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1947
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDFGGTFVFGGGTKLTV L rHC + LC-66 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGTVTIDHYPY NO: 1948
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGRFVFGGGTKLTV L rHC + LC-69 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1949
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSFVFGGGTKLTV L rHC + LC-75 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIWHYPY NO: 1950
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGSFVFGGGTKLTV L rHC + LC-83 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1951
WFQQKPGQAPRTLISDTIYKESWTPARFAGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-93 SEQ ID
QAVVTQEPSLTVSQGGTVTLTCGSSTGDVTIDHYPY NO: 1952
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYRGNMVFGGGTKLTV L rHC + LC-94 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1953
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-99 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIDHYPY NO: 1954
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGAFVFGGGTKLTV L rHC + LC-A10 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1955
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGRLVFGGGTKLTV L rHC + LC-A12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1956
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-A2 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1957
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYDGTFVFGGGTKLTV L rHC + LC-A3 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTSEHYPY NO: 1958
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L rHC + LC-A8 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1959
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYRGTMVFGGGTKLTV L rHC + LC-B1 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGTVTIDHYPY NO: 1960
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-B12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1961
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYNGTFVFGGGTKLTV L rHC + LC-B4 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1962
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGRFVFGGGTKLTV L rHC + LC-C1 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTMWHYPY NO: 1963
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGRFVFGGGTKLTV L rHC + LC-C11 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1964
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGNLVFGGGTKLTV L rHC + LC-C12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1965
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGAFVFGGGTKLTV L rHC + LC-C2 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY
NO: 1966 WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDFGGTFVFGGGTKLTV L rHC + LC-C3 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1967
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-C4 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGPVTIDHYPY NO: 1968
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGRFVFGGGTKLTV L rHC + LC-C6 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1969
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFNGGNVVFGGGTKLTV L rHC + LC-C7 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1970
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFFGGTVVFGGGTKLTV L rHC + LC-C8 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGRVTIDHYPY NO: 1971
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDFGGTFVFGGGTKLTV L rHC + LC-D10 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTIDHYPY NO: 1972
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYEGTFVFGGGTKLTV L rHC + LC-D11 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1973
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-D3 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1974
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-D6 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTIEHYPY NO: 1975
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L rHC + LC-E11 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTVDHYPY NO: 1976
WFQQKPGQAPRTLISDTTDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-E3 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTTDHYPY NO: 1977
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTLVFGGGTKLTV L rHC + LC-E5 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGNVTIDHYPY NO: 1978
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-E6 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1979
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTLVFGGGTKLTV L rHC + LC-E8 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1980
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDDGGTFVFGGGTKLTV L rHC + LC-E9 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1981
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-F10 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1982
WFQQKPGQAPRTLIFDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGTVVFGGGTKLTV L rHC + LC-F12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1983
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTLVFGGGTKLTV L rHC + LC-F3 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTIDHYPY NO: 1984
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGAFVFGGGTKLTV L rHC + LC-F4 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1985
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYEGTFVFGGGTKLTV L rHC + LC-G1 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTAGHYPY NO: 1986
WFQQKPGQAPRTLISDTIDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGAFVFGGGTKLTV L rHC + LC-G11 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTIGHYPY NO: 1987
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYRGTMVFGGGTKLTV L rHC + LC-G12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGEVTIDHYPY NO: 1988
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-G2 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGHYPY NO: 1989
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-G6 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIEHYPY NO: 1990
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDFGGTFVFGGGTKLTV L rHC + LC-G7 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIGHYPY NO: 1991
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-H1 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1993
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGNFVFGGGTKLTV L rHC + LC-H12 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIDHYPY NO: 1994
WFQQKPGQAPRTLISDTDDKPSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTFVFGGGTKLTV L rHC + LC-H3 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1995
WFQQKPGQAPRTLISDTNDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGRFVFGGGTKLTV L rHC + LC-H4 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTIEHYPY NO: 1996
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLFYGGNVVFGGGTKLTV L rHC + LC-H6 SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTIDHYPY NO: 1997
WFQQKPGQAPRTLISDTDDKHSWTPARFSGSLLGGK
AALTLSGAQPEDEAEYYCLLDYGGTVVFGGGTKLTV L
[0580] The tables below provide a list of AE10-6 clones which were
converted into IgG proteins for characterization, both VH (Table 8)
and VL (Table 9) sequences.
TABLE-US-00009 TABLE 8 VH sequences of IgG converted clones
Sequence Protein region SEQ ID NO 123456789012345678901234567890
AM1 VH SEQ ID NO: 1998 EVQLVQSGAEVKKPGASVKVSCKVSGYTLSE
LSMHWVRQAPGKGLEWMGGFDPEVGELIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDTDGYWEKFFQHWGQGTLVTVSS AM1 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 1998 AM1 VH CDR-H2 Residues 50-66 GFDPEVGELIYAQKFQG of
SEQ ID NO.: 1998 AM1 VH CDR-H3 Residues 99-110 DTDGYWEKFFQH of SEQ
ID NO.: 1998 AM2 VH SEQ ID NO: 1999 EVQLVQSGAEVKKPGASVKVSCKVSGYTLTE
LSMHWVRQAPGKGLEWMGGFDPEAGETIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDGELYWYKFFQHWGQGTLVTVSS AM2 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 1999 AM2 VH CDR-H2 Residues 50-66 GFDPEAGETIYAQKFQG of
SEQ ID NO.: 1999 AM2 VH CDR-H3 Residues 99-110 GFDPEAGETIYAQKFQG of
SEQ ID NO.: 1999 AM3 VH SEQ ID NO: 2000
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTE LSMHWVRQAPGKGLEWMGGFDPEDGETIYAQ
KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV YYCATDSAGYWYKFFQHWGQGTLVTVSS AM3 VH
CDR-H1 Residues 31-35 ELSMH of SEQ ID NO.: 2000 AM3 VH CDR-H2
Residues 50-66 GFDPEDGETIYAQKFQG of SEQ ID NO.: 2000 AM3 VH CDR-H3
Residues 99-110 DSAGYWYKFFQH of SEQ ID NO.: 2000 AM4 VH SEQ ID NO:
2001 EVQLVQSGAEVKKPGASVKVSCKVSGYTLTE
LSMHWVRQAPGKGLEWMGGFDPEHGETIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDSVSFWEKFFQHWGQGTLVTVSS AM4 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 2001 AM4 VH CDR-H2 Residues 50-66 GFDPEHGETIYAQKFQG of
SEQ ID NO.: 2001 AM4 VH CDR-H3 Residues 99-110 DSVSFWEKFFQH of SEQ
ID NO.: 2001 AM5 VH SEQ ID NO: 2002 EVQLVQSGAEVKKPGASVKVSCKVSGYTLSE
LSMHWVRQAPGKGLEWMGGFDPEGGETIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDAGGYWYKFFQHWGQGTLVTVSS AM5 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 2002 AM5 VH CDR-H2 Residues 50-66 GFDPEGGETIYAQKFQG of
SEQ ID NO.: 2002 AM5 VH CDR-H3 Residues 99-110 DAGGYWYKFFQH of SEQ
ID NO.: 2002 AM6 VH SEQ ID NO: 2003 EVQLVQSGAEVKKPGASVKVSCKVSGYTLSE
LSMHWVRQAPGKGLEWMGGFDPEGGETIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDTEGYWEKFFQHWGQGTLVTVSS AM6 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 2003 AM6 VH CDR-H2 Residues 50-66 GFDPEGGETIYAQKFQG of
SEQ ID NO.: 2003 AM6 VH CDR-H3 Residues 99-110 DTEGYWEKFFQH of SEQ
ID NO.: 2003 AM7 VH SEQ ID NO: 2004 EVQLVQSGAEVKKPGASVKVSCKVSGYTLTE
LSMHWVRQAPGKGLEWMGGFDPEDGETIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDTVGYWEKFFQHWGQGTLVTVSS AM7 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 2004 AM7 VH CDR-H2 Residues 50-66 GFDPEDGETIYAQKFQG of
SEQ ID NO.: 2004 AM7 VH CDR-H3 Residues 99-110 DTVGYWEKFFQH of SEQ
ID NO.: 2004 AM8 VH SEQ ID NO: 2005 EVQLVQSGAEVKKPGASVKVSCKVSGYTLSE
LSIHWVRQAPGKGLEWMGGFDPEDGETIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDSAGYWYKFFQHWGQGTLVTVSS AM8 VH CDR-H1 Residues 31-35 ELSIH of
SEQ ID NO.: 2005 AM8 VH CDR-H2 Residues 50-66 GFDPEDGETIYAQKFQG of
SEQ ID NO.: 2005 AM8 VH CDR-H3 Residues 99-110 DSAGYWYKFFQH of SEQ
ID NO.: 2005 AM9 VH SEQ ID NO: 2006 EVQLVQSGAEVKKPGASVKVSCKVSGYTLSE
LSMHWVRQAPGKGLEWMGGFDPEVGELIYAQ KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV
YYCATDAGGFWYKFFQHWGPGTLVTVSS AM9 VH CDR-H1 Residues 31-35 ELSMH of
SEQ ID NO.: 2006 AM9 VH CDR-H2 Residues 50-66 GFDPEVGELIYAQKFQG of
SEQ ID NO.: 2006 AM9 VH CDR-H3 Residues 99-110 DAGGFWYKFFQH of SEQ
ID NO.: 2006 AM10 VH SEQ ID NO: 2007
EVQLVQSGAEVKKPGASVKVSCKVSGYTLTE LSMHWVRQAPGKGLEWMGGFDPEDGETIYAQ
KFQGRVTMTEDTSTDTAYMELSSLRSEDTAV YYCATDSEGYWEKYFQHWGRGTLVTVSS AM10
VH CDR-H1 Residues 31-35 ELSMH of SEQ ID NO.: 2007 AM10 VH CDR-H2
Residues 50-66 GFDPEDGETIYAQKFQG of SEQ ID NO.: 2007 AM10 VH CDR-H3
Residues 99-110 DSEGYWEKYFQH of SEQ ID NO.: 2007
TABLE-US-00010 TABLE 9 VL sequences of IgG converted clones
Sequence Protein region SEQ ID NO 123456789012345678901234567890
AM1 VL SEQ ID NO: 2008 QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGTFVFGGGTKLTVL AM1 VL CDR-L1 Residues 23-36 GSSTGAVTIDHYPY of SEQ
ID NO.: 2008 AM1 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2008 AM1 VL CDR-L3 Residues 101-109 LLDYGGTFV of SEQ ID NO.: 2008
AM2 VL SEQ ID NO: 2009 QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTI
WHYPYWFQQKPGQAPRTLISDTNDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGSFVFGGGTKLTVL AM2 VL CDR-L1 Residues 23-36 GSSTGAVTIWHYPY of SEQ
ID NO.: 2009 AM2 VL CDR-L2 Residues 52-58 DTNDKHS of SEQ ID NO.:
2009 AM2 VL CDR-L3 Residues 101-109 LLDYGGSFV of SEQ ID NO.: 2009
AM3 VL SEQ ID NO: 2010 QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGTFVFGGGTKLTVL AM3 VL CDR-L1 Residues 23-36 GSSTGAVTIDHYPY of SEQ
ID NO.: 2010 AM3 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2010 AM3 VL CDR-L3 Residues 101-109 LLDYGGTFV of SEQ ID NO.: 2010
AM4 VL SEQ ID NO: 2011 QAVVTQEPSLTVSPGGTVTLTCGSSTGSVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGSFVFGGGTKLTVL AM4 VL CDR-L1 Residues 23-36 GSSTGSVTIDHYPY of SEQ
ID NO.: 2011 AM4 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2011 AM4 VL CDR-L3 Residues 101-109 LLDYGGSFV of SEQ ID NO.: 2011
AM5 VL SEQ ID NO: 2012 QAVVTQEPSLTVSPGGTVTLTCGSSTGTVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLF
YGGTVVFGGGTKLTVL AM5 VL CDR-L1 Residues 23-36 GSSTGTVTIDHYPY of SEQ
ID NO.: 2012 AM5 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2012 AM5 VL CDR-L3 Residues 101-109 LLFYGGTVV of SEQ ID NO.: 2012
AM6 VL SEQ ID NO: 2013 QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGTFVFGGGTKLTVL AM6 VL CDR-L1 Residues 23-36 GSSTGAVTIDHYPY of SEQ
ID NO.: 2013 AM6 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2013 AM6 VL CDR-L3 Residues 101-109 LLDYGGTFV of SEQ ID NO.: 2013
AM7 VL SEQ ID NO: 2014 QAVVTQEPSLTVSPGGTVTLTCGSSTGTVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGRFVFGGGTKLTVL AM7 VL CDR-L1 Residues 23-36 GSSTGTVTIDHYPY of SEQ
ID NO.: 2014 AM7 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2014 AM7 VL CDR-L3 Residues 101-109 LLDYGGRFV of SEQ ID NO.: 2014
AM8 VL SEQ ID NO: 2015 QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLD
YGGSFVFGGGTKLTVL AM8 VL CDR-L1 Residues 23-36 GSSTGAVTIDHYPY of SEQ
ID NO.: 2015 AM8 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID NO.:
2015 AM8 VL CDR-L3 Residues 101-109 LLDYGGSFV of SEQ ID NO.: 2015
AM9 VL SEQ ID NO: 2016 QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTS
GHYPYWFQQKPGQAPRTLISDTNDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLF
YGGTVVFGGGTKLTVL AM9 VL CDR-L1 Residues 23-36 GSSTGAVTSGHYPY of SEQ
ID NO.: 2016 AM9 VL CDR-L2 Residues 52-58 DTNDKHS of SEQ ID NO.:
2016 AM9 VL CDR-L3 Residues 101-109 LLFYGGTVV of SEQ ID NO.: 2016
AM10 VL SEQ ID NO: 2017 QAVVTQEPSLTVSPGGTVTLTCGSSTGDVTI
DHYPYWFQQKPGQAPRTLISDTDDKHSWTPA RFSGSLLGGKAALTLSGAQPEDEAEYYCLLF
YGGTLVFGGGTKLTVL AM10 VL CDR-L1 Residues 23-36 GSSTGDVTIDHYPY of
SEQ ID NO.: 2017 AM10 VL CDR-L2 Residues 52-58 DTDDKHS of SEQ ID
NO.: 2017 AM10 VL CDR-L3 Residues 101-109 LLFYGGTLV of SEQ ID NO.:
2017
[0581] Heavy and light chain pairs were prepared as follows in
Table 10:
TABLE-US-00011 TABLE 10 Heavy and light chain pairs of AE10-6
affinity matured clones Clone name HC LC Protein name rHC + LC S4
AE10-6 AM1 AE10-6 AM1 AE10-6-AM1 clone 10 rHC + LC S4 AE10-6 AM2
AE10-6 AM2 AE10-6-AM2 clone 102 rHC + LC S4 AE10-6 AM3 AE10-6 AM3
AE10-6-AM3 clone 105 rHC + LC S4 AE10-6 AM4 AE10-6 AM4 AE10-6-AM4
clone 114 rHC + LC S4 AE10-6 AM5 AE10-6 AM5 AE10-6-AM5 clone 117
rHC + LC S4 AE10-6 AM6 AE10-6 AM6 AE10-6-AM6 clone 119 rHC + LC S4
AE10-6 AM7 AE10-6 AM7 AE10-6-AM7 clone 131 rHC + LC S4 AE10-6 AM8
AE10-6 AM8 AE10-6-AM8 clone 135 HC M2S5 AE10-6 AM9 AE10-6 AM9
AE10-6-AM9 clone 21 LC M2S5 AE10-6 AM10 AE10-6 AM10 AE10-6-AM10
clone 12
Example 1.1.3
Affinity Maturation of the Fully Human Anti-Human Sclerostin
Binding Protein MSL10
[0582] The MSL10 human binding protein to human TNF was affinity
matured by in vitro display technology. The VH and VL sequence of
the parental MSL10 antibody are provided below.
TABLE-US-00012 Parental MSL10 VH (SEQ ID NO: 2018)
EVQLVESGGGLVQPGGSLRLSCTASGFTFDDYALHWVRQAPGKGLEWVS
GISWHGDFIDYADSVKGRFTISRDNSKNTLYLQMNGLRVEDMAIYYCAG
NNRGYGGLDVWGQGTTVTVSS Parental MSL1.1 VL (SEQ ID NO: 2019)
QSGLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLI
YSNNQRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCAAWDDSLNG SYVFGGGTKLTVL
[0583] To improve the affinity of MSL10 to Sclerostin, hypermutated
CDR residues were identified from other human antibody sequences in
the IgBLAST database that also shared high identity to human
germlines. The corresponding MSL10 CDR residues were then subjected
to limited mutagenesis by PCR with primers having low degeneracy at
these positions to create two antibody libraries in the scFv format
suitable for surface display.
[0584] The tables below provides a list of amino acid sequences of
VH (Table 14) and VL (Table 15) regions of affinity matured fully
human Sclerostin antibodies derived from MSL10. Amino acid residues
of individual CDRs of each VH sequence are indicated in bold.
TABLE-US-00013 TABLE 14 VH sequences of affinity matured MSL10
variants SEQ ID Clone NO: VH MSL10-AM1 SEQ ID NO: 2020
EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA
PGKGLEWVSGINWEGDDIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNSRGYGGLDVWGQGTTVTVSS MSL10-AM2 VH SEQ ID NO:
2021 EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA
PGKGLEWVSGIGWEDDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNSRGYGGLDVWGQGTTVTVSS MSL10-AM3 VH SEQ ID NO:
2022 EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA
PGKGLEWVSGIGWDEDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10-AM4 VH SEQ ID NO:
2023 EVQLVESGGGLVQPGRSLRLSCAASGFTFEDYALHWVRQA
PGKGLEWVSGIGWDDDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10-AM5 VH SEQ ID NO:
2024 EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA
PGKGLEWVSGISWHGDFIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10-AM6 VH SEQ ID NO:
2025 EVQLVESGGGLVQPGRSLRLSCAASGFTFDQYALHWVRQA
PGKGLEWVSGINWDGDYIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNIRGYGGLDVWGQGTTVTVSS MSL10-AM7 VH SEQ ID NO:
2026 EVQLVESGGGLVQPGRSLRLSCAASGFTFEDYALHWVRQA
PGKGLEWVSGIGWNDDEIDYADSVKGRFTISRDNSKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10-AM8 VH SEQ ID NO:
2027 EVQLVESGGGLVQPGRSLRLSCAASGFTFEDYALHWVRQA
PGKGLEWVSGIGWDRDFIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNKIGYGGLDVWGQGTTVTVSS MSL10-AM9 VH SEQ ID NO:
2028 EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA
PGKGLEWVSGIGWDDDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10-AM10 SEQ ID NO: 2029
EVQLVESGGGLVQPGRSLRLSCAASGFTFSDYALHWVRQA VH
PGKGLEWVSGISWYGDDIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNIRGYGGLDVWGQGTTVTVSS MSL10-AM11 SEQ ID NO: 2030
EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA VH
PGKGLEWVSGISWHGDFIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10- SEQ ID NO: 2031
EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA AM1.2VH
PGKGLEWVSGINWEGDDIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNSRGYGGLDVWGQGTTVTVSS MSL10- SEQ ID NO: 2032
EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA AM2.2VH
PGKGLEWVSGIGWEDDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNSRGYGGLDVWGQGTTVTVSS MSL10- SEQ ID NO: 2033
EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYALHWVRQA AM3.2VH
PGKGLEWVSGIGWDEDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS MSL10- SEQ ID NO: 2034
EVQLVESGGGLVQPGRSLRLSCAASGFTFEDYALHWVRQA AM4.2VL
PGKGLEWVSGIGWDDDMIDYADSVKGRFTISRDNAKNSLY
LQMNSLRVEDTALYYCAGNNRGYGGLDVWGQGTTVTVSS
TABLE-US-00014 TABLE 15 VL sequences of affinity matured MSL10
variants SEQ ID Clone NO: VH MSL10-AM1 VL SEQ ID NO: 2035
QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNTVNWYQQL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDNLESYVFGGGTKLTVL MSL10-AM2 VL SEQ ID NO: 2036
QSVLTQPPSASGTPGQRVTISCSGSSSNIGGNTVNWYQQL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLEGSYVFGGGTKLTVL MSL10-AM3 VL SEQ ID NO: 2037
QSVLTQPPSASGTPGQRVTISCSGSWSNIGSNTVNWYQQL
PGTAPKLLIYNNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLSGEYVFGGGTKLTVL MSL10-AM4 VL SEQ ID NO: 2038
QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQL
PGTAPKLLIYNNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLDSYVFGGGTKLTVL MSL10-AM5 VL SEQ ID NO: 2039
QSVLTQPPSASGTPGQRVTISCSGSWSNIGGNTVNWYQQL
PGTAPKLLIYNNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDTLEGSYVFGGGTKLTVL MSL10-AM6 VL SEQ ID NO: 2040
QSVLTQPPSASGTPGQRVTISCSGSWSNIGGNTVNWYQQL
PGTAPKLLIYNNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLDGEYVFGGGTKLTVL MSL10-AM7 VL SEQ ID NO: 2041
QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDELELYVFGGGTKLTVL MSL10-AM8 VL SEQ ID NO: 2042
QSVLTQPPSASGTPGQRVTISCSGSSSNIGTNTVNWYQQL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDQLEAYVFGGGTKLTVL MSL10-AM9 VL SEQ ID NO: 2043
QSVLTQPPSASGTPGQRVTISCSGSSSNIGTNTVNWYQQL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDRLDEYVFGGGTKLTVL MSL10-AM10 SEQ ID NO: 2044
QSVLTQPPSASGTPGQRVTISCSGSWSNIGSNTVNWYQQL VL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLDGAYVFGGGTKLTVL MSL10-AM11 SEQ ID NO: 2045
QSVLTQPPSASGTPGQRVTISCSGSSSNIGGNTVNWYQQL VL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDILESYVFGGGTKLTVL MSL10- SEQ ID NO: 2046
QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNTVNWYQQL AM1.2VL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDNLESYVFGGGTKLTVLG MSL10- SEQ ID NO: 2047
QSVLTQPPSASGTPGQRVTISCSGSSSNIGGNTVNWYQQL AM2.2VL
PGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLEGSYVFGGGTKLTVLG MSL10- SEQ ID NO: 2048
QSVLTQPPSASGTPGQRVTISCSGSWSNIGSNTVNWYQQL AM3.2VL
PGTAPKLLIYNNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLSGEYVFGGGTKLTVLG MSL10- SEQ ID NO: 2049
QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQL AM4.2VL
PGTAPKLLIYNNNQRPSGVPDRFSGSKSGTSASLAISGLQ
SEDEADYYCAAWDDSLDSYVFGGGTKLTVLG
[0585] The table below (Table 16) provides a list of amino acid
sequences of VH and VL of a fully human SOST antibody MSL10 Amino
acid residues of individual CDRs of each VH and VL sequence are
indicated in bold.
