U.S. patent application number 13/591027 was filed with the patent office on 2013-07-04 for dual-specific il-1a/ il-1b antibodies.
This patent application is currently assigned to ABBOTT LABORATORIES. The applicant listed for this patent is Carrie Enever, Steven Grant, Chung-ming Hsieh, Yuliya Kutskova, Bradford L. McRae, John E. Memmott, Michael Roguska, Ian Tomlinson, Mihriban Tuna. Invention is credited to Carrie Enever, Steven Grant, Chung-ming Hsieh, Yuliya Kutskova, Bradford L. McRae, John E. Memmott, Michael Roguska, Ian Tomlinson, Mihriban Tuna.
Application Number | 20130171146 13/591027 |
Document ID | / |
Family ID | 39589136 |
Filed Date | 2013-07-04 |
United States Patent
Application |
20130171146 |
Kind Code |
A1 |
Hsieh; Chung-ming ; et
al. |
July 4, 2013 |
DUAL-SPECIFIC IL-1A/ IL-1B ANTIBODIES
Abstract
The invention provides an isolated, dual-specific antibody, or
an antigen-binding portion thereof, which is specific for human
IL-1.alpha. and human IL-1.beta.. The dual specific antibodies of
the invention also neutralize both human IL-1.alpha. and human
IL-1.beta.. The invention also provides domain antibodies (dAbs)
specific for human IL-1.alpha. and human IL-1.beta..
Inventors: |
Hsieh; Chung-ming; (Newton,
MA) ; McRae; Bradford L.; (Northborough, MA) ;
Kutskova; Yuliya; (Northborough, MA) ; Memmott; John
E.; (Framingham, MA) ; Roguska; Michael;
(Ashland, MA) ; Tomlinson; Ian; (Cambridge,
GB) ; Enever; Carrie; (Cambridge, GB) ; Grant;
Steven; (Cambrdige, GB) ; Tuna; Mihriban;
(Cambridge, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Hsieh; Chung-ming
McRae; Bradford L.
Kutskova; Yuliya
Memmott; John E.
Roguska; Michael
Tomlinson; Ian
Enever; Carrie
Grant; Steven
Tuna; Mihriban |
Newton
Northborough
Northborough
Framingham
Ashland
Cambridge
Cambridge
Cambrdige
Cambridge |
MA
MA
MA
MA
MA |
US
US
US
US
US
GB
GB
GB
GB |
|
|
Assignee: |
ABBOTT LABORATORIES
Abbott Park
IL
|
Family ID: |
39589136 |
Appl. No.: |
13/591027 |
Filed: |
August 21, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12006068 |
Dec 28, 2007 |
8324350 |
|
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13591027 |
|
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60878165 |
Dec 29, 2006 |
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Current U.S.
Class: |
424/136.1 ;
435/320.1; 435/328; 435/69.6; 530/387.3; 530/389.2; 536/23.4;
536/23.53 |
Current CPC
Class: |
A61P 37/00 20180101;
C07K 16/245 20130101; A61K 39/3955 20130101; C07K 16/468 20130101;
C07K 2317/569 20130101; C07K 2317/92 20130101; A61K 45/06
20130101 |
Class at
Publication: |
424/136.1 ;
530/387.3; 536/23.53; 435/328; 435/320.1; 435/69.6; 536/23.4;
530/389.2 |
International
Class: |
C07K 16/46 20060101
C07K016/46; A61K 45/06 20060101 A61K045/06; C07K 16/24 20060101
C07K016/24; A61K 39/395 20060101 A61K039/395 |
Claims
1. An isolated, dual-specific antibody, or an antigen-binding
portion thereof, with the following characteristics: a) dissociates
from human IL-1.alpha. with a K.sub.D of 3.times.10.sup.-7 M or
less; b) dissociates from human IL-1.beta. with a K.sub.D of
5.times.10.sup.-5 M or less; and c) does not bind mouse IL-1.alpha.
or mouse IL-1.beta..
2. The antibody, or antigen-binding portion, of claim 1, which has
characteristics selected from the group consisting of: (a)
neutralizes human IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 900 nM or less; (b) neutralizes IL-1.alpha. in a
standard in vitro assay with an ND.sub.50 of 10 nM or less; (c)
neutralizes IL-1a in a standard in vitro assay with an ND.sub.50 of
10 nM or less and neutralizes IL-1.alpha. in a standard MRC5 in
vitro assay with an ND.sub.50 of 10 nM or less; (d) neutralizes
human IL-1.beta. in a standard in vitro assay with an ND.sub.50 of
800 nM or less or 200 nM or less; (e) neutralizes human IL-1.beta.
in a standard in vitro assay with an ND.sub.50 of 800 nM or less or
200 nM or less; and neutralizes IL-1.beta. in a standard in vitro
MRC5 assay with an ND.sub.50 of 200 nM or less; (f) neutralizes
human IL-1.beta. in a standard in vitro assay with an ND.sub.50 of
800 nM or less or 200 nM or less; and neutralizes IL-1.alpha. in a
standard in vitro assay with an ND.sub.50 of 10 nM or less; (g)
neutralizes human IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 900 nM or less, and neutralizes human IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 800 nM or less; (h)
neutralizes human IL-1.alpha. in a standard in vitro MRC5 assay
with an ND.sub.50 of 900 nM or less, and/or neutralizes human
IL-1.beta. in a standard MRC5 in vitro assay with an ND.sub.50 of
800 nM or less; (i) dissociates from IL-1.alpha. with a K.sub.D
selected from the group consisting of 1.times.10.sup.-8M or less;
1.times.10.sup.-9 M or less; 40-86 nM or less; 20-42 nM or less;
32-42 nM or less; 7-12 nM or less; 3.0.times.10.sup.-7 M or less;
1.1.times.10.sup.-7 M or less; 6.1.times.10.sup.-8M or less;
6.times.10.sup.-8M or less; 4.2.times.10.sup.-8M or less;
1.3.times.10.sup.-8M or less; and 1.1.times.10.sup.-9 M or less;
(j) dissociates from IL-1.beta. with a K.sub.D selected from the
group consisting of 5.4.times.10.sup.-5 M or less;
2.8.times.10.sup.-6M or less; 1.3.times.10.sup.-6M or less;
9.3.times.10.sup.-7 M or less; 2.times.10.sup.-7 M or less;
1.1.times.10.sup.-7 M or less; and 2.8.times.10.sup.-8M or less;
and (k) dissociates from IL-1.beta. with a K.sub.D selected from
the group consisting of 5.4.times.10.sup.-5 M or less;
2.8.times.10.sup.-6M or less; 1.3.times.10.sup.-6M or less;
9.3.times.10.sup.-7 M or less; 2.times.10.sup.-7 M or less;
1.1.times.10.sup.-7 M or less; and 2.8.times.10.sup.-8M or less;
and dissociates from IL-1.alpha. with a K.sub.D selected from the
group consisting of 1.times.10.sup.-8M or less; 1.times.10.sup.-9 M
or less; 40-86 nM or less; 20-42 nM or less; 32-42 nM or less; 7-12
nM or less; 3.0.times.10.sup.-7 M or less; 1.1.times.10.sup.-7 M or
less; 6.1.times.10.sup.-8M or less; 6.times.10.sup.-8M or less;
4.2.times.10.sup.-8M or less; 1.3.times.10.sup.-8M or less; and
1.1.times.10.sup.-9 M or less.
3-12. (canceled)
13. The antibody, or antigen-binding portion, of claim 1,
comprising (a) a heavy chain variable region comprising
complementary determining regions (CDRs) as set forth in an amino
acid sequence selected from the group consisting of SEQ ID NO: 16
(ABT1-96); SEQ ID NO: 52 (ABT2-108); SEQ ID NO: 44 (ABT2-65); SEQ
ID NO: 16 (ABT1-96); SEQ ID NO: 32 (ABT2-13); SEQ ID NO: 20
(ABT1-98); SEQ ID NO: 4 (ABT1-6-23); and a light chain variable
region comprising CDRs as set forth in an amino acid sequence
selected from the group consisting of SEQ ID NO: 36 (ABT2-42); SEQ
ID NO: 24 (ABT1-122); SEQ ID NO: 28 (ABT1-141); SEQ ID NO: 40
(ABT2-46); SEQ ID NO: 12 (ABT1-95); SEQ ID NO: 48 (ABT2-76); or (b)
at least two heavy chain variable regions each comprising CDRs as
set forth in an amino acid sequence selected from the group
consisting of SEQ ID NO: 16 (ABT1-96); SEQ ID NO: 32 (ABT2-13); SEQ
ID NO: 4 (ABT1-6-23); SEQ ID NO: 40 (ABT2-46).
14. (canceled)
15. An isolated antibody, or an antigen-binding portion thereof,
having dual-specificity for human IL-1.alpha. and human IL-1.beta.
comprising a variable light chain comprising complementary
determining regions (CDRs) as set forth in an amino acid sequence
selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 24,
and SEQ ID NO: 28, or a variable heavy chain comprising CDRS as set
forth in an amino acid sequence selected from the group consisting
of SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 16, and SEQ ID NO:
20.
16. The antibody, or antigen-binding portion thereof, of claim 15
which has characteristics selected from the group consisting of:
(a) dissociates from human IL-1.beta. with a K.sub.D selected from
the group consisting of 5.times.10.sup.-5 M or less;
5.4.times.10.sup.-5M or less; 2.8.times.10.sup.-6 M or less;
1.3.times.10.sup.-6 M or less; 9.3.times.10.sup.-7 M or less;
2.times.10.sup.-7 M or less; 1.1.times.10.sup.-7 M or less; and
2.8.times.10.sup.-8 M or less; (b) neutralizes human IL-1.beta. in
a standard in vitro assay with an ND.sub.50 of 800 nM or less; (c)
neutralizes human IL-1.beta. in a standard in vitro assay with an
ND.sub.50 of 800 nM or less; and neutralizes human IL-1.beta. in a
standard in vitro MRC5 assay with an ND.sub.50 of 800 nM or less;
(d) neutralizes IL-1.beta. in a standard in vitro assay with an
ND.sub.50 of 200 nM or less; (e) neutralizes IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 200 nM or less; and
neutralizes IL-1.beta. in a standard in vitro MRC5 assay with an
ND.sub.50 of 200 nM or less; and (f) dissociates from human
IL-1.beta. with a K.sub.D of 5.times.10.sup.-5 M or less and
neutralizes human IL-1.beta. in a standard in vitro MRC5 assay with
an ND.sub.50 of 800 nM or less.
17-21. (canceled)
22. The antibody, or an antigen-binding portion thereof, of claim
15, further comprising either a variable light chain comprising
CDRs as set forth in an amino acid sequence selected from the group
consisting of SEQ ID NO: 36, SEQ ID NO: 40, and SEQ ID NO: 48, or a
variable heavy chain comprising CDRs as set forth in an amino acid
sequence selected from the group consisting of SEQ ID NO: 32, SEQ
ID NO: 44, SEQ ID NO: 52.
23. An isolated antibody, or an antigen-binding portion thereof,
having dual-specificity for human IL-1.alpha. and human IL-1.beta.
comprising either a variable light chain comprising CDRs as set
forth in an amino acid sequence selected from the group consisting
of SEQ ID NO: 36, SEQ ID NO: 40, and SEQ ID NO: 48, or a variable
heavy chain comprising CDRs as set forth in an amino acid sequence
selected from the group consisting of SEQ ID NO: 32, SEQ ID NO: 44,
SEQ ID NO: 52.
24. The antibody, or antigen-binding portion thereof, of claim 23,
which has characteristics selected from the group consisting of:
(a) dissociates from IL-1.alpha. with a K.sub.D selected from the
group consisting of 3.0.times.10.sup.-7 M or less;
1.1.times.10.sup.-7 M or less; 6.times.10.sup.-8 M or less;
4.2.times.10.sup.-8 M or less; 1.times.10.sup.-8 M or less;
1.times.10.sup.-9M or less; (b) dissociates from human IL-1.alpha.
with a K.sub.D of 1.times.10.sup.-7 M or less and neutralizes human
IL-1.alpha. in a standard in vitro assay with an ND.sub.50 of 900
nM or less; (c) dissociates from human IL-1.alpha. with a K.sub.D
of 1.times.10.sup.-7 M or less and neutralizes human IL-1.alpha. in
a standard in vitro assay with an ND.sub.50 of 900 nM or less; and
neutralizes human IL-1.alpha. in a standard in vitro MRC5 assay
with an ND.sub.50 of 900 nM or less; (d) dissociates from human
IL-1.alpha. with a K.sub.D of 1.times.10.sup.-7 M or less and
neutralizes human IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 900 nM or less; neutralizes human IL-1.alpha. in a
standard in vitro MRC5 assay with an ND.sub.50 of 900 nM or less;
and neutralizes IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 10 nM or less; (e) dissociates from human IL-1.alpha.
with a K.sub.D of 1.times.10.sup.-7 M or less and neutralizes human
IL-1.alpha. in a standard in vitro assay with an ND.sub.50 of 900
nM or less; neutralizes human IL-1.alpha. in a standard in vitro
MRC5 assay with an ND.sub.50 of 900 nM or less; neutralizes
IL-1.alpha. in a standard in vitro assay with an ND.sub.50 of 10 nM
or less; and neutralizes IL-1.alpha. in a standard in vitro MRC5
assay with an ND.sub.50 of 10 nM or less.
25-28. (canceled)
29. The isolated antibody, or an antigen-binding portion thereof,
of claim 23, further comprising a variable light chain comprising
CDRs as set forth in an amino acid sequence selected from the group
consisting of SEQ ID NO: 12, SEQ ID NO: 24, and SEQ ID NO: 28, or a
variable heavy chain comprising CDRs as set forth in an amino acid
sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID
NO: 8, SEQ ID NO: 16, and SEQ ID NO: 20.
30. An isolated antibody, or an antigen-binding portion thereof,
having dual-specificity for human IL-1.alpha. and human IL-1.beta.,
comprising an IL-1.alpha. antigen binding region having either a
light chain variable sequence comprising a CDR3 selected from the
group consisting of SEQ ID NO: 11, SEQ ID NO: 23, and SEQ ID NO: 27
or a heavy chain variable sequence comprising a CDR3 selected from
the group consisting of SEQ ID NO: 3, SEQ ID NO: 7, SEQ ID NO: 15,
SEQ ID NO: 19, and an IL-1.beta. antigen binding region having
either a light chain variable sequence comprising a CDR3 selected
from the group consisting of SEQ ID NO: 35, SEQ ID NO: 39, and SEQ
ID NO: 47 or a heavy chain variable sequence comprising a CDR3
selected from the group consisting of SEQ ID NO: 31, SEQ ID NO: 43,
and SEQ ID NO: 51.