TABLE-US-00015 TABLE 16 List Of Amino Acid Sequences Of VH And VL
Regions Of Fully Human SOST Antibodies Sequence Protein region SEQ
ID NO. 123456789012345678901234567890 MSL10 SOST SEQ ID NO: 3
EVQLVESGGGLVQPGGSLRLSCTASGFTFDDYA VH
LHWVRQAPGKGLEWVSGISWHGDFIDYADSVKG RFTISRDNSKNTLYLQMNGLRVEDMAIYYCAGN
NRGYGGLDVWGQGTTVTVSS MSL10CDR- Residues 31-35 DYALH H1 of SEQ ID
NO: 3 MSL10CDR- Residues 50-66 GISWHGDFIDYADSVKG H2 of SEQ ID NO: 3
MSL10CDR- Residues 99-108 NNRGYGGLDV H3 of SEQ ID NO: 3 MSL10 VL
SOST SEQ ID NO: 4 QSGLTQPPSASGTPGQRVTISCSGSSSNIGSNT
VNWYQQLPGTAPKLLIYSNNQRPSGVPDRFSGS KSGTSASLAISGLQSEDEADYYCAAWDDSLNGS
YVFGTGTKVTVLG MSL10CDR- Residues 23-35 SGSSSNIGSNTVN L1 of SEQ ID
NO: 4 MSL10CDR- Residues 51-57 SNNQRPS L2 of SEQ ID NO: 4 MSL10CDR-
Residues 90-101 AAWDDSLNGSYV L3 of SEQ ID NO: 4 MSL17 SOST SEQ ID
NO: 5 EVQLLESGGGLVKPGRSLRLSCVAYGFSLTGYS VH
MNWVRQAPGKGLEWVSSISPNDTYRHYADSVKG RFTISRDNAKNSLYLQMNSLRAEDTAVYYCARD
TNYRDSGGYFYDVFDIWGQGTMVTVSS MSL17 Residues 31-35 GYSMN CDR-H1 of
SEQ ID NO: 5 MSL17 Residues 50-66 SISPNDTYRHYADSVKG CDR-H2 of SEQ
ID NO: 5 MSL17 Residues 99-115 DTNYRDSGGYFYDVFDI CDR-H3 of SEQ ID
NO: 5 MSL17 SOST SEQ ID NO: 6 SYELTQPPSVSVAPGETARVTCEGNNIGNKGVH VL
WYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNS GNTATLTISRVEAGDDADYYCQVWDSSSDVFGS
GTKVTVLG MSL17 Residues 23-33 EGNNIGNKGVH CDR-L1 of SEQ ID NO: 6
MSL17 Residues 49-55 DDSDRPS CDR-L2 of SEQ ID NO: 6 MSL17 Residues
88-96 QVWDSSSDV CDR-L3 of SEQ ID NO: 6 MSL9-8 SOST SEQ ID NO: 7
EVQLVESGGGLVQPGGSLRLSCAASGFRFTDYW VH
MTWVRQAPGKGPEWVANINEDGSKKHYADSVKD RFIISRDNAKKSLSLQMKRMRAEDTAVYYCAAD
LNPHWLVGWGQGTLVTVSS MSL9-8 Residues 31-35 DYWMT CDR-H1 of SEQ ID
NO: 7 MSL9-8 Residues 50-66 NINEDGSKKHYADSVKD CDR-H2 of SEQ ID NO:
7 MSL9-8 Residues 99-107 DLNPHWLVG CDR-H3 of SEQ ID NO: 7 MSL9-8
SOST SEQ ID NO: 8 QPVLTQPPSVSVAPGKTARITCGGNNIGSRRVH VL
WYQQKPGQAPVLVVYDDNDRPSGIPERFSGSKS GNTATLTISRVEAGDEADYYCQVWHSGRVFGTG
TKVTVLG MSL9-8 Residues 23-33 GGNNIGSRRVH CDR-L1 of SEQ ID NO: 8
MSL9-8 Residues 49-55 DDNDRPS CDR-L2 of SEQ ID NO: 8 MSL9-8
Residues 88-95 QVWHSGRV CDR-L3 of SEQ ID NO: 8 MSK9 SOST SEQ ID NO:
9 EVQLLESGGGLVQPGGSLRLSCAASGFTFRSYW VH
MSWVRQAPGKGLEWVASIKQDGSKKHYADSVKD RFIISRDNAKKSLSLQMKRMRAEDTAVYYCAAD
LNPHWLVGWGQGTLVTVSS MSK9 Residues 31-35 SYWMS CDR-H1 of SEQ ID NO:
9 MSK9 Residues 50-66 SIKQDGSKKHYADSVKD CDR-H2 of SEQ ID NO: 9 MSK9
Residues 99-107 DLNPHWLVG CDR-H3 of SEQ ID NO: 9 MSK9 SOST SEQ ID
NO: 10 DIVMTQTPLSLPVTPGEPASISCRSSQSLLHSN VL
GYNYLDWYLQKPGQSPQLLIYLGSKRASGVPDR FSGSGSGTDFTLKISRVGAEDVGVYYCMQGLQT
PKAFGPGTKVDIKR MSK9 Residues 24-39 RSSQSLLHSNGYNYLD CDR-L1 of SEQ
ID NO: 10 MSK9 Residues 55-61 LGSKRAS CDR-L2 of SEQ ID NO: 10 MSK9
Residues 94-112 MQGLQTPKA CDR-L3 of SEQ ID NO: 10 MSK13 SOST SEQ ID
NO: 11 EMQLVQSGAEVKKPGASVKVSCKASGYTFSGYY VH
MNWVRQAPGQGLEWMGWINPYSGAATYAQDFQG RITVTRDTSISTGYMELTRLTSTDTAVYYCARG
GTITGASWYFDVWGRGTLVTVSS MSK13 Residues 31-35 GYYMN CDR-H1 of SEQ ID
NO: 11 MSK13 Residues 50-66 WINPYSGAATYAQDFQG CDR-H2 of SEQ ID NO:
11 MSK13 Residues 99-111 GGTITGASWYFDV CDR-H3 of SEQ ID NO: 11
MSK13 SOST SEQ ID NO: 12 DIVMTQTPLSLPVTLGQPASISCRSSQSLVHSD VL
GNTYLNWFQQRPGQSPRRLIYKVSNRDSGVPDR FSGSGSGTDFTLKISRVEAEDVGVYYCMQGTHW
PQLTFGGGTKVEIKR MSK13 Residues 24-39 RSSQSLVHSDGNTYLN CDR-L1 of SEQ
ID NO: 12 MSK13 Residues 55-61 KVSNRDS CDR-L2 of SEQ ID NO: 12
MSK13 Residues 94-113 MQGTHWPQLT CDR-L3 of SEQ ID NO: 12 MSK21 SOST
SEQ ID NO: 13 EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNI VH
VAWNWIRQSPSRGLEWLGRTYYTSKWFNQYAMS VKSRITINPDTSKNQVSLKLSSVTAADTAAYYC
ARVGGTYDFWSGYYRPYYYGMDVWGQGTMVTVS S MSK21 Residues 31-37 SNIVAWN
CDR-H1 of SEQ ID NO: 13 MSK21 Residues 52-69 RTYYTSKWFNQYAMSVKS
CDR-H2 of SEQ ID NO: 13 MSK21 Residues VGGTYDFWSGYYRPYYYGMDV CDR-H3
102-122 of SEQ ID NO: 13 MSK21 SOST SEQ ID NO: 14
DIRLTQSPSSLSASIGDTVTISCRSSQPINTHL VL
NWFRQLPGRAPELLIYGSSSLHTGVPSRFSGSG SGTDFTLTITSLQRGDFLTYYCQQTHRLPITFG
QGTRLDIKR MSK21 Residues 24-34 RSSQPINTHLN CDR-L1 of SEQ ID NO: 14
MSK21 Residues 50-56 GSSSLHT CDR-L2 of SEQ ID NO: 14 MSK21 Residues
89-97 QQTHRLPIT CDR-L3 of SEQ ID NO: 14
[0586] The sequences of the individual CDRs of the VH and VL
regions of the fully human SOST antibodies in the above table can
be aligned to provide consensus CDR sequences such as those in the
table below (Table 17).
TABLE-US-00016 TABLE 17 Consensus Sequence of SOST Antibody CDRs
CDR Sequence Consensus region Identifier Sequence CDR- SEQ ID
X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7 H1 NO: 15 D
- - Y A L H (MSL10) G - - Y S M N (MSL17) D - - Y W M T (MSL9-8) S
- - Y W M S (MSK-9) G - - Y Y M N (MSK-13) S N I V A W N (MSK-21)
CDR- SEQ ID X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7
X.sub.8 X.sub.9 X.sub.10 X.sub.11 X.sub.12 X.sub.13 X.sub.14
X.sub.15 X.sub.16 X.sub.17 X.sub.18 H2 NO: 16 G I S W H G D F I - D
Y A D S V K G (MSL10) S I S P N D T Y R - H Y A D S V K G (MSL17) N
I N E D G S K K - H Y A D S V K D (MSL9-8) S I K Q D G S K K - H Y
A D S V K D (MSK-9) W I N P Y S G A A - T Y A Q D F Q G (MSK-13) R
T Y Y T S K W F N Q Y A M S V K S (MSK-21) CDR- SEQ ID X.sub.1
X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7 X.sub.8 X.sub.9
X.sub.10 X.sub.11 X .sub.12 X.sub.13 X.sub.14 X.sub.15 X.sub.16
X.sub.17 X.sub.18 X.sub.19 X.sub.20 X.sub.21 H3 NO: 17 - - N N R G
Y G G - - - - - - - - - L D V (MSL10) D T N Y R D S G G Y F Y - - -
- D V F D I (MSL17) D L N P H W L V G (MSL9-8) D L N P H W L V G
(MSK-9) G G T I T G A S W Y - - - - - - - - F D V (MSK-13) V G G T
Y D F W S G Y Y R P Y Y Y G M D V (MSK-21) CDR- SEQ ID X.sub.1
X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7 X.sub.8 X.sub.9
X.sub.10 X.sub.11 X.sub.12 X.sub.13 X.sub.14 X.sub.15 X.sub.16 L1
NO: 18 S G S S - - - S N I G S N T V N (MSL10) E G N - - - - - N I
G N K G V H (MSL17) G G N - - - - - N I G S R R V H (MSL9-8) R S S
Q S L L H S N G Y N Y L D (MSK-9) R S S Q S L V H S D G N T Y L N
(MSK-13) R S S Q - - - - - P I N T H L N (MSK-21) CDR- SEQ ID
X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7 L2 NO: 19 S
N N Q R P S (MSL10) D D S D R P S (MSL17) D D N D R P S (MSL9-8) L
G S K R A S (MSK-9) K V S N R D S (MSK-13) G S S S L H T (MSK-21)
CDR- SEQ ID X.sub.1 X.sub.2 X.sub.3 X.sub.4 X.sub.5 X.sub.6 X.sub.7
X.sub.8 X.sub.9 X.sub.10 X.sub.11 X.sub.12 L3 NO: 20 A A W D D S L
N G S Y V (MSL10) Q V W D S S - - - S D V (MSL17) Q V W H S G - - -
- R V (MSL9-8) M Q G L Q T P - - - K A (MSK-9) M Q G T H W P - - Q
L T (MSK-13) Q Q - T H R L P I T (MSK-21)
"CDRH1" sequences: "MSL10" residues 31-35 of SEQ ID NO: 3, "MSL17"
residues 31-35 of SEQ ID NO: 5, "MSL9-8" residues 31-35 of SEQ ID
NO: 7, "MSK-9" residues 31-35 of SEQ ID NO: 9, "MSK-13" residues
31-35 of SEQ ID NO: 11 and "MSK-21" residues 31-35 of SEQ ID NO:
13. "CDRH2" sequences: "MSL10" residues 50-66 of SEQ ID NO: 3,
"MSL17" residues 50-66 of SEQ ID NO: 5, "MSL9-8" residues 50-66 of
SEQ ID NO: 7, "MSK-9" residues 50-66 of SEQ ID NO: 9, "MSK-13"
residues 50-66 of SEQ ID NO: 11 and "MSK-21" residues 52-69 of SEQ
ID NO: 13. "CDRH3" sequences: "MSL10" residues 99-108 of SEQ ID NO:
3, "MSL17" residues 99-115 of SEQ ID NO: 5, "MSL9-8" residues
99-107 of SEQ ID NO: 7, "MSK-9" residues 99-107 of SEQ ID NO: 9,
"MSK-13" residues 99-111 of SEQ ID NO: 11 and "MSK-21" residues
102-122 of SEQ ID NO: 13. "CDRL1" sequences: "MSL10" residues 23-35
of SEQ ID NO: 4, "MSL17" residues 23-33 of SEQ ID NO: 6, "MSL9-8"
residues 23-33 of SEQ ID NO: 8, "MSK-9" residues 24-39 of SEQ ID
NO: 10, "MSK-13" residues 24-39 of SEQ ID NO: 12 and "MSK-21"
residues 24-34 of SEQ ID NO: 14. "CDRL2" sequences: "MSL10"
residues 51-57 of SEQ ID NO: 4, "MSL17" residues 49-55 of SEQ ID
NO: 6, "MSL9-8" residues 49-55 of SEQ ID NO: 8, "MSK-9" residues
55-61 of SEQ ID NO: 10, "MSK-13" residues 55-61 of SEQ ID NO: 12
and "MSK-21" residues 50-56 of SEQ ID NO: 14. "CDRL3" sequences:
"MSL10" residues 90-101 of SEQ ID NO: 4, "MSL17" residues 88-96 of
SEQ ID NO: 6, "MSL9-8" residues 88-95 of SEQ ID NO: 8, "MSK-9"
residues 94-112 of SEQ ID NO: 10, "MSK-13" residues 94-113 of SEQ
ID NO: 12 and "MSK-21" residues 89-97 of SEQ ID NO: 14.
Example 1.2
Functional Characterization of Human SOST Antibodies
Example 1.2.1
Sclerostin Enzyme-Linked Immunosorbent Assay (ELISA) Protocols
Direct Bind ELISA
[0587] The following protocol was used to characterize the binding
of SOST antibodies to human SOST by enzyme-linked immunosorbent
assay (ELISA). ELISA plates were coated with 50 .mu.l per well of
goat anti-mouse IgG-Fc at 2 .mu.g/ml overnight at 4.degree. C.
(Jackson cat#115-005-164). Plates were washed 3 times with
PBS/Tween. 50 .mu.l antibody diluted to 1 .mu.g/ml in PBS/0.1% BSA
was added to appropriate wells and incubated for 1 hour at room
temperature (RT). Plates were washed 3 times with PBS/Tween. 50
.mu.l of serial diluted biotin-SOST was added to appropriate wells
and incubated for 1 hour at RT. Plates were washed 3 times with
PBS/Tween. 50 .mu.l of Streptavidin (Thermo Scientific cat#21126)
diluted 1:10,000 in PBS/0.1% BSA was added to the appropriate wells
and incubated for 1 hour at RT. Plates were washed 3 times with
PBS/Tween. 50 .mu.l of TMB (Zymed cat#002023) was added to the
appropriate wells and the reaction allowed to proceed for 1 minute.
The reaction was stopped with 50 .mu.l 2N H.sub.2SO.sub.4 and
absorbance was read at 450 nm.
Anti-Human Fc Capture ELISA
[0588] Antibody specific to anti-human Fc was diluted in 0.2 molar
sodium bicarbonate buffer (pH 9.4) to 1 .mu.g/ml. Plates were
coated with 100 .mu.l per well at RT for 2 hours. Blocking of
additional binding capacity was conducted at RT for 1 hour by
addition of 200 .mu.l of 5% non-fat dry milk in PBS to each well.
After washing plate, 100 .mu.l of 0.5 .mu.g/ml of diluted
individual antibodies were added to each well in duplicate. Plates
were incubated for 1 hour at RT and washed. 100 .mu.l of 1:6
serially diluted biotin rh (rcyno/rrat/rmouse) sclerostin from 100
nM to 0.001 nM was added to each well and plate was incubated at RT
for one hour. After washing, 1:10,000 diluted SA-HRP was added at
120 .mu.l per well. A 15 minute incubation with SA-HRP at RT was
followed by a wash. Finally, 120 .mu.l of TMB substrate from
Invitrogen (CAT#00-2023 Lot#425820A) was added to each well and
color allowed to develop for 10 minutes. The reaction was stopped
with 60 .mu.l of 2N sulfuric acid. The plate was read at 450 nm,
data collected and analyzed.
Anti-Biotin Capture ELISA
[0589] Goat anti-biotin (Sigma CAT B3640-1 MG) was diluted in 0.2 M
sodium bicarbonate buffer (Pierce CAT 28382 Lot IA 109342) pH 9.4
to a concentration of 1 .mu.g/ml. The plate was coated at 100 .mu.l
per well at RT for 2 hours. Blocking of additional binding capacity
was conducted at RT for 1 hour by addition of 200 .mu.l of 5%
non-fat dry milk in PBS to each well. After washing, 100 .mu.l of 2
.mu.g/ml of biotin rh (rcyno/r rat/rmouse) sclerostin was added to
each well. Samples were incubated at RT for 1 hour. After washing
the plate, 100 .mu.l of 1:6 serially diluted antibodies was added
to the wells starting at 25 .mu.g/ml. Samples were incubated at RT
for one hour and the plate washed. 1:5000 diluted anti human Fc-HRP
(Jackson CAT#109-036-098) was added at 120 .mu.l per well and
incubated at RT for 30 minutes. After washing, 120 .mu.l of TMB
substrate was added to each well (Invitrogen CAT#00-2023
Lot#425820A). Color was developed for 5 to 10 minutes. The reaction
was stopped with 60 .mu.l of 2N sulfuric acid. The plate was read
at 450 nm, and data collected and analyzed. Several sclerostin
specific antibodies were identified using this format (Table
11).
TABLE-US-00017 TABLE 11 Sclerostin antibodies Antibody name EC50,
hu scl (nM) MSL10 0.4267 (capture) MSL17 0.7783 (capture) MSL9-8
12.34 (capture) MSK9 +(direct) MSK13 +(direct) MSK21 +(direct)
Example 1.2.2
TopFlash Wnt Pathway Neutralization Assay
[0590] The following protocol was used to assess sclerostin
neutralizing properties of mAbs and DVD-binding proteins via
restoration of Wnt pathway activity inhibited by sclerostin. HEK
293A (Invitrogen, cat#:51-0036, lot#737470) cells were stably
transduced with TopFlash Lentivirus (SA Biosciences, cat CLS
018L-1) and a selected 1G6 clone was further infected with Wnt-1
lentivirus (Origene Cat# SC303644), resulting in clones that
co-express Luciferase and Wnt-1. One double stable clone (clone
#14) has been maintained in culture medium: DMEM (Invitrogen
Cat#11965-092) with 10% Qualified FBS (Invitrogen Cat#26140-079),
Pen-Strep (Invitrogen Cat#15140-122), L-glutamine (Invitrogen
Cat#25030-081 2 mM final), Sodium Pyruvate (Invitrogen
Cat#11360-070 final 1 mM) and 2.5 .mu.g/ml Puromycin (Invivogen
Cat#ant-pr-1) in T75 flasks until 80-90% confluent on day of assay.
Assay is performed in assay medium: culture medium without
puromycin. Human sclerostin (Abbott), cyno sclerostin (Abbott),
murine sclerostin and rat sclerostin were aliquoted and stored
frozen at -80.degree. C. On day 1 clone#14 cells are plated at
10,000 cells per well in 50 ul assay medium in black-sided, clear
bottomed tissue culture treated 96 well plates (Costar #3603) and
incubated at 37.degree. C. overnight (20-24 hrs). The next day (day
2) the sclerostin stock is diluted to 60 nM (4.times.) for human
and cyno and 200 nM (4.times.) for mouse and rat, in the assay
medium. Anti-sclerostin antibodies are diluted, usually starting at
800 nM (4.times.) in the assay medium. Media is removed from plates
with cells and replaced with 50 .mu.l/well of fresh assay medium.
Cells are next incubated for 1 hr with 25 .mu.l of 60 nM (4.times.)
for human and cyno sclerostin or 25 .mu.l of 200 nM for mouse and
rat sclerostin. After this anti-Sclerostin antibodies (25 .mu.l of
4.times.) are added to cells and plates are incubated overnight at
37.degree. C. (20-24 hrs). The final volume in each well is 100
.mu.l. The following day (day3) cells are washed once with 200
.mu.l of PBS (RT). Promega Luciferase Kit #E1501 is used for cell
lysis and Luciferase read out. Briefly, 5.times. cell lysis reagent
(Promega, cat #E153A) is diluted with milliQ water to 1.times. and
20 .mu.l is added to each well. To ensure a complete lysis, plate
is rotated 500 rpm for 20 min. 100 .mu.l of Luciferase assay
reagent (1 vial cat #E151A substrate+10 ml cat #E152A assay buffer)
is added to each well. Plate is read on TopCount machine (Program:
Luciferase 96, Assay 17:1 sec/well read).
[0591] AE10-6 AM2, AE10-6 AM3, AE10-6 AM5, AE10-6 AM6, AE10-6 AM7,
and AE10-6 AM8 neutralized recombinant human sclerostin with IC50
of 1.5-7.7 nM, recombinant cynomolgus monkey sclerostin with IC50
of 4.7-21.1 nM. AE10-6 AM3, AE10-6 AM7, and AE10-6 AM8 neutralized
recombinant mouse sclerostin with IC50 of 9.6-11.7 nM recombinant
rat sclerostin with IC50 of 12.1 to 15.7 nM. MSL10 AM1-11
antibodies neutralized recombinant human sclerostin with IC50 of
3-23.3 nM. MSL10 AM6, MSL10 AM7, MSL10 AM8, MSL10 AM9, MSL10 AM10,
MSL10 AM11 neutralized recombinant cynomolgus monkey sclerostin
with IC50 of 13.5-29.7 nM.
Example 1.2.3
Affinity Measurement of Sclerostin Antibodies by Surface Plasmon
Resonance
[0592] The binding of antibodies or anti-sclerostin-TNF DVD to
purified recombinant human, cynomolgus monkey (cyno), rat and mouse
sclerostin and TNF.alpha. was determined by surface plasmon
resonance-based measurements with a Biacore T100 or T200
instruments (GE Healthcare Life Sciences, Piscataway, N.J., USA)
using running HBS-EP+ (10 mM HEPES [pH 7.4], 150 mM NaCl, 3 mM
EDTA, and 0.005% surfactant P20) containing additional 150 mM NaCl,
and/or 10 mg/ml carboxymethyl dextran, 0.1 mg/ml BSA at 25.degree.
C. All chemicals were obtained from GE Healthcare Life Sciences,
Piscataway, N.J., USA or otherwise from a different source as
described in the text. Approximately 10,000 RU (or 3000 RU when CM3
chip was used) of goat anti-mouse or anti-human IgG (Fc.gamma.)
fragment specific polyclonal antibody (Pierce Biotechnology Inc,
Rockford, Ill.) diluted in 10 mM sodium acetate (pH 4.5) was
directly immobilized across a CM5 (or CM3) research grade biosensor
chip using a standard amine coupling kit according to
manufacturer's instructions and procedures at 15 .mu.g/ml.
Unreacted moieties on the biosensor surface were blocked with
ethanolamine. Modified carboxymethyl dextran surface in flowcells
2, 3 and 4 were used as a reaction surface. Modified carboxymethyl
dextran in flow cell 1 was used as the reference surface.
Biaevaluation T100 software version 2.0.2, GE Healthcare Life
Sciences was used to simultaneously fit association and
dissociation phases of all injections (using 1:1 fit analysis with
local Rmax). Purified antibodies were diluted in running buffer for
capture across goat anti-mouse or anti-human IgG specific reaction
surfaces. Antibodies to be captured as a ligand (1 .mu.g/ml) were
injected over reaction matrices at a flow rate of 10 .mu.l/minute.
The association and dissociation rate constants, kon (unit M-1 s-1)
and koff (unit s-1) were determined under a continuous flow rate of
50 .mu.l/minute. Rate constants were derived by making kinetic
binding measurements at 6 to 8 different antigen concentrations
ranging from 0.39-50 nM for sclerostin antigens and 0.195-25 nM for
TNF. At the end of each cycle the surfaces were regenerated with 10
s injection of 50 mM NaOH or/and by 10 s injection of 10 mM Glycine
pH1.5 at a flow rate of 100 .mu.l/min. Instrument appropriate
Biaevaluation software, GE Healthcare Life Sciences was used to
simultaneously fit association and dissociation constants as well
as K.sub.D (using 1:1 global fit analysis with local R.sub.max).
Binding was recorded as a function of time and kinetic rate
constants are calculated. In this assay, on-rates as fast as
10.sup.7 M.sup.-1 s.sup.-1 and off-rates as slow as 10.sup.-6
s.sup.-1 can be measured. In cases when the off rate for certain
antibodies/DVD-binding proteins was slower than 10.sup.-6 s.sup.-1,
k.sub.d was assumed to be lesser or equal to 1*10.sup.-6 s.sup.-1
and K.sub.D value was calculated under assumption of in such cases.
was done by dividing the on-rate by 1*10.sup.-6 and was given as
"at least".
[0593] The affinity and kinetic rates anti-sclerostin MSL10 clones
for human and cyno sclerostin are provided in Table 12 below, while
the affinity and kinetic rates of AE10-6 clones for all sclerostin
species (human, cyno, rat and mouse) are provided in Table 13 (the
MSL10 clones did not bind mouse and rat sclerostin).
TABLE-US-00018 TABLE 12 Affinity and kinetic rates of
anti-sclerostin MSL 10 clones for human and cyno SOST name antigen
k.sub.a, M.sup.-1s.sup.-1 k.sub.d, s.sup.-1 K.sub.D, M MSL10-.1 hu
sclerostin 2.80E+04 7.80E-05 2.80E-09 MSL10-AM1 hu sclerostin
5.20E+04 1.60E-05 3.00E-10 MSL10-AM2 hu sclerostin 1.20E+05
7.60E-06 6.50E-11 MSL10-AM3 hu sclerostin 8.40E+04 7.60E-05
9.00E-10 MSL10-AM4 hu sclerostin 1.40E+05 7.30E-04 5.30E-09
MSL10-AM5 hu sclerostin 7.70E+04 7.40E-05 9.70E-10 MSL10-AM6 hu
sclerostin 1.70E+05 5.50E-04 3.40E-09 MSL10-AM7 hu sclerostin
1.70E+05 7.20E-05 4.20E-10 MSL10-AM8 hu sclerostin 1.10E+05
7.70E-05 7.30E-10 MSL10-AM9 hu sclerostin 9.60E+04 5.20E-06
5.40E-11 MSL10-AM10 hu sclerostin 1.30E+05 3.90E-05 2.90E-10
MSL10-AM11 hu sclerostin 8.80E+04 7.40E-05 8.50E-10 MSL10-AM2.2 hu
sclerostin 3.40E+05 1.10E-04 3.30E-10 MSL10-AM3.2 hu sclerostin
2.80E+05 2.50E-04 9.00E-10 MSL10-.1 cyno sclerostin 3.40E+04
9.90E-05 2.90E-09 MSL10-AM1 cyno sclerostin 5.40E+04 2.30E-05
4.20E-10 MSL10-AM2 cyno sclerostin 1.10E+05 8.80E-05 7.70E-10
MSL10-AM3 cyno sclerostin 7.70E+04 1.00E-04 1.30E-09 MSL10-AM4 cyno
sclerostin 1.30E+05 7.30E-04 5.60E-09 MSL10-AM5 cyno sclerostin
8.20E+04 8.70E-05 1.10E-09 MSL10-AM2.2 cyno sclerostin 6.00E+05
6.70E-05 1.10E-10 MSL10-AM3.2 cyno sclerostin 3.00E+05 1.00E-04
3.60E-10
TABLE-US-00019 TABLE 13 Affinity and kinetic rates of
anti-sclerostin AE10-6 clones for human and cyno SOST name antigen
k.sub.a, M.sup.-1s.sup.-1 k.sub.d, s.sup.-1 K.sub.D, M AE10-6-AM2
hu sclerostin 3.40E+06 2.70E-05 8.10E-12 AE10-6-AM3 hu sclerostin
7.00E+06 3.80E-06 5.40E-13 AE10-6-AM5 hu sclerostin 4.30E+06
1.30E-05 3.00E-12 AE10-6-AM6 hu sclerostin 9.10E+06 1.20E-05
1.30E-12 AE10-6-AM7 hu sclerostin 7.10E+06 2.10E-06 3.00E-13
AE10-6-AM8 hu sclerostin 7.40E+06 9.70E-06 1.30E-12 AE10-6-AM2 Cyno
sclerostin 3.60E+06 2.60E-05 7.10E-12 AE10-6-AM3 Cyno sclerostin
6.90E+06 1.60E-06 2.40E-13 AE10-6-AM5 Cyno sclerostin 5.20E+06
7.60E-06 1.50E-12 AE10-6-AM6 Cyno sclerostin 7.90E+06 7.00E-06
8.80E-13 AE10-6-AM7 Cyno sclerostin 7.00E+06 <1e-7 <1.4E-13
AE10-6-AM8 Cyno sclerostin 7.70E+06 3.00E-06 3.90E-13 AE10-6-AM2 ms
sclerostin 2.10E+07 2.00E-03 9.40E-11 AE10-6-AM3 ms sclerostin
1.00E+07 5.50E-05 5.50E-12 AE10-6-AM5 ms sclerostin 7.00E+06
1.10E-04 1.60E-11 AE10-6-AM6 ms sclerostin 3.20E+07 8.70E-04
2.70E-11 AE10-6-AM7 ms sclerostin 1.30E+07 1.20E-04 9.30E-12
AE10-6-AM8 ms sclerostin 1.10E+07 8.20E-05 7.40E-12 AE10-6-AM2 rat
sclerostin 7.60E+06 8.80E-04 1.20E-10 AE10-6-AM3 rat sclerostin
1.90E+06 1.90E-05 1.00E-11 AE10-6-AM5 rat sclerostin 1.30E+06
7.50E-05 5.60E-11 AE10-6-AM6 rat sclerostin 1.90E+06 1.80E-04
9.80E-11 AE10-6-AM7 rat sclerostin 1.80E+06 4.60E-05 2.60E-11
AE10-6-AM8 rat sclerostin 1.80E+06 5.90E-05 3.40E-11
Example 2
Pharmacokinetic Analysis of SOST Antibodies in Rat
[0594] Pharmacokinetic studies of human anti-human SOST antibodies
are carried out in Sprague Dawley rats. Male rats are dosed
intravenously with a single dose of 4 mg/kg of antibody proteins
and serum samples are analyzed using antigen capture based
chemiluminescent MSD (Meso Scale Discovery) method. Pharmacokinetic
parameters are calculated by non-compartmental analysis using
WinNonlin.
Example 2.2.1
Preparation of Rat Serum
[0595] Surgically altered (jugular vein cannulated, JVC) and
regular male Sprague-Dawley Rats (approximately seven weeks old,
weighing 240-390 grams) are purchased from Charles River
Laboratories (Wilmington, Mass.). The animals are housed in rooms
maintained at constant temperature and humidity under 12 hour
light/dark cycle, fed with normal rodent chow and are allowed food
and water ad libitum. Hydration and clinical conditions of the
animals are monitored daily.
[0596] Blood samples are collected (0.2 mL) at various timepoints,
allowed to clot for 30 minutes at room temperature, and centrifuged
for 8 minutes at 13,200 rpm. Serum is transferred to eppendorf
tubes and stored frozen at -80.degree. C.
Example 2.2.2
MSD Assay Used to Quantify SOST Antibody in PK Serum Samples
[0597] MSD streptavidin plates (Meso Scale Discovery) are washed
with phosphate buffered saline containing 0.05% Tween-20 (diluted
from 10.times.PBS, Abbott Bioresearch Center, Media Room,
Worcester, Mass. and Tween-20, Sigma, St. Louis, Mo.). Plates are
blocked with 150 .mu.L/well blocking solution (MSD Block, Meso
Scale Discovery, diluted to 3% final concentration in PBS) for 1
hour, covered, with shaking (600 rpm) at room temperature.
[0598] Prior to analysis, rat serum samples are thawed on ice,
mixed gently, and centrifuged at 14,000 rpm for 3 minutes at
4.degree. C. in an eppendorf centrifuge. Standard curve and control
samples are prepared in rat serum. Study samples, standard curve
samples, blanks, and quality control samples are incubated in
solution in a separate 2 mL deep well 96-well plate (Corning,
Corning, N.Y.) 1:1:1=V:V:V with biotinylated human SOST (0.1 ug/mL
in assay buffer) and sulfo-tagged goat anti-human IgG (Meso Scale
Discovery, 1 .mu.g/mL in assay buffer) for 1 hour at room
temperature. The samples are then transferred to the MSD plates and
incubated for an additional hour with shaking (600 rpm) at room
temperature. The MSD plates are washed and developed with 2.times.
Read Buffer (Meso Scale Discovery). Chemiluminiscence is measured
within ten minutes on the MSD Sector Imager 6000.
[0599] Standard curves are analyzed using four-parameter logistic
fit and sample concentrations are calculated by XLfit4 software
version 2.2.1 Build 16, (Microsoft Corporation, Redmond, Wash.).
Pharmacokinetic parameters are calculated for each animal using
Winonlin software version 5.0.1 (Pharsight Corporation, Mountain
View, Calif.) by noncompartmental analysis.
Example 3
Generation of TNF/SOST DVD-Binding Proteins
Example 3.1
Construction of TNF/SOST DVD-Binding Protein DNA Constructs
[0600] Anti-TNF antibody variable domains from human antibody D2E7,
deimmunized D2E7, mouse or humanized antibody MAK199, and mouse or
humanized antibody MAK195 were combined with multiple SOST antibody
variable domains by overlapping PCR amplification with intervening
linker DNA sequences. The amplified PCR products were subcloned
into expression vectors suitable for transient expression in HEK293
cells and the open reading frame regions are confirmed by
sequencing before DVD-binding protein expression.
Example 3.2
Expression and Production of TNF/SOST DVD-Binding Proteins
[0601] All DVD-binding protein cDNA constructs were sequenced,
expanded in E. coli and DNA purified using Qiagen Hispeed Maxi
Preps (CAT#12662, QIAGEN). DVD-binding protein DNA was transfected
into log phase 293E cells (0.5.times.10.sup.6/ml, viability>95%)
by mixing PEI and DNA @ 2:1 ratio with 0.2 .mu.g/ml heavy chain DNA
and 0.3 .mu.g/ml light chain DNA. DNA:PEI complex was formed at
room temperature in TC hood for fifteen minutes before adding to
293E cells. Twenty four later, 0.5% TN1 was added to 293E cells. At
day five, supernatants were collected for human IgG1 titer
measurement. Cell supernatant was harvested at day seven and
filtered through a 0.2 .mu.M PES filter. Supernatant was purified
by using Protein A Sepharose Affinity Chromatography according to
the manufacturer's instruction. Purified DVD-binding proteinss were
eluted from the column by 0.1 M glycine (pH 2.99), buffered by 2 M
tris-HCl or phosphate and/or dialyzed into 15 mM histidine buffer
(pH 6.0) immediately. The binding proteins were quantitated by A280
and analyzed by mass spectrometry and SEC.
Example 3.3
Sequences of TNF/SOST DVD-Binding Constructs
[0602] Amino acid sequence of heavy chain and light chain of
DVD-binding proteins capable of binding human TNF and SOST were
determined. The amino acid sequences of variable heavy chains,
variable light chains, and constant regions of TNF/SOST DVD-binding
proteins are provided in the table below (Table 18).
TABLE-US-00020 TABLE 18 Sequences Of Variable And Constant Regions
Of TNF/SOST DVD-Binding Proteins DVD- Binding Protein Outer Inner
Variable Variable Variable Domain Domain Domain Sequence Sequence
Name Name Name Identifier 123456789012345678901234567890 DVD-
AB247VH AB387VH SEQ ID NO: 21 EIQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSGGGGSGGGGSEVQLVESGGGL DVD2014H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
22 EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 23 GGGGSGGGGS VH SEQ ID NO: 24
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 25 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB247VL AB387VL SEQ ID NO: 26
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYF Light
CQQGNTLPPTFGQGTKLEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2014L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 27
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYF CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 28 GGSGGGGSG VL SEQ ID NO: 29 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 30
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB247VH SEQ ID NO: 31 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSEIQLVQSGSELKKPGA DVD2015H
SVKVSCKASGYTFTNYGMNWVRQAPGQGL EWMGWINTYTGEPTYADDFKGRFVFSLDT
SVSTAYLQISSLKAEDTAVYFCARKFLTT VVVTDYAMDYWGQGTTVTVSS VH SEQ ID NO:
32 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 33 GGGGSGGGGS VH SEQ ID NO: 34
EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 35 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB247VL SEQ ID NO: 36
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQSPSSLSASVGDRVTITCR DVD2015L
ASQDISNYLNWYQQKPGKTVKLLIYYTSR LQSGVPSRFSGSGSGTDYTLTISSLQPED
FATYFCQQGNTLPPTFGQGTKLEIKR VL SEQ ID NO: 37
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 38 GGSGGGGSG VL SEQ ID NO: 39 DIQMTQSPSSLSASVGDRVTITCRASQDI
SNYLNWYQQKPGKTVKLLIYYTSRLQSGV PSRFSGSGSGTDYTLTISSLQPEDFATYF
CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO: 40
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB247VH
AB387VH SEQ ID NO: 41 EIQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSASTKGPEVQLVESGGGLVQPG DVD2016H
GSLRLSCTASGFTFDDYALHWVRQAPGKG LEWVSGISWHGDFIDYADSVKGRFTISRD
NSKNTLYLQMNGLRVEDMAIYYCAGNNRG YGGLDVWGQGTTVTVSS VH SEQ ID NO: 42
EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 43 ASTKGP VH SEQ ID NO: 44
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 45 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB247VL AB387VL SEQ ID NO: 46
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYF Light
CQQGNTLPPTFGQGTKLEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2016L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 47
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYF CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 48 TVAAP VL SEQ ID NO: 49 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 50
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB247VH SEQ ID NO: 51 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEIQLVQSGSELKKPGASVKV DVD2017H
SCKASGYTFTNYGMNWVRQAPGQGLEWMG WINTYTGEPTYADDFKGRFVFSLDTSVST
AYLQISSLKAEDTAVYFCARKFLTTVVVT DYAMDYWGQGTTVTVSS VH SEQ ID NO: 52
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 53 ASTKGP VH SEQ ID NO: 54
EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 55 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB247VL SEQ ID NO: 56
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASQ DVD2017L
DISNYLNWYQQKPGKTVKLLIYYTSRLQS GVPSRFSGSGSGTDYTLTISSLQPEDFAT
YFCQQGNTLPPTFGQGTKLEIKR VL SEQ ID NO: 57
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG
Linker SEQ ID NO: 58 QPKAAP VL SEQ ID NO: 59
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYF CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO:
60 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB247VH
AB387VH SEQ ID NO: 61 EIQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSASTKGPEVQLVESGGGLVQPG DVD2018H
GSLRLSCTASGFTFDDYALHWVRQAPGKG LEWVSGISWHGDFIDYADSVKGRFTISRD
NSKNTLYLQMNGLRVEDMAIYYCAGNNRG YGGLDVWGQGTTVTVSS VH SEQ ID NO: 62
EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 63 ASTKGP VH SEQ ID NO: 64
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 65 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB247VL AB387VL SEQ ID NO: 66
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYF Light
CQQGNTLPPTFGQGTKLEIKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2018L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 67
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYF CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 68 TVAAPSVFIFPP VL SEQ ID NO: 69 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 70
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB247VH SEQ ID NO: 71 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEIQLVQSGSELKKPGASVKV DVD2019H
SCKASGYTFTNYGMNWVRQAPGQGLEWMG WINTYTGEPTYADDFKGRFVFSLDTSVST
AYLQISSLKAEDTAVYFCARKFLTTVVVT DYAMDYWGQGTTVTVSS VH SEQ ID NO: 72
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 73 ASTKGP VH SEQ ID NO: 74
EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 75 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB247VL SEQ ID NO: 76
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQSPSSLSASVGDRVT DVD2019L
ITCRASQDISNYLNWYQQKPGKTVKLLIY YTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK R VL SEQ ID NO: 77
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 78 QPKAAPSVTLFPP VL SEQ ID NO: 79
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYF CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO:
80 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB247VH
AB387VH SEQ ID NO: 81 EIQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSASTKGPSVFPLAPEVQLVESG DVD2020H
GGLVQPGGSLRLSCTASGFTFDDYALHWV RQAPGKGLEWVSGISWHGDFIDYADSVKG
RFTISRDNSKNTLYLQMNGLRVEDMAIYY CAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
NO: 82 EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 83 ASTKGPSVFPLAP VH SEQ ID NO: 84
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 85 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB247VL AB387VL SEQ ID NO: 86
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYF Light
CQQGNTLPPTFGQGTKLEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2020L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 87
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYF CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 88 TVAAP VL SEQ ID NO: 89 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 90
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB247VH SEQ ID NO: 91 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPEIQLVQSGSELKK DVD2021H
PGASVKVSCKASGYTFTNYGMNWVRQAPG QGLEWMGWINTYTGEPTYADDFKGRFVFS
LDTSVSTAYLQISSLKAEDTAVYFCARKF LTTVVVTDYAMDYWGQGTTVTVSS VH SEQ ID
NO: 92 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 93 ASTKGPSVFPLAP VH SEQ ID NO: 94
EIQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 95 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB247VL SEQ ID NO: 96
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASQ DVD2021L
DISNYLNWYQQKPGKTVKLLIYYTSRLQS GVPSRFSGSGSGTDYTLTISSLQPEDFAT
YFCQQGNTLPPTFGQGTKLEIKR VL SEQ ID NO: 97
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 98 QPKAAP VL SEQ ID NO: 99 DIQMTQSPSSLSASVGDRVTITCRASQDI
SNYLNWYQQKPGKTVKLLIYYTSRLQSGV PSRFSGSGSGTDYTLTISSLQPEDFATYF
CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO: 100
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB365VH
AB387VH SEQ ID NO: 101 EVQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSGGGGSGGGGSEVQLVESGGGL DVD2022H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
102 EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 103 GGGGSGGGGS VH SEQ ID NO: 104
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 105 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB365VL AB387VL SEQ ID NO: 106
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYY Light
CQQGNTLPPTFGQGTKLEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2022L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 107
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 108 GGSGGGGSG VL SEQ ID NO: 109 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 110
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB365VH SEQ ID NO: 111 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSEVQLVQSGSELKKPGA DVD2023H
SVKVSCKASGYTFTNYGMNWVRQAPGQGL EWMGWINTYTGEPTYADDFKGRFVFSLDT
SVSTAYLQISSLKAEDTAVYFCARKFLTT VVVTDYAMDYWGQGTTVTVSS VH SEQ ID NO:
112 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 113 GGGGSGGGGS VH SEQ ID NO: 114
EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 115 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB365VL SEQ ID NO: 116
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQSPSSLSASVGDRVTITCR DVD2023L
ASQDISNYLNWYQQKPGKAPKLLIYYTSR LQSGVPSRFSGSGSGTDYTLTISSLQPED
FATYYCQQGNTLPPTFGQGTKLEIKR VL SEQ ID NO: 117
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 118 GGSGGGGSG VL SEQ ID NO: 119
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO:
120 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB365VH
AB387VH SEQ ID NO: 121 EVQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSASTKGPEVQLVESGGGLVQPG DVD2024H
GSLRLSCTASGFTFDDYALHWVRQAPGKG LEWVSGISWHGDFIDYADSVKGRFTISRD
NSKNTLYLQMNGLRVEDMAIYYCAGNNRG YGGLDVWGQGTTVTVSS VH SEQ ID NO: 122
EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 123 ASTKGP VH SEQ ID NO: 124
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 125 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB365VL AB387VL SEQ ID NO: 126
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYY Light
CQQGNTLPPTFGQGTKLEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2024L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 127
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 128 TVAAP VL SEQ ID NO: 129 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 130
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB365VH SEQ ID NO: 131 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVQSGSELKKPGASVKV DVD2025H
SCKASGYTFTNYGMNWVRQAPGQGLEWMG WINTYTGEPTYADDFKGRFVFSLDTSVST
AYLQISSLKAEDTAVYFCARKFLTTVVVT DYAMDYWGQGTTVTVSS VH SEQ ID NO: 132
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 133 ASTKGP VH SEQ ID NO: 134
EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 135 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB365VL SEQ ID NO: 136
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASQ DVD2025L
DISNYLNWYQQKPGKAPKLLIYYTSRLQS GVPSRFSGSGSGTDYTLTISSLQPEDFAT
YYCQQGNTLPPTFGQGTKLEIKR VL SEQ ID NO: 137
QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ ID NO: 138 QPKAAP VL SEQ ID
NO: 139 DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO:
140 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB365VH
AB387VH SEQ ID NO: 141 EVQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSASTKGPEVQLVESGGGLVQPG DVD2026H
GSLRLSCTASGFTFDDYALHWVRQAPGKG LEWVSGISWHGDFIDYADSVKGRFTISRD
NSKNTLYLQMNGLRVEDMAIYYCAGNNRG YGGLDVWGQGTTVTVSS VH SEQ ID NO: 142
EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 143 ASTKGP VH SEQ ID NO: 144
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 145 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB365VL AB387VL SEQ ID NO: 146
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYY Light
CQQGNTLPPTFGQGTKLEIKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2026L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 147
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 148 TVAAPSVFIFPP VL SEQ ID NO: 149
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID
NO: 150 QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB365VH SEQ ID NO: 151 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVQSGSELKKPGASVKV DVD2027H
SCKASGYTFTNYGMNWVRQAPGQGLEWMG WINTYTGEPTYADDFKGRFVFSLDTSVST
AYLQISSLKAEDTAVYFCARKFLTTVVVT DYAMDYWGQGTTVTVSS VH SEQ ID NO: 152
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 153 ASTKGP VH SEQ ID NO: 154
EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 155 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB365VL SEQ ID NO: 156
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQSPSSLSASVGDRVT DVD2027L
ITCRASQDISNYLNWYQQKPGKAPKLLIY YTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK R VL SEQ ID NO: 157
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 158 QPKAAPSVTLFPP VL SEQ ID NO: 159
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO:
160 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB365VH
AB387VH SEQ ID NO: 161 EVQLVQSGSELKKPGASVKVSCKASGYTF Binding
TNYGMNWVRQAPGQGLEWMGWINTYTGEP Protein TYADDFKGRFVFSLDTSVSTAYLQISSLK
Heavy AEDTAVYFCARKFLTTVVVTDYAMDYWGQ Variable
GTTVTVSSASTKGPSVFPLAPEVQLVESG DVD2028H
GGLVQPGGSLRLSCTASGFTFDDYALHWV RQAPGKGLEWVSGISWHGDFIDYADSVKG
RFTISRDNSKNTLYLQMNGLRVEDMAIYY CAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
NO: 162 EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS Linker SEQ ID NO: 163 ASTKGPSVFPLAP VH SEQ ID NO: 164
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 165 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB365VL AB387VL SEQ ID NO: 166
DIQMTQSPSSLSASVGDRVTITCRASQDI Binding SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYTLTISSLQPEDFATYY Light
CQQGNTLPPTFGQGTKLEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2028L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 167
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR Linker SEQ ID
NO: 168 TVAAP VL SEQ ID NO: 169 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 170
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB365VH SEQ ID NO: 171 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPEVQLVQSGSELKK DVD2029H
PGASVKVSCKASGYTFTNYGMNWVRQAPG QGLEWMGWINTYTGEPTYADDFKGRFVFS
LDTSVSTAYLQISSLKAEDTAVYFCARKF LTTVVVTDYAMDYWGQGTTVTVSS VH SEQ ID
NO: 172 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 173 ASTKGPSVFPLAP VH SEQ ID NO: 174
EVQLVQSGSELKKPGASVKVSCKASGYTF TNYGMNWVRQAPGQGLEWMGWINTYTGEP
TYADDFKGRFVFSLDTSVSTAYLQISSLK AEDTAVYFCARKFLTTVVVTDYAMDYWGQ
GTTVTVSS CH SEQ ID NO: 175 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB365VL SEQ ID NO: 176
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASQ DVD2029L
DISNYLNWYQQKPGKAPKLLIYYTSRLQS GVPSRFSGSGSGTDYTLTISSLQPEDFAT
YYCQQGNTLPPTFGQGTKLEIKR
VL SEQ ID NO: 177 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ ID NO: 178 QPKAAP VL SEQ ID
NO: 179 DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLQSGV
PSRFSGSGSGTDYTLTISSLQPEDFATYY CQQGNTLPPTFGQGTKLEIKR CL SEQ ID NO:
180 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB388VH
AB387VH SEQ ID NO: 181 QIQLVQSGPELKKPGETVMISCKASGYTF Binding
TNYGMNWVKQAPGKGLKWMGWINTYTGEP Protein TYADDFKGRFAFSLETSASTAYLQINNLK
Heavy NEDTATYFCARKFLTTVVVTDYAMDYWGQ Variable
GTSVTVSSGGGGSGGGGSEVQLVESGGGL DVD2030H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
182 QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS Linker SEQ ID NO: 183 GGGGSGGGGS VH SEQ ID NO: 184
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 185 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB388VL AB387VL SEQ ID NO: 186
DIQMTQTTSSLSASLGDRVTISCRASQDI Binding SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYSLTISNLEQEDIATYF Light
CQQGNTLPPTFGVGTKLELKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2030L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 187
DIQMTQTTSSLSASLGDRVTISCRASQDI SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYSLTISNLEQEDIATYF CQQGNTLPPTFGVGTKLELKR Linker SEQ ID
NO: 188 GGSGGGGSG VL SEQ ID NO: 189 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 190
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB388VH SEQ ID NO: 191 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSQIQLVQSGPELKKPGE DVD2031H
TVMISCKASGYTFTNYGMNWVKQAPGKGL KWMGWINTYTGEPTYADDFKGRFAFSLET
SASTAYLQINNLKNEDTATYFCARKFLTT VVVTDYAMDYWGQGTSVTVSS VH SEQ ID NO:
192 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 193 GGGGSGGGGS VH SEQ ID NO: 194
QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS CH SEQ ID NO: 195 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB388VL SEQ ID NO: 196
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQTTSSLSASLGDRVTISCR DVD2031L
ASQDISNYLNWYQQKPDGTVKLLIYYTSR LQSGVPSRFSGSGSGTDYSLTISNLEQED
IATYFCQQGNTLPPTFGVGTKLELKR VL SEQ ID NO: 197
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 198 GGSGGGGSG VL SEQ ID NO: 199
DIQMTQTTSSLSASLGDRVTISCRASQDI SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYSLTISNLEQEDIATYF CQQGNTLPPTFGVGTKLELKR CL SEQ ID NO:
200 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB388VH
AB387VH SEQ ID NO: 201 QIQLVQSGPELKKPGETVMISCKASGYTF Binding
TNYGMNWVKQAPGKGLKWMGWINTYTGEP Protein TYADDFKGRFAFSLETSASTAYLQINNLK
Heavy NEDTATYFCARKFLTTVVVTDYAMDYWGQ Variable
GTSVTVSSASTKGPEVQLVESGGGLVQPG DVD2032H
GSLRLSCTASGFTFDDYALHWVRQAPGKG LEWVSGISWHGDFIDYADSVKGRFTISRD
NSKNTLYLQMNGLRVEDMAIYYCAGNNRG YGGLDVWGQGTTVTVSS VH SEQ ID NO: 202
QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS Linker SEQ ID NO: 203 ASTKGP VH SEQ ID NO: 204
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 205 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB388VL AB387VL SEQ ID NO: 206
DIQMTQTTSSLSASLGDRVTISCRASQDI Binding SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYSLTISNLEQEDIATYF Light
CQQGNTLPPTFGVGTKLELKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2032L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 207
DIQMTQTTSSLSASLGDRVTISCRASQDI SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYSLTISNLEQEDIATYF CQQGNTLPPTFGVGTKLELKR Linker SEQ ID
NO: 208 TVAAP VL SEQ ID NO: 209 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 210
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB388VH SEQ ID NO: 211 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPQIQLVQSGPELKKPGETVMI DVD2033H
SCKASGYTFTNYGMNWVKQAPGKGLKWMG WINTYTGEPTYADDFKGRFAFSLETSAST
AYLQINNLKNEDTATYFCARKFLTTVVVT DYAMDYWGQGTSVTVSS VH SEQ ID NO: 212
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 213 ASTKGP VH SEQ ID NO: 214
QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS CH SEQ ID NO: 215 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB388VL SEQ ID NO: 216
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQTTSSLSASLGDRVTISCRASQ DVD2033L
DISNYLNWYQQKPDGTVKLLIYYTSRLQS GVPSRFSGSGSGTDYSLTISNLEQEDIAT
YFCQQGNTLPPTFGVGTKLELKR VL SEQ ID NO: 217
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 218 QPKAAP VL SEQ ID NO: 219 DIQMTQTTSSLSASLGDRVTISCRASQDI
SNYLNWYQQKPDGTVKLLIYYTSRLQSGV PSRFSGSGSGTDYSLTISNLEQEDIATYF
CQQGNTLPPTFGVGTKLELKR CL SEQ ID NO: 220
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB388VH
AB387VH SEQ ID NO: 221 QIQLVQSGPELKKPGETVMISCKASGYTF Binding
TNYGMNWVKQAPGKGLKWMGWINTYTGEP Protein TYADDFKGRFAFSLETSASTAYLQINNLK
Heavy NEDTATYFCARKFLTTVVVTDYAMDYWGQ Variable
GTSVTVSSASTKGPEVQLVESGGGLVQPG DVD2034H
GSLRLSCTASGFTFDDYALHWVRQAPGKG LEWVSGISWHGDFIDYADSVKGRFTISRD
NSKNTLYLQMNGLRVEDMAIYYCAGNNRG YGGLDVWGQGTTVTVSS VH SEQ ID NO: 222
QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS Linker SEQ ID NO: 223 ASTKGP VH SEQ ID NO: 224
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 225 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB388VL AB387VL SEQ ID NO: 226
DIQMTQTTSSLSASLGDRVTISCRASQDI Binding SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYSLTISNLEQEDIATYF Light
CQQGNTLPPTFGVGTKLELKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2034L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 227
DIQMTQTTSSLSASLGDRVTISCRASQDI SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYSLTISNLEQEDIATYF CQQGNTLPPTFGVGTKLELKR Linker SEQ ID
NO: 228 TVAAPSVFIFPP VL SEQ ID NO: 229
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID
NO: 230 QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB388VH SEQ ID NO: 231 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPQIQLVQSGPELKKPGETVMI DVD2035H
SCKASGYTFTNYGMNWVKQAPGKGLKWMG WINTYTGEPTYADDFKGRFAFSLETSAST
AYLQINNLKNEDTATYFCARKFLTTVVVT DYAMDYWGQGTSVTVSS VH SEQ ID NO: 232
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 233 ASTKGP VH SEQ ID NO: 234
QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS CH SEQ ID NO: 235 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB388VL SEQ ID NO: 236
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQTTSSLSASLGDRVT DVD2035L
ISCRASQDISNYLNWYQQKPDGTVKLLIY YTSRLQSGVPSRFSGSGSGTDYSLTISNL
EQEDIATYFCQQGNTLPPTFGVGTKLELK R VL SEQ ID NO: 237
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 238 QPKAAPSVTLFPP VL SEQ ID NO: 239
DIQMTQTTSSLSASLGDRVTISCRASQDI SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYSLTISNLEQEDIATYF CQQGNTLPPTFGVGTKLELKR CL SEQ ID NO:
240 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB388VH
AB387VH SEQ ID NO: 241 QIQLVQSGPELKKPGETVMISCKASGYTF Binding
TNYGMNWVKQAPGKGLKWMGWINTYTGEP Protein TYADDFKGRFAFSLETSASTAYLQINNLK
Heavy NEDTATYFCARKFLTTVVVTDYAMDYWGQ Variable
GTSVTVSSASTKGPSVFPLAPEVQLVESG DVD2036H
GGLVQPGGSLRLSCTASGFTFDDYALHWV RQAPGKGLEWVSGISWHGDFIDYADSVKG
RFTISRDNSKNTLYLQMNGLRVEDMAIYY CAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
NO: 242 QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS Linker SEQ ID NO: 243 ASTKGPSVFPLAP VH SEQ ID NO: 244
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 245 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB388VL AB387VL SEQ ID NO: 246
DIQMTQTTSSLSASLGDRVTISCRASQDI Binding SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
Protein PSRFSGSGSGTDYSLTISNLEQEDIATYF Light
CQQGNTLPPTFGVGTKLELKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2036L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 247
DIQMTQTTSSLSASLGDRVTISCRASQDI SNYLNWYQQKPDGTVKLLIYYTSRLQSGV
PSRFSGSGSGTDYSLTISNLEQEDIATYF CQQGNTLPPTFGVGTKLELKR Linker SEQ ID
NO: 248 TVAAP VL SEQ ID NO: 249 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 250
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB388VH SEQ ID NO: 251 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPQIQLVQSGPELKK DVD2037H
PGETVMISCKASGYTFTNYGMNWVKQAPG KGLKWMGWINTYTGEPTYADDFKGRFAFS
LETSASTAYLQINNLKNEDTATYFCARKF LTTVVVTDYAMDYWGQGTSVTVSS VH SEQ ID
NO: 252 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 253 ASTKGPSVFPLAP VH SEQ ID NO: 254
QIQLVQSGPELKKPGETVMISCKASGYTF TNYGMNWVKQAPGKGLKWMGWINTYTGEP
TYADDFKGRFAFSLETSASTAYLQINNLK NEDTATYFCARKFLTTVVVTDYAMDYWGQ
GTSVTVSS CH SEQ ID NO: 255 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB388VL SEQ ID NO: 256
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQTTSSLSASLGDRVTISCRASQ
DVD2037L DISNYLNWYQQKPDGTVKLLIYYTSRLQS
GVPSRFSGSGSGTDYSLTISNLEQEDIAT YFCQQGNTLPPTFGVGTKLELKR VL SEQ ID NO:
257 QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 258 QPKAAP VL SEQ ID NO: 259 DIQMTQTTSSLSASLGDRVTISCRASQDI
SNYLNWYQQKPDGTVKLLIYYTSRLQSGV PSRFSGSGSGTDYSLTISNLEQEDIATYF
CQQGNTLPPTFGVGTKLELKR CL SEQ ID NO: 260
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB213VH
AB387VH SEQ ID NO: 261 QVQLKESGPGLVAPSQSLSITCTVSGFSL Binding
TDYGVNWVRQPPGKGLEWLGMIWGDGSTD Protein YDSTLKSRLSISKDNSKSQIFLKMNSLQT
Heavy DDTARYYCAREWHHGPVAYWGQGTLVTVS Variable
AGGGGSGGGGSEVQLVESGGGLVQPGGSL DVD2038H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 262
QVQLKESGPGLVAPSQSLSITCTVSGFSL TDYGVNWVRQPPGKGLEWLGMIWGDGSTD
YDSTLKSRLSISKDNSKSQIFLKMNSLQT DDTARYYCAREWHHGPVAYWGQGTLVTVS A
Linker SEQ ID NO: 263 GGGGSGGGGS VH SEQ ID NO: 264
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 265 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB213VL AB387VL SEQ ID NO: 266
DIVMTQSHKFMSTTVGDRVSITCKASQAV Binding SSAVAWYQQKPGQSPKLLIYWASTRHTGV
Protein PDRFTGSGSVTDFTLTIHNLQAEDLALYY Light
CQQHYSTPFTFGSGTKLEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2038L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 267
DIVMTQSHKFMSTTVGDRVSITCKASQAV SSAVAWYQQKPGQSPKLLIYWASTRHTGV
PDRFTGSGSVTDFTLTIHNLQAEDLALYY CQQHYSTPFTFGSGTKLEIKR Linker SEQ ID
NO: 268 GGSGGGGSG VL SEQ ID NO: 269 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 270
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB213VH SEQ ID NO: 271 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSQVQLKESGPGLVAPSQ DVD2039H
SLSITCTVSGFSLTDYGVNWVRQPPGKGL EWLGMIWGDGSTDYDSTLKSRLSISKDNS
KSQIFLKMNSLQTDDTARYYCAREWHHGP VAYWGQGTLVTVSA VH SEQ ID NO: 272
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 273 GGGGSGGGGS VH SEQ ID NO: 274
QVQLKESGPGLVAPSQSLSITCTVSGFSL TDYGVNWVRQPPGKGLEWLGMIWGDGSTD
YDSTLKSRLSISKDNSKSQIFLKMNSLQT DDTARYYCAREWHHGPVAYWGQGTLVTVS A CH
SEQ ID NO: 275 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB213VL SEQ ID NO: 276
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIVMTQSHKFMSTTVGDRVSITCK DVD2039L
ASQAVSSAVAWYQQKPGQSPKLLIYWAST RHTGVPDRFTGSGSVTDFTLTIHNLQAED
LALYYCQQHYSTPFTFGSGTKLEIKR VL SEQ ID NO: 277
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 278 GGSGGGGSG VL SEQ ID NO: 279
DIVMTQSHKFMSTTVGDRVSITCKASQAV SSAVAWYQQKPGQSPKLLIYWASTRHTGV
PDRFTGSGSVTDFTLTIHNLQAEDLALYY CQQHYSTPFTFGSGTKLEIKR CL SEQ ID NO:
280 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB213VH
AB387VH SEQ ID NO: 281 QVQLKESGPGLVAPSQSLSITCTVSGFSL Binding
TDYGVNWVRQPPGKGLEWLGMIWGDGSTD Protein YDSTLKSRLSISKDNSKSQIFLKMNSLQT
Heavy DDTARYYCAREWHHGPVAYWGQGTLVTVS Variable
AASTKGPEVQLVESGGGLVQPGGSLRLSC DVD2040H
TASGFTFDDYALHWVRQAPGKGLEWVSGI SWHGDFIDYADSVKGRFTISRDNSKNTLY
LQMNGLRVEDMAIYYCAGNNRGYGGLDVW GQGTTVTVSS VH SEQ ID NO: 282
QVQLKESGPGLVAPSQSLSITCTVSGFSL TDYGVNWVRQPPGKGLEWLGMIWGDGSTD
YDSTLKSRLSISKDNSKSQIFLKMNSLQT DDTARYYCAREWHHGPVAYWGQGTLVTVS A
Linker SEQ ID NO: 283 ASTKGP VH SEQ ID NO: 284
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 285 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB213VL AB387VL SEQ ID NO: 286
DIVMTQSHKFMSTTVGDRVSITCKASQAV Binding SSAVAWYQQKPGQSPKLLIYWASTRHTGV
Protein PDRFTGSGSVTDFTLTIHNLQAEDLALYY Light
CQQHYSTPFTFGSGTKLEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2040L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 287
DIVMTQSHKFMSTTVGDRVSITCKASQAV SSAVAWYQQKPGQSPKLLIYWASTRHTGV
PDRFTGSGSVTDFTLTIHNLQAEDLALYY CQQHYSTPFTFGSGTKLEIKR Linker SEQ ID
NO: 288 TVAAP VL SEQ ID NO: 289 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 290
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB213VH SEQ ID NO: 291 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPQVQLKESGPGLVAPSQSLSI DVD2041H
TCTVSGFSLTDYGVNWVRQPPGKGLEWLG MIWGDGSTDYDSTLKSRLSISKDNSKSQI
FLKMNSLQTDDTARYYCAREWHHGPVAYW GQGTLVTVSA VH SEQ ID NO: 292
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 293 ASTKGP VH SEQ ID NO: 294
QVQLKESGPGLVAPSQSLSITCTVSGFSL TDYGVNWVRQPPGKGLEWLGMIWGDGSTD