31. The antibody, or antigen-binding portion thereof, of claim 30,
wherein (a) the light chain variable sequence of the IL-1.alpha.
antigen binding region further comprises a CDR2 selected from the
group consisting of SEQ ID NO: 10, SEQ ID NO: 22, and SEQ ID NO:
26; (b) the heavy chain variable sequence of the IL-1.alpha.
antigen binding region further comprises a CDR2 selected from the
group consisting of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 14, and
SEQ ID NO: 18; (c) the light chain variable sequence of the
IL-1.alpha. antigen binding region further comprises a CDR2
selected from the group consisting of SEQ ID NO: 10, SEQ ID NO: 22,
and SEQ ID NO: 26; and the light chain variable sequence of the
IL-1.alpha. antigen binding region further comprises a CDR1
selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 21,
and SEQ ID NO: 25; (d) the heavy chain variable sequence of the
IL-1.alpha. antigen binding region further comprises a CDR2
selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 6,
SEQ ID NO: 14, and SEQ ID NO: 18; and the heavy chain variable
sequence of the IL-1.alpha. antigen binding region further
comprises a CDR1 selected from the group consisting of SEQ ID NO:
1, SEQ ID NO: 5, SEQ ID NO: 13, and SEQ ID NO: 17; or (e) the light
chain variable sequence of the IL-1.beta. antigen binding region
further comprises a CDR2 selected from the group consisting of SEQ
ID NO: 34, SEQ ID NO: 38, and SEQ ID NO: 46; (f) the heavy chain
variable sequence of the IL-1.beta. antigen binding region further
comprises a CDR2 selected from the group consisting of SEQ ID NO:
30, SEQ ID NO: 43, and SEQ ID NO: 50; (g) the light chain variable
sequence of the IL-1.beta. antigen binding region further comprises
a CDR2 selected from the group consisting of SEQ ID NO: 34, SEQ ID
NO: 38, and SEQ ID NO: 46; and the light chain variable sequence of
the IL-1.alpha. antigen binding region further comprises a CDR1
selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 37,
and SEQ ID NO: 45; or (h) the heavy chain variable sequence of the
IL-1.beta. antigen binding region further comprises a CDR2 selected
from the group consisting of SEQ ID NO: 30, SEQ ID NO: 43, and SEQ
ID NO: 50; and the heavy chain variable sequence of the IL-1.alpha.
antigen binding region further comprises a CDR1 selected from the
group consisting of SEQ ID NO: 29, SEQ ID NO: 41, and SEQ ID NO:
49.
32-38. (canceled)
39. A dual-specific, isolated antibody, or antigen-binding portion
thereof, comprising an IL-1.alpha. antigen binding region and an
IL-1.beta. antigen binding region, wherein the antibody, or
antigen-binding portion thereof, comprises a heavy chain variable
region and a light chain variable region combination selected from
the group consisting of a) a heavy chain variable region comprising
CDRs as set forth in SEQ ID NO: 16 (ABT1-96) and a light chain
variable region comprising CDRs as set forth in SEQ ID NO: 40
(ABT2-46); b) light chain variable region comprising CDRs as set
forth in SEQ ID NO: 24 (ABT1-122) and a heavy chain variable region
comprising CDRs as set forth in SEQ ID NO: 52 (ABT2-108); c) a
light chain variable region comprising CDRs as set forth in SEQ ID
NO: 28 (ABT1-141) and a heavy chain variable region comprising CDRs
as set forth in SEQ ID NO: 52 (ABT2-108); d) a light chain variable
region comprising CDRs as set forth in SEQ ID NO: 28 (ABT1-141) and
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 44 (ABT2-65); e) a heavy chain variable region comprising
CDRs as set forth in SEQ ID NO: 16 (ABT1-96) and a light chain
variable region comprising CDRs as set forth in SEQ ID NO: 36
(ABT2-42); f) a light chain variable region comprising CDRs as set
forth in SEQ ID NO: 12 (ABT1-95) and a heavy chain variable region
comprising CDRs as set forth in SEQ ID NO: 32 (ABT2-13); g) a light
chain variable region comprising CDRs as set forth in SEQ ID NO: 24
(ABT1-122) and a heavy chain variable region comprising CDRs as set
forth in SEQ ID NO: 44 (ABT2-65); and h) a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 20 (ABT1-98) and
a light chain variable region comprising CDRs as set forth in SEQ
ID NO: 48 (ABT2-76).
40. The antibody, or antigen-binding portion of claim 1, which has
an IgG1 or an IgG4 heavy chain constant region.
41. The antibody, or antigen-binding portion of claim 1, which is
an antibody fragment selected from the group consisting of a Fab, a
Fab', a Fab.sub.2, a Fab'.sub.2, an Fd, an Fd', a single chain Fv
(scFv), an scFv.sub.a, and a domain antibody (dAb).
42. The antibody, or antigen-binding portion of claim 1, wherein
the antibody, or antigen-binding portion thereof, is human.
43. A pharmaceutical composition comprising the antibody, or
antigen-binding portion thereof, of claim 1, and a pharmaceutically
acceptable carrier.
44. The pharmaceutical composition of claim 43, which further
comprises at least one additional therapeutic agent for treating a
disorder in which IL-1.alpha./IL-1.beta. activity is
detrimental.
45. A method for inhibiting human IL-1.alpha./IL-1.beta. activity
comprising contacting human IL-1.alpha. and IL-1.beta. with the
antibody, or antigen-binding portion thereof, of claim 1 such that
human IL-1.alpha./IL-1.beta. activity is inhibited.
46. A method for inhibiting human IL-1.alpha./IL-1.beta. activity
in a human subject suffering from a disorder in which
IL-1.alpha./IL-1.beta. activity is detrimental, comprising
administering to the human subject the antibody, or antigen-binding
portion thereof, of claim 1 such that human IL-1.alpha./IL-1.beta.
activity in the human subject is inhibited.
47. The method of claim 46, wherein the disorder in which
IL-1.alpha./IL-1.beta. activity is detrimental is selected from the
group consisting of an autoimmune disease, an intestinal disorder,
a skin disorder, a neurological disorder, a metabolic disorder,
rheumatoid arthritis, Crohn's disease, multiple sclerosis, insulin
dependent diabetes, mellitus, and psoriasis.
48. (canceled)
49. The method of claim 48, wherein the antibody, or antigen
binding portion thereof, is administered to the subject with an
additional therapeutic agent.
50. A single domain antibody (dAb) comprising an amino acid
sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID
NO: 8, SEQ ID NO: 12, SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 24,
SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 36, SEQ ID NO: 40, SEQ ID
NO: 44, SEQ ID NO: 48, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54,
SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID
NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63,
SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID
NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72,
SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID
NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81,
SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID
NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90,
SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID
NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99,
SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ
ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID
NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO:
112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO:
116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:
120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO:
124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO:
128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO:
132, and SEQ ID NO: 133.
51. An isolated nucleic acid encoding a light chain CDR3 domain
selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 23,
and SEQ ID NO: 27.
52. The isolated nucleic acid of claim 51, which encodes an
antibody light chain variable region.
53. The isolated nucleic acid of claim 52, which encodes a CDR2
domain selected from the group consisting of SEQ ID NO: 10, SEQ ID
NO: 22, and SEQ ID NO: 26; and/or a CDR1 domain selected from the
group consisting of SEQ ID NO: 9, SEQ ID NO: 21, and SEQ ID NO:
25.
54. (canceled)
55. An isolated nucleic acid encoding a heavy chain CDR3 domain
selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 7,
SEQ ID NO: 15, and SEQ ID NO: 19.
56. The isolated nucleic acid of claim 55, which encodes an
antibody heavy chain variable region.
57. The isolated nucleic acid of claim 56, which encodes a CDR2
domain selected from the group consisting of SEQ ID NO: 2, SEQ ID
NO: 6, SEQ ID NO: 14, and SEQ ID NO: 18; and/or a CDR1 domain
selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 5,
SEQ ID NO: 13, and SEQ ID NO: 17.
58. (canceled)
59. An isolated nucleic acid encoding a light chain CDR3 domain
selected from the group consisting of SEQ ID NO: 35, SEQ ID NO: 39,
and SEQ ID NO: 47.
60. The isolated nucleic acid of claim 59, which encodes an
antibody light chain variable region.
61. The isolated nucleic acid of claim 60, which encodes a CDR2
domain selected from the group consisting of SEQ ID NO: 34, SEQ ID
NO: 38, and SEQ ID NO: 46; and/or a CDR1 domain selected from the
group consisting of SEQ ID NO: 33, SEQ ID NO: 37, and SEQ ID NO:
45.
62. (canceled)
63. An isolated nucleic acid encoding a heavy chain CDR3 domain
selected from the group consisting of SEQ ID NO: 31, SEQ ID NO: 43,
and SEQ ID NO: 51.
64. The isolated nucleic acid of claim 63, which encodes an
antibody heavy chain variable region.
65. The isolated nucleic acid of claim 64, which encodes a CDR2
domain selected from the group consisting of SEQ ID NO: 30, SEQ ID
NO: 42, and SEQ ID NO: 50; and/or a CDR1 domain selected from the
group consisting of SEQ ID NO: 29, SEQ ID NO: 41, and SEQ ID NO:
49.
66. (canceled)
67. An isolated nucleic acid (a) encoding an antibody light chain
variable region comprising the amino acid sequence selected from
the group consisting of SEQ ID NO: 12, SEQ ID NO: 24, SEQ ID NO:
28, SEQ ID NO: 36, SEQ ID NO: 40, SEQ ID NO: 48, SEQ ID NO: 94, SEQ
ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO:
99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103,
SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ
ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID
NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO:
116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO:
120, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO:
124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO:
128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO:
132, and SEQ ID NO: 133; (b) encoding an antibody heavy chain
variable region comprising the amino acid sequence selected from
the group consisting of SEQ ID NO: 4, SEQ ID NO 8, SEQ ID NO: 16,
SEQ ID NO: 20, SEQ ID NO: 32, SEQ ID NO: 44, SEQ ID NO: 52, SEQ ID
NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58,
SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID
NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67,
SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID
NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76,
SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID
NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85,
SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID
NO: 90, SEQ ID NO: 91, and SEQ ID NO: 92; or (c) selected from the
group consisting of SEQ ID NO: 134 to SEQ ID NO: 843.
68-69. (canceled)
70. A recombinant expression vector comprising a nucleic acid of
claim 67.
71. A polypeptide comprising an amino acid sequence encoded by the
nucleic acid sequence of claim 67(c).
72. An expression vector comprising: a) an episomal origin of
replication; b) an insertion site for inserting a nucleic acid
sequence encoding a gene of interest; c) a stuffer sequence at the
insertion site; and d) a nucleic acid encoding an antibody heavy or
light chain constant region.
73. The expression vector of claim 72, wherein the stuffer sequence
comprises (i) a restriction enzyme site(s) selected from the group
consisting of a) NruI, FspAI, or a combination thereof at the 5'
end of the stuffer sequence; b) AfeI, SnaBI, BsiWI, HpaI, SalI, or
a combination thereof at the 3' end of the stuffer sequence; and c)
at least one restriction enzyme site from (a) and at least one
restriction enzyme site from (b); and/or (ii) a nucleic acid
sequence having at least 80% identity to the sequence set forth at
nucleotides 124 to 1100 of SEQ ID NO: 844.
74. (canceled)
75. The expression vector of claim 72, wherein the antibody heavy
chain constant region is murine or human.
76. The expression vector of claim 72, wherein the antibody
constant region is selected from the group consisting of murine or
human lambda, human kappa, human IgG3, IgA, IgE, IgM, murine IgG2b,
murine IgG3, and an Fc domain.
77. The expression vector of claim 72, wherein the gene of interest
is an antibody heavy or light chain variable region.
78. The expression vector of claim 72, comprising a sequence
selected from the group consisting of SEQ ID NO: 844, SEQ ID NO:
845, SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848, SEQ ID NO:
849, SEQ ID NO: 850, SEQ ID NO: 851, SEQ ID NO: 852, SEQ ID NO:
853, SEQ ID NO: 854, and SEQ ID NO: 855.
79. A host cell into which the recombinant expression vector of
claim 72 has been introduced.
80. A method of synthesizing a human antibody that binds human
IL-1.alpha. and IL-1.beta. comprising culturing the host cell of
claim 78 in a culture medium until a human antibody that binds
human IL-1.alpha. and IL-1.beta. is synthesized by the cell.
Description
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Appln.
No. 60/878,165, filed on Dec. 29, 2006, which is incorporated in
its entirety herein.
REFERENCE TO JOINT RESEARCH AGREEMENT
[0002] Contents of this application are under a joint research
agreement entered into by and between Domantis Limited and Abbott
Laboratories on Oct. 25, 2002, and directed to recombinantly
engineered antibodies to IL-1.
BACKGROUND OF THE INVENTION
[0003] Cytokines, such as interleukin-1 (IL-1) and tumor necrosis
factor (TNF), are molecules produced by a variety of cells, such as
monocytes and macrophages, which have been identified as mediators
of inflammatory processes. Interleukin-1 is a cytokine with a wide
range of biological and physiological effects, including fever,
prostaglandin synthesis (in e.g., fibroblasts, muscle and
endothelial cells), T-lymphocyte activation, and interleukin 2
production.
[0004] cDNAs coding for two distinct forms of IL-1 have been
isolated and expressed; these cDNAs represent two different gene
products, termed IL-1.beta. (Auron et al. (1984) Proc. Natl. Acad.
Sci. USA 81:7909) and IL-1.alpha. (Lomedico et al. (1984) Nature
312:458). IL-1.beta. is the predominant form produced by human
monocytes both at the mRNA and protein level. The two forms of
human IL-1 share only 26% amino acid homology. Despite their
distinct polypeptide sequences, the two forms of IL-1 have
structural similarities (Auron et al. (1985) J. Mol. Cell Immunol.
2:169), in that the amino acid homology is confined to discrete
regions of the IL-1 molecule.
[0005] The two forms of IL-1 possess similar biological properties,
including induction of fever, slow wave sleep, and neutrophilia, T-
and B-lymphocyte activation, fibroblast proliferation, cytotoxicity
for certain cells, induction of collagenases, synthesis of hepatic
acute phase proteins, and increased production of colony
stimulating factors and collagen. As such, both forms of IL-1 have
been implicated in the pathophysiology of a variety of human
diseases and disorders, including autoimmune and inflammatory
diseases, e.g., multiple sclerosis (Brosnan et al. (1995).
Neurology 45(6 suppl 6):
[0006] Antibodies capable of binding of IL-1.alpha. and IL-1.beta.
have been described in the art (see US Publication No. 20030040083
and PCT publication WO 07/063,308). However, there exists a need
for improved therapeutics to IL-1.alpha. and IL-1.beta..
SUMMARY OF THE INVENTION
[0007] High levels of IL-1.alpha. and IL-1.beta. are major concerns
in the management and treatment of inflammatory disease. The
invention provides a therapeutic means with which to inhibit both
IL-1.alpha. and IL-1.beta. by providing compositions and methods
for treating disease associated with increased levels of
IL-1.alpha./IL-1.beta., particularly inflammatory disorders.
[0008] The invention includes an isolated, dual-specific antibody,
or an antigen-binding portion thereof, which dissociates from human
IL-1.alpha. with a K.sub.D of 3.times.10.sup.-7 M or less;
dissociates from human IL-1.beta. with a K.sub.D of
5.times.10.sup.-5 M or less; and does not bind mouse IL-1.alpha. or
mouse IL-1.beta..
[0009] In one embodiment, the antibody, or antigen-binding portion,
neutralizes human IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 900 nM or less.