YDSTLKSRLSISKDNSKSQIFLKMNSLQT DDTARYYCAREWHHGPVAYWGQGTLVTVS A CH
SEQ ID NO: 295 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB213VL SEQ ID NO: 296
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA
Variable APDIVMTQSHKFMSTTVGDRVSITCKASQ DVD2041L
AVSSAVAWYQQKPGQSPKLLIYWASTRHT GVPDRFTGSGSVTDFTLTIHNLQAEDLAL
YYCQQHYSTPFTFGSGTKLEIKR VL SEQ ID NO: 297
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 298 QPKAAP VL SEQ ID NO: 299 DIVMTQSHKFMSTTVGDRVSITCKASQAV
SSAVAWYQQKPGQSPKLLIYWASTRHTGV PDRFTGSGSVTDFTLTIHNLQAEDLALYY
CQQHYSTPFTFGSGTKLEIKR CL SEQ ID NO: 300
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB390VH
AB387VH SEQ ID NO: 301 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVESGGGLVQP DVD2042H
GGSLRLSCTASGFTFDDYALHWVRQAPGK GLEWVSGISWHGDFIDYADSVKGRFTISR
DNSKNTLYLQMNGLRVEDMAIYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID NO: 302
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 303 GGGGSGGGGS VH SEQ ID NO: 304
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 305 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB390VL AB387VL SEQ ID NO: 306
DIQMTQSPSSLSASVGDRVTITCRASGGI Binding RNYLGWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2042L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 307
DIQMTQSPSSLSASVGDRVTITCRASGGI RNYLGWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 308 GGGGSGGGGS VL SEQ ID NO: 309 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 310
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB390VH SEQ ID NO: 311 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSEVQLVESGGGLVQPGR DVD2043H
SLRLSCAASGFTFDDYAMHWVRQAPGKGL EWVSAITWNSGHIDYADSVEGRFTISRDN
AKNSLYLQMNSLRAEDTAVYYCAKVSYLS TASSLDYWGQGTLVTVSS VH SEQ ID NO: 312
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 313 GGGGSGGGGS VH SEQ ID NO: 314
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 315 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB390VL SEQ ID NO: 316
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQSPSSLSASVGDRVTITCR DVD2043L
ASGGIRNYLGWYQQKPGKAPKLLIYAAST LQSGVPSRFSGSGSGTDFTLTISSLQPED
VATYYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 317
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 318 GGSGGGGSG VL SEQ ID NO: 319
DIQMTQSPSSLSASVGDRVTITCRASGGI RNYLGWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
320 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB390VH
AB387VH SEQ ID NO: 321 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2044H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 322
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 323 ASTKGP VH SEQ ID NO: 324
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 325 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB390VL AB387VL SEQ ID NO: 326
DIQMTQSPSSLSASVGDRVTITCRASGGI Binding RNYLGWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2044L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 327
DIQMTQSPSSLSASVGDRVTITCRASGGI RNYLGWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 328 TVAAP VL SEQ ID NO: 329 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 330
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB390VH SEQ ID NO: 331 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2045H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 332
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 333 ASTKGP VH SEQ ID NO: 334
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 335 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB390VL SEQ ID NO: 336
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY
Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASG DVD2045L
GIRNYLGWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 337
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 338 QPKAAP VL SEQ ID NO: 339 DIQMTQSPSSLSASVGDRVTITCRASGGI
RNYLGWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 340
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB390VH
AB387VH SEQ ID NO: 341 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2046H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 342
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 343 ASTKGP VH SEQ ID NO: 344
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 345 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB390VL AB387VL SEQ ID NO: 346
DIQMTQSPSSLSASVGDRVTITCRASGGI Binding RNYLGWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2046L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 347
DIQMTQSPSSLSASVGDRVTITCRASGGI RNYLGWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 348 TVAAPSVFIFPP VL SEQ ID NO: 349
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID
NO: 350 QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB390VH SEQ ID NO: 351 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2047H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 352
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 353 ASTKGP VH SEQ ID NO: 354
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 355 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB390VL SEQ ID NO: 356
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQSPSSLSASVGDRVT DVD2047L
ITCRASGGIRNYLGWYQQKPGKAPKLLIY AASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDVATYYCQRYNRAPYTFGQGTKVEIK R VL SEQ ID NO: 357
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 358 QPKAAPSVTLFPP VL SEQ ID NO: 359
DIQMTQSPSSLSASVGDRVTITCRASGGI RNYLGWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
360 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB390VH
AB387VH SEQ ID NO: 361 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPSVFPLAPEVQLVESGGGL DVD2048H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
362 EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 363 ASTKGPSVFPLAP VH SEQ ID NO: 364
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 365 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB390VL AB387VL SEQ ID NO: 366
DIQMTQSPSSLSASVGDRVTITCRASGGI Binding RNYLGWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2048L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 367
DIQMTQSPSSLSASVGDRVTITCRASGGI RNYLGWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 368 TVAAP VL SEQ ID NO: 369 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 370
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB390VH SEQ ID NO: 371 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPEVQLVESGGGLVQ DVD2049H
PGRSLRLSCAASGFTFDDYAMHWVRQAPG KGLEWVSAITWNSGHIDYADSVEGRFTIS
RDNAKNSLYLQMNSLRAEDTAVYYCAKVS YLSTASSLDYWGQGTLVTVSS VH SEQ ID NO:
372 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 373 ASTKGPSVFPLAP VH SEQ ID NO: 374
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 375 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB390VL SEQ ID NO: 376
QSGLTQPPSASGTPGQRVTISCSGSSSNI
Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG Protein
VPDRFSGSKSGTSASLAISGLQSEDEADY Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA
Variable APDIQMTQSPSSLSASVGDRVTITCRASG DVD2049L
GIRNYLGWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 377
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 378 QPKAAP VL SEQ ID NO: 379 DIQMTQSPSSLSASVGDRVTITCRASGGI
RNYLGWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 380
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB391VH
AB387VH SEQ ID NO: 381 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVESGGGLVQP DVD2050H
GGSLRLSCTASGFTFDDYALHWVRQAPGK GLEWVSGISWHGDFIDYADSVKGRFTISR
DNSKNTLYLQMNGLRVEDMAIYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID NO: 382
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 383 GGGGSGGGGS VH SEQ ID NO: 384
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 385 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB391VL AB387VL SEQ ID NO: 386
DIQMTQSPSSLSASVGDRVTITCRASQSI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2050L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 387
DIQMTQSPSSLSASVGDRVTITCRASQSI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 388 GGSGGGGSG VL SEQ ID NO: 389 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 390
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB391VH SEQ ID NO: 391 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSEVQLVESGGGLVQPGR DVD2051H
SLRLSCAASGFTFDDYAMHWVRQAPGKGL EWVSAITWNSGHIDYADSVEGRFTISRDN
AKNSLYLQMNSLRAEDTAVYYCAKVSYLS TASSLDYWGQGTLVTVSS VH SEQ ID NO: 392
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 393 GGGGSGGGGS VH SEQ ID NO: 394
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 395 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB391VL SEQ ID NO: 396
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQSPSSLSASVGDRVTITCR DVD2051L
ASQSIRNYLSWYQQKPGKAPKLLIYAAST LQSGVPSRFSGSGSGTDFTLTISSLQPED
VATYYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 397
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 398 GGSGGGGSG VL SEQ ID NO: 399
DIQMTQSPSSLSASVGDRVTITCRASQSI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
400 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB391VH
AB387VH SEQ ID NO: 401 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2052H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 402
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 403 ASTKGP VH SEQ ID NO: 404
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 405 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB391VL AB387VL SEQ ID NO: 406
DIQMTQSPSSLSASVGDRVTITCRASQSI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2052L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 407
DIQMTQSPSSLSASVGDRVTITCRASQSI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 408 TVAAP VL SEQ ID NO: 409 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 410
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB391VH SEQ ID NO: 411 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2053H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 412
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 413 ASTKGP VH SEQ ID NO: 414
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 415 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK
DVD- AB387VL AB391VL SEQ ID NO: 416 QSGLTQPPSASGTPGQRVTISCSGSSSNI
Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG Protein
VPDRFSGSKSGTSASLAISGLQSEDEADY Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA
Variable APDIQMTQSPSSLSASVGDRVTITCRASQ DVD2053L
SIRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 417
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 418 QPKAAP VL SEQ ID NO: 419 DIQMTQSPSSLSASVGDRVTITCRASQSI
RNYLSWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 420 DVD- AB391VH AB387VH SEQ ID
NO: 421 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2054H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 422
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 423 ASTKGP VH SEQ ID NO: 424
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 425 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB391VL AB387VL SEQ ID NO: 426
DIQMTQSPSSLSASVGDRVTITCRASQSI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2054L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 427
DIQMTQSPSSLSASVGDRVTITCRASQSI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 428 TVAAPSVFIFPP VL SEQ ID NO: 429
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID
NO: 430 QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB391VH SEQ ID NO: 431 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2055H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 432
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 433 ASTKGP VH SEQ ID NO: 434
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 435 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB391VL SEQ ID NO: 436
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQSPSSLSASVGDRVT DVD2055L
ITCRASQSIRNYLSWYQQKPGKAPKLLIY AASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDVATYYCQRYNRAPYTFGQGTKVEIK R VL SEQ ID NO: 437
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 438 QPKAAPSVTLFPP VL SEQ ID NO: 439
DIQMTQSPSSLSASVGDRVTITCRASQSI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
440 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB391VH
AB387VH SEQ ID NO: 441 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPSVFPLAPEVQLVESGGGL DVD2056H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
442 EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 443 ASTKGPSVFPLAP VH SEQ ID NO: 444
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 445 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB391VL AB387VL SEQ ID NO: 446
DIQMTQSPSSLSASVGDRVTITCRASQSI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2056L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 447
DIQMTQSPSSLSASVGDRVTITCRASQSI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 448 TVAAP VL SEQ ID NO: 449 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 450
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB391VH SEQ ID NO: 451 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPEVQLVESGGGLVQ DVD2057H
PGRSLRLSCAASGFTFDDYAMHWVRQAPG KGLEWVSAITWNSGHIDYADSVEGRFTIS
RDNAKNSLYLQMNSLRAEDTAVYYCAKVS YLSTASSLDYWGQGTLVTVSS VH SEQ ID NO:
452 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 453 ASTKGPSVFPLAP VH SEQ ID NO: 454
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 455 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB391VL SEQ ID NO: 456
QSGLTQPPSASGTPGQRVTISCSGSSSNI
Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG Protein
VPDRFSGSKSGTSASLAISGLQSEDEADY Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA
Variable APDIQMTQSPSSLSASVGDRVTITCRASQ DVD2057L
SIRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 457
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 458 QPKAAP VL SEQ ID NO: 459 DIQMTQSPSSLSASVGDRVTITCRASQSI
RNYLSWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 460
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB392VH
AB387VH SEQ ID NO: 461 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVESGGGLVQP DVD2058H
GGSLRLSCTASGFTFDDYALHWVRQAPGK GLEWVSGISWHGDFIDYADSVKGRFTISR
DNSKNTLYLQMNGLRVEDMAIYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID NO: 462
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 463 GGGGSGGGGS VH SEQ ID NO: 464
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 465 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB392VL AB387VL SEQ ID NO: 466
DIQMTQSPSSLSASVGDRVTITCRASRGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2058L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 467
DIQMTQSPSSLSASVGDRVTITCRASRGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 468 GGSGGGGSG VL SEQ ID NO: 469 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 470
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB392VH SEQ ID NO: 471 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSEVQLVESGGGLVQPGR DVD2059H
SLRLSCAASGFTFDDYAMHWVRQAPGKGL EWVSAITWNSGHIDYADSVEGRFTISRDN
AKNSLYLQMNSLRAEDTAVYYCAKVSYLS TASSLDYWGQGTLVTVSS VH SEQ ID NO: 472
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 473 GGGGSGGGGS VH SEQ ID NO: 474
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 475 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB392VL SEQ ID NO: 476
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQSPSSLSASVGDRVTITCR DVD2059L
ASRGIRNYLSWYQQKPGKAPKLLIYAAST LQSGVPSRFSGSGSGTDFTLTISSLQPED
VATYYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 477
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 478 GGSGGGGSG VL SEQ ID NO: 479
DIQMTQSPSSLSASVGDRVTITCRASRGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
480 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB392VH
AB387VH SEQ ID NO: 481 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2060H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 482
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 483 ASTKGP VH SEQ ID NO: 484
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 485 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB392VL AB387VL SEQ ID NO: 486
DIQMTQSPSSLSASVGDRVTITCRASRGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2060L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 487
DIQMTQSPSSLSASVGDRVTITCRASRGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 488 TVAAP VL SEQ ID NO: 489 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 490
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB392VH SEQ ID NO: 491 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2061H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 492
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 493 ASTKGP VH SEQ ID NO: 494
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 495 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK
DVD- AB387VL AB392VL SEQ ID NO: 496 QSGLTQPPSASGTPGQRVTISCSGSSSNI
Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG Protein
VPDRFSGSKSGTSASLAISGLQSEDEADY Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA
Variable APDIQMTQSPSSLSASVGDRVTITCRASR DVD2061L
GIRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 497
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 498 QPKAAP VL SEQ ID NO: 499 DIQMTQSPSSLSASVGDRVTITCRASRGI
RNYLSWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 500
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB392VH
AB387VH SEQ ID NO: 501 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2062H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 502
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 503 ASTKGP VH SEQ ID NO: 504
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 505 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB392VL AB387VL SEQ ID NO: 506
DIQMTQSPSSLSASVGDRVTITCRASRGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2062L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 507
DIQMTQSPSSLSASVGDRVTITCRASRGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 508 TVAAPSVFIFPP VL SEQ ID NO: 509
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID
NO: 510 QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB392VH SEQ ID NO: 511 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2063H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 512
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 513 ASTKGP VH SEQ ID NO: 514
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 515 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB392VL SEQ ID NO: 516
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQSPSSLSASVGDRVT DVD2063L
ITCRASRGIRNYLSWYQQKPGKAPKLLIY AASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDVATYYCQRYNRAPYTFGQGTKVEIK R VL SEQ ID NO: 517
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 518 QPKAAPSVTLFPP VL SEQ ID NO: 519
DIQMTQSPSSLSASVGDRVTITCRASRGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
520 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB392VH
AB387VH SEQ ID NO: 521 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPSVFPLAPEVQLVESGGGL DVD2064H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
522 EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 523 ASTKGPSVFPLAP VH SEQ ID NO: 524
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 525 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB392VL AB387VL SEQ ID NO: 526
DIQMTQSPSSLSASVGDRVTITCRASRGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2064L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 527
DIQMTQSPSSLSASVGDRVTITCRASRGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 528 TVAAP VL SEQ ID NO: 529 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 530
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB392VH SEQ ID NO: 531 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPEVQLVESGGGLVQ DVD2065H
PGRSLRLSCAASGFTFDDYAMHWVRQAPG KGLEWVSAITWNSGHIDYADSVEGRFTIS
RDNAKNSLYLQMNSLRAEDTAVYYCAKVS YLSTASSLDYWGQGTLVTVSS VH SEQ ID NO:
532 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 533 ASTKGPSVFPLAP VH SEQ ID NO: 534
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 535 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB392VL SEQ ID NO: 536
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASR DVD2065L
GIRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 537
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 538 QPKAAP VL SEQ ID NO: 539 DIQMTQSPSSLSASVGDRVTITCRASRGI
RNYLSWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 540
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB393VH
AB387VH SEQ ID NO: 541 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVESGGGLVQP DVD2066H
GGSLRLSCTASGFTFDDYALHWVRQAPGK GLEWVSGISWHGDFIDYADSVKGRFTISR
DNSKNTLYLQMNGLRVEDMAIYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID NO: 542
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 543 GGGGSGGGGS VH SEQ ID NO: 544
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 545 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB393VL AB387VL SEQ ID NO: 546
DIQMTQSPSSLSASVGDRVTITCRASHGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRGGSGGGGS Variable
GQSGLTQPPSASGTPGQRVTISCSGSSSN DVD2066L
IGSNTVNWYQQLPGTAPKLLIYSNNQRPS GVPDRFSGSKSGTSASLAISGLQSEDEAD
YYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 547
DIQMTQSPSSLSASVGDRVTITCRASHGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 548 GGSGGGGSG VL SEQ ID NO: 549 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 550
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB393VH SEQ ID NO: 551 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSGGGGSGGGGSEVQLVESGGGLVQPGR DVD2067H
SLRLSCAASGFTFDDYAMHWVRQAPGKGL EWVSAITWNSGHIDYADSVEGRFTISRDN
AKNSLYLQMNSLRAEDTAVYYCAKVSYLS TASSLDYWGQGTLVTVSS VH SEQ ID NO: 552
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 553 GGGGSGGGGS VH SEQ ID NO: 554
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 555 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB393VL SEQ ID NO: 556
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGGGSG Variable
GGGSGDIQMTQSPSSLSASVGDRVTITCR DVD2067L
ASHGIRNYLSWYQQKPGKAPKLLIYAAST LQSGVPSRFSGSGSGTDFTLTISSLQPED
VATYYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 557
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 558 GGSGGGGSG VL SEQ ID NO: 559
DIQMTQSPSSLSASVGDRVTITCRASHGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
560 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB393VH
AB387VH SEQ ID NO: 561 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2068H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 562
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 563 ASTKGP VH SEQ ID NO: 564
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 565 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB393VL AB387VL SEQ ID NO: 566
DIQMTQSPSSLSASVGDRVTITCRASHGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2068L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 567
DIQMTQSPSSLSASVGDRVTITCRASHGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 568 TVAAP VL SEQ ID NO: 569 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 570
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB393VH SEQ ID NO: 571 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2069H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 572
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 573 ASTKGP VH SEQ ID NO: 574
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 575 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK DVD- AB387VL AB393VL SEQ
ID NO: 576 QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG Protein VPDRFSGSKSGTSASLAISGLQSEDEADY
Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASH DVD2069L
GIRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 577
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 578 QPKAAP VL SEQ ID NO: 579 DIQMTQSPSSLSASVGDRVTITCRASHGI
RNYLSWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 580
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB393VH
AB387VH SEQ ID NO: 581 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVESGGGLVQPGGSL DVD2070H
RLSCTASGFTFDDYALHWVRQAPGKGLEW VSGISWHGDFIDYADSVKGRFTISRDNSK
NTLYLQMNGLRVEDMAIYYCAGNNRGYGG LDVWGQGTTVTVSS VH SEQ ID NO: 582
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 583 ASTKGP VH SEQ ID NO: 584
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 585 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB393VL AB387VL SEQ ID NO: 586
DIQMTQSPSSLSASVGDRVTITCRASHGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPSVF Variable
IFPPQSGLTQPPSASGTPGQRVTISCSGS DVD2070L
SSNIGSNTVNWYQQLPGTAPKLLIYSNNQ RPSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 587
DIQMTQSPSSLSASVGDRVTITCRASHGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 588 TVAAPSVFIFPP VL SEQ ID NO: 589
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID
NO: 590 QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB393VH SEQ ID NO: 591 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPEVQLVESGGGLVQPGRSLRL DVD2071H
SCAASGFTFDDYAMHWVRQAPGKGLEWVS AITWNSGHIDYADSVEGRFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAKVSYLSTASS LDYWGQGTLVTVSS VH SEQ ID NO: 592
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 593 ASTKGP VH SEQ ID NO: 594
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 595 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB387VL AB393VL SEQ ID NO: 596
QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
Protein VPDRFSGSKSGTSASLAISGLQSEDEADY Light
YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APSVTLFPPDIQMTQSPSSLSASVGDRVT DVD2071L
ITCRASHGIRNYLSWYQQKPGKAPKLLIY AASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDVATYYCQRYNRAPYTFGQGTKVEIK R VL SEQ ID NO: 597
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 598 QPKAAPSVTLFPP VL SEQ ID NO: 599
DIQMTQSPSSLSASVGDRVTITCRASHGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO:
600 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB393VH
AB387VH SEQ ID NO: 601 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPSVFPLAPEVQLVESGGGL DVD2072H
VQPGGSLRLSCTASGFTFDDYALHWVRQA PGKGLEWVSGISWHGDFIDYADSVKGRFT
ISRDNSKNTLYLQMNGLRVEDMAIYYCAG NNRGYGGLDVWGQGTTVTVSS VH SEQ ID NO:
602 EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 603 ASTKGPSVFPLAP VH SEQ ID NO: 604
EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS CH
SEQ ID NO: 605 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB393VL AB387VL SEQ ID NO: 606
DIQMTQSPSSLSASVGDRVTITCRASHGI Binding RNYLSWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPQSG Variable
LTQPPSASGTPGQRVTISCSGSSSNIGSN DVD2072L
TVNWYQQLPGTAPKLLIYSNNQRPSGVPD RFSGSKSGTSASLAISGLQSEDEADYYCA
AWDDSLNGSYVFGTGTKVTVLG VL SEQ ID NO: 607
DIQMTQSPSSLSASVGDRVTITCRASHGI RNYLSWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 608 TVAAP VL SEQ ID NO: 609 QSGLTQPPSASGTPGQRVTISCSGSSSNI
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG VPDRFSGSKSGTSASLAISGLQSEDEADY
YCAAWDDSLNGSYVFGTGTKVTVLG CL SEQ ID NO: 610
QPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPS
KQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS DVD- AB387VH
AB393VH SEQ ID NO: 611 EVQLVESGGGLVQPGGSLRLSCTASGFTF Binding
DDYALHWVRQAPGKGLEWVSGISWHGDFI Protein DYADSVKGRFTISRDNSKNTLYLQMNGLR
Heavy VEDMAIYYCAGNNRGYGGLDVWGQGTTVT Variable
VSSASTKGPSVFPLAPEVQLVESGGGLVQ DVD2073H
PGRSLRLSCAASGFTFDDYAMHWVRQAPG KGLEWVSAITWNSGHIDYADSVEGRFTIS
RDNAKNSLYLQMNSLRAEDTAVYYCAKVS YLSTASSLDYWGQGTLVTVSS VH SEQ ID NO:
612 EVQLVESGGGLVQPGGSLRLSCTASGFTF DDYALHWVRQAPGKGLEWVSGISWHGDFI
DYADSVKGRFTISRDNSKNTLYLQMNGLR VEDMAIYYCAGNNRGYGGLDVWGQGTTVT VSS
Linker SEQ ID NO: 613 ASTKGPSVFPLAP VH SEQ ID NO: 614
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
CH SEQ ID NO: 615 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK DVD- AB387VL AB393VL SEQ
ID NO: 616 QSGLTQPPSASGTPGQRVTISCSGSSSNI Binding
GSNTVNWYQQLPGTAPKLLIYSNNQRPSG Protein VPDRFSGSKSGTSASLAISGLQSEDEADY
Light YCAAWDDSLNGSYVFGTGTKVTVLGQPKA Variable
APDIQMTQSPSSLSASVGDRVTITCRASH DVD2073L
GIRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVAT
YYCQRYNRAPYTFGQGTKVEIKR VL SEQ ID NO: 617
QSGLTQPPSASGTPGQRVTISCSGSSSNI GSNTVNWYQQLPGTAPKLLIYSNNQRPSG
VPDRFSGSKSGTSASLAISGLQSEDEADY YCAAWDDSLNGSYVFGTGTKVTVLG Linker SEQ
ID NO: 618 QPKAAP VL SEQ ID NO: 619 DIQMTQSPSSLSASVGDRVTITCRASHGI
RNYLSWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR CL SEQ ID NO: 620
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB017VH
AB235VH SEQ ID NO: 621 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVQSGAEVKKP DVD2074H
GASVKVSCKASGYTFTDYNMHWVRQAPGQ GLEWMGEINPNSGGAGYNQKFKGRVTMTT
DTSTSTAYMELRSLRSDDTAVYYCARLGY DDIYDDWYFDVWGQGTTVTVSS VH SEQ ID NO:
622 EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 623 GGGGSGGGGS VH SEQ ID NO: 624
EVQLVQSGAEVKKPGASVKVSCKASGYTF TDYNMHWVRQAPGQGLEWMGEINPNSGGA
GYNQKFKGRVTMTTDTSTSTAYMELRSLR SDDTAVYYCARLGYDDIYDDWYFDVWGQG TTVTVSS
CH SEQ ID NO: 625 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB017VL AB235VL SEQ ID NO: 626
DIQMTQSPSSLSASVGDRVTITCRASQGI Binding RNYLAWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRGGSGGGGS Variable
GDIQMTQSPSSLSASVGDRVTITCRASQD DVD2074L
ISNYLNWYQQKPGKAPKLLIYYTSRLLSG VPSRFSGSGSGTDFTLTISSLQPEDFATY
YCQQGDTLPYTFGGGTKVEIKR VL SEQ ID NO: 627
DIQMTQSPSSLSASVGDRVTITCRASQGI RNYLAWYQQKPGKAPKLLIYAASTLQSGV
PSRFSGSGSGTDFTLTISSLQPEDVATYY CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
NO: 628 GGSGGGGSG VL SEQ ID NO: 629 DIQMTQSPSSLSASVGDRVTITCRASQDI
SNYLNWYQQKPGKAPKLLIYYTSRLLSGV PSRFSGSGSGTDFTLTISSLQPEDFATYY
CQQGDTLPYTFGGGTKVEIKR CL SEQ ID NO: 630
TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- AB235VH
AB017VH SEQ ID NO: 631 EVQLVESGGGLVQPGRSLRLSCAASGFTF Binding
DDYAMHWVRQAPGKGLEWVSAITWNSGHI Protein DYADSVEGRFTISRDNAKNSLYLQMNSLR
Heavy AEDTAVYYCAKVSYLSTASSLDYWGQGTL Variable
VTVSSASTKGPEVQLVQSGAEVKKPGASV DVD2075H
KVSCKASGYTFTDYNMHWVRQAPGQGLEW MGEINPNSGGAGYNQKFKGRVTMTTDTST
STAYMELRSLRSDDTAVYYCARLGYDDIY DDWYFDVWGQGTTVTVSS VH SEQ ID NO: 632
EVQLVESGGGLVQPGRSLRLSCAASGFTF DDYAMHWVRQAPGKGLEWVSAITWNSGHI
DYADSVEGRFTISRDNAKNSLYLQMNSLR AEDTAVYYCAKVSYLSTASSLDYWGQGTL VTVSS
Linker SEQ ID NO: 633 ASTKGP VH SEQ ID NO: 634
EVQLVQSGAEVKKPGASVKVSCKASGYTF TDYNMHWVRQAPGQGLEWMGEINPNSGGA
GYNQKFKGRVTMTTDTSTSTAYMELRSLR SDDTAVYYCARLGYDDIYDDWYFDVWGQG TTVTVSS
CH SEQ ID NO: 635 ASTKGPSVFPLAPSSKSTSGGTAALGCLV
KDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNH
KPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYK
CKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIA
VEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK DVD- AB235VL AB017VL SEQ ID NO: 636
DIQMTQSPSSLSASVGDRVTITCRASQGI Binding RNYLAWYQQKPGKAPKLLIYAASTLQSGV
Protein PSRFSGSGSGTDFTLTISSLQPEDVATYY Light
CQRYNRAPYTFGQGTKVEIKRTVAAPDIQ Variable
MTQSPSSLSASVGDRVTITCRASQDISNY DVD2075L
LNWYQQKPGKAPKLLIYYTSRLLSGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQQ
GDTLPYTFGGGTKVEIKR VL SEQ ID NO: 637 DIQMTQSPSSLSASVGDRVTITCRASQGI
RNYLAWYQQKPGKAPKLLIYAASTLQSGV PSRFSGSGSGTDFTLTISSLQPEDVATYY
CQRYNRAPYTFGQGTKVEIKR Linker SEQ ID NO: 638 TVAAP VL SEQ ID NO: 639
DIQMTQSPSSLSASVGDRVTITCRASQDI SNYLNWYQQKPGKAPKLLIYYTSRLLSGV
PSRFSGSGSGTDFTLTISSLQPEDFATYY CQQGDTLPYTFGGGTKVEIKR CL SEQ ID NO:
640 TVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQ
DSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC DVD- MAK199-
AE10-6 SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO:
641 FANYGIIWVRQAPGQGLEWMGWINTYTG Protein
KPTYAQKFQGRVTMTTDTSTSTAYMELS Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD
Variable YWGQGTTVTVSSGGGGSGGGGSEVQLVQ MAK199-
SGAEVKKPGASVKVSCKVSGYTLTELSM 1-GS- HWVRQAPGKGLEWMGGFDPEDGETIYAQ
AE10-6 KFQGRVTMTEDTSTDTAYMELSSLRSED AM7
TAVYYCATDTVGYWEKFFQHWGQGTLVT VSS VH SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 642 FANYGIIWVRQAPGQGLEWMGWINTYTG
KPTYAQKFQGRVTMTTDTSTSTAYMELS SLRSEDTAVYYCARKLFTTMDVTDNAMD
YWGQGTTVTVSS Linker SEQ ID GGGGSGGGGS NO: 643 VH SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 644 LTELSMHWVRQAPGKGLEWMGGFDPEDG
ETIYAQKFQGRVTMTEDTSTDTAYMELS SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 645
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- MAK199- AE10-6 SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO: 646
ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein GVPSRFSGSGSGTDFTLTISSLQPEDFA
Light TYFCQQGNTWPPTFGQGTKLEIKRGGSG Variable
GGGSGQAVVTQEPSLTVSPGGTVTLTCG MAK199- SSTGTVTIDHYPYWFQQKPGQAPRTLIS
1-GS- DTDDKHSWTPARFSGSLLGGKAALTLSG AE10-6
AQPEDEAEYYCLLDYGGRFVFGGGTKLT AM7 VLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 647 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 648 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 649
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 650 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK199- AE10-6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO: 651
FANYGIIWVRQAPGQGLEWMGWINTYTG Protein KPTYAQKFQGRVTMTTDTSTSTAYMELS
Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD Variable
YWGQGTTVTVSSASTKGPEVQLVQSGAE MAK199- VKKPGASVKVSCKVSGYTLTELSMHWVR
1-SS- QAPGKGLEWMGGFDPEDGETIYAQKFQG AE10-6
RVTMTEDTSTDTAYMELSSLRSEDTAVY AM7 YCATDTVGYWEKFFQHWGQGTLVTVSS VH SEQ
ID EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 652
FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID ASTKGP NO:
653 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 654
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 655
VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
656 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRTVAA
Variable PQAVVTQEPSLTVSPGGTVTLTCGSSTG MAK199-
TVTIDHYPYWFQQKPGQAPRTLISDTDD 1-SS- KHSWTPARFSGSLLGGKAALTLSGAQPE
AE10-6 DEAEYYCLLDYGGRFVFGGGTKLTVLG AM7 VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 657 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 658 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 659
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 660 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK195- AE10-6 SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT Binding 21 VH AM7 VH NO: 661
FSNYGVTWVRQAPGKGLEWVSMIWADGS Protein THYASSVKGRFTISRDNSKNTLYLQMNS
Heavy LRAEDTAVYYCAREWQHGPVAYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVQSGAEVKK MAK195- PGASVKVSCKVSGYTLTELSMHWVRQAP
21-GS- GKGLEWMGGFDPEDGETIYAQKFQGRVT AE10-6
MTEDTSTDTAYMELSSLRSEDTAVYYCA AM7 TDTVGYWEKFFQHWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 662 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 663 VH SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 664 LTELSMHWVRQAPGKGLEWMGGFDPEDG
ETIYAQKFQGRVTMTEDTSTDTAYMELS SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 665