[0010] In one embodiment, the antibody, or antigen-binding portion,
neutralizes human IL-1.beta. in a standard in vitro assay with an
ND.sub.50 of 800 nM or less.
[0011] In one embodiment, the antibody, or antigen-binding portion,
neutralizes human IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 900 nM or less, and neutralizes human IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 800 nM or less.
[0012] In one embodiment, the antibody, or antigen-binding portion,
neutralizes human IL-1.alpha. in a standard in vitro MRC5 assay
with an ND.sub.50 of 900 nM or less, and/or neutralizes human
IL-1.beta. in a standard MRC5 in vitro assay with an ND.sub.50 of
800 nM or less.
[0013] In one embodiment, the antibody, or antigen-binding portion,
dissociates from IL-1.alpha. with a K.sub.D of 1.times.10.sup.-8 M
or less; dissociates from IL-1.alpha. with a K.sub.D of
1.times.10.sup.-9 M or less; dissociates from IL-1.alpha. with a
K.sub.D of 40-86 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 20-42 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 32-42 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 7-12 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 3.0.times.10.sup.-7 M or less; dissociates from
IL-1.alpha. with a K.sub.D of 1.1.times.10.sup.-7 M or less;
dissociates from IL-1.alpha. with a K.sub.D of 6.1.times.10.sup.-8
M or less; dissociates from IL-1.alpha. with a K.sub.D of
6.times.10.sup.-8 M or less; dissociates from IL-1.alpha. with a
K.sub.D of 4.2.times.10.sup.-8 M or less; dissociates from
IL-1.alpha. with a K.sub.D of 1.3.times.10.sup.-8 M or less; or
dissociates from IL-1.alpha. with a K.sub.D of 1.1.times.10.sup.-9
M or less.
[0014] In one embodiment, the antibody, or antigen-binding portion
thereof, dissociates from IL-1.beta. with a K.sub.D of
5.4.times.10.sup.-5 M or less; dissociates from IL-1.beta. with a
K.sub.D of 2.8.times.10.sup.-6 M or less; dissociates from
IL-1.beta. with a K.sub.D of 1.3.times.10.sup.-6 M or less;
dissociates from IL-1.beta. with a K.sub.D of 9.3.times.10.sup.-7 M
or less; dissociates from IL-1.beta. with a K.sub.D of
2.times.10.sup.-7 M or less; dissociates from IL-1.beta. with a
K.sub.D of 1.1.times.10.sup.-7 M or less; or dissociates from
IL-1.beta. with a K.sub.D of 2.8.times.10.sup.-8 M or less.
[0015] In one embodiment, the antibody, or antigen-binding portion
thereof, neutralizes IL-1.alpha. in a standard in vitro assay with
an ND.sub.50 of 10 nM or less. In one embodiment, the antibody, or
antigen-binding portion thereof, neutralizes IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 200 nM or less. In one
embodiment, the antibody, or antigen-binding portion thereof,
neutralizes IL-1.alpha. in a standard MRC5 in vitro assay with an
ND.sub.50 of 10 nM or less. In one embodiment, the antibody, or
antigen-binding portion thereof, neutralizes IL-1.beta. in a
standard in vitro MRC5 assay with an ND.sub.50 of 200 nM or
less.
[0016] In another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a heavy chain variable
region comprising complementary determining regions (CDRs) as set
forth in SEQ ID NO: 16 (ABT1-96) and a light chain variable region
comprising CDRs as set forth in SEQ ID NO: 36 (ABT2-42).
[0017] In still another embodiment, the antibody, or
antigen-binding portion thereof, of the invention comprises a light
chain variable region comprising CDRs as set forth in SEQ ID NO: 24
(ABT1-122) and a heavy chain variable region comprising CDRs as set
forth in SEQ ID NO: 52 (ABT2-108).
[0018] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention comprises a light chain variable region
comprising CDRs as set forth in SEQ ID NO: 28 (ABT1-141) and a
heavy chain variable region comprising CDRs as set forth in SEQ ID
NO: 52 (ABT2-108).
[0019] In yet another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 16 (ABT1-96) and
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 32 (ABT2-13).
[0020] In still another embodiment, the antibody, or
antigen-binding portion thereof, of the invention comprises a heavy
chain variable region comprising CDRs as set forth in SEQ ID NO: 4
(ABT1-6-23) and a second heavy chain variable region comprising
CDRs as set forth in SEQ ID NO: 40 (ABT2-46).
[0021] In another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a light chain variable
region comprising CDRs as set forth in SEQ ID NO: 28 (ABT1-141) and
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 44 (ABT2-65).
[0022] In another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 16 (ABT1-96) and
a light chain variable region comprising CDRs as set forth in SEQ
ID NO: 40 (ABT2-46).
[0023] In another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a light chain variable
region comprising CDRs as set forth in SEQ ID NO: 12 (ABT1-95) and
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 32 (ABT2-13).
[0024] In yet another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a light chain variable
region comprising CDRs as set forth in SEQ ID NO: 24 (ABT1-122) and
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 44 (ABT2-65).
[0025] In another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: (ABT1-98) and a
light chain variable region comprising CDRs as set forth in SEQ ID
NO: 48 (ABT2-76).
[0026] In another embodiment, the antibody, or antigen-binding
portion thereof, of the invention comprises a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 4 (ABT1-6-23) and
a second heavy chain variable region comprising CDRs as set forth
in SEQ ID NO: 32 (ABT2-13).
[0027] The invention also includes an isolated antibody, or an
antigen-binding portion thereof, having dual-specificity for human
IL-1.alpha. and human IL-1.beta. comprising a variable light chain
comprising complementary determining regions (CDRs) as set forth in
an amino acid sequence selected from the group consisting of SEQ ID
NO: 12, SEQ ID NO: 24, and SEQ ID NO: 28, or a variable heavy chain
comprising CDRS as set forth in an amino acid sequence selected
from the group consisting of SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO:
16, and SEQ ID NO: 20.
[0028] In one embodiment, the antibody, or antigen-binding portion
thereof, dissociates from human IL-1.beta. with a K.sub.D of
5.times.10.sup.-5 M or less; dissociates from IL-1.beta. with a
K.sub.D of 5.4.times.10.sup.-5 M or less; dissociates from
IL-1.beta. with a K.sub.D of 2.8.times.10.sup.-6 M or less;
dissociates from IL-1.beta. with a K.sub.D of 1.3.times.10.sup.-6 M
or less; dissociates from IL-1.beta. with a K.sub.D of
9.3.times.10.sup.-7 M or less; dissociates from IL-1.beta. with a
K.sub.D of 2.times.10.sup.-7 M or less; dissociates from IL-1.beta.
with a K.sub.D of 1.1.times.10.sup.-7 M or less; or dissociates
from IL-1.beta. with a K.sub.D of 2.8.times.10.sup.-8 M or
less.
[0029] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention neutralizes human IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 800 nM or less;
neutralizes human IL-1.beta. in a standard in vitro MRC5 assay with
an ND.sub.50 of 800 nM or less; neutralizes IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 200 nM or less;
neutralizes IL-1.beta. in a standard in vitro MRC5 assay with an
ND.sub.50 of 200 nM or less.
[0030] The invention also includes an antibody, or antigen-binding
portion thereof, which dissociates from human IL-1.beta. with a
K.sub.D of 5.times.10.sup.-5 M or less and neutralizes human
IL-1.beta. in a standard in vitro MRC5 assay with an ND.sub.50 of
800 nM or less.
[0031] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention comprises variable light chain CDRs as
set forth in an amino acid sequence selected from the group
consisting of SEQ ID NO: 36, SEQ ID NO: 40, and SEQ ID NO: 48, or
variable heavy chain comprising CDRS as set forth in an amino acid
sequence selected from the group consisting of SEQ ID NO: 32, SEQ
ID NO: 44, SEQ ID NO: 52.
[0032] The invention provides an isolated antibody, or an
antigen-binding portion thereof, having dual-specificity for human
IL-1.alpha. and human IL-1.beta. comprising a variable light chain
comprising CDRs as set forth in an amino acid sequence selected
from the group consisting of SEQ ID NO: 36, SEQ ID NO: 40, and SEQ
ID NO: 48, or a variable heavy chain comprising CDRS as set forth
in an amino acid sequence selected from the group consisting of SEQ
ID NO: 32, SEQ ID NO: 44, SEQ ID NO: 52.
[0033] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention dissociates from IL-1.alpha. with a
K.sub.D of 3.0.times.10.sup.-7 M or less; dissociates from
IL-1.alpha. with a K.sub.D of 1.1.times.10.sup.-7 M or less;
dissociates from IL-1.alpha. with a K.sub.D of 6.times.10.sup.-8 M
or less; dissociates from IL-1.alpha. with a K.sub.D of
4.2.times.10.sup.-8 M or less; dissociates from IL-1.alpha. with a
K.sub.D of 1.times.10.sup.-8 M or less; or dissociates from
IL-1.alpha. with a K.sub.D of 1.times.10.sup.-9 M or less.
[0034] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention dissociates from human IL-1.alpha. with a
K.sub.D of 1.times.10.sup.-7 M or less and neutralizes human
IL-1.alpha. in a standard in vitro assay with an ND.sub.50 of 900
nM or less. In one embodiment, the antibody, or antigen-binding
portion thereof, neutralizes human IL-1.alpha. in a standard in
vitro MRC5 assay with an ND.sub.50 of 900 nM or less. In another
embodiment, the antibody, or antigen-binding portion thereof,
neutralizes IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 10 nM or less. In still another embodiment, the
antibody, or antigen-binding portion thereof, neutralizes
IL-1.alpha. in a standard in vitro MRC5 assay with an ND.sub.50 of
10 nM or less.
[0035] In one embodiment of the invention, the isolated antibody,
or an antigen-binding portion thereof, comprises a variable light
chain comprising CDRs as set forth in an amino acid sequence
selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 24,
and SEQ ID NO: 28, or a variable heavy chain comprising CDRS as set
forth in an amino acid sequence selected from the group consisting
of SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 16, and SEQ ID NO:
20.
[0036] The invention describes an isolated antibody, or an
antigen-binding portion thereof, having dual-specificity for human
IL-1.alpha. and human IL-1.beta., comprising an IL-1.alpha. antigen
binding region with a light chain variable sequence comprising a
CDR3 selected from the group consisting of SEQ ID NO: 11, SEQ ID
NO: 23, and SEQ ID NO: 27 or a heavy chain variable sequence
comprising a CDR3 selected from the group consisting of SEQ ID NO:
3, SEQ ID NO: 7, SEQ ID NO: 15, SEQ ID NO: 19, and an IL-1.beta.
antigen binding region with a light chain variable sequence
comprising a CDR3 selected from the group consisting of SEQ ID NO:
35, SEQ ID NO: 39, and SEQ ID NO: 47 or a heavy chain variable
sequence comprising a CDR3 selected from the group consisting of
SEQ ID NO: 31, SEQ ID NO: 43, and SEQ ID NO: 51.
[0037] In one embodiment, the light chain variable sequence of the
IL-1.alpha. antigen binding region further comprises a CDR2
selected from the group consisting of SEQ ID NO: 10, SEQ ID NO: 22,
and SEQ ID NO: 26. In another embodiment, the light chain variable
sequence of the IL-1.alpha. antigen binding region further
comprises CDR1 selected from the group consisting of SEQ ID NO: 9,
SEQ ID NO: 21, and SEQ ID NO: 25. In one embodiment, the light
chain variable sequence of the IL-1.beta. antigen binding region
further comprises a CDR2 selected from the group consisting of SEQ
ID NO: 34, SEQ ID NO: 38, and SEQ ID NO: 46. In another embodiment,
the light chain variable sequence of the IL-1.alpha..antigen
binding region further comprises CDR1 selected from the group
consisting of SEQ ID NO: 33, SEQ ID NO: 37, and SEQ ID NO: 45.
[0038] In another embodiment, the heavy chain variable sequence of
the IL-1.alpha. antigen binding region further comprises a CDR2
selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 6,
SEQ ID NO: 14, and SEQ ID NO: 18. In one embodiment, the heavy
chain variable sequence of the IL-1.alpha. antigen binding region
further comprises CDR1 selected from the group consisting of SEQ ID
NO: 1, SEQ ID NO: 5, SEQ ID NO: 13, and SEQ ID NO: 17. In one
embodiment, the heavy chain variable sequence of the
IL-1.beta..antigen binding region further comprises a CDR2 selected
from the group consisting of SEQ ID NO: 30, SEQ ID NO: 43, and SEQ
ID NO: 50. In one embodiment, the heavy chain variable sequence of
the IL-1.alpha..antigen binding region further comprises CDR1
selected from the group consisting of SEQ ID NO: 29, SEQ ID NO: 41,
and SEQ ID NO: 49.
[0039] The invention also provides a dual-specific, isolated
antibody, or antigen-binding portion thereof, comprising an
IL-1.alpha. antigen binding region and an IL-1.beta. antigen
binding region, wherein the antibody, or antigen-binding portion
thereof, comprises a heavy chain variable region and a light chain
variable region combination selected from the group consisting of a
heavy chain variable region comprising CDRs as set forth in SEQ ID
NO: 16 (ABT1-96) and a light chain variable region comprising CDRs
as set forth in SEQ ID NO: 40 (ABT2-46); light chain variable
region comprising CDRs as set forth in SEQ ID NO: 24 (ABT1-122) and
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 52 (ABT2-108); a light chain variable region comprising CDRs
as set forth in SEQ ID NO: 28 (ABT1-141) and a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 52 (ABT2-108); a
light chain variable region comprising CDRs as set forth in SEQ ID
NO: 28 (ABT1-141) and a heavy chain variable region comprising CDRs
as set forth in SEQ ID NO: 44 (ABT2-65); a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 16 (ABT1-96) and
a light chain variable region comprising CDRs as set forth in SEQ
ID NO: 36 (ABT2-42); a light chain variable region comprising CDRs
as set forth in SEQ ID NO: 12 (ABT1-95) and a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 32 (ABT2-13); a
light chain variable region comprising CDRs as set forth in SEQ ID
NO: 24 (ABT1-122) and a heavy chain variable region comprising CDRs
as set forth in SEQ ID NO: 44 (ABT2-65); and a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 20 (ABT1-98) and
a light chain variable region comprising CDRs as set forth in SEQ
ID NO: 48 (ABT2-76).
[0040] In one embodiment, the invention provides antibody, or
antigen-binding portion thereof, comprising an IL-1.alpha. antigen
binding region and/or an IL-1.beta. antigen binding region, wherein
the antibody comprises SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 12,
SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 24, SEQ ID NO: 28, SEQ ID
NO: 32, SEQ ID NO: 36, SEQ ID NO: 40, SEQ ID NO: 44, SEQ ID NO: 48,
SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID
NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60,
SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63 SEQ ID NO: 64, SEQ ID
NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69,
SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID
NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,
SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID
NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,
SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID
NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96,
SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID
NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO:
105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:
109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO: 113,
SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ
ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID
NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:
126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO:130,
SEQ ID NO: 131, SEQ ID NO: 132, and SEQ ID NO: 133, or CDRs from
said sequences.