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- MAK195- AE10-6 SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL Binding 21 VL AM7 VL NO: 666
VSSAVAWYQQKPGKAPKLLIYWASARHT Protein GVPSRFSGSGSGTDFTLTISSLQPEDFA
Light TYYCQQHYKTPFTFGQGTKLEIKGGSGG Variable
GGSGQAVVTQEPSLTVSPGGTVTLTCGS MAK195- STGTVTIDHYPYWFQQKPGQAPRTLISD
21-GS- TDDKHSWTPARFSGSLLGGKAALTLSGA AE10-6
QPEDEAEYYCLLDYGGRFVFGGGTKLTV AM7 LG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 667 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIK Linker SEQ ID
GGSGGGGSG NO: 668 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 669
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 670 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK195- AE10-6 SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT Binding 21 VH AM7 VH NO: 671
FSNYGVTWVRQAPGKGLEWVSMIWADGS Protein THYASSVKGRFTISRDNSKNTLYLQMNS
Heavy LRAEDTAVYYCAREWQHGPVAYWGQGTL Variable
VTVSSASTKGPEVQLVQSGAEVKKPGAS MAK195- VKVSCKVSGYTLTELSMHWVRQAPGKGL
21-SS- EWMGGFDPEDGETIYAQKFQGRVTMTED AE10-
TSTDTAYMELSSLRSEDTAVYYCATDTV 6AM7 GYWEKFFQHWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 672 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 673 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT
NO: 674 LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 675 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD- MAK195-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL Binding 21 VL AM7 VL NO:
676 VSSAVAWYQQKPGKAPKLLIYWASARHT Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYYCQQHYKTPFTFGQGTKLEIKRTVAA
Variable PQAVVTQEPSLTVSPGGTVTLTCGSSTG MAK195-
TVTIDHYPYWFQQKPGQAPRTLISDTDD 21-SS- KHSWTPARFSGSLLGGKAALTLSGAQPE
AE10-6 DEAEYYCLLDYGGRFVFGGGTKLTVLG AM7 VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 677 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 678 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 679
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 680 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK199- AE10-6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO: 681
FANYGIIWVRQAPGQGLEWMGWINTYTG Protein KPTYAQKFQGRVTMTTDTSTSTAYMELS
Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD Variable
YWGQGTTVTVSSGGGGSGGGGSEVQLVQ MAK199- SGAEVKKPGASVKVSCKVSGYTLTELSM
1-GS- HWVRQAPGKGLEWMGGFDPEDGETIYAQ AE10-6
KFQGRVTMTEDTSTDTAYMELSSLRSED AM7 QL TAVYYCATDTVGYWEKFFQHWGQGTLVT
VSS VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 682
FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID GGGGSGGGGS
NO: 683 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 684
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH QL SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 685 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDQLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVLHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
686 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRGGSG
Variable GGGSGQAVVTQEPSLTVSPGGTVTLTCG MAK199-
SSTGTVTIDHYPYWFQQKPGQAPRTLIS 1-GS- DTDDKHSWTPARFSGSLLGGKAALTLSG
AE10-6 AQPEDEAEYYCLLDYGGRFVFGGGTKLT AM7 QL VLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 687 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 688 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 689
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 690 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK199- AE10-6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO: 691
FANYGIIWVRQAPGQGLEWMGWINTYTG Protein KPTYAQKFQGRVTMTTDTSTSTAYMELS
Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD Variable
YWGQGTTVTVSSASTKGPEVQLVQSGAE MAK199- VKKPGASVKVSCKVSGYTLTELSMHWVR
1-SS- QAPGKGLEWMGGFDPEDGETIYAQKFQG AE10-6
RVTMTEDTSTDTAYMELSSLRSEDTAVY AM7 QL YCATDTVGYWEKFFQHWGQGTLVTVSS VH
SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 692
FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID ASTKGP NO:
693 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 694
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH QL SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 695 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDQLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVLHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
696 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRTVAA
Variable PQAVVTQEPSLTVSPGGTVTLTCGSSTG MAK199-
TVTIDHYPYWFQQKPGQAPRTLISDTDD 1-SS- KHSWTPARFSGSLLGGKAALTLSGAQPE
AE10-6 DEAEYYCLLDYGGRFVFGGGTKLTVLG AM7 QL VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 697 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 698 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 699
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 700 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK195- AE10-6 SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT Binding 21 VH AM3 VH NO: 701
FSNYGVTWVRQAPGKGLEWVSMIWADGS Protein THYASSVKGRFTISRDNSKNTLYLQMNS
Heavy LRAEDTAVYYCAREWQHGPVAYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVQSGAEVKK MAK195- PGASVKVSCKVSGYTLTELSMHWVRQAP
1-GS- GKGLEWMGGFDPEDGETIYAQKFQGRVT AE10-6
MTEDTSTDTAYMELSSLRSEDTAVYYCA AM3 TDSAGYWYKFFQHWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 702 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 703 VH SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 704 LTELSMHWVRQAPGKGLEWMGGFDPEDG
ETIYAQKFQGRVTMTEDTSTDTAYMELS SLRSEDTAVYYCATDSAGYWYKFFQHWG QGTLVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 705
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- MAK195- AE10-6 SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL Binding 21 VL AM3 VL NO: 706
VSSAVAWYQQKPGKAPKLLIYWASARHT Protein GVPSRFSGSGSGTDFTLTISSLQPEDFA
Light TYYCQQHYKTPFTFGQGTKLEIKGGSGG Variable
GGSGQAVVTQEPSLTVSPGGTVTLTCGS MAK195- STGAVTIDHYPYWFQQKPGQAPRTLISD
21-GS- TDDKHSWTPARFSGSLLGGKAALTLSGA AE10-6
QPEDEAEYYCLLDYGGTFVFGGGTKLTV AM3 LG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 707 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIK Linker SEQ ID
GGSGGGGSG NO: 708 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGA NO: 709
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGTFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 710 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- AE10-6 MAK195- SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYT Binding AM7 VH 21 VH NO: 711
LTELSMHWVRQAPGKGLEWMGGFDPEDG Protein ETIYAQKFQGRVTMTEDTSTDTAYMELS
Heavy SLRSEDTAVYYCATDTVGYWEKFFQHWG Variable
QGTLVTVSSGGGGSGGGGSEVQLVESGG AE10-6 GLVQPGGSLRLSCAASGFTFSNYGVTWV
AM7-GS- RQAPGKGLEWVSMIWADGSTHYASSVKG MAK195-
RFTISRDNSKNTLYLQMNSLRAEDTAVY 21 YCAREWQHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 712 LTELSMHWVRQAPGKGLEWMGGFDPEDG
ETIYAQKFQGRVTMTEDTSTDTAYMELS SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 713 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 714 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 715
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- AE10-6 MAK195/ SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGT Binding AM7 VL 21 VL NO: 716
VTIDHYPYWFQQKPGQAPRTLISDTDDK Protein HSWTPARFSGSLLGGKAALTLSGAQPED
Light EAEYYCLLDYGGRFVFGGGTKLTVLGGG Variable
SGGGGSGDIQMTQSPSSLSASVGDRVTI AE10-6 TCRASQLVSSAVAWYQQKPGKAPKLLIY
AM7-GS- WASARHTGVPSRFSGSGSGTDFTLTISS MAK195-
LQPEDFATYYCQQHYKTPFTFGQGTKLE 21 IKR VL SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 717 VTIDHYPYWFQQKPGQAPRTLISDTDDK
HSWTPARFSGSLLGGKAALTLSGAQPED EAEYYCLLDYGGRFVFGGGTKLTVLG Linker SEQ
ID GGSGGGGSG NO: 718 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL NO: 719
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
720 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD- MAK199- AE10-6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO: 721
FANYGIIWVRQAPGQGLEWMGWINTYTG Protein KPTYAQKFQGRVTMTTDTSTSTAYMELS
Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD Variable
YWGQGTTVTVSSGGGGSGGGGSEVQLVQ MAK199- SGAEVKKPGASVKVSCKVSGYTLTELSM
1-GS- HWVRQAPGKGLEWMGGFDPEDGETIYAQ AE10-6
KFQGRVTMTEDTSTDTAYMELSSLRSED AM7 wtQL TAVYYCATDTVGYWEKFFQHWGQGTLVT
VSS VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 722
FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID GGGGSGGGGS
NO: 723 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 724
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH wt QL SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 725 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDQLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVLHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
726 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRGGSG
Variable GGGSGQAVVTQEPSLTVSPGGTVTLTCG MAK199-
SSTGTVTIDHYPYWFQQKPGQAPRTLIS 1-GS- DTDDKHSWTPARFSGSLLGGKAALTLSG
AE10-6 AQPEDEAEYYCLLDYGGRFVFGGGTKLT AM7 wtQL VLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 727 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 728 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 729
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 730 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS
DVD- MAK199- AE10-6 SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1
VH AM7 VH NO: 731 FANYGIIWVRQAPGQGLEWMGWINTYTG Protein
KPTYAQKFQGRVTMTTDTSTSTAYMELS Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD
Variable YWGQGTTVTVSSASTKGPEVQLVQSGAE MAK199-
VKKPGASVKVSCKVSGYTLTELSMHWVR 1-SS- QAPGKGLEWMGGFDPEDGETIYAQKFQG
AE10-6 RVTMTEDTSTDTAYMELSSLRSEDTAVY AM7 wtQL
YCATDTVGYWEKFFQHWGQGTLVTVSS VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT
NO: 732 FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID ASTKGP NO:
733 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 734
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH wt QL SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 735 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDQLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVLHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
736 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRTVAA
Variable PQAVVTQEPSLTVSPGGTVTLTCGSSTG MAK199-
TVTIDHYPYWFQQKPGQAPRTLISDTDD 1-SS- KHSWTPARFSGSLLGGKAALTLSGAQPE
AE10-6 DEAEYYCLLDYGGRFVFGGGTKLTVLG AM7 wtQL VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 737 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 738 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 739
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 740 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK199- AE10-6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO: 741
FANYGIIWVRQAPGQGLEWMGWINTYTG Protein KPTYAQKFQGRVTMTTDTSTSTAYMELS
Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD Variable
YWGQGTTVTVSSGGGGSGGGGSEVQLVQ MAK199- SGAEVKKPGASVKVSCKVSGYTLTELSM
1-GS- HWVRQAPGKGLEWMGGFDPEDGETIYAQ AE10-6
KFQGRVTMTEDTSTDTAYMELSSLRSED AM7 wt TAVYYCATDTVGYWEKFFQHWGQGTLVT
VSS VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 742
FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID GGGGSGGGGS
NO: 743 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 744
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH wt SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 745 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
746 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRGGSG
Variable GGGSGQAVVTQEPSLTVSPGGTVTLTCG MAK199-
SSTGTVTIDHYPYWFQQKPGQAPRTLIS 1-GS- DTDDKHSWTPARFSGSLLGGKAALTLSG
AE10-6 AQPEDEAEYYCLLDYGGRFVFGGGTKLT AM7 wt VLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 747 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 748 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 749
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 750 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK199- AE10-6 SEQ ID
EVQLVQSGAEVKKPGASVKVSCKASGYT Binding 1 VH AM7 VH NO: 751
FANYGIIWVRQAPGQGLEWMGWINTYTG Protein KPTYAQKFQGRVTMTTDTSTSTAYMELS
Heavy SLRSEDTAVYYCARKLFTTMDVTDNAMD Variable
YWGQGTTVTVSSASTKGPEVQLVQSGAE MAK199- VKKPGASVKVSCKVSGYTLTELSMHWVR
1-SS- QAPGKGLEWMGGFDPEDGETIYAQKFQG AE10-6
RVTMTEDTSTDTAYMELSSLRSEDTAVY AM7 wt YCATDTVGYWEKFFQHWGQGTLVTVSS VH
SEQ ID EVQLVQSGAEVKKPGASVKVSCKASGYT NO: 752
FANYGIIWVRQAPGQGLEWMGWINTYTG KPTYAQKFQGRVTMTTDTSTSTAYMELS
SLRSEDTAVYYCARKLFTTMDVTDNAMD YWGQGTTVTVSS Linker SEQ ID ASTKGP NO:
753 VH SEQ ID EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 754
LTELSMHWVRQAPGKGLEWMGGFDPEDG ETIYAQKFQGRVTMTEDTSTDTAYMELS
SLRSEDTAVYYCATDTVGYWEKFFQHWG QGTLVTVSS CH wt SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 755 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD- MAK199-
AE10-6 SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQD Binding 1 VL AM7 VL NO:
756 ISQYLNWYQQKPGKAPKLLIYYTSRLQS Protein
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYFCQQGNTWPPTFGQGTKLEIKRTVAA
Variable PQAVVTQEPSLTVSPGGTVTLTCGSSTG MAK199-
TVTIDHYPYWFQQKPGQAPRTLISDTDD 1-SS- KHSWTPARFSGSLLGGKAALTLSGAQPE
AE10-6 DEAEYYCLLDYGGRFVFGGGTKLTVLG AM7 wt VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQD NO: 757 ISQYLNWYQQKPGKAPKLLIYYTSRLQS
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYFCQQGNTWPPTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 758 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 759
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 760 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- MAK195- AE10-6 SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT Binding 21 VH AM8 VH NO: 761
FSNYGVTWVRQAPGKGLEWVSMIWADGS Protein THYASSVKGRFTISRDNSKNTLYLQMNS
Heavy LRAEDTAVYYCAREWQHGPVAYWGQGTL Variable
VTVSSGGGGSGGGGSEVQLVQSGAEVKK MAK195- PGASVKVSCKVSGYTLSELSIHWVRQAP
21-GS- GKGLEWMGGFDPEDGETIYAQKFQGRVT AE10-6
MTEDTSTDTAYMELSSLRSEDTAVYYCA AM8 TDSAGYWYKFFQHWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 762 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 763 VH SEQ ID
EVQLVQSGAEVKKPGASVKVSCKVSGYT NO: 764 LSELSIHWVRQAPGKGLEWMGGFDPEDG
ETIYAQKFQGRVTMTEDTSTDTAYMELS SLRSEDTAVYYCATDSAGYWYKFFQHWG QGTLVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 765
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- MAK- AE10-6 SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL Binding 195-21 AM8 VL NO: 766
VSSAVAWYQQKPGKAPKLLIYWASARHT Protein VL
GVPSRFSGSGSGTDFTLTISSLQPEDFA Light TYYCQQHYKTPFTFGQGTKLEIKGGSGG
Variable GGSGQAVVTQEPSLTVSPGGTVTLTCGS MAK195-
STGAVTIDHYPYWFQQKPGQAPRTLISD 21-GS- TDDKHSWTPARFSGSLLGGKAALTLSGA
AE10-6 QPEDEAEYYCLLDYGGSFVFGGGTKLTV AM8 LG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 767 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIK Linker SEQ ID GGSGGGGSG NO: 768 VL SEQ ID
QAVVTQEPSLTVSPGGTVTLTCGSSTGA NO: 769 VTIDHYPYWFQQKPGQAPRTLISDTDDK
HSWTPARFSGSLLGGKAALTLSGAQPED EAEYYCLLDYGGSFVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 770 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD- D2E7 VH
AE10-6 SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT Binding AM7 VH NO: 771
FDDYAMHWVRQAPGKGLEWVSAITWNSG Protein HIDYADSVEGRFTISRDNAKNSLYLQMN
Heavy SLRAEDTAVYYCAKVSYLSTASSLDYWG Variable
QGTLVTVSSGGGGSGGGGSEVQLVQSGA D2E7-GS- EVKKPGASVKVSCKVSGYTLTELSMHWV
AE10-6 RQAPGKGLEWMGGFDPEDGETIYAQKFQ AM7
GRVTMTEDTSTDTAYMELSSLRSEDTAV YYCATDTVGYWEKFFQHWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 772 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 773 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 774 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 775
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- D2E7 VL AE10-6 SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG Binding AM7 VL NO: 776
IRNYLAWYQQKPGKAPKLLIYAASTLQS Protein GVPSRFSGSGSGTDFTLTISSLQPEDVA
Light TYYCQRYNRAPYTFGQGTKVEIKRGGSG Variable
GGGSGQAVVTQEPSLTVSPGGTVTLTCG D2E7-GS- SSTGTVTIDHYPYWFQQKPGQAPRTLIS
AE10-6 DTDDKHSWTPARFSGSLLGGKAALTLSG AM7
AQPEDEAEYYCLLDYGGRFVFGGGTKLT VLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 777 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 778 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 779
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 780 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD- D2E7 VH AE10-6 SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT Binding AM7 VH NO: 781
FDDYAMHWVRQAPGKGLEWVSAITWNSG Protein HIDYADSVEGRFTISRDNAKNSLYLQMN
Heavy SLRAEDTAVYYCAKVSYLSTASSLDYWG Variable
QGTLVTVSSGGGGSGGGGSEVQLVQSGA di D2E7- EVKKPGASVKVSCKVSGYTLTELSMHWV
GS-AE10- RQAPGKGLEWMGGFDPEDGETIYAQKFQ 6 AM7
GRVTMTEDTSTDTAYMELSSLRSEDTAV YYCATDTVGYWEKFFQHWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 782 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 783 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 784 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 785
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD- diD2E7ss AE10-6 SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS Binding VL AM7 VL NO: 786
IRNYLSWYQQKPGKAPKLLIYAASTLQS Protein GVPSRFSGSGSGTDFTLTISSLQPEDVA
Light TYYCQRYNRAPYTFGQGTKVEIKRGGSG Variable
GGGSGQAVVTQEPSLTVSPGGTVTLTCG diD2E7- SSTGTVTIDHYPYWFQQKPGQAPRTLIS
GS-AE10- DTDDKHSWTPARFSGSLLGGKAALTLSG 6 AM7
AQPEDEAEYYCLLDYGGRFVFGGGTKLT VLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 787 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 788 VL SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGT NO: 789
VTIDHYPYWFQQKPGQAPRTLISDTDDK HSWTPARFSGSLLGGKAALTLSGAQPED
EAEYYCLLDYGGRFVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 790 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2577H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 791
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFDDYALHWV RQAPGKGLEWVSGINWEGDDIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 792 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 793 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 794 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 795
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2577L AB438VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG AM1.2VL NO: 796
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGRNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDNLESYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 797 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 798 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 799
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 800 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2578H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 801
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGINWEGDDIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 802 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 803 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 804 FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 805 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK
DVD2578L AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM1.2VL
NO: 806 IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG
NO: 807 IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 808 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 809 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 810 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2579H AB438VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 811
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGINWEGDDIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 812
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 813 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 814
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 815 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2579L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM1.2VL NO: 816
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 817 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 818 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 819
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 820 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2580H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 821
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGINWEGDDIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 822 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 823 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 824 FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 825 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2580L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM1.2VL NO: 826
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 827 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 828 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 829
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 830 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2581H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 831
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGINWEGDDIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 832
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 833 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 834
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 835 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2581L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM1.2VL NO: 836
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG
NO: 837 IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 838 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 839 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 840 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2582H AB438VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 841
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFDDYALHWV RQAPGKGLEWVSGIGWEDDMIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 842 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 843 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 844 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG
TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 845
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2582L AB438VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG AM2.2VL NO: 846
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGGNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLEGSYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 847 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 848 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 849
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 850 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2583H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 851
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWEDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 852 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 853 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 854 FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 855 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2583L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM2.2VL NO: 856
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 857 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 858 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 859
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 860 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2584H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 861
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWEDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 862
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 863 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 864
FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 865 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2584L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM2.2VL NO: 866
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 867 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 868 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 869
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 870 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2585H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 871
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWEDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 872 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 873 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 874 FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 875 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2585L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM2.2VL NO: 876
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 877 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 878 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 879
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 880 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2586H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 881
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWEDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S
VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 882
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 883 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 884
FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 885 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2586L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM2.2VL NO: 886
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 887 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 888 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 889
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 890 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2587H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 891
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFDDYALHWV RQAPGKGLEWVSGIGWDEDMIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 892 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 893 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 894 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 895
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2587L AB438VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG AM3.2VL NO: 896
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSWSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLSGEYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 897 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 898 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 899
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 900 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2588H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 901
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWDEDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 902 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 903 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 904 FDDYALHWVRQAPGKGLEWVSGIGWDED MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 905 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2588L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM3.2VH NO: 906
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 907 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 908 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 909
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 910 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2589H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 911
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWDEDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 912
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 913 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 914
FDDYALHWVRQAPGKGLEWVSGIGWDED MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 915 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2589L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM3.2VL NO: 916
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 917 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 918 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 919
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 920 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2590H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 921
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWDEDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 922 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 923 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 924 FDDYALHWVRQAPGKGLEWVSGIGWDED MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 925 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2590L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM3.2VL NO: 926
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 927 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 928 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 929
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 930 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2591H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 931
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWDEDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 932
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 933 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 934
FDDYALHWVRQAPGKGLEWVSGIGWDED MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 935 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2591L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM3.2VL NO: 936
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 937 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 938 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 939
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 940 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2592H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 941
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFEDYALHWV RQAPGKGLEWVSGIGWDDDMIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 942 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 943 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 944 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 945
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2592L AB438VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG AM4.2VL NO: 946
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLDSYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 947 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 948 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 949
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 950 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2593H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 951
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFEDYALHWVRQAP GKGLEWVSGIGWDDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 952 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 953 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 954 FEDYALHWVRQAPGKGLEWVSGIGWDDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 955 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2593L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM4.2VH NO: 956
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG
NO: 957 IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 958 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 959 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 960 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2594H AB438VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 961
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFEDYAL HWVRQAPGKGLEWVSGIGWDDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 962
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 963 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 964
FEDYALHWVRQAPGKGLEWVSGIGWDDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 965 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2594L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM4.2VL NO: 966
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 967 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 968 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 969
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 970 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2595H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 971
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFEDYALHWVRQAP GKGLEWVSGIGWDDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 972 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 973 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 974 FEDYALHWVRQAPGKGLEWVSGIGWDDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 975 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2595L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM4.2VL NO: 976
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQG NO: 977 IRNYLAWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 978 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 979
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 980 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2596H AB438VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 981
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFEDYAL HWVRQAPGKGLEWVSGIGWDDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 982
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 983 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 984
FEDYALHWVRQAPGKGLEWVSGIGWDDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 985 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2596L
AB438VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG AM4.2VL NO: 986
IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQG
NO: 987 IRNYLAWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 988 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 989 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 990 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2597H AB439VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 991
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFDDYALHWV RQAPGKGLEWVSGINWEGDDIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 992 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 993 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 994 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 995
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2597L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM1.2VL NO: 996
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGRNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDNLESYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 997 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 998 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 999