[0041] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention has an IgG1 or an IgG4 heavy chain
constant region.
[0042] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention is an antibody fragment selected from the
group consisting of a Fab, a Fab', a Fab2, a Fab'2, an Fd, an Fd',
a single chain Fv (scFv), an scFv.sub.a, and a domain antibody
(dAb).
[0043] In one embodiment, the antibody, or antigen-binding portion
thereof, of the invention is human.
[0044] The invention also provides a pharmaceutical composition
comprising the antibody, or antigen-binding portion thereof, of the
invention, and a pharmaceutically acceptable carrier.
[0045] In one embodiment, the pharmaceutical composition of the
invention further comprises at least one additional therapeutic
agent for treating a disorder in which IL-1.alpha./IL-1.beta.
activity is detrimental.
[0046] The invention also provides a method for inhibiting human
IL-1.alpha./IL-1.beta. activity comprising contacting human
IL-1.alpha. and IL-1.beta. with the antibody, or antigen-binding
portion thereof, of the invention such that human
IL-1.alpha./IL-1.beta. activity is inhibited.
[0047] The invention includes a method for inhibiting human
IL-1.alpha./IL-1.beta. activity in a human subject suffering from a
disorder in which IL-1.alpha./IL-1.beta. activity is detrimental,
comprising administering to the human subject the antibody, or
antigen-binding portion thereof, of the invention such that human
IL-1.alpha./IL-1.beta. activity in the human subject is
inhibited.
[0048] In one embodiment, the disorder in which
IL-1.alpha./IL-1.beta. activity is detrimental is selected from the
group consisting of an autoimmune disease, an intestinal disorder,
a skin disorder, a neurological disorder, and a metabolic
disorder.
[0049] In one embodiment, the disorder in which
IL-1.alpha./IL-1.beta. activity is detrimental is selected from the
group consisting of rheumatoid arthritis, Crohn's disease, multiple
sclerosis, insulin dependent diabetes, mellitus, and psoriasis.
[0050] In one embodiment, the antibody, or antigen binding portion
thereof, of the invention is administered to the subject with an
additional therapeutic agent.
[0051] The invention describes a single domain antibody (dAb)
comprising an amino acid sequence selected from the group
consisting of SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 12, SEQ ID NO:
16, SEQ ID NO: 20, SEQ ID NO: 24, SEQ ID NO: 28, SEQ ID NO: 32, SEQ
ID NO: 36, SEQ ID NO: 40, SEQ ID NO: 44, SEQ ID NO: 48, SEQ ID NO:
52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ
ID NO: 57, --SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID
NO: 61, SEQ ID NO: 62, SEQ ID NO: 63 SEQ ID NO: 64, SEQ ID NO: 65,
SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID
NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74,
SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID
NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83,
SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID
NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92,
SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID
NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID NO:
101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO:
105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:
109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO: 113,
SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ
ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID
NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:
126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO:130,
SEQ ID NO: 131, SEQ ID NO: 132, and SEQ ID NO: 133.
[0052] The invention also describes an isolated nucleic acid
encoding a light chain CDR3 domain selected from the group
consisting of SEQ ID NO: 11, SEQ ID NO: 23, and SEQ ID NO: 27. In
one embodiment, the nucleic acid of the invention encodes an
antibody light chain variable region. In one embodiment, the
isolated nucleic acid of the invention comprises a CDR2 domain of
the light chain variable region comprises the amino acid sequence
selected from the group consisting of SEQ ID NO: 10, SEQ ID NO: 22,
and SEQ ID NO: 26. In one embodiment, the isolated nucleic acid of
the invention comprises the CDR1 domain of the light chain variable
region comprises the amino acid sequence selected from the group
consisting of SEQ ID NO: 9, SEQ ID NO: 21, and SEQ ID NO: 25.
[0053] The invention also describes an isolated nucleic acid
encoding encoding a heavy chain CDR3 domain selected from the group
consisting of SEQ ID NO: 3, SEQ ID NO: 7, SEQ ID NO: 15, and SEQ ID
NO: 19. In one embodiment, the nucleic acid encodes an antibody
heavy chain variable region. In one embodiment, the CDR2 domain of
the heavy chain variable region comprises the amino acid sequence
selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 6,
SEQ ID NO: 14, and SEQ ID NO: 18. In one embodiment, the CDR1
domain of the heavy chain variable region comprises the amino acid
sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID
NO: 5, SEQ ID NO: 13, and SEQ ID NO: 17.
[0054] The invention also provides an isolated nucleic acid
encoding a light chain CDR3 domain selected from the group
consisting of SEQ ID NO: 35, SEQ ID NO: 39, and SEQ ID NO: 47. In
one embodiment, the nucleic acid encodes an antibody light chain
variable region. In one embodiment, the CDR2 domain of the light
chain variable region comprises the amino acid sequence selected
from the group consisting of SEQ ID NO: 34, SEQ ID NO: 38, and SEQ
ID NO: 46. In one embodiment, the CDR1 domain of the light chain
variable region comprises the amino acid sequence selected from the
group consisting of SEQ ID NO: 33, SEQ ID NO: 37, and SEQ ID NO:
45.
[0055] The invention provides an isolated nucleic acid encoding a
heavy chain CDR3 domain selected from the group consisting of SEQ
ID NO: 31, SEQ ID NO: 43, and SEQ ID NO: 51. In one embodiment, the
nucleic acid encodes an antibody heavy chain variable region. In
one embodiment, the CDR2 domain of the heavy chain variable region
comprises the amino acid sequence selected from the group
consisting of SEQ ID NO: 30, SEQ ID NO: 42, and SEQ ID NO: 50. In
one embodiment, the CDR1 domain of the heavy chain variable region
comprises the amino acid sequence selected from the group
consisting of SEQ ID NO: 29, SEQ ID NO: 41, and SEQ ID NO: 49.
[0056] The invention describes an isolated nucleic acid encoding a
CDR3 domain comprising an amino acid sequence selected from the
group consisting of SEQ ID NO: 3, SEQ ID NO 7, SEQ ID NO: 11, SEQ
ID NO: 15, SEQ ID NO: 19, SEQ ID NO: 23, SEQ ID NO: 27, SEQ ID NO:
31, SEQ ID NO: 35, SEQ ID NO: 39, SEQ ID NO: 43, SEQ ID NO: 47, and
SEQ ID NO: 51.
[0057] The invention also provides an isolated nucleic acid
encoding an antibody light chain variable region comprising the
amino acid sequence selected from the group consisting of SEQ ID
NO: 12, SEQ ID NO: 24, SEQ ID NO: 28, SEQ ID NO: 36, SEQ ID NO: 40,
and SEQ ID NO: 48. In one embodiment, the nucleic acid encodes the
antibody light chain variable region and an antibody light chain
constant region
[0058] The invention also provides an isolated nucleic acid
encoding an antibody heavy chain variable region comprising the
amino acid sequence selected from the group consisting of SEQ ID
NO: 4, SEQ ID NO 8, SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 32,
SEQ ID NO: 44, and SEQ ID NO: 52. In one embodiment, the nucleic
acid encodes the antibody heavy chain variable region and an
antibody heavy chain constant region.
[0059] The invention also provides a recombinant expression vector
encoding an antibody light chain having a variable region
comprising an amino acid sequence selected from the group
consisting of SEQ ID NO: 12, SEQ ID NO: 24, and SEQ ID NO: 28; and
an antibody heavy chain having a variable region comprising an
amino acid sequence selected from the group consisting of SEQ ID
NO: 32, SEQ ID NO: 44, and SEQ ID NO: 52.
[0060] The invention also provides a recombinant expression vector
encoding an antibody heavy chain having a variable region
comprising an amino acid sequence selected from the group
consisting of SEQ ID NO: 4, SEQ ID NO: 8, and SEQ ID NO: 16; and an
antibody light chain having a variable region comprising an amino
acid sequence selected from the group consisting of SEQ ID NO: 36,
SEQ ID NO: 40, and SEQ ID NO: 48.
[0061] The nucleic acid and amino acid sequences described herein
are also included in the invention. In one embodiment, sequences
which are at least 80%, 85%, 90%, 95%, 98%, or 99% identical to
those described herein are included in invention. In one
embodiment, the invention provides an isolated nucleic acid
encoding an antibody light chain variable region comprising the
amino acid sequence selected from the group consisting of SEQ ID
NO: 12, SEQ ID NO: 24, SEQ ID NO: 28, SEQ ID NO: 36, SEQ ID NO: 40,
SEQ ID NO: 48, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID
NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:100, SEQ ID NO:
101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO:
105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO:
109, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO:112, SEQ ID NO: 113,
SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ
ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, SEQ ID
NO: 122, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO:
126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO:130,
SEQ ID NO: 131, SEQ ID NO: 132, and SEQ ID NO: 133. In one
embodiment, the invention provides an isolated nucleic acid
encoding an antibody heavy chain variable region comprising the
amino acid sequence selected from the group consisting of SEQ ID
NO: 4, SEQ ID NO 8, SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 32,
SEQ ID NO: 44, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID
NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60,
SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63 SEQ ID NO: 64, SEQ ID
NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69,
SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID
NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78,
SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID
NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87,
SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, and SEQ
ID NO: 92. Also included in the invention are nucleic acids set
forth in SEQ ID NO: 134 to 843. Polypeptides encoded by the nucleic
acids of SEQ ID NOs: 134 to 843 are also included in the
invention.
[0062] The invention provides a recombinant expression vector
comprising a stuffer sequence and a nucleic acid sequence encoding
a heavy or light chain constant region. In one embodiment, the gene
of interest is an antibody heavy or light chain variable region.
Such vectors may be used to express full length heavy or light
chains of an antibody, by cloning a heavy or light chain variable
sequence into the insertion site of the vector, which is operably
linked to a constant region. In another embodiment, the vector
comprises an antibody heavy chain constant region is murine or
human, e.g., murine or human lambda, human kappa human IgG3, IgA,
IgE, IgM, murine IgG2b, murine IgG3, and an Fc domain. In a further
embodiment, the antibody heavy constant region further comprises an
alanine mutation at position 234, 235, 237, or any combination
thereof. IN still another embodiment, the antibody light chain
constant region is either a human kappa isotype or a human lambda
isotype. In still another embodiment, the antibody heavy constant
region is selected from the group consisting of gamma1, z, a;
gamma1, z, non-a; gamma2, n+; gamma2, n-; and gamma 4.
[0063] In one embodiment, the vector of the invention may be used
to express a fusion protein, e.g., an Fc fusion protein.
[0064] The invention also provides an expression vector comprising
an episomal origin of replication; an insertion site for inserting
a nucleic acid sequence encoding a gene of interest; a stuffer
sequence at the insertion site; and a nucleic acid encoding an
antibody heavy or light chain constant region. In one embodiment,
the stuffer sequence comprises the restriction enzyme sites NruI,
FspAI, or a combination thereof at the 5' end of the stuffer
sequence. In another embodiment, the stuffer sequence comprises the
restriction enzyme sites MeI, SnaBI, BsiWI, HpaI, SalI, or a
combination thereof at the 3' end of the stuffer sequence. In one
embodiment, the stuffer sequence comprises a nucleic acid sequence
having at least 80%, 90%, 95%, 98%, or 99% identity to the sequence
set forth at nucleotides 124 to 1100 of SEQ ID NO: 844.
[0065] The vector of the invention may also include certain
features desirable for protein expression. For example, the vector
may includes an episomal origin of replication is an SV40 origin of
replication. In one embodiment, the vector comprises a promoter
operably linked to the insertion site, wherein the promoter is an
EF-1a promoter.
[0066] In one embodiment, the invention provides a nucleic acid
comprising the recombinant expression vector, e.g., SEQ ID NOs: 844
to 855. Sequences having 80%, 85%, 90%, 95%, 99% identity to the
sequences described in SEQ ID NO: 844, SEQ ID NO: 845, SEQ ID NO:
846, SEQ ID NO: 847, SEQ ID NO: 848, SEQ ID NO: 849, SEQ ID NO:
850, SEQ ID NO: 851; SEQ ID NO: 852, SEQ ID NO: 853, SEQ ID NO:
854, and SEQ ID NO: 855 are also included in the invention. The
invention also provides a pEF-BOS recombinant expression vector
comprising a stuffer sequence which is inbetween an upstream signal
sequence and a downstream Ig constant region sequence.
[0067] The invention also includes a host cell into which the
recombinant expression vector of the invention has been
introduced.
[0068] The invention further provides a method of synthesizing a
human antibody that binds human IL-1.alpha. and IL-1.beta.
comprising culturing the host cell of the invention in a culture
medium until a human antibody that binds human IL-1.alpha. and
IL-1.beta. is synthesized by the cell.
BRIEF DESCRIPTION OF THE DRAWINGS
[0069] FIG. 1 shows a schematic of a plasmid map of
pYDs-TEV-2-108/1-122.
[0070] FIG. 2 shows the sequence randomization of HCDRs in the 30
small libraries.
[0071] FIG. 3 shows a schematic and sequences used for CDR
recombination of clone 2-108. Bold amino acids highlight
differences between clone 2-108 and the affinity matured
clones.
[0072] FIG. 4 shows K.sub.D on yeast surface of single CDR affinity
matured and parental 2-108 clones.
[0073] FIG. 5 shows IL-1.beta. neutralization potency of single CDR
affinity matured vs parental 2-108 clones as IgG.
[0074] FIG. 6 shows K.sub.D on yeast surface of CDR recombination
vs. single CDR affinity matured 2-108 clones.
[0075] FIG. 7 shows a plasmid map (7A) and schematic representation
of Fab expression on yeast surfaces (7B).
[0076] FIGS. 8A and 8B show Fab library sorting for ABT2-108
affinity maturation.
[0077] FIG. 9 shows analysis of 2-108 clones on yeast surface.
[0078] FIG. 10 shows sorting results for ABT 2-122 affinity
maturation.
[0079] FIG. 11 shows the plasmid map of
pYDs-TEV-2-108-538x/1-95-15.
[0080] FIG. 12 provides a diagram of the yeast CDR spiking
libraries construction.
[0081] FIG. 13 shows ABT-95-15 CDR recombination library
construction.
[0082] FIG. 14 provides results of shows K.sub.D analysis of
ABT2-108-538x/ABT1-95-15 scFv on yeast surface.
[0083] FIG. 15 provides off-rate analysis if the single CDR mutants
on yeast surface.
[0084] FIG. 16 describe an on rate and off rate analysis of
affinity-matured ABT1-95-15 clones on yeast surface.
[0085] FIG. 17 describes an analysis of affinity-matured ABT1-95-15
clones on yeast surface.
[0086] FIGS. 18A and 18B show the design and use of mouse and human
pBOS templates, respectively.
[0087] FIG. 19 describes a representative plasmid map of an scFv
fusion construct (pBOS-1-98/2-108-538x-scFc).
[0088] FIG. 20 shows combined MRC5 neutralisation data for
IL-1.alpha. and IL-1.beta. of the ABT1-95-A2 and ABT2-65-166 dAb
molecules and the IgG resulting from the combination of these two
dAb variable domains.