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1000 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2598H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1001
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGINWEGDDIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1002 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1003 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1004 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1005
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2598L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM1.2VL NO: 1006
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS
NO: 1007 IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 1008 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1009 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1010 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2599H AB439VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1011
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGINWEGDDIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1012
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1013 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1014
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1015 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2599L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM1.2VL NO: 1016
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1017 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1018 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1019 IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1020 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2600H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1021
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGINWEGDDIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1022 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1023 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1024 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1025
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2600L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM1.2VL NO: 1026
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1027 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1028 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1029 IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1030 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2601H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1031
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGINWEGDDIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1032
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1033 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1034
FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1035
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2601L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM1.2VL NO: 1036
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS
NO: 1037 IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 1038 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1039 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1040 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2602H AB439VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 1041
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFDDYALHWV RQAPGKGLEWVSGIGWEDDMIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1042 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 1043 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1044 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1045
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2602L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM2.2VL NO: 1046
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGGNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLEGSYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1047 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 1048 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1049
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1050 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2603H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 1051
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWEDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1052 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1053 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1054 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1055
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2603L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM2.2VL NO: 1056
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1057 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 1058 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1059
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1060 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2604H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 1061
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWEDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1062
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1063 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1064
FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1065 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2604L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM2.2VL NO: 1066
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1067 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1068 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1069 IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1070 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2605H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 1071
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWEDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA
GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO:
1072 FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGP NO:
1073 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1074
FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1075 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2605L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM2.2VL NO: 1076
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1077 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1078 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1079 IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1080 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2606H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM2.2VH NO: 1081
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWEDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNSRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1082
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1083 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1084
FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1085 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2606L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM2.2VL NO: 1086
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1087 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 1088 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1089
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1090 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2607H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 1091
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFDDYALHWV RQAPGKGLEWVSGIGWDEDMIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1092 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 1093 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1094 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1095
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2607L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM3.2VL NO: 1096
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSWSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLSGEYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1097 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 1098 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1099
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1100 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2608H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 1101
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWDEDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1102 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1103 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1104 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1105
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2608L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM3.2VH NO: 1106
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1107 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 1108 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1109
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG
CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1110
ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2609H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 1111
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWDEDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1111
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1112 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1113
FDDYALHWVRQAPGKGLEWVSGIGWDED MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1114 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2609L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM3.2VL NO: 1115
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1117 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1118 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO:
1119 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1120 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2610H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 1121
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWDEDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1122 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1123 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1124 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1125
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2610L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM3.2VL NO: 1126
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1127 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1128 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO:
1129 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1130 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2611H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM3.2VH NO: 1131
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFDDYAL HWVRQAPGKGLEWVSGIGWDEDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1132
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1133 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1134
FDDYALHWVRQAPGKGLEWVSGIGWDED MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1135 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2611L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM3.2VL NO: 1136
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1137 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAP NO: 1138 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1139
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1140 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2612H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 1141
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSGGGGSGGGGSEVQLVESGG
GLVQPGRSLRLSCAASGFTFEDYALHWV RQAPGKGLEWVSGIGWDDDMIDYADSVK
GRFTISRDNAKNSLYLQMNSLRVEDTAL YYCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1142 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID GGGGSGGGGS NO: 1143 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1144 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1145
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2612L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM4.2VL NO: 1146
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLDSYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1147 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
GGSGGGGSG NO: 1148 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1149
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1150 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2613H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 1151
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFEDYALHWVRQAP GKGLEWVSGIGWDDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1152 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1153 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1154 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1155
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2613L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM4.2VH NO: 1156
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS
NO: 1157 IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 1158 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1159 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1160 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2614H AB439VH
MSL10- SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 1161
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFEDYAL HWVRQAPGKGLEWVSGIGWDDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1162
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1163 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1164
FEDYALHWVRQAPGKGLEWVSGIGWDDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1165 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2614L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM4.2VL NO: 1166
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1167 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1168 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1169 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1170 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2615H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 1171
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPEVQLVESGGGLVQ
PGRSLRLSCAASGFTFEDYALHWVRQAP GKGLEWVSGIGWDDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1172 FDDYAMHWVRQAPGKGLEWVSAITWNSG
HIDYADSVEGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS
Linker SEQ ID ASTKGP NO: 1173 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1174 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1175
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2615L AB439VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS AM4.2VL NO: 1176
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQS NO: 1177 IRNYLSWYQQKPGKAPKLLIYAASTLQS
GVPSRFSGSGSGTDFTLTISSLQPEDVA TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1178 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1179 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1180 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2616H AB439VH MSL10- SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM4.2VH NO: 1181
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSSASTKGPSVFPLAPEVQLVE
SGGGLVQPGRSLRLSCAASGFTFEDYAL HWVRQAPGKGLEWVSGIGWDDDMIDYAD
SVKGRFTISRDNAKNSLYLQMNSLRVED TALYYCAGNNRGYGGLDVWGQGTTVTVS S VH SEQ
ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1182
FDDYAMHWVRQAPGKGLEWVSAITWNSG HIDYADSVEGRFTISRDNAKNSLYLQMN
SLRAEDTAVYYCAKVSYLSTASSLDYWG QGTLVTVSS Linker SEQ ID ASTKGPSVFPLAP
NO: 1183 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1184
FEDYALHWVRQAPGKGLEWVSGIGWDDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1185 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2616L
AB439VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS AM4.2VL NO: 1186
IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQS
NO: 1187 IRNYLSWYQQKPGKAPKLLIYAASTLQS GVPSRFSGSGSGTDFTLTISSLQPEDVA
TYYCQRYNRAPYTFGQGTKVEIKR Linker SEQ ID TVAAP NO: 1188 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1189 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1190 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2617H AB436VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1191
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGINWEGDDIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1192 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1193 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1194 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1195
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2617L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM1.2VL NO: 1196
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGRNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDNLESYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1197 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1198 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1199
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1200 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2618H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1201
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGINWEGDDIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1202 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1203 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1204 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1205
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2618L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM1.2VL NO: 1206
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL
NO: 1207 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1208 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1209 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1210 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2619H AB436VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1211
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGINWEGDDIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1212 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1213 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1214 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1215
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2619L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM1.2VL NO: 1216
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1217 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1218 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1219 IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1220 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2620H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1221
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGINWEGDDIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1222 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1223 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT
NO: 1224 FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1225 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2620L
AB436VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL AM1.2VL NO: 1226
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1227 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1228 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1229 IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1230 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2621H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1231
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGINWEGDDIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1232 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1233 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1234 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1235
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2621L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM1.2VL NO: 1236
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL
NO: 1237 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1238 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1239 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1240 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2622H AB436VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1241
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWEDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1242 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1243 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1244 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1245
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2622L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM2.2VL NO: 1246
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGGNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLEGSYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1247 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1248 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1249
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1250 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2623H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1251
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWEDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1252 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1253 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1254 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1255
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2623L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM2.2VL NO: 1256
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1257 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1258 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1259
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1260 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2624H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1261
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWEDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY
YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT
NO: 1262 FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS Linker SEQ ID ASTKGPSVFPLAP NO:
1263 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1264
FDDYALHWVRQAPGKGLEWVSGIGWEDD MIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1265 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2624L
AB436VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL AM2.2VL NO: 1266
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1267 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1268 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1269 IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1270 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2625H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1271
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWEDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1272 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1273 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1274 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1275
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2625L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM2.2VL NO: 1276
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1277 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1278 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1279 IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1280 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2626H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1281
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWEDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1282 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1283 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1284 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1285
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2626L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM2.2VL NO: 1286
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1287 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1288 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1289
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL
NO: 1290 VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2627H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1291
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWDEDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1292 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1293 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1294 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1295
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2627L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM3.2VL NO: 1296
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSWSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLSGEYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1297 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR
Linker SEQ ID GGSGGGGSG NO: 1298 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1299 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1300 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2628H AB436VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1301
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWDEDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1302 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1303 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1304 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1305
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2628L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM3.2VL NO: 1306
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1307 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1308 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1309
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1310 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2629H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1311
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWDEDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1312 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1313 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1314 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1315
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2629L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM3.2VL NO: 1316
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1317 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1318 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO:
1319 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1320 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2630H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1321
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWDEDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1322 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1323 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1324 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1325
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2630L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM3.2VL NO: 1326
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1327 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1328 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO:
1329 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1330 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2631H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1331
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWDEDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1332 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1333 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1334 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1335
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2631L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM3.2VL NO: 1336
VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKRTVAA
PQSVLTQPPSASGTPGQRVTISCSGSWS NIGSNTVNWYQQLPGTAPKLLIYNNNQR
PSGVPDRFSGSKSGTSASLAISGLQSED EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ
ID DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1337
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1338 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1339 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1340 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2632H AB436VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1341
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFEDYALHWVRQAP GKGLEWVSGIGWDDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1342 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1343 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1344 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1345
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2632L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM4.2VL NO: 1346
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLDSYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1347 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1348 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1349
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1350 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2633H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1351
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFEDYALHWVRQAPGKGL EWVSGIGWDDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1352 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1353 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1354 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1355
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2633L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM4.2VL NO: 1356
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL
NO: 1357 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1358 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1359 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1360 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2634H AB436VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1361
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFEDYALHWVR QAPGKGLEWVSGIGWDDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1362 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1363 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1364 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1365
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2634L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM4.2VL NO: 1366
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1367 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1368 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1369 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1370 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2635H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1371
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFEDYALHWVRQAPGKGL EWVSGIGWDDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1372 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1373 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1374 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1375
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2635L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM4.2VL NO: 1376
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1377 VSSAVAWYQQKPGKAPKLLIYWASARHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1378 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1379 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1380 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2636H AB436VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1381
FSNYGVTWVRQAPGKGLEWVSMIWADGS THYASSVKGRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFEDYALHWVR QAPGKGLEWVSGIGWDDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1382 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1383 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1384 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1385
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2636L AB436VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQL AM4.2VL NO: 1386
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL
NO: 1387 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1388 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1389 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1390 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2637H AB437VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1391
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGINWEGDDIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1392 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1393 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1394 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1395
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2637L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM1.2VL NO: 1396
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGRNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDNLESYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1397 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1398 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1399
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1400 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2638H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1401
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGINWEGDDIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1402 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1403 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1404 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1405
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2638L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM1.2VL NO: 1406
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL
NO: 1407 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1408 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1409 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1410 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2639H AB437VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1411