[0089] FIGS. 21A to 21D show sequences of ABT1-95, ABT1-122,
ABT1-141, and ABT2-65, respectively, clones as maturation has
progressed.
[0090] FIG. 22 shows simultaneous binding of rhIL-1 alpha followed
by rhIL-1 beta to ABT 108-620x/ABT1-95-A3-showing non interferring
independent binding
[0091] FIG. 23 shows IL-6 inhibition using an in vivo study to
determine the efficacy of a dual specific antibody for inhibiting
IL-1.alpha. and IL-1.beta. activity. Key for figure reads top of
key to left of figure, e.g., hIgG (250 .mu.g) coincides to days 7,
4, and 1 beginning at left of x axis.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions
[0092] In order that the present invention may be more readily
understood, certain terms are first defined.
[0093] The term "IL-1" as used herein refers to interleukin-1. The
term "IL-1" is intended to include both IL-1.alpha. and
IL-1.beta..
[0094] The term "antibody" as referred to herein includes whole
antibodies and any antigen binding fragment (i.e., "antigen-binding
portion") or single chains thereof. An "antibody" refers to a
glycoprotein comprising at least two heavy (H) chains and two light
(L) chains inter-connected by disulfide bonds, or an antigen
binding portion thereof. Each heavy chain is comprised of a heavy
chain variable region (abbreviated herein as V.sub.H) 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 V.sub.L)
and a light chain constant region. The light chain constant region
is comprised of one domain, CL. The V.sub.H and V.sub.L 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 V.sub.H and V.sub.L 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. The variable regions
of the heavy and light chains contain a binding domain that
interacts with an antigen. The constant regions of the antibodies
may mediate the binding of the immunoglobulin to host tissues or
factors, including various cells of the immune system (e.g.,
effector cells) and the first component (Clq) of the classical
complement system.
[0095] The term "antigen-binding portion" of an antibody (or simply
"antibody portion"), as used herein, refers to one or more
fragments of an antibody that retain the ability to specifically
bind to an antigen (e.g., IL-1.alpha., IL-1.beta.). It has been
shown that the antigen-binding function of an antibody can be
performed by fragments of a full-length antibody. 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 V.sub.L, V.sub.H, 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 V.sub.H and C.sub.H1 domains;
(iv) a Fv fragment consisting of the V.sub.L and V.sub.H domains of
a single atm of an antibody, (v) a dAb fragment (Ward et al, (1989)
Nature 341:544-546), which consists of a V.sub.H or V.sub.L domain;
and (vi) an isolated complementarity determining region (CDR).
Furthermore, although the two domains of the Fv fragment, VL and
V.sub.H, 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
e.g., 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. These antibody fragments
are obtained using conventional techniques known to those with
skill in the art, and the fragments are screened for utility in the
same manner as are intact antibodies. In one embodiment if the
invention, the antibody fragment is selected from the group
consisting of a Fab, an Fd, an Fd', a single chain Fv (scFv), an
scFv.sub.a, and a domain antibody (dAb).
[0096] Still further, an antibody or antigen-binding portion
thereof may be part of a larger immunoadhesion molecules, 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 Antibodies and 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.
[0097] Two antibody domains are "complementary" where they belong
to families of structures which form cognate pairs or groups or are
derived from such families and retain this feature. For example, a
VH domain and a VL domain of an antibody are complementary; two VH
domains are not complementary, and two VL domains are not
complementary. Complementary domains may be found in other members
of the immunoglobulin superfamily, such as the V.alpha. and V.beta.
(or gamma and delta) domains of the T-cell receptor
[0098] The term "domain" refers to a folded protein structure which
retains its tertiary structure independently of the rest of the
protein. Generally, domains are responsible for discrete functional
properties of proteins, and in many cases may be added, removed or
transferred to other proteins without loss of function of the
remainder of the protein and/or of the domain. By single antibody
variable domain is meant a folded polypeptide domain comprising
sequences characteristic of antibody variable domains. It therefore
includes complete antibody variable domains and modified variable
domains, for example in which one or more loops have been replaced
by sequences which are not characteristic of antibody variable
domains, or antibody variable domains which have been truncated or
comprise N- or C-terminal extensions, as well as folded fragments
of variable domains which retain at least in part the binding
activity and specificity of the full-length domain.
[0099] Variable domains of the invention may be combined to form a
group of domains; for example, complementary domains may be
combined, such as VL domains being combined with VH domains.
Non-complementary domains may also be combined. Domains may be
combined in a number of ways, involving linkage of the domains by
covalent or non-covalent means.
[0100] A "dAb" or "domain antibody" refers to a single antibody
variable domain (V.sub.H or V.sub.L) polypeptide that specifically
binds antigen.
[0101] An "isolated dual-specific antibody", or an "isolated
antibody," as used herein, is intended to refer to an antibody that
is substantially free of other antibodies having different
antigenic specificities (e.g., an isolated antibody that
specifically binds IL-1.alpha./IL-1.beta. that is substantially
free of antibodies that specifically bind antigens other than
IL-1.alpha.). Moreover, an isolated antibody may be substantially
free of other cellular material and/or chemicals.
[0102] As referred to herein, the terms "dual-specific antibody" or
"an antibody having dual-specificity" means an antibody comprising
two antigen-binding sites or regions, a first binding site or
region having affinity for a first antigen or epitope and a second
binding site or region having binding affinity for a second antigen
or epitope that is distinct from the first. In one embodiment, the
heavy chain variable domain comprises one antigen binding region,
i.e., IL-1.alpha. or IL-1.beta., and the light chain variable
domain comprises a further antigen binding region, i.e.,
IL-1.alpha. or IL-1.beta., such that the antibody has dual-epitope
specificity for IL-1.alpha. and IL-1.beta.. In addition, the
invention also includes within its scope dual-specific
V.sub.H/V.sub.L combinations and V.sub.L/V.sub.L combinations.
[0103] As used herein, the term "antigen binding region" or
"antigen binding site" refers to the portion(s) of an antibody
molecule, or antigen binding portion thereof, which contains the
amino acid residues that interact with an antigen and confers on
the antibody its specificity and affinity for the antigen. In one
embodiment, the antibody of the invention comprises two antigen
binding regions, one which is specific for IL-1.alpha. and another
which is specific for IL-1.beta..
[0104] The term "epitope" is meant to refer to that portion of any
molecule capable of being recognized by and bound by an antibody at
one or more of the antibody's antigen binding regions. In the
context of the present invention, first and second "epitopes" are
understood to be epitopes which are not the same and are not bound
by a single monospecific antibody, or antigen-binding portion
thereof. In the invention, the first and second epitopes are
advantageously on different antigens, IL-1.alpha. or IL-1.beta..
Likewise, the first and second antigens are advantageously not the
same.
[0105] A "neutralizing antibody," as used herein, is intended to
refer to an antibody whose binding to a particular antigen(s),
i.e., IL-1.alpha. or IL-1.beta., results in inhibition of the
biological activity of the antigen. Inhibition of the biological
activity of IL-1.alpha. or IL-1.beta. can be assessed by measuring
one or more indicators of IL-1.alpha. or IL-1.beta. activity, such
as IL-1.alpha. or IL-1.beta.-induced cytotoxicity (either in vitro
or in vivo), or IL-1.alpha. or IL-1.beta. binding to IL-1.alpha. or
IL-1.beta. receptors. These indicators of IL-1.alpha./IL-1.beta.
activity can be assessed by one or more standard in vitro or in
vivo assays known in the art (for example see Example 5). In one
embodiment, the ability of a dual-specific antibody to neutralize
both IL-1.alpha. and IL-1.beta. activity is assessed by inhibition
of IL-1.alpha. and IL-1.beta. in human embryonic lung fibroblasts,
MRC-5 cells.
[0106] The "ND.sub.50" value of an antibody, or antigen-binding
portion thereof, refers to the concentration of antibody resulting
in a one-half maximal inhibition of the given cytokine activity on
a responsive cell line.
[0107] As used herein, the term "EC50" is defined as the
concentration of an antibody, or antigen-binding portion thereof,
that results in 50% of a measured biological effect. For example,
the EC50 of a therapeutic agent having a measurable biological
effect may comprise the value at which the agent displays 50% of
the biological effect. The term "IC50" is defined as the
concentration of an antibody, or antigen-binding portion thereof,
that results in 50% inhibition of a measured effect.
[0108] The term "surface plasmon resonance", as used herein, 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; Johnson et al. (1995) J. Mol. Recognit.
8:125-131; and Johanson et al. (1991) Anal. Biochem.
198:268-277.
[0109] The term "K.sub.off", as used herein, is intended to refer
to the off rate constant for dissociation of an antibody from the
antibody/antigen complex.
[0110] The term "K.sub.d" or "K.sub.D" as used herein, is intended
to refer to the dissociation constant of a particular
antibody-antigen interaction, which is obtained from the ratio of
k.sub.d to k.sub.a (i.e., k.sub.d/k.sub.a) and is expressed as a
molar concentration (M). K.sub.D values for antibodies can be
determined using methods well established in the art. A preferred
method for determining the K.sub.D of an antibody is by using
surface plasmon resonance, preferably using a biosensor system such
as a Biacore.RTM. system.
[0111] A "monoclonal antibody" as used herein is intended to refer
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. Furthermore, in contrast
to polyclonal antibody preparations that typically include
different antibodies directed against different determinants
(epitopes), each monoclonal antibody 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.
[0112] The phrase "recombinant antibody" refers to 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, antibodies isolated from a
recombinant, combinatorial antibody library, 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) or antibodies prepared, expressed, created or
isolated by any other means that involves splicing of particular
immunoglobulin gene sequences (such as human immunoglobulin gene
sequences) to other DNA sequences. Examples of recombinant
antibodies include chimeric, CDR-grafted and humanized
antibodies.
[0113] The term "human antibody" refers to antibodies having
variable and constant regions corresponding to, or derived from,
human germline immunoglobulin sequences as described by, for
example, Kabat et al. (See Kabat, et al. (1991) Sequences of
Proteins of Immunological Interest, Fifth Edition, U.S. Department
of Health and Human Services, NTH Publication No. 91-3242). The
human antibodies of the invention, however, 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.
[0114] Recombinant human antibodies of the invention have variable
regions, and may also include constant regions, derived from human
germline immunoglobulin sequences (See Kabat et al. (1991)
Sequences of Proteins of Immunological Interest, Fifth Edition,
U.S. Department of Health and Human Services, NIH Publication No.
91-3242). 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. In certain embodiments, however, such recombinant
antibodies are the result of selective mutagenesis or backmutation
or both.
[0115] The term "backmutation" refers to a process in which some or
all of the somatically mutated amino acids of a human antibody are
replaced with the corresponding germline residues from a homologous
germline antibody sequence. The heavy and light chain sequences of
a human antibody of the invention are aligned separately with the
germline sequences in the VBASE database to identify the sequences
with the highest homology. Differences in the human antibody of the
invention are returned to the germline sequence by mutating defined
nucleotide positions encoding such different amino acid. The role
of each amino acid thus identified as candidate for backmutation
should be investigated for a direct or indirect role in antigen
binding and any amino acid found after mutation to affect any
desirable characteristic of the human antibody should not be
included in the final human antibody. To minimize the number of
amino acids subject to backmutation those amino acid positions
found to be different from the closest germline sequence but
identical to the corresponding amino acid in a second germline
sequence can remain, provided that the second germline sequence is
identical and colinear to the sequence of the human antibody of the
invention for at least 10, preferably 12 amino acids, on both sides
of the amino acid in question. Backmuation may occur at any stage
of antibody optimization.
[0116] The term "chimeric antibody" refers to antibodies which
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.
[0117] The term "CDR-grafted antibody" refers to antibodies which
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.
[0118] The term "humanized antibody" refers to antibodies which
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.
[0119] The term "nucleic acid molecule", as used herein, is
intended to include DNA molecules and RNA molecules. A nucleic acid
molecule may be single-stranded or double-stranded, but preferably
is double-stranded DNA.
[0120] The term "isolated nucleic acid molecule", as used herein in
reference to nucleic acids encoding antibodies or antibody portions
(e.g., VH, VL, CDR3) that bind IL-1.alpha. and IL-1.beta., is
intended to refer to a nucleic acid molecule in which the
nucleotide sequences encoding the antibody or antibody portion are
free of other nucleotide sequences encoding antibodies or antibody
portions that bind antigens other than hTNF.alpha., which other
sequences may naturally flank the nucleic acid in human genomic
DNA. Thus, for example, an isolated nucleic acid of the invention
encoding a VH region of an anti-TNF.alpha. antibody contains no
other sequences encoding other VH regions that bind antigens other
than IL-1.alpha. and IL-1.beta..
[0121] The term "vector", as used herein, 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.,
vectors may have a bacterial origin of replication and/or an
episomal origin of replication (generally derived from a viral
sequence, e.g., SV40). 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, the invention is
intended to include such other forms of expression vectors, such as
viral vectors (e.g., replication defective retroviruses,
adenoviruses and adeno-associated viruses), which serve equivalent
functions.
[0122] As used herein, the term "episomally replicating vector" or
"episomal vector" refers to a vector which is typically and very
preferably not integrated into the genome of the host cell, but
exists in parallel. An episomally replicating vector, as used
herein, is replicated during the cell cycle and in the course of
this replication the vector copies are distributed statistically in
the resulting cells depending on the number of the copies present
before and after cell division. Preferably, the episomally
replicating vector may take place in the nucleus of the host cell,
and preferably replicates during S-phase of the cell cycle.
Moreover, the episomally replicating vector is replicated at least
once, i.e. one or multiple times, in the nucleus of the host cell
during S-phase of the cell cycle. In a very preferred embodiment,
the episomally replicating vector is replicated once in the nucleus
of the host cell during S-phase of the cell cycle.
[0123] As used herein, the terms "origin of replication sequences"
or "origin of replication," used interchangeably herein, refer to
sequences which, when present in a vector, initiate replication. An
origin of replication may be recognized by a replication initiation
factor or, alternatively, by a DNA helicase.
[0124] The "gene of interest" as used herein, refers to an
exogenous DNA sequence which is added to the vector of the
invention. The gene of interest, for example, may comprise a coding
sequence which can be either spaced by introns or which is a cDNA
encoding the open reading frame. The region of the vector to which
the gene of interest is cloned is referred to herein as an
"insertion site." Preferably, the gene of interest comprises a
portion of the antibody or fusion protein that is expressed using a
vector of the invention. For example, the heavy chain variable
region of the antibody ABT2-108, i.e., the gene of interest, may
cloned into the vector of the invention which comprises a heavy
chain constant region
[0125] In one embodiment of the invention, the vector comprises an
antibody light or heavy chain constant region which is 3' to the
insertion site for the gene of interest and is operably linked
thereto. Thus, in one embodiment, the gene of interest is a
variable region of a light or heavy chain of an antibody which is
operably linked to the antibody light or heavy chain constant
region encoded in the vector of the invention.
[0126] The term "promoter" includes any nucleic acid sequence
sufficient to direct transcription in a eukaryotic cell, including
inducible promoters, repressible promoters and constitutive
promoters. Typically a promoter includes elements that are
sufficient to render promoter-dependent gene expression
controllable in a cell type-specific, tissue-specific or
temporal-specific manner, or inducible by external signals or
agents. Such elements can be located in the 5' or 3' or intron
sequence regions of a particular gene. Ordinarily, gene expression
will be constitutive, although regulatable promoters can be
employed in the present invention if desired. Gene expression can
also be controlled by transcription-regulation using heat, light,
or metals, such as by the use of metallothionine genes or heat
shock genes.
[0127] "Upstream" and "downstream" are terms used to describe the
relative orientation between two elements present in a nucleotide
sequence or vector. An element that is "upstream" of another is
located in a position closer to the 5' end of the sequence (i.e.,
closer to the end of the molecule that has a phosphate group
attached to the 5' carbon of the ribose or deoxyribose backbone if
the molecule is linear) than the other element. An element is said
to be "downstream" when it is located in a position closer to the
3' end of the sequence (i.e., the end of the molecule that has an
hydroxyl group attached to the 3' carbon of the ribose or
deoxyribose backbone in the linear molecule) when compared to the
other element.
[0128] As used herein, the term "stuffer sequence" refers to a
nucleic acid sequence, preferably in a vector, which is flanked by
restriction enzyme sites at both the 5' and 3' ends. The stuffer
sequence is located in a vector at the insertion site for the
nucleic acid encoding the gene of interest. During the cloning
process, the stuffer sequence is digested away from the vector
using the appropriate restriction enzymes, and the nucleic acid
encoding the gene of interest is ligated or homologously recombined
into the vector at the former position of the stuffer sequence.
Preferably, the stuffer sequence is large enough to provide
sufficient distance between the 5' and 3' restriction enzyme sites
so that the restriction enzyme can efficiently cut the vector. In
addition, it is preferred that the length of the stuffer sequence
is different than the size of the nucleic acid encoding the gene of
interest, e.g., a stuffer sequence of about 300 base pairs or less
or about 400 base pairs or more may be used for a nucleic acid
encoding the gene of interest which is about 350 base pairs. In
another embodiment, the stuffer sequence is about 1 kb in size.
[0129] The term "recombinant host cell" (or simply "host cell"), as
used herein, is intended to refer to a cell into which a
recombinant expression vector has been introduced. It should be
understood that such terms are intended to refer not only to the
particular subject cell but 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" as used herein.
[0130] The term "repertoire" refers to a collection of diverse
variants, for example polypeptide variants which differ in their
primary sequence. A library used in the present invention will
encompass a repertoire of polypeptides comprising at least 1000
members.
[0131] The term "library" refers to a mixture of heterogeneous
polypeptides or nucleic acids. The library is composed of members,
each of which have a single polypeptide or nucleic acid sequence.
To this extent, library is synonymous with repertoire. Sequence
differences between library members are responsible for the
diversity present in the library. The library may take the form of
a simple mixture of polypeptides or nucleic acids, or may be in the
form of organisms or cells, for example bacteria, viruses, animal
or plant cells and the like, transformed with a library of nucleic
acids. Preferably, each individual organism or cell contains only
one or a limited number of library members. Advantageously, the
nucleic acids are incorporated into expression vectors, in order to
allow expression of the polypeptides encoded by the nucleic acids.
In a preferred aspect, therefore, a library may take the form of a
population of host organisms, each organism containing one or more
copies of an expression vector containing a single member of the
library in nucleic acid form which can be expressed to produce its
corresponding polypeptide member. Thus, the population of host
organisms has the potential to encode a large repertoire of
genetically diverse polypeptide variants.
[0132] Various aspects of the invention are described in further
detail in the following subsections.
II. IL-1.alpha./IL-1.beta. Dual-Specific Antibodies
[0133] The invention provides isolated, dual-specific antibodies,
or antigen binding portions thereof, that bind human IL-1.alpha.
and IL-1.beta. with high affinity and neutralizing capacity. In a
preferred embodiment, the antibody, or antigen-binding fragment
thereof, is human. Preferably, the antibodies of the invention are
recombinant, neutralizing anti-IL-1.alpha. and anti-IL-1.beta.. In
one embodiment, the invention provides an isolated, dual-specific
antibody, or an antigen-binding portion thereof, which dissociates
from human IL-1.alpha. with a K.sub.D of 1.times.10.sup.-7 M or
less and neutralizes human IL-1.alpha. in a standard in vitro MRC5
assay with an ND.sub.50 of 900 nM or less, and dissociates from
human IL-1.beta. with a K.sub.D of 5.times.10.sup.-5 M or less, and
neutralizes human IL-1.beta. in a standard in vitro MRC5 assay with
an ND.sub.50 of 800 nM or less. In a preferred embodiment, the
IL-1.alpha./IL-1.beta. dual-specific antibody, or antigen-binding
portion thereof, binds human IL-1.alpha. and IL-1.beta.
(hIL-1.alpha. and hIL-1.beta.), but does not bind mouse IL-1.alpha.
or mouse IL-1.beta.. In the case that the variable domains are
selected from V-gene repertoires selected for instance using phage
display technology as herein described, then these variable domains
can comprise a universal framework region, such that is they may be
recognised by a specific generic ligand as herein defined. The use
of universal frameworks, generic ligands and the like is described
in WO99/20749. In the present invention, reference to phage display
includes the use of both phage and/or phagemids.
[0134] The dual-specific antibodies of the invention may comprise
variable regions which are derived from antibodies directed against
IL-1.alpha. and IL-1.beta.. Alternatively, the antibodies of the
invention may be derived from a repertoire of single antibody
domains such as those expressed on the surface of filamentous
bacteriophage. Selection may be performed as described below in
section III and in the Examples provided herein.
[0135] The invention provides single domain antibodies which are
have neutralizing and affinity properties specific for IL-1.alpha.
or IL-1.beta., as summarized in Tables 8 and 61 to 64, as well as
the Examples.
[0136] Dual-specific antibodies, or antigen-binding portions
thereof, according to the present invention preferably comprise
combinations of heavy and light chain domains. For example, the
dual specific antibody, or antigen-binding portion thereof, may
comprise a V.sub.H domain and a V.sub.L domain, which may be linked
together in the form of an scFv. In addition, the antibody, or
antigen-binding portion thereof, may comprise one or more C.sub.H
or C.sub.L domains. For example, the antibody, or antigen-binding
portion thereof, may comprise a C.sub.H1 domain, C.sub.H2 or
C.sub.H3 domain, and/or a C.sub.L domain, C.mu.1, C.mu.2, C.mu.3 or
C.mu.4 domains, or any combination thereof. A hinge region domain
may also be included. Such combinations of domains may, for
example, mimic natural antibodies, such as IgG or IgM, or fragments
thereof, such as Fv, scFv, Fab or F(ab').sub.2 molecules. Other
structures, such as a single arm of an IgG molecule comprising
V.sub.H, V.sub.L, C.sub.H1 and C.sub.L domains, are envisaged.
[0137] Preferably, the dual specific antibody, or antigen-binding
portion thereof, of the invention comprises only two variable
domains although several such antibodies, or antigen-binding
portions thereof, may be incorporated together into the same
protein, for example two such antibodies, or antigen-binding
portions thereof, can be incorporated into an IgG or a multimeric
immunoglobulin, such as IgM. Alternatively, in another embodiment a
plurality of dual specific ligands are combined to form a multimer.
For example, two different dual specific ligands are combined to
create a tetra-specific molecule.
[0138] It will be appreciated by one skilled in the art that the
light and heavy variable regions of a dual-specific antibodies, or
antigen-binding portions thereof, produced according to the methods
described herein and known in the art, may be on the same
polypeptide chain, or alternatively, on different polypeptide
chains. In the case that the variable regions are on different
polypeptide chains, then they may be linked via a linker, generally
a flexible linker (such as a polypeptide chain), a chemical linking
group, or any other method known in the art.
[0139] In one embodiment, the present invention provides dual
specific antibodies, or antigen-binding portions thereof, which
comprise at least two complementary variable domains, e.g., a VH
and a VL domain. In another embodiment, the present invention
provides dual specific antibodies, or antigen-binding portions
thereof, which comprise at least two non-complementary variable
domains. For example, the antibody, or antigen-binding portion
thereof, may comprise a pair of VH domains or a pair of VL domains.
Advantageously, the domains are of non-camelid origin; preferably
they are human domains or comprise human framework regions (FWs)
and one or more heterologous CDRs. CDRs and framework regions are
those regions of an immunoglobulin variable domain as defined in
the Kabat database of Sequences of Proteins of Immunological
Interest.
[0140] Dual-specific antibodies of the invention may include
variable heavy and light chain amino acid regions described in SEQ
ID NOs: 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, and 52 (see
Table 61 and Example 3 for a summary). Dula specific antibodies may
also include any of the variable heavy and light chain amino acid
regions described in SEQ ID NOs: 53 to 133.
[0141] Pairings of the single domain antibodies described in SEQ ID
NOs: 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, and 52, as well
as any of the variable sequences described in SEQ ID NOs: 53 to
133, are also included in the invention. Examples of dual-specific
IL-1.alpha. and IL-1.beta. antibodies are described in the
Examples, including Example 5 and Table 9. In one embodiment, the
invention provides a dual-specific, isolated antibody, or
antigen-binding portion thereof, comprising an IL-1.alpha. antigen
binding region and an IL-1.beta. antigen binding region, wherein
the antibody, or antigen-binding portion thereof, comprises a heavy
chain variable region and a light chain variable region combination
comprising a heavy chain variable region comprising CDRs as set
forth in SEQ ID NO: 16 (ABT1-96) and a light chain variable region
comprising CDRs as set forth in SEQ ID NO: 40 (ABT2-46); a light
chain variable region comprising CDRs as set forth in SEQ ID NO: 24
(ABT1-122) and a heavy chain variable region comprising CDRs as set
forth in SEQ ID NO: 52 (ABT2-108); a light chain variable region
comprising CDRs as set forth in SEQ ID NO: 28 (ABT1-141) and a
heavy chain variable region comprising CDRs as set forth in SEQ ID
NO: 52 (ABT2-108); a light chain variable region comprising CDRs as
set forth in SEQ ID NO: 28 (ABT1-141) and a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 44 (ABT2-65); a
heavy chain variable region comprising CDRs as set forth in SEQ ID
NO: 16 (ABT1-96) and a light chain variable region comprising CDRs
as set forth in SEQ ID NO: 36 (ABT2-42); a light chain variable
region comprising CDRs as set forth in SEQ ID NO: 12 (ABT1-95) and
a heavy chain variable region comprising CDRS as set forth in SEQ
ID NO: 32 (ABT2-13); a light chain variable region comprising CDRS
as set forth in SEQ ID NO: 24 (ABT1-122) and a heavy chain variable
region comprising CDRs as set forth in SEQ ID NO: 44 (ABT2-65); or
a heavy chain variable region comprising CDRs as set forth in SEQ
ID NO: 20 (ABT1-98) and a light chain variable region comprising
CDRs as set forth in SEQ ID NO: 48 (ABT2-76).
[0142] It is known in the art that antibody heavy and light chain
CDR3 domains play an important role in the binding
specificity/affinity of an antibody for an antigen. Accordingly, in
another aspect, the invention pertains to antibodies, including
human antibodies, that have slow dissociation kinetics for
association with either IL-1.alpha. or IL-1.beta. and that have
light and heavy chain CDR3 domains that structurally are identical
to or related to those identified herein, including those CDR3
domains described in SEQ ID NOs: 11, 23, 27, 3, 7, 15, and 19.
[0143] In one aspect, the invention pertains to a dual-specific
antibody, or antigen-binding portion thereof, of the invention is
preferably selected to have desirable binding kinetics (e.g., high
affinity, low dissociation, slow off-rate, strong neutralizing
activity) for IL-1.alpha. and IL-1.beta., to which the antibody
specifically binds. For example, the dual-specific antibody, or
portion thereof, may bind IL-1.alpha. and IL-1.beta. with a
k.sub.off rate constant of 0.1 s.sup.4 or less, more preferably a
k.sub.off rate constant of 1.times.10.sup.-2 s.sup.-1 or less, even
more preferably a k.sub.off rate constant of 1.times.10.sup.-3
s.sup.-1 or less, even more preferably a k.sub.off rate constant of
1.times.10.sup.-4 s.sup.-1 or less, or even more preferably a
k.sub.off rate constant of 1.times.10.sup.-5 s.sup.-1 or less, as
determined by surface plasmon resonance. In one embodiment, the
isolated, dual-specific antibody, or an antigen-binding portion
thereof, dissociates from human IL-1.alpha. with a K.sub.D of about
1.times.10.sup.-7 M to about 1.times.10.sup.-8 M or less and
dissociates from human IL-1.beta. with a K.sub.D of about
5.times.10.sup.-5 M to about 1.times.10.sup.-9 M or less. Ranges
intermediate to the above recited constants, e.g.,
4.0.times.10.sup.-8 M, are also intended to be part of this
invention. For example, ranges of values using a combination of any
of the above recited values as upper and/or lower limits are
intended to be included.
[0144] In one embodiment, the antibody, or antigen-binding portion
thereof, dissociates from IL-1.beta. with a K.sub.D of
5.4.times.10.sup.-5 M or less; dissociates from IL-1.beta. with a
K.sub.D of 2.8.times.10.sup.-6 M or less; dissociates from
IL-1.beta. with a K.sub.D of 1.3.times.10.sup.-6 M or less;
dissociates from IL-1.beta. with a K.sub.D of 9.3.times.10.sup.-7 M
or less; dissociates from IL-1.beta. with a K.sub.D of
2.times.10.sup.-7 M or less; dissociates from IL-1.beta. with a
K.sub.D of 1.1.times.10.sup.-7 M or less; or dissociates from
IL-1.beta. with a K.sub.D of 2.8.times.10.sup.-8 M or less. Ranges
intermediate to the above recited constants, e.g., K.sub.D of
1.7.times.10.sup.-7 M or less, are also intended to be part of this
invention. For example, ranges of values using a combination of any
of the above recited values as upper and/or lower limits are
intended to be included.
[0145] In one embodiment, the antibody, or antigen-binding portion,
dissociates from IL-1.alpha. with a K.sub.D of 1.times.10.sup.-8 M
or less; dissociates from IL-1.alpha. with a K.sub.D of
1.times.10.sup.-9 M or less; dissociates from IL-1.alpha. with a
K.sub.D of 40-86 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 20-42 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 32-42 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 7-12 nM or less; dissociates from IL-1.alpha. with a
K.sub.D of 3.0.times.10.sup.-7 M or less; dissociates from
IL-1.alpha. with a K.sub.D of 1.1.times.10.sup.-7 M or less;
dissociates from IL-1.alpha. with a K.sub.D of 6.1.times.10.sup.-8
M or less; dissociates from IL-1.alpha. with a K.sub.D of
6.times.10.sup.-8 M or less; dissociates from IL-1.alpha. with a
K.sub.D of 4.2.times.10.sup.-8 M or less; dissociates from
IL-1.alpha. with a K.sub.D of 1.3.times.10.sup.-8 M or less; or
dissociates from IL-1.alpha. with a K.sub.D of 1.1.times.10.sup.-9
M or less. Ranges intermediate to the above recited constants,
e.g., K.sub.D of 1.7.times.10.sup.-7 M or less, are also intended
to be part of this invention. For example, ranges of values using a
combination of any of the above recited values as upper and/or
lower limits are intended to be included.