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGINWEGDDIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1412 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP
NO: 1413 VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1414
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1415 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2639L
AB437VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL AM1.2VL NO: 1416
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1417 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1418 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1419 IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1420 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2640H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1421
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGINWEGDDIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1422 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1423 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1424 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1425
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2640L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM1.2VL NO: 1426
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGRNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDNLESYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1427 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1428 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1429 IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1430 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2641H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM1.2VH NO: 1431
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGINWEGDDIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1432 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1433 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1434 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1435
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2641L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM1.2VL NO: 1436
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGRNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDNLESYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL
NO: 1437 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1438 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1439 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1440 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2642H AB437VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1441
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWEDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1442 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1443 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1444 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1445
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2642L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM2.2VL NO: 1446
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGGNTVNWYQQLPGTAPKLLIYS NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLEGSYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1447 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1448 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1449
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1450 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2643H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1451
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWEDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1452 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1453 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1454 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1455
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2643L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM2.2VL NO: 1456
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1457 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1458 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1459
IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1460 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2644H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1461
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWEDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1462 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1463 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1464 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1465
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2644L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM2.2VL NO: 1466
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1467 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1468 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1469 IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1470 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2645H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1471
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWEDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNSR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1472 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1473 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1474 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1475
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2645L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM2.2VL NO: 1476
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGGNTVNWYQQLPGTAPKLL IYSNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLEGSYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1477 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1478 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1479 IGGNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1480 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2646H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM2.2VH NO: 1481
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWEDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNSRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1482 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1483 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1484 FDDYALHWVRQAPGKGLEWVSGIGWEDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1485
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2646L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM2.2VL NO: 1486
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGGNTVNWYQQLPGTAPKLLIYSNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLEGSYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1487 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1488 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1489
IGGNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLEGSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1490 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2647H AB437VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1491
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFDDYALHWVRQAP GKGLEWVSGIGWDEDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1492 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1493 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1494 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1495
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2647L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM3.2VL NO: 1496
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSWSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLSGEYVFGGGTK LTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1497 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1498 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1499
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1500 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2648H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1501
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWDEDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1502 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1503 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1504 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1505
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2648L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM3.2VL NO: 1506
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1507 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1508 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1509
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1510 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2649H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1511
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWDEDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1512 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1513 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1514 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1515
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2649L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM3.2VL NO: 1516
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1517 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1518 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO:
1519 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1520 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2650H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1521
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFDDYALHWVRQAPGKGL EWVSGIGWDEDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1522 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1523 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1524 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1525
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2650L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM3.2VL NO: 1526
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSWSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLSGEYVFGG GTKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1527 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1528 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO:
1529 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1530 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2651H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM3.2VH NO: 1531
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFDDYALHWVR QAPGKGLEWVSGIGWDEDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1532 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1533 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1534 FDDYALHWVRQAPGKGLEWVSGIGWDED
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1535
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2651L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM3.2VL NO: 1536
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSWS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLSGEYVFGGGTKLTVL G VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1537 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAP NO: 1538 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSWSN NO: 1539
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLSGEYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1540 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2652H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1541
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSGGGGSGGGGSEVQLVESGGGLVQ
PGRSLRLSCAASGFTFEDYALHWVRQAP GKGLEWVSGIGWDDDMIDYADSVKGRFT
ISRDNAKNSLYLQMNSLRVEDTALYYCA GNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1542 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID GGGGSGGGGS NO: 1543 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1544 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1545
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2652L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM4.2VL NO: 1546
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRGGSG GGGSGQSVLTQPPSASGTPGQRVTISCS
GSSSNIGSNTVNWYQQLPGTAPKLLIYN NNQRPSGVPDRFSGSKSGTSASLAISGL
QSEDEADYYCAAWDDSLDSYVFGGGTKL TVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1547 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
GGSGGGGSG NO: 1548 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1549
IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1550 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2653H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1551
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFEDYALHWVRQAPGKGL EWVSGIGWDDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1552 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1553 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1554 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1555
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2653L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM4.2VL NO: 1556
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL
NO: 1557 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1558 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1559 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1560 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2654H AB437VH
MSL10- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1561
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFEDYALHWVR QAPGKGLEWVSGIGWDDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1562 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1563 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1564 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1565
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2654L
AB437VL MSL10- SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL AM4.2VL NO: 1566
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1567 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1568 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1569 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1570 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2655H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1571
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPEVQLVESGGGLVQPGRS
LRLSCAASGFTFEDYALHWVRQAPGKGL EWVSGIGWDDDMIDYADSVKGRFTISRD
NAKNSLYLQMNSLRVEDTALYYCAGNNR GYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1572 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGP NO: 1573 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1574 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1575
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2655L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM4.2VL NO: 1576
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PSVFIFPPQSVLTQPPSASGTPGQRVTI
SCSGSSSNIGSNTVNWYQQLPGTAPKLL IYNNNQRPSGVPDRFSGSKSGTSASLAI
SGLQSEDEADYYCAAWDDSLDSYVFGGG TKLTVLG VL SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1577 VSSAVAWYQQKPGKAPKLLIYWASTLHT
GVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID
TVAAPSVFIFPP NO: 1578 VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN NO:
1579 IGSNTVNWYQQLPGTAPKLLIYNNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID QPKAAPSVTLFPPSSEELQANKATLVCL
NO: 1580 ISDFYPGAVTVAWKADSSPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSY
SCQVTHEGSTVEKTVAPTECS DVD2656H AB437VH MSL10- SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT AM4.2VH NO: 1581
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSSASTKGPSVFPLAPEVQLVESGGG
LVQPGRSLRLSCAASGFTFEDYALHWVR QAPGKGLEWVSGIGWDDDMIDYADSVKG
RFTISRDNAKNSLYLQMNSLRVEDTALY YCAGNNRGYGGLDVWGQGTTVTVSS VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1582 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1583 VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1584 FEDYALHWVRQAPGKGLEWVSGIGWDDD
MIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNNRGYGGLDVWGQG TTVTVSS
CH SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1585
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2656L AB437VL MSL10- SEQ ID
DIQMTQSPSSLSASVGDRVTITCKASQL AM4.2VL NO: 1586
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKRTVAA PQSVLTQPPSASGTPGQRVTISCSGSSS
NIGSNTVNWYQQLPGTAPKLLIYNNNQR PSGVPDRFSGSKSGTSASLAISGLQSED
EADYYCAAWDDSLDSYVFGGGTKLTVLG VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL
NO: 1587 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Linker SEQ ID TVAAP NO: 1588 VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1589 IGSNTVNWYQQLPGTAPKLLIYNNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDSLDSYVFGGGTKLTVLG CL SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCL NO: 1590 ISDFYPGAVTVAWKADSSPVKAGVETTT
PSKQSNNKYAASSYLSLTPEQWKSHRSY SCQVTHEGSTVEKTVAPTECS DVD2657H MSL10-
AB436VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1591
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSGGGGSGGGGSEVQLVESGGGL
VQPGGSLRLSCAASGFTFSNYGVTWVRQ APGKGLEWVSMIWADGSTHYASSVKGRF
TISRDNSKNTLYLQMNSLRAEDTAVYYC AREWQHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1592 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID GGGGSGGGGS NO: 1593 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1594 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1595
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2657L MSL10- AB436VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1596
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGG GSGGGGSGDIQMTQSPSSLSASVGDRVT
ITCRASQLVSSAVAWYQQKPGKAPKLLI YWASARHTGVPSRFSGSGSGTDFTLTIS
SLQPEDFATYYCQQHYKTPFTFGQGTKL EIKR VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1597 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID GGSGGGGSG NO: 1598 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL NO:
1599 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1600 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2658H MSL10- AB436VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1601
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPEVQLVESGGGLVQPG
GSLRLSCAASGFTFSNYGVTWVRQAPGK GLEWVSMIWADGSTHYASSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCAREW QHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1602 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGP NO: 1603 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1604
FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1605
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2658L MSL10- AB436VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1606
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPDIQMTQSPSSLSASVGDRVTITC
RASQLVSSAVAWYQQKPGKAPKLLIYWA SARHTGVPSRFSGSGSGTDFTLTISSLQ
PEDFATYYCQQHYKTPFTFGQGTKLEIK R VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1607 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAP NO: 1608 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1609
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1610 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2659H MSL10- AB436VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1611
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPSVFPLAPEVQLVESG
GGLVQPGGSLRLSCAASGFTFSNYGVTW VRQAPGKGLEWVSMIWADGSTHYASSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAV YYCAREWQHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1612 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1613 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1614 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1615
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2659L MSL10- AB436VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1616
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPSVTLFPPDIQMTQSPSSLSASVG
DRVTITCRASQLVSSAVAWYQQKPGKAP KLLIYWASARHTGVPSRFSGSGSGTDFT
LTISSLQPEDFATYYCQQHYKTPFTFGQ GTKLEIKR VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1617 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAPSVTLFPP NO: 1618 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL
NO: 1619 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1620 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2660H MSL10- AB436VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1621
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPEVQLVESGGGLVQPG
GSLRLSCAASGFTFSNYGVTWVRQAPGK GLEWVSMIWADGSTHYASSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCAREW QHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1622 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGP NO: 1623 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1624 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1625
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2660L MSL10- AB436VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1626
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPSVTLFPPDIQMTQSPSSLSASVG
DRVTITCRASQLVSSAVAWYQQKPGKAP KLLIYWASARHTGVPSRFSGSGSGTDFT
LTISSLQPEDFATYYCQQHYKTPFTFGQ GTKLEIKR VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1627 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAPSVTLFPP NO: 1628 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL
NO: 1629 VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1630 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2661H MSL10- AB436VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1631
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPSVFPLAPEVQLVESG
GGLVQPGGSLRLSCAASGFTFSNYGVTW VRQAPGKGLEWVSMIWADGSTHYASSVK
GRFTISRDNSKNTLYLQMNSLRAEDTAV YYCAREWQHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1632 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1633 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1634 FSNYGVTWVRQAPGKGLEWVSMIWADGS
THYASSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1635
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2661L MSL10- AB436VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1636
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPDIQMTQSPSSLSASVGDRVTITC
RASQLVSSAVAWYQQKPGKAPKLLIYWA SARHTGVPSRFSGSGSGTDFTLTISSLQ
PEDFATYYCQQHYKTPFTFGQGTKLEIK R VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1637 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAP NO: 1638 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQL NO: 1639
VSSAVAWYQQKPGKAPKLLIYWASARHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYKTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1640 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2662H MSL10- AB437VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1641
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSGGGGSGGGGSEVQLVESGGGL
VQPGGSLRLSCAASGFTFSNYGVEWVRQ APGKGLEWVSGIWADGSTHYADTVKSRF
TISRDNSKNTLYLQMNSLRAEDTAVYYC
AREWQHGPVAYWGQGTLVTVSS VH SEQ ID EVQLVESGGGLVQPGRSLRLSCAASGFT NO:
1642 FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS Linker SEQ ID GGGGSGGGGS NO:
1643 VH SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1644
FSNYGVEWVRQAPGKGLEWVSGIWADGS THYADTVKSRFTISRDNSKNTLYLQMNS
LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1645 VKDYFPEPVTVSWNSGALTSGVHTFPAV
LQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPC
PAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPV
LDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK DVD2662L MSL10-
AB437VL SEQ ID QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1646
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGG GSGGGGSGDIQMTQSPSSLSASVGDRVT
ITCKASQLVSSAVAWYQQKPGKAPKLLI YWASTLHTGVPSRFSGSGSGTDFTLTIS
SLQPEDFATYYCQQHYRTPFTFGQGTKL EIKR VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1647 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID GGSGGGGSG NO: 1648 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL NO:
1649 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1650 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2663H MSL10- AB437VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1651
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPEVQLVESGGGLVQPG
GSLRLSCAASGFTFSNYGVEWVRQAPGK GLEWVSGIWADGSTHYADTVKSRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCAREW QHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1652 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGP NO: 1653 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1654 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1655
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2663L MSL10- AB437VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1656
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPDIQMTQSPSSLSASVGDRVTITC
KASQLVSSAVAWYQQKPGKAPKLLIYWA STLHTGVPSRFSGSGSGTDFTLTISSLQ
PEDFATYYCQQHYRTPFTFGQGTKLEIK R VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1657 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAP NO: 1658 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1659
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1660 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2664H MSL10- AB437VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1661
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPSVFPLAPEVQLVESG
GGLVQPGGSLRLSCAASGFTFSNYGVEW VRQAPGKGLEWVSGIWADGSTHYADTVK
SRFTISRDNSKNTLYLQMNSLRAEDTAV YYCAREWQHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1662 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1663 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1664 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1665
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2664L MSL10- AB437VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1666
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPSVTLFPPDIQMTQSPSSLSASVG
DRVTITCKASQLVSSAVAWYQQKPGKAP KLLIYWASTLHTGVPSRFSGSGSGTDFT
LTISSLQPEDFATYYCQQHYRTPFTFGQ GTKLEIKR VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1667 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAPSVTLFPP NO: 1668 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL
NO: 1669 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1670 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2665H MSL10- AB437VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1671
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPEVQLVESGGGLVQPG
GSLRLSCAASGFTFSNYGVEWVRQAPGK GLEWVSGIWADGSTHYADTVKSRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCAREW QHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1672 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGP NO: 1673 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1674 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1675
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2665L MSL10- AB437VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1676
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPSVTLFPPDIQMTQSPSSLSASVG
DRVTITCKASQLVSSAVAWYQQKPGKAP KLLIYWASTLHTGVPSRFSGSGSGTDFT
LTISSLQPEDFATYYCQQHYRTPFTFGQ GTKLEIKR VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1677 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAPSVTLFPP NO: 1678 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL
NO: 1679 VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR
Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO: 1680
NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC DVD2666H MSL10- AB437VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT AM1.2VH NO: 1681
FDDYALHWVRQAPGKGLEWVSGINWEGD DIDYADSVKGRFTISRDNAKNSLYLQMN
SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSSASTKGPSVFPLAPEVQLVESG
GGLVQPGGSLRLSCAASGFTFSNYGVEW VRQAPGKGLEWVSGIWADGSTHYADTVK
SRFTISRDNSKNTLYLQMNSLRAEDTAV YYCAREWQHGPVAYWGQGTLVTVSS VH SEQ ID
EVQLVESGGGLVQPGRSLRLSCAASGFT NO: 1682 FDDYALHWVRQAPGKGLEWVSGINWEGD
DIDYADSVKGRFTISRDNAKNSLYLQMN SLRVEDTALYYCAGNSRGYGGLDVWGQG TTVTVSS
Linker SEQ ID ASTKGPSVFPLAP NO: 1683 VH SEQ ID
EVQLVESGGGLVQPGGSLRLSCAASGFT NO: 1684 FSNYGVEWVRQAPGKGLEWVSGIWADGS
THYADTVKSRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAREWQHGPVAYWGQGTL VTVSS CH
SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCL NO: 1685
VKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTP
EVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDW
LNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK DVD2666L MSL10- AB437VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN AM1.2VL NO: 1686
IGRNTVNWYQQLPGTAPKLLIYSNNQRP SGVPDRFSGSKSGTSASLAISGLQSEDE
ADYYCAAWDDNLESYVFGGGTKLTVLGQ PKAAPDIQMTQSPSSLSASVGDRVTITC
KASQLVSSAVAWYQQKPGKAPKLLIYWA STLHTGVPSRFSGSGSGTDFTLTISSLQ
PEDFATYYCQQHYRTPFTFGQGTKLEIK R VL SEQ ID
QSVLTQPPSASGTPGQRVTISCSGSSSN NO: 1687 IGRNTVNWYQQLPGTAPKLLIYSNNQRP
SGVPDRFSGSKSGTSASLAISGLQSEDE ADYYCAAWDDNLESYVFGGGTKLTVLG Linker SEQ
ID QPKAAP NO: 1688 VL SEQ ID DIQMTQSPSSLSASVGDRVTITCKASQL NO: 1689
VSSAVAWYQQKPGKAPKLLIYWASTLHT GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQHYRTPFTFGQGTKLEIKR Ck SEQ ID TVAAPSVFIFPPSDEQLKSGTASVVCLL NO:
1690 NNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVY
ACEVTHQGLSSPVTKSFNRGEC
Example 3.4
Binding Affinity of DVD-Igs
[0603] For measuring simultaneous binding of two antigens to DVD,
DVD-Igs were captured onto a goat anti huIgG FC at 5 .mu.l/min for
1 min. The sclerostin at 50 nM was then injected over the captured
surface at 5 .mu.l/min for 10 min, to saturate DVD binding,
followed by coinjection of 50 nM TNF at 5 .mu.l/min for 10 min to
check for TNF binding to the DVD, which has sclerostin already
bound to it. The surfaces were regenerated with 10 s injection of
50 mM NaOH followed by 10 s injection of 10 mM Glycine pH1.5 at a
flow rate of 100 .mu.l/min. The same experiment was repeated in the
reverse sequence of antigens where TNF was injected first followed
by coinjection of sclerostin. Exemplary binding profiles are
provided for MAK199-1-GS-AE10-6AM7 DVD-Ig (FIG. 7A) and DVD2623
(FIG. 7B).
[0604] The affinity and kinetic parameters of the DVD-Ig binding to
SOST and TNF are provided in Table 19 below. The detailed
description of the method used is provided in section
TABLE-US-00021 TABLE 19 Affinity and kinetic parameters of the
DVD-Ig binding to sclerostin and TNF name antigen k.sub.a,
M.sup.-1s.sup.-1 k.sub.d, s.sup.-1 K.sub.D, M DVD2603 hu sclerostin
1.40E+05 <1E-06 <7.1E-12 DVD2658 hu sclerostin 2.70E+05
<1E-06 <3.7E-12 DVD2657 hu sclerostin 2.80E+05 4.90E-06
1.70E-11 DVD2605 hu sclerostin 5.70E+04 3.70E-05 6.40E-10 DVD2606
hu sclerostin 9.90E+04 6.50E-05 6.60E-10 DVD2645 hu sclerostin
8.50E+04 6.30E-05 7.50E-10 DVD2643 hu sclerostin 7.60E+04 6.30E-05
8.30E-10 DVD2642 hu sclerostin 6.00E+04 6.00E-05 1.00E-09 DVD2646
hu sclerostin 1.10E+05 1.20E-04 1.10E-09 DVD2628 hu sclerostin
6.40E+04 9.30E-05 1.50E-09 DVD2631 hu sclerostin 7.90E+04 1.40E-04
1.80E-09 DVD2650 hu sclerostin 5.10E+04 1.30E-04 2.60E-09 DVD2651
hu sclerostin 4.30E+04 1.20E-04 2.70E-09 DVD2648 hu sclerostin
8.30E+04 3.30E-04 4.00E-09 DVD2647 hu sclerostin 4.60E+04 2.10E-04
4.60E-09 DVD2607 hu sclerostin 5.4E+04 1.9E-04 3.5E-09 DVD2608 hu
sclerostin 7.4E+04 1.5E-04 2.1E-09 DVD2610 hu sclerostin 5.3E+04
9.2E-05 1.7E-09 DVD2611 hu sclerostin 5.7E+04 7.1E-05 1.2E-09
DVD2623 hu sclerostin 7.5E+04 <1E-06 <1.3E-11 DVD2627 hu
sclerostin 4.5E+04 4.3E-05 9.6E-10 DVD2630 hu sclerostin 3.7E+04
4.8E-05 1.3E-09 DVD2603 cyno 1.70E+05 1.00E-04 6.00E-10 sclerostin
DVD2658 cyno 5.00E+05 3.00E-05 6.10E-11 sclerostin DVD2657 cyno
3.90E+05 5.00E-05 1.30E-10 sclerostin DVD2605 cyno 8.90E+04
9.60E-06 1.10E-10 sclerostin DVD2606 cyno 2.00E+05 2.00E-05
9.60E-11 sclerostin DVD2645 cyno 1.30E+05 2.80E-05 2.10E-10
sclerostin DVD2643 cyno 1.70E+05 <1E-06 <5.9E-12 sclerostin
DVD2642 cyno 1.50E+05 <1E-06 <6.7E-12 sclerostin DVD2646 cyno
2.30E+05 6.60E-05 2.90E-10 sclerostin DVD2628 cyno 9.00E+04
2.80E-04 3.10E-09 sclerostin DVD2631 cyno 1.10E+05 2.40E-04
2.30E-09 sclerostin DVD2650 cyno 1.10E+05 1.50E-04 1.40E-09
sclerostin DVD2651 cyno 7.80E+04 2.00E-04 2.60E-09 sclerostin
DVD2648 cyno 9.30E+04 1.20E-04 1.30E-09 sclerostin DVD2647 cyno
7.30E+04 2.30E-04 3.10E-09 sclerostin DVD2607 cyno 9.90E+04
1.40E-04 1.50E-09 sclerostin DVD2608 cyno 1.20E+05 2.10E-04
1.50E-09 sclerostin DVD2610 cyno 7.30E+04 8.60E-05 1.20E-09
sclerostin DVD2611 cyno 1.10E+05 1.70E-04 1.50E-09 sclerostin
DVD2623 cyno 9.90E+04 <1e-6 <1.0e-11 sclerostin DVD2627 cyno
5.40E+04 1.40E-04 2.60E-09 sclerostin DVD2630 cyno 4.60E+04
1.20E-05 2.50E-10 sclerostin DVD2603 huTNF 1.10E+06 1.10E-04
1.10E-10 DVD2658 huTNF 1.00E+05 5.30E-05 5.20E-10 DVD2657 huTNF
1.70E+05 1.20E-04 7.20E-10 DVD2605 huTNF 1.30E+06 5.20E-05 4.10E-11
DVD2606 huTNF 1.20E+06 4.80E-05 4.00E-11 DVD2645 huTNF 2.20E+06
3.30E-05 1.50E-11 DVD2643 huTNF 2.40E+06 3.50E-05 1.50E-11 DVD2642
huTNF 2.70E+06 3.30E-05 1.20E-11 DVD2646 huTNF 2.40E+06 3.00E-05
1.20E-11 DVD2628 huTNF 4.80E+06 6.30E-05 1.30E-11 DVD2650 huTNF
1.80E+06 3.50E-05 2.00E-11 DVD2651 huTNF 2.30E+06 3.80E-05 1.70E-11
DVD2648 huTNF 1.80E+06 5.60E-05 3.10E-11 DVD2647 huTNF 2.40E+06
3.40E-05 1.40E-11 DVD2603 huTNF 4.20E+06 3.00E-05 7.00E-12 DVD2607
huTNF 1.10E+05 4.70E-05 4.20E-10 DVD2608 huTNF 7.00E+04 1.50E-04
2.20E-09 DVD2610 huTNF 8.70E+04 3.30E-05 3.80E-10 DVD2611 huTNF
9.00E+04 2.40E-05 2.70E-10 DVD2623 huTNF 2.10E+05 4.70E-05 2.30E-10
DVD2627 huTNF 1.90E+05 3.40E-05 1.80E-10 DVD2630 huTNF 2.00E+05
2.90E-05 1.40E-10 MAK195-21-SS- hu sclerostin 6.8E+05 <1E-06
<1.5E-12 AE106-AM7 MAK195-21-GS- hu sclerostin 2.2E+06 7.9E-06
3.6E-12 AE106-AM8 MAK195-21-GS- hu sclerostin 2.1E+06 1.7E-05
8.2E-12 AE106-AM3 MAK195-21-GS- hu sclerostin 2.0E+06 2.0E-05
1.0E-11 AE106-AM7 AE10-6AM7-GS- hu sclerostin 6.8E+06 <1E-06
<1.5E-13 MAK195-21 D2E7-GS-AE10- hu sclerostin 1.2E+06 <1E-06
<8.3E-13 6AM7 diD2E7ss-GS- hu sclerostin 1.1E+06 2.1E-07 1.9E-13
AE10-6AM7 MAK199-1-GS- hu sclerostin 5.9E+06 2.3E-06 3.9E-13 AM10-6
AM7 MAK199-1-GS- hu sclerostin 6.1E+06 <1e-06 <1.6e-13
AE10-6AM7QL MAK199-1-GS- cyno 9.9E+06 <1e-6 <1e-13
AE10-6AM7QL sclerostin MAK199-1-GS- cyno 9.3E+06 <1e-6
<1.1e-13 AE10-6AM7 sclerostin MAK199-1-SS- hu sclerostin 1.6E+05
8.3E-06 5.2E-11 AE10-6AM7 MAK195-21-SS- huTNF 7.3E+06 6.0E-05
8.2E-12 AE10-6AM7 MAK195-21-GS- huTNF 4.3E+06 4.8E-05 1.1E-11
AE10-6AM7 MAK195-21-GS- huTNF 4.2E+06 4.7E-05 1.1E-11 AE10-6AM3
MAK195-21-GS- huTNF 4.0E+06 4.8E-05 1.2E-11 AE10-6AM8 AE10-6AM7-GS-
huTNF 1.0E+06 4.8E-05 4.7E-11 MAK195-21 D2E7GSAE106AM7 huTNF
5.8E+06 4.3E-05 7.3E-12 diD2E7ssGSAE106 huTNF 2.4E+06 6.2E-05
2.6E-11 AM7 MAK199- huTNF 2.1E+06 5.8E-05 2.7E-11 1GSAM10-6 AM7
MAK199- huTNF 4.2E+06 3.0E-05 7.0E-12 1GSAE106AM7QL MAK199-1-SS-
huTNF 2.6E+06 4.4E-05 1.7E-11 AE106AM7
Example 3.5
TopFlash Wnt Pathway Sclerostin Neutralization Assay
[0605] The assay that demonstrates neutralization of sclerostin
inhibition of Wnt pathway was performed as described above.
DVD2603, DVD2605, DVD2606, DVD2607, DVD2608, DVD2610, DVD2611,
DVD2623, DVD2627, DVD2628, DVD2630, DVD2631, DVD2642, DVD2643,
DVD2645, DVD2646, DVD2647, DVD2648, DVD2650, DVD2651, DVD2657,
DVD2658 inhibited recombinant human sclerostin with IC50 of 12-24.4
nM. MAK199-1-GS-AE10-6 AM7, MAK199-1-SS-AE10-6 AM7,
MAK199-1-GS-AE10-6 AM7 QL, MAK195/21-GS-AE10-6 AM3,
MAK195/21-GS-AE10-6 AM8, diD2E7ss-GS-AE10-6 AM7, D2E7-GS-AE10-6
AM7, AE10-6 AM7-GS MAK195/21 inhibited recombinant human sclerostin
with IC50 of 1.7-6.6 nM and recombinant cynomolgus monkey
sclerostin with IC50 of 2-9.1 nM. MAK199-1-GS-AE10-6 AM7,
MAK199-1-SS-AE10-6 AM7, MAK195/21-GS-AE10-6 AM7,
MAK195/21-SS-AE10-6 AM7, MAK195/21-GS-AE10-6 AM3,
MAK195/21-GS-AE10-6 AM8, diD2E7ss-GS-AE10-6 AM7, D2E7-GS-AE10-6
AM7, AE10-6 AM7-GS MAK195/21 inhibited recombinant mouse sclerostin
with IC50 of 9.4-27.6 nM.
Example 3.6
Neutralization of Recombinant TNF-.alpha.
[0606] TNF-.alpha. neutralizing potency of the DVD-Igs was
evaluated in L929 Neutralization of recombinant TNF assay.
Semi-confluent L929 mouse fibroblast cells (ATCC, cat#CCL-1.TM.)
were grown and harvested using 0.25% tryspin (Gibco,
cat#25200-056). The cells were washed with PBS (Gibco,
cat#14190-144), counted and resuspended at 5.times.10.sup.5
cells/mL in complete assay media consisting of RPMI 1640 (Gibco,
cat#21870), 10% Fetal Bovine Serum (Hyclone, cat#SH30070.03), 1%
L-Glutamine (Gibco, cat#25030), 1% Sodium Pyruvate (Gibco,
cat#113670), 1% Non-Essential Aminos (Gibco, cat#11140), 50
units/mL Penicillin/50 .mu.g/mL Streptomycin (Gibco, cat#15140), 55
.mu.M of 2-BME (Mercaptoethanol-Gibco, cat#21985), and a 2.times.
concentration of 2 .mu.g/mL actinomycin D (Sigma, cat#A1410). The
cells were seeded in a 96-well plate (Costar, cat#3599) at a volume
of 100 .mu.L corresonding to 5.times.10.sup.4 cells/well.
Monoclonal antibodies (mAb), DVD-Igs.TM. and control IgG were
diluted to a 4.times. concentration in assay media and serial
dilutions were performed. Recombinant TNF-.alpha. was diluted to a
4.times. concentration of 600 pg/mL in assay media. All antibody
samples (mAbs or DVD-Igs.TM.) were pre-incubated with recombinant
TNF-.alpha. at a 1:1 volume ratio and allowed to incubate for 1
hour at 37.degree. C., 5% CO.sub.2. In some instances, recombinant
human sclerostin was also added to the pre-incubation step. For
these conditions, 8.times. dilutions of DVD-Ig.TM., recombinant
TNF-.alpha. (1200 ng/mL) and human sclerostin (400 nM) and 50 .mu.L
of complete assay media were pre-incubated for 1 hour at 37.degree.
C., 5% CO.sub.2.
[0607] One hundred .mu.L of the mAb or DVD-Ig.TM./recombinant
TNF-.alpha. solution was added to the plated cells at 100 .mu.L for
a 1.times. final concentration of 150 pg/mL of recombinant
TNF-.alpha., mAb or DVD-Ig.TM. and 1 .mu.g/mL actinomycin D. In
cases where human sclersotin was added, 100 .mu.L of the
DVD-Ig.TM./recombinant TNF-.alpha./human sclerostin/media solution
was added to the plated cells at 100 .mu.L for a 1.times. final
concentration of 150 pg/mL of recombinant TNF-.alpha., mAb or
DVD-Ig.TM., human sclersotin (50 nM) and 1 .mu.g/mL actinomycin D.
Plates were incubated overnight (16-24 hours) at 37.degree. C., 5%
CO.sub.2. To quantitate viability, 100 .mu.L was removed from the
wells and 10 .mu.L of WST-1 reagent (Roche, cat#11644807001) was
added. Plates were incubated at 37.degree. C., 5% CO.sub.2 for an
additional 3-5 hours and read on a Spectromax 190 ELISA plate
reader at OD 420-600 nm. Neutralization data was plotted using
GraphPad Prism. Reported IC.sub.50 values were calculated using
GraphPad Prism sigmoidal curve dose software and are provided in
Table 20 below.
TABLE-US-00022 TABLE 20 Neutralization of Recombinant TNF-.alpha.
Human TNFa Rhesus/cynoTNFa IC50, pM IC50, pM DVD2607 19.5 10.5
DVD2608 64 43 DVD2610 9.5 11 DVD2611 27.5 16.5 DVD2630 9.5 6
DVD2627 8.5 9 DVD2623 20.5 28.5 DVD2657 1128 ND DVD2658 1746 ND
MAK195-21-GS-AE10-6 AM7 19.5 ND MAK195-21-GS-AE10-6 AM3 44.9 ND
MAK195-21-GS-AE10-6 AM8 28.2 ND MAK195-21-SS-AE10-6 AM7 27.7 ND
MAK199-1-GS-AE10-6 AM7 67.7 42 MAK199-1-SS-AE10-6 AM7 58.1 56.9
diD2E7ss-GS-AE10-6 AM7 46.3 35.2 D2E7-GS-AE10-6AM7 38.8 28.2 AE10-6
AM7-GS-MAK195-21 1233 >2000 DVD2580 8 ND DVD2581 11 ND DVD2577 2
ND DVD2578 6 ND
In order to demonstrate that binding of sclerostin to the DVD-Igs
does not affect TNF-.alpha. neutralizing properties of the
molecules, DVD-Igs were first pre-incubated with 50 nM SOST at
least for 30 min and then L929 assay was performed with 50 nM SOST
throught the experiment. The results of this experiment are
provided are in Table 21 below.
TABLE-US-00023 TABLE 21 Example of comparison of TNF-a
neutralization with and without 50 nM SOST Human TNFa IC50, pM
Human TNFa IC50, pM with 50 nM SOST without 50 nM SOST MAK199-1-GS-
36 25 AE10-6 AM7 MAK199-1-GS- 32 33 AE10-6 AM7 QL DVD2623 46 56
Example 4
Pharmacokinetic (PK) Analysis for TNF/Sclerostin DVD Ig
[0608] Male Sprague-Dawley JVC rats weighing approximately 280 g
received a single dose in the jugular vein with the DVD-Ig or the
parental antibody at 5 mg/kg. Serum samples were collected from the
tail vein at 15 minutes, 4 and 24 hours, and 2, 3, 7, 10, 14, 21
and 28 days after dosing. Serum samples were frozen at -80.degree.
C. until analysis.
[0609] Male CD1 mice weighing approximately 28 g received a single
dose via tail vein with the DVD-Ig or the parental antibody at 5
mg/kg. Whole blood samples were collected from the tail vein at 1
and 24 hours, and 4, 7, 10, 14 and 21 days after dosing. Samples
were frozen at -80.degree. C. until analysis.
[0610] Bioanalysis of samples was performed using an MSD assay.
Briefly, strepatavidin plates were coated with biotinylated human
antigen and incubated overnight. Plates were blocked, and diluted
serum samples (final serum concentration was 1%) were added to the
wells, and Sulfo-tag-labeled goat anti-human IgG was used for
detection. Pharmacokinetic parameters for each animal were
calculated with WinNonlin software Version 5.2.1 by
non-compartmental analysis using linear trapezoidal fit. The PK
parameters obtained in mouse and rat are provided in Tables 22 and
23 below, respectively.
TABLE-US-00024 TABLE 22 PK Parameters in Mouse Compound T.sub.1/2
(day) V.sub.ss (mL/kg) Cl (mL/hr/kg) MAK195/24-GS-MSL10- 15 114
0.21 AM2 DVD2623 14.1 257 0.56 MAK199-1-GS-AE10-6 AM7 15.3 84 0.16
MAK199-1-GS-AE10-6 12.3 105 0.26 AM7 QL
TABLE-US-00025 TABLE 23 PK Parameters in Rat V.sub.ss Compound
T.sub.1/2 (day) (mL/kg) Cl (mL/hr/kg) MAK195/24-GS-MSL10-AM2 13.3
68 0.16 DVD2603 8.7 80 0.3 D2E7-GS-MSL10 11.5 67 0.21 DVD2578 10.2
79 0.24 DVD2580 7.4 65 0.26 DVD2581 10.6 84 0.26 MAK199-1-GS-AE10-6
AM7 6.8 58 0.34 MAK199-1-GS-AE10-6 AM7 QL 5.6 58 0.33
Example 5
Evaluation of DVD Domains for In Vivo Efficacy
[0611] The anti-sclerostin domains of the DVDs were evaluated for
in vivo efficacy using an acute PD model. All DVD-Igs tested
induced increased bone formation based on the increase in the bone
biomarker, PINP. Naive DBA/1 male mice, greater than 10 weeks of
age, were treated with a single dose of DVD-Ig, i.p. Three days
post injection, serum was collected via cardiac puncture and tested
for levels of the bone formation biomarker, PINP, by ELISA. All
DVD-Igs tested induced increased bone formation based on the
increases in serum levels of the bone biomarker, PINP. See Table
24.
TABLE-US-00026 TABLE 24 Percent Increase in PINP % increase PR# (30
mg/kg) MAK195-21-GS- 31 AE10-6 AM7 DVD2623 24 MAK195/21-GS- 57
AE10-6AM8 MAK199-1-SS- 52 AE10-6 AM7 MAK199-1-GS- 38 AE10-6 AM7
[0612] The anti-TNF domains of the DVDs were evaluated for in vivo
efficacy in a human TNF/D-Galactosamine lethality model in which
naive C57Bl/6 female mice were treated with the DVD, i.p. Eighteen
hours post dose, animals were challenged with 0.5 .mu.g/mouse TNF
and 20 mg/mouse D-Galactosamine intraperitoneally and animals were
monitored twice daily for survival. TNF domains in all DVD-Igs
tested were active and protected mice from TNF induced lethality.
See Table 25.
TABLE-US-00027 TABLE 25 Percent Survival PR# 1.0 mg/kg 03. mg/kg
0.1 mg/kg 0.03 mg/kg MAK199-1- 100 100 70 0 SS-AE10-6 AM7
MAK195/21- 90 80 60 40 SS-AE10- 6AM7 MAK199-1- 100 100 90 20
GS-AE10-6 AM7 DVD2623 100 90 30 10 DVD2603 100 100 50 30
Efficacy of Anti-TNF, Anti-SOST, and Combined Anti-TNF and
Anti-SOST Therapies in an Induced Arthritis Mouse Model
[0613] Anti-TNF, anti-SOST, and combined anti-TNF and anti-SOST
therapies were evaluated for their efficacy in preventing arthritis
in a mouse model of induced arthritis. Mice were immunized with
collagen at day 0, boosted with an intraperitoneal injection of
Zymosan on day 21, and selected for clinical signs of arthritis
from days 24-28. The mice were then treated starting at the day of
enrollment with an anti-TNF therapeutic (anti-TNF, 12 mg/kg,
subcutaneous injection, twice per week), an anti-sclerostin
therapeutic (30 mg/kg, subcutaneous injection, twice per week), or
the two therapeutics combined. Mice were evaluated changes in paw
thickness over a period of 20 days, and on day 44-48, serum and
bones were collected for endpoint analysis. As shown in FIG. 2A,
mice treated with either the anti-TNF or the combined therapies
exhibited less paw swelling than the control group treated with
0.9% saline. In addition, combined treatment with anti-TNF and
anti-sclerostin therapies maintained or increased bone thickness in
both the ankle and spine, respectively, as measured by micro-CT
(FIGS. 2B and 2C).
[0614] Combined neutralization of TNF and SOST was tested in a late
therapeutic mouse collagen induced arthritis (CIA) model in which
therapy began five days after the onset of inflammation, a
timepoint at which moderate bone loss has occurred. DBA/1 mice were
immunized with collagen on day 0, and boosted with zymosan on day
21. From day 24-28 mice were enrolled in the study based on first
clinical signs of arthritis. Five days after enrollment, mice were
dosed with 12 mg/kg of an anti-mouse TNF mAb, 30 mg/kg of an
anti-sclerostin mAb, or a combination of both mAbs twice weekly for
two weeks. Inflammation was monitored via paw swelling using
calipers. Micro CT analysis of ankles and spines was used to
evaluate changes in bone volume/density at study termination. A
group of immunized mice was sacrificed on day 5 after the first
signs of inflammation and the amount of bone lost at this timepoint
was assessed using micro-computed tomography (.mu.CT). As
demonstrated in (FIG. 4a) treatment with anti-TNF treatment is
ineffective at preventing inflammation when treatment starts 5 days
after the onset of disease. Treatment with either anti-TNF or
anti-sclerostin mAbs alone did not prevent bone loss from the
arthritic ankle, but the combination of both mAbs protected from
further bone loss at this site (FIG. 4b). In contrast, while TNF
inhibtion did not protect skeletal bone, bone was restored to
levels seen in naive animals in the spine of animals treated with
anti-sclerostin mAb, either alone or in combination with anti-TNF
mAb (FIG. 4c)
[0615] Since TNF inhibition is ineffective at controlling
inflammation in the mouse CIA model, an alternative
anti-inflammatory mechanism was used to test the ability of
sclerostin inhibition to restore bone in the arthritic joint.
Combined IL-1a and IL-1b inhibition blocks inflammation when dosing
begins five days after the onset of inflammation so a study was
done in which mice were immunized with collagen at day 0, boosted
with an intraperitoneal injection of Zymosan on day 21, and
selected for clinical signs of arthritis from days 24-28. The mice
were treated twice weekly starting five days after the onset of
inflammation with anti-IL1.alpha. therapeutic (9 mg/kg, ip) and
anti-IL1.beta. (4.5 mg/kg, ip), anti-sclerostin (30 mg/kg, ip), or
a combination of all three mAbs. Paw swelling was monitored during
the 20 days of treatment, after which serum and bones were
collected for endpoint analysis. The anti-inflammatory activity
achieved with the combined inhibition of IL-la and IL-lb is
demonstrated in FIG. 5a. Micro CT analysis of the arthritic ankle
showed that blockade of inflammation combined with sclerostin
inhibition allowed restoration of bone in the arthritic ankle (FIG.
5b). Analysis of trabecular bone of the lumbar spines of these
animals showed that sclerostin inhibition was able to restore
skeletal bone alone or in combination with inflammation blockade
(FIG. 5c).
Example 6
Efficacy of Anti-TNF, Anti-Sclerostin, and Combined Anti-TNF and
Anti-Sclerostin Therapies in a Mouse Model of Crohn's Disease
[0616] Skeletal bone loss is a common co-morbidity in patients with
Crohn's disease. We tested anti-sclerostin mAb, anti-TNF mAb, and
the combination of both Abs in a mouse model of Crohns disease and
measured changes in trabecular bone in the lumbar spine.
[0617] Splenocytes from female BALB/c mice (Taconic) were collected
by mechanical disruption and enriched for CD4.sup.+ cells using a
negative selection magnetic bead kit (StemCell). The purified
CD4.sup.+ cells were stained with anti-GITR (glucocorticoid induced
TNF family receptor) labeled with fluorescein (R&D Systems).
Cells were analyzed by FACS (MoFlow Legacy) and the lowest 40%
GITR-staining cells were collected. Post sorting analysis showed
the population to be CD4.sup.+GITR.sup.- with 95-98% purity. Cells
were washed, re-suspended and 5.times.10.sup.5 cells were injected
i.p. into female CB-17 scid mice (Taconic). Recipient mice were
monitored 2-3 times/week for body weight, presence of loose or
bloody stool, rectal prolapse, and general condition. A cohort of
animals was sacrificed on Day 27 (day of treatment initiation) for
histological analysis of colons and .mu.CT examination of spines to
determine baseline disease. Treatment consisted of twice weekly
injections of vehicle (PBS), anti-TNF (15 mg/kg ip),
anti-sclerostin (30 mg/kg ip), or both anti-TNF and
anti-sclerostin, and continued until the study was terminated on
Day 57 post cell injection. Animals were sacrificed; serum was
collected and stored at -20.degree. C. until used for determination
of antibody levels. Colons were collected, flushed with PBS to
remove fecal material, weighed and measured, placed in histology
cassettes using the swiss roll technique, and stored in 10%
formalin until processing. Following paraffin embedding, sections
were stained with hemotoxylin and eosin, and scored for the
presence of inflammation, crypt damage, mucosal ulcers, and extent
of disease. There were 8 or 9 animals per group Animals losing
greater than 20% of initial body weight were euthanized in
accordance with IACUC guidelines. Additionally, spines were
collected and trabecular bone in the L5 vertebrae was analyzed
using .mu.CT. Anti-TNF alone or in combination with anti-sclerostin
mAb inhibited intestinal inflammation while anti-sclerostin mAb did
not (FIG. 5a). In contrast, anti-sclerostin mAb treatment, alone or
in combination with anti-TNF was able to restore bone lost in the
lumbar spine to levels seen in a naive animal (FIG. 5b).
INCORPORATION BY REFERENCE
[0618] Techniques well known in the field of molecular biology,
drug delivery, immunology, molecular biology and cell biology are
incorporated by reference in their entirety. These techniques
include, but are not limited to, techniques described in the
following publications: Ausubel et al. (eds.), Current Protocols in
Molecular Biology, John Wiley & Sons, NY (1993); Ausubel, F. M.
et al. eds., Short Protocols In Molecular Biology (4th Ed. 1999)
John Wiley & Sons, NY. (ISBN 0-471-32938-X). Controlled Drug
Bioavailability Drug Product Design and Performance, Smolen and
Ball (eds.), Wiley, New York (1984); Giege, R. and Ducruix, A.
Barrett, Crystallization of Nucleic Acids and Proteins, a Practical
Approach, 2nd ed., pp. 20 1-16, Oxford University Press, New York,
N.Y., (1999); Goodson, in Medical Applications of Controlled
Release, vol. 2, pp. 115-138 (1984); Hammerling, et al., in:
Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier,
N.Y., 1981; Harlow et al., Antibodies: A Laboratory Manual, (Cold
Spring Harbor Laboratory Press, 2nd ed. 1988); Kabat et al.,
Sequences of Proteins of Immunological Interest (National
Institutes of Health, Bethesda, Md. (1987) and (1991); Kabat, E.
A., et al. (1991) Sequences of Proteins of Immunological Interest,
Fifth Edition, U.S. Department of Health and Human Services, NIH
Publication No. 91-3242; Kontermann and Dubel eds., Antibody
Engineering (2001) Springer-Verlag. New York. 790 pp. (ISBN
3-540-41354-5); Kriegler, Gene Transfer and Expression, A
Laboratory Manual, Stockton Press, NY (1990); Lu and Weiner eds.,
Cloning and Expression Vectors for Gene Function Analysis (2001)
BioTechniques Press. Westborough, Mass. 298 pp. (ISBN
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and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Old, R. W.
& S. B. Primrose, Principles of Gene Manipulation: An
Introduction To Genetic Engineering (3d Ed. 1985) Blackwell
Scientific Publications, Boston. Studies in Microbiology; V.2:409
pp. (ISBN 0-632-01318-4); Sambrook, J. et al. eds., Molecular
Cloning: A Laboratory Manual (2d Ed. 1989) Cold Spring Harbor
Laboratory Press, NY. Vols. 1-3 (ISBN 0-87969-309-6); Sustained and
Controlled Release Drug Delivery Systems, J. R. Robinson, ed.,
Marcel Dekker, Inc., New York, 1978; Winnacker, E. L. From Genes To
Clones: Introduction To Gene Technology (1987) VCH Publishers, N.Y.
(translated by Horst Ibelgaufts). 634 pp. (ISBN 0-89573-614-4).
[0619] Further, the contents of all cited references (including
literature references, patents, patent applications, and websites)
that maybe cited throughout this application are hereby expressly
incorporated by reference in their entirety for any purpose, as are
the references cited therein.
EQUIVALENTS
[0620] That which is provided may be embodied in other specific
forms without departing from the spirit or essential
characteristics thereof. The foregoing embodiments described herein
are therefore to be considered in all respects illustrative rather
than limiting. Scope is thus indicated by the appended claims
rather than by the foregoing description, and all changes that come
within the meaning and range of equivalency of the claims are
therefore intended to be embraced herein.
Sequence CWU 0 SQTB SEQUENCE LISTING The patent application
contains a lengthy "Sequence Listing" section. A copy of the
"Sequence Listing" is available in electronic form from the USPTO
web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20150266977A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
0 SQTB SEQUENCE LISTING The patent application contains a lengthy
"Sequence Listing" section. A copy of the "Sequence Listing" is
available in electronic form from the USPTO web site
(http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20150266977A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
* * * * *
References