[0146] It should be noted that the aforementioned affinity
properties may also apply to single domain antibodies of the
invention which bind and neutralize IL-1.alpha. or IL-1.beta..
Surface plasmon resonance analysis may be used for determining
kinetic values, including K.sub.D and k.sub.off values.
[0147] The IL-1.alpha./IL-1.beta. dual-specific antibody, or
antigen-binding portion thereof, of the invention may exhibit a
strong capacity to neutralize both hIL-1.alpha. and hIL-1.beta.
activity, as determined using a standard in vitro or in vivo assay
(see also Examples, including Example 5). For example, in one
embodiment if the invention, IL-1.alpha./IL-1.beta. antibodies
neutralize human IL-1.alpha. in a standard in vitro assay with
ND.sub.50 values of about 900 nM to about 10 nM or less. The
antibodies of the invention are also able to neutralize human
IL-1.beta. in a standard in vitro assay with ND.sub.50 values of
about 800 nM to about 200 nM or less. Alternatively or
additionally, a dual-specific antibody, or antigen binding portion
thereof, may inhibit the activity of one, and more preferably both,
of IL-1.alpha. and/or IL-1.beta. with an ND.sub.50 of
1.times.10.sup.-6 M or less, even more preferably with an ND.sub.50
of 1.times.10.sup.-4 M or less, even more preferably with an
ND.sub.50 of 1.times.10.sup.-8 M or less, even more preferably with
an ND.sub.50 of 1.times.10.sup.-9M or less, even more preferably
with an ND.sub.50 of 1.times.10.sup.-10 M or less, or even more
preferably with an ND.sub.50 of 1.times.10.sup.-11 M or less. In
one embodiment, the antibody, or antigen-binding portion,
neutralizes human IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 900 nM or less, and/or neutralizes human IL-1.beta. in
a standard in vitro assay with an ND.sub.50 of 800 nM or less. In
one embodiment, the antibody, or antigen-binding portion thereof,
neutralizes IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 10 nM or less. In one embodiment, the antibody, or
antigen-binding portion thereof, neutralizes IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 200 nM or less. In one
embodiment, the antibody, or antigen-binding portion thereof,
neutralizes IL-1.alpha. in a standard in vitro assay with an
ND.sub.50 of 10 nM or less. In one embodiment, the antibody, or
antigen-binding portion thereof, neutralizes IL-1.beta. in a
standard in vitro assay with an ND.sub.50 of 200 nM or less. Ranges
intermediate to the above recited values, e.g., ND.sub.50 of 80 nM
or less, ND.sub.50 of 4.0.times.10.sup.-10 M or less, are also
intended to be part of this invention. For example, ranges of
values using a combination of any of the above recited values as
upper and/or lower limits are intended to be included.
[0148] In one embodiment, neutralization properties of an antibody,
or antigen-binding portion thereof, may be determined using an in
vitro MRC-5 cell assay.
[0149] It should be noted that the aforementioned neutralization
properties may also apply to single domain antibodies of the
invention which bind and neutralize IL-1.alpha. or IL-1.beta..
[0150] In yet another embodiment, an antibody of the invention
comprises heavy and light chain variable regions comprising amino
acid sequences that are homologous to the amino acid sequences of
the preferred antibodies described herein, and wherein the
antibodies retain the desired functional properties of the
IL-1.alpha./IL-1.beta. dual-specific antibodies of the invention.
For example, the invention provides an isolated antibody, or
antigen binding portion thereof, comprising a heavy chain variable
region and a light chain variable region, wherein (a) the heavy
chain variable region comprises an amino acid sequence that is at
least 80% homologous to an amino acid sequence selected from the
group consisting of SEQ ID NOs: 4, 16, 20, 32, 44, 52, or 53 to 92;
(b) the light chain variable region comprises an amino acid
sequence that is at least 80% homologous to an amino acid sequence
selected from the group consisting of SEQ ID NOs: 8, 12, 24, 28,
36, 40, 48, or 93 to 133; and (c) the antibody specifically binds
to IL-1.alpha./IL-1.beta. and exhibits at least one of the
functional properties described herein, preferably several of the
functional properties described herein. Also included in the
invention is an isolated antibody, or antigen binding portion
thereof, comprising two heavy (or two light) chain variable regions
that specifically binds to IL-1.alpha./IL-1.beta..
[0151] In other embodiments, the VH and/or VL amino acid sequences
may be 85%, 90%, 95%, 96%, 97%, 98% or 99% homologous to the
sequences set forth herein, including SEQ ID NOs: 52 to 133. An
antibody having VH and VL regions having high (i.e., 80% or
greater) homology to the VH and VL regions of the sequences set
forth above, can be obtained by mutagenesis (e.g., site-directed or
PCR-mediated mutagenesis) of nucleic acid molecules encoding SEQ ID
NOs: 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, and 52 to 133,
followed by testing of the encoded altered antibody for retained
function (i.e., affinity and neutralization properties) using the
functional assays described herein.
[0152] In another embodiment, the invention provides nucleic acid
sequences which may be 85%, 90%, 95%, 96%, 97%, 98% or 99%
homologous to the sequences set forth herein, including SEQ ID NOs:
134 to 843.
[0153] As used herein, the percent homology between two amino acid
sequences is equivalent to the percent identity between the two
sequences. The percent identity between the two sequences is a
function of the number of identical positions shared by the
sequences (i.e., % homology=# of identical positions/total# of
positions.times.100), taking into account the number of gaps, and
the length of each gap, which need to be introduced for optimal
alignment of the two sequences. The comparison of sequences and
determination of percent identity between two sequences can be
accomplished using a mathematical algorithm, as described in the
non-limiting examples below.
[0154] The percent identity between two amino acid sequences can be
determined using the algorithm of Meyers and Miller (Comput. Appl.
Biosci., 4:11-17 (1988)) which has been incorporated into the ALIGN
program (version 2.0), using a PAM120 weight residue table, a gap
length penalty of 12 and a gap penalty of 4. In addition, the
percent identity between two amino acid sequences can be determined
using the Needleman and Wunsch (J. Mol. Biol. 48:444-453 (1970))
algorithm which has been incorporated into the GAP program in the
GCG software package (available at http://www.gcg.com), using
either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of
16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or
6.
[0155] Additionally or alternatively, the protein sequences of the
present invention can further be used as a "query sequence" to
perform a search against public databases to, for example, identify
related sequences. Such searches can be performed using the XBLAST
program (version 2.0) of Altschul et al. (1990) J. Mol. Biol.
215:403-10. BLAST protein searches can be performed with the XBLAST
program, score=50, wordlength=3 to obtain amino acid sequences
homologous to the antibody molecules of the invention. To obtain
gapped alignments for comparison purposes, Gapped BLAST can be
utilized as described in Altschul et al. (1997) Nucleic Acids Res.
25(17):3389-3402. When utilizing BLAST and Gapped BLAST programs,
the default parameters of the respective programs (e.g., XBLAST and
NBLAST) can be used. See http://www.ncbi.nlm.nih.gov.
[0156] In certain embodiments, an antibody of the invention
comprises a heavy chain variable region comprising CDR1, CDR2 and
CDR3 sequences and a light chain variable region comprising CDR1,
CDR2 and CDR3 sequences, wherein one or more of these CDR sequences
comprise specified amino acid sequences based on the preferred
antibodies described herein, or conservative modifications thereof,
and wherein the antibodies retain the desired functional properties
of the anti-IL-1.alpha./IL-1.beta. dual-specific antibodies of the
invention.
[0157] The skilled artisan will appreciate that substitution of
other amino acids within the CDR3 domains identified herein may be
possible while still retaining the low off rate constant of the
antibody, in particular substitutions with conservative amino
acids. Accordingly, the invention provides an isolated antibody, or
antigen binding portion thereof, comprising a heavy chain variable
region comprising CDR1, CDR2, and CDR3 sequences and a light chain
variable region comprising CDR1, CDR2, and CDR3 sequences, wherein:
(a) the heavy chain variable region CDR3 sequence comprises an
amino acid sequence selected from the group consisting of amino
acid sequences of SEQ ID NOs: 3, 7, 15, 19, 31, 43, and 51, and
conservative modifications thereof; (b) the light chain variable
region CDR3 sequence comprises an amino acid sequence selected from
the group consisting of amino acid sequences of SEQ ID NOs: 11, 23,
27, 35, 39, and 47, and conservative modifications thereof; and (c)
the antibody specifically binds to IL-1.alpha./IL-1.beta. and
exhibits at least one of the functional properties described
herein, more preferably several of the functional properties
described herein, i.e., high affinity and neutralizing for both
IL-1.alpha./IL-1.beta..
[0158] In a further embodiment, the heavy chain variable region
CDR2 sequence comprises an amino acid sequence selected from the
group consisting of amino acid sequences of SEQ ID NOs: 2, 6, 14,
18, 30, 42, and 50, and conservative substitutions thereof; and the
light chain variable region CDR2 sequence comprises an amino acid
sequence selected from the group consisting of amino acid sequences
of SEQ ID NOs: 10, 22, 26, 34, 38, and 46, and conservative
modifications thereof. In a still further embodiment, the heavy
chain variable region CDR1 sequence comprises an amino acid
sequence selected from the group consisting of amino acid sequences
of SEQ ID NOs: 1, 5, 13, 17, 29, 41, and 49, and conservative
modifications thereof; and the light chain variable region CDR1
sequence comprises an amino acid sequence selected from the group
consisting of amino acid sequences of SEQ ID NOs: 9, 21, 25, 33,
37, and 45, and conservative modifications thereof.
[0159] A "conservative amino acid substitution" or a "conservative
substitution", as used herein, is one in which one amino acid
residue is replaced with another amino acid residue having a
similar side chain. Families of amino acid residues having similar
side chains have been defined in the art, including basic side
chains (e.g., lysine, arginine, histidine), acidic side chains
(e.g., aspartic acid, glutamic acid), uncharged polar side chains
(e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine,
cysteine), nonpolar side chains (e.g., alanine, valine, leucine,
isoleucine, proline, phenylalanine, methionine, tryptophan),
beta-branched side chains (e.g., threonine, valine, isoleucine) and
aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,
histidine). Substitutions and modifications can be introduced into
an antibody of the invention by standard techniques known in the
art, such as site-directed mutagenesis and PCR-mediated
mutagenesis. Thus, one or more amino acid residues within the CDR
regions of an antibody of the invention can be replaced with other
amino acid residues from the same side chain family and the altered
antibody can be tested for retained function, e.g., affinity or
neutralization characteristics, using the functional assays
described herein.
[0160] Accordingly, another embodiment of the invention pertains to
an isolated dual-specific antibody, or antigen binding portion
thereof, comprising a heavy chain variable region comprising CDR1,
CDR2, and CDR3 sequences comprising an amino acid sequence selected
from the group consisting of SEQ ID NOs: 1, 5, 13, 17, 29, 41, and
49, SEQ ID NOs: 2, 6, 14, 18, 30, 42, and 50 and SEQ ID NOs: 3, 7,
15, 19, 31, 43, and 51, respectively, and a light chain variable
region comprising CDR1, CDR2, and CDR3 sequences comprising an
amino acid sequence selected from the group consisting of SEQ ID
NOs: .delta. 9, 21, 25, 33, 37, and 45, SEQ ID NOs: 10, 22, 26, 34,
38, and 46 and SEQ ID NOs: 11, 23, 27, 35, 39, and 47,
respectively. Thus, such antibodies contain the VH and/or VL CDR
sequences of antibodies described herein, yet may contain different
framework sequences from these antibodies.
[0161] Preferred human framework regions are those encoded by
germline gene segments DP47 and DPK9. Advantageously, FW1, FW2 and
FW3 of a VH or VL domain have the sequence of FW1, FW2 or FW3 from
DP47 or DPK9. The human frameworks may optionally contain
mutations, for example up to about 5 amino acid changes or up to
about 10 amino acid changes collectively in the human frameworks
used in the ligands of the invention.
[0162] Such framework sequences can be obtained from public DNA
databases or published references that include germline antibody
gene sequences. For example, germline DNA sequences for human heavy
and light chain variable region genes can be found in the "VBase"
human germline sequence database (available on the Internet at
www.mrc-cpe.cam.ac.uk/vbase), as well as in Kabat et al. (1991)
Sequences of Proteins of Immunological Interest, Fifth Edition,
U.S. Department of Health and Human Services, NIH Publication No.
91-3242; Tomlinson, I. M., et al. (1992) "The Repertoire of Human
Germline VH Sequences Reveals about Fifty Groups of VH Segments
with Different Hypervariable Loops" J. Mol. Biol. 227:776-798; and
Cox et al., (1994) "A Directory of Human Germ-line VH Segments
Reveals a Strong Bias in their Usage" Eur. J. Immunol. 24:827-836;
the contents of each of which are expressly incorporated herein by
reference. Preferred framework sequences for use in the antibodies
of the invention are those that are structurally similar to the
framework sequences used by selected antibodies of the invention.
The VH CDR1, 2 and 3 sequences of SEQ ID NOs: 1, 2, 3, 5, 6, 7, 13,
14, 15, 17, 18, 19, 29, 30, 31, 41, 42, 43 49, 50, and 51 and the
VL CDR1, 2 and 3 sequences of SEQ ID NOs: 9, 10, 11, 21, 22, 23,
25, 26, 27, 33, 34, 35, 37, 38, 39, 45, 46, and 47, can be grafted
onto framework regions that have the same sequence as that found in
the germline immunoglobulin gene from which the framework sequence
derive, or the CDR sequences can be grafted onto framework regions
that contain one or more mutations as compared to the germline
sequences. For example, it has been found that in certain instances
it is beneficial to mutate residues within the framework regions to
maintain or enhance the antigen binding ability of the antibody
(see e.g., U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and
6,180,370 to Queen et al).
[0163] In one embodiment, an antibody of the invention may be
prepared using an antibody having one or more of the V.sub.H and/or
V.sub.L sequences disclosed herein as starting material to engineer
a modified antibody, which modified antibody may have altered
properties from the starting antibody. An antibody can be
engineered by modifying one or more residues within one or both
variable regions (i.e., V.sub.H and/or V.sub.L), for example within
one or more CDR regions and/or within one or more framework
regions. Additionally or alternatively, an antibody can be
engineered by modifying residues within the constant region(s), for
example to alter the effector function(s) of the antibody.
[0164] One type of variable region engineering that can be
performed is CDR grafting. Antibodies interact with target antigens
predominantly through amino acid residues that are located in the
six heavy and light chain complementarity determining regions
(CDRs). For this reason, the amino acid sequences within CDRs are
more diverse between individual antibodies than sequences outside
of CDRs. Because CDR sequences are responsible for most
antibody-antigen interactions, it is possible to express
recombinant antibodies that mimic the properties of specific
naturally occurring antibodies by constructing expression vectors
that include CDR sequences from the specific naturally occurring
antibody grafted onto framework sequences from a different antibody
with different properties (see, e.g., Riechmann et al. (1998)
Nature 332:323-327; Jones. et al. (1986) Nature 321:522-525; Queen
et al. (1989) Proc. Natl. Acad. See. U.S.A. 86:10029-10033; U.S.
Pat. No. 5,225,539 to Winter, and U.S. Pat. Nos. 5,530,101;
5,585,089; 5,693,762 and 6,180,370 to Queen et al.)
[0165] Another type of variable region modification is to mutate
amino acid residues within the VH and/or VL CDR1, CDR2 and/or CDR3
regions to thereby improve one or more binding properties (e.g.,
affinity) of the antibody of interest. Site-directed mutagenesis or
PCR-mediated mutagenesis can be performed to introduce the
mutation(s) and the effect on antibody binding, or other functional
property of interest, can be evaluated in in vitro or in vivo
assays as described herein and provided in the Examples. Preferably
conservative modifications (as discussed above) are introduced. The
mutations may be amino acid substitutions, additions or deletions,
but are preferably substitutions. Moreover, typically no more than
five residues are altered within a CDR region are altered.
[0166] Engineered antibodies of the invention include those in
which modifications have been made to framework residues within
V.sub.H and/or V.sub.L, e.g. to improve the properties of the
antibody. Typically such framework modifications are made to
decrease the immunogenicity of the antibody. For example, one
approach is to "backmutate" one or more framework residues to the
corresponding germline sequence. More specifically, an antibody
that has undergone somatic mutation may contain framework residues
that differ from the germline sequence from which the antibody is
derived. Such residues can be identified by comparing the antibody
framework sequences to the germline sequences from which the
antibody is derived. To return the framework region sequences to
their germline configuration, the somatic mutations can be
"backmutated" to the germline sequence by, for example,
site-directed mutagenesis or PCR-mediated mutagenesis.
[0167] Another type of framework modification involves mutating one
or more residues within the framework region, or even within one or
more CDR regions, to remove T cell epitopes to thereby reduce the
potential immunogenicity of the antibody. This approach is also
referred to as "deimmunization" and is described in further detail
in U.S. patent Publication No. 20030153043 by Carr et al.
[0168] In addition or alternative to modifications made within the
framework or CDR regions, dual-specific antibodies of the invention
may be engineered to include modifications within the Fc region,
typically to alter one or more functional properties of the
antibody, such as serum half-life, complement fixation, Fc receptor
binding, and/or antigen-dependent cellular cytotoxicity.
Furthermore, an antibody of the invention may be chemically
modified (e.g., one or more chemical moieties can be attached to
the antibody) or be modified to alter it's glycosylation, again to
alter one or more functional properties of the antibody. Each of
these embodiments is described in further detail below. The
numbering of residues in the Fc region is that of the EU index of
Kabat.
[0169] In one embodiment, the hinge region of CH1 is modified such
that the number of cysteine residues in the hinge region is
altered, e.g., increased or decreased. This approach is described
further in U.S. Pat. No. 5,677,425 by Bodmer et al. The number of
cysteine residues in the hinge region of CH1 is altered to, for
example, facilitate assembly of the light and heavy chains or to
increase or decrease the stability of the dual-specific
antibody.
[0170] In another embodiment, the Fc hinge region of the antibody
of the invention is mutated to decrease the biological half life of
the antibody. More specifically, one or more amino acid mutations
are introduced into the CH2-CH3 domain interface region of the
Fe-hinge fragment such that the antibody has impaired
Staphylococcyl protein A (SpA) binding relative to native Fc-hinge
domain SpA binding. This approach is described in further detail in
U.S. Pat. No. 6,165,745 by Ward et al.
[0171] In another embodiment, the antibody of the invention is
modified to increase its biological half life. Various approaches
are possible. For example, one or more of the following mutations
can be introduced: T252L, T254S, T256F, as described in U.S. Pat.
No. 6,277,375 to Ward. Alternatively, to increase the biological
half life, the antibody may be altered within the CH1 or CL region
to contain a salvage receptor binding epitope taken from two loops
of a CH2 domain of an Fc region of an IgG, as described in U.S.
Pat. Nos. 5,869,046 and 6,121,022 by Presta et al.
[0172] In yet other embodiments, the Fc region is altered by
replacing at least one amino acid residue with a different amino
acid residue to alter the effector function(s) of the dual-specific
antibody. For example, one or more amino acids selected from amino
acid residues 234, 235, 236, 237, 297, 318, 320 and 322 can be
replaced with a different amino acid residue such that the antibody
has an altered affinity for an effector antibody but retains the
antigen-binding ability of the parent antibody. The effector
antibody to which affinity is altered can be, for example, an Fc
receptor or the C1 component of complement. This approach is
described in further detail in U.S. Pat. Nos. 5,624,821 and
5,648,260, both by Winter et at.
[0173] In another example, one or more amino acids selected from
amino acid residues 329, 331 and 322 can be replaced with a
different amino acid residue such that the dual-specific antibody
has altered Clq binding and/or reduced or abolished complement
dependent cytotoxicity (CDC). This approach is described in further
detail in U.S. Pat. No. 6,194,551 by Idusogie et al.
[0174] In another example, one or more amino acid residues within
amino acid positions 231 and 239 are altered to thereby alter the
ability of the antibody to fix complement. This approach is
described further in PCT Publication WO 94/29351 by Bodmer et
al.
[0175] In yet another example, the Fc region is modified to
increase the ability of the dual-specific antibody to mediate
antibody dependent cellular cytotoxicity (ADCC) and/or to increase
the affinity of the antibody for an Fey receptor by modifying one
or more amino acids at the following positions: 238, 239, 248, 249,
252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 276, 278,
280, 283, 285, 286, 289, 290, 292, 293, 294, 295, 296, 298, 301,
303, 305, 307, 309, 312, 315, 320, 322, 324, 326, 327, 329, 330,
331, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, 388,
389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or 439. This
approach is described further in PCT Publication WO 00/42072 by
Presta. Moreover, the binding sites on human IgG1 for Fc.gamma.RI,
Fc.gamma.RII, Fc.gamma.RIII and FcRn have been mapped and variants
with improved binding have been described (see Shields et al.,
(2001) J. Biol. Chem. 276:6591-6604). Specific mutations at
positions 256, 290, 298, 333, 334 and 339 were shown to improve
binding to Fc.gamma.RIII. Additionally, the following combination
mutants were shown to improve Fc.gamma./RIII binding: T256A/S298A,
S298A/E333A, S298A/K224A and S298A/E333A/K334A.
[0176] In still another embodiment, the glycosylation of an
antibody 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 framework 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 U.S. Pat. Nos. 5,714,350 and 6,350,861 by Co
et al.
[0177] Additionally or alternatively, an antibody can be made that
has an altered type of glycosylation, such as a hypofucosylated
antibody having reduced amounts of fucosyl residues 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 have been described in the art and can be
used as host cells in which to express recombinant antibodies of
the invention to thereby produce an antibody with altered
glycosylation. For example, EP 1,176,195 by Hanai et at describes a
cell line with a functionally disrupted FUT8 gene, which encodes a
fucosyl transferase, such that antibodies expressed in such a cell
line exhibit hypofircosylation. PCT Publication WO 03/035835 by
Presta describes a variant CHO cell line, Lec13 cells, with reduced
ability to attach fucose to Asn(297)-linked carbohydrates, also
resulting in hypofucosylation of antibodies expressed in that host
cell (see also Shields et at (2002) J. Biol. Chem.
277:26733-26740). PCT Publication WO 99/54342 by Umana et at
describes cell lines engineered to express glycoprotein-modifying
glycosyl transferases (e.g.,
beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) such that
antibodies expressed in the engineered cell lines exhibit increased
bisecting GlcNac structures which results in increased ADCC
activity of the antibodies (see also Umana et at (1999) Nat.
Biotech. 17:176-180).
[0178] Another modification of the antibodies herein that is
contemplated by the invention is pegylation. An antibody can be
pegylated to, for example, increase the biological (e.g., serum)
half life of the antibody. To pegylate an antibody, the antibody,
or fragment thereof, typically is reacted with polyethylene glycol
(PEG), such as a reactive ester or aldehyde derivative of PEG,
under conditions in which one or more PEG groups become attached to
the antibody or antibody fragment. Preferably, the pegylation is
carried out via an acylation reaction or an alkylation reaction
with a reactive PEG molecule (or an analogous reactive
water-soluble polymer). As used herein, the term "polyethylene
glycol" is intended to encompass any of the forms of PEG that have
been used to derivatize other proteins, such as mono (C1-C10)
alkoxy- or aryloxy-polyethylene glycol or polyethylene
glycol-maleimide. In certain embodiments, the antibody to be
pegylated is an aglycosylated antibody. Methods for pegylating
proteins are known in the art and can be applied to the antibodies
of the invention. See for example, EP 0 154 316 by Nishimura et al.
and EP 0 401 384 by Ishikawa et al.
[0179] In a further aspect, the present invention provides a
composition comprising a dual-specific antibody, or antigen-binding
portion thereof, obtainable by a method described herein or known
in the art, and a pharmaceutically acceptable carrier, diluent or
excipient.
[0180] Moreover, the present invention provides a method for the
treatment and/or prevention of disease using a dual-specific
antibody, or antigen-binding portion thereof, or a composition
according to the present invention.
III. Methods of Making IL-1.alpha./IL-1.beta. Dual-Specific
Antibodies
[0181] Dual-specific antibodies of the invention may be prepared
according to established techniques used in the field of antibody
engineering. Techniques for the preparation of antibodies, and in
particular dual-specific antibodies, are, for example, described in
the following reviews and the references cited therein: Winter
& Milstein, (1991) Nature 349:293-299; Plueckthun (1992)
Immunological Reviews 130:151-188; Wright et al., (1992) Crti. Rev.
Immuno1.12:125-168; Holliger, P. & Winter, G. (1993) Curr. Op.
Biotechn. 4, 446-449; Carter, et al. (1995) J. Hematother. 4,
463-470; Chester, & Hawkins (1995) Trends Biotechn. 13,
294-300; Hoogenboom, H. R. (1997) Nature Biotechnol. 15, 125-126;
Fearon, D. (1997) Nature Biotechnol. 15, 618-619; Pluckthun, A.
& Pack, P. (1997) Immunotechnology 3, 83-105; Carter &
Merchant (1997) Curr. Opin. Biotechnol. 8, 449-454; Holliger &
Winter (1997) Cancer Immunol. Immunother. 45, 128-130.
[0182] The invention provides for the selection of variable domains
against two different antigens or epitopes, i.e., IL-1.alpha. or
IL-1.beta., and subsequent combination of the variable domains into
a dual-specific antibody, or antigen-binding portion thereof.
[0183] The techniques employed for selection of the IL-1.alpha. or
IL-1.beta.-specific variable domains employ libraries and selection
procedures which are known in the art. Natural libraries (Marks et
al. (1991) J. Mol. Biol., 222: 581; Vaughan et al. (1996) Nature
Biotech., 14: 309) which use rearranged V genes harvested from
human B cells are well known to those skilled in the art. Synthetic
libraries (Hoogenboom & Winter (1992) J. Mol. Biol., 227: 381;
Barbas et al. (1992) Proc. Natl. Acad. Sci. USA, 89: 4457; Nissim
et al. (1994) EMBO J., 13: 692; Griffiths et al. (1994) EMBO J.,
13: 3245; De Kruif et al. (1995) J. Mol. Biol., 248: 97) are
prepared by cloning immunoglobulin V genes, usually using PCR
Errors in the PCR process can lead to a high degree of
randomisation. VH and/or VL libraries may be selected against
target antigens or epitopes separately, in which case single domain
binding is directly selected for, or together.
[0184] A preferred method for making a dual-specific antibody
according to the present invention comprises using a selection
system in which a repertoire of variable domains is selected for
binding to a first antigen or epitope (i.e., IL-1.alpha. or
IL-1.beta.) and a repertoire of variable domains is selected for
binding to a second antigen or epitope (i.e., IL-1.alpha. or
IL-1.beta.). The selected variable first and second variable
domains are then combined and the dual-specific antibody selected
for binding to both first and second antigen or epitope.
[0185] In one embodiment, the antibody or antigen-binding portion
thereof, is identified through a method comprising the general
steps of: (a) selecting a first variable domain by its ability to
bind to a first epitope (i.e., IL-1.alpha. or IL-1.beta.), (b)
selecting a second variable region by its ability to bind to a
second epitope (i.e., IL-1.alpha. or IL-1.beta.), (c) combining the
variable domains; and (d) selecting the antibody, or
antigen-binding portion thereof, by its ability to bind to said
first epitope and to said second epitope.
[0186] In a preferred embodiment of the invention, the variable
regions are selected from single domain V gene repertoires.
Generally the repertoire of single antibody domains is displayed on
the surface of filamentous bacteriophage. In a preferred embodiment
each single antibody domain is selected by binding of a phage
repertoire to antigen.
[0187] In a preferred embodiment of the invention each single
variable domain may be selected for binding to its target antigen
or epitope in the absence of a complementary variable region. In an
alternative embodiment, the single variable domains may be selected
for binding to its target antigen or epitope in the presence of a
complementary variable region. Thus the first single variable
domain may be selected in the presence of a third complementary
variable domain, and the second variable domain may be selected in
the presence of a fourth complementary variable domain. The
complementary third or fourth variable domain may be the natural
cognate variable domain having the same specificity as the single
domain being tested, or a non-cognate complementary domain--such as
a "dummy" variable domain.
Library Vector Systems
[0188] A variety of selection systems are known in the art which
are suitable for use in the present invention. Examples of such
systems are described below.
[0189] Phage display technology (see, e.g., Smith (1985) Science
228:1315; Scott and Smith (1990) Science 249:386; McCafferty at al.
(1990) Nature 348: 552) provides an approach for the selection of
antibody polypeptides which bind a desired target from among large,
diverse repertoires of antibody polypeptides. Phage display
libraries may contain synthetic libraries whereby germline V gene
segments are `rearranged` in vitro (Hoogenboom and Winter (1992) J
Mol Bio 227:381; Nissim et al. (1994) EMBO J., 13: 692; Griffiths
et al. (1994) EMBO J., 13: 3245; De Kruif et al. (1995) J. Mol.
Biol., 248: 97) or where synthetic CDRs are incorporated into a
single rearranged V gene (Barbas et al. (1992) PNAs 89:4457). Most
often, the antibody polypeptides displayed on the phage comprise
antigen-binding antibody fragments. In one embodiment, the antibody
polypeptides displayed on the phage are single domains (dAbs).
[0190] Bacteriophage lambda expression systems may be screened
directly as bacteriophage plaques or as colonies of lysogens, both
as previously described (Huse et al. (1989) Science, 246: 1275;
Caton and Koprowski (1990) Proc. Natl. Acad. Sci. U.S.A., 87;
Mullinax et al. (1990) Proc. Natl. Acad. Sci. U.S.A., 87: 8095;
Persson et al. (1991) Proc. Natl. Acad. Sci. U.S.A., 88: 2432) and
are of use in the invention. Whilst such expression systems can be
used to screen up to 10.sup.6 different members of a library, they
are not really suited to screening of larger numbers (greater than
10.sup.6 members).
[0191] Of particular use in the construction of libraries are
selection display systems, which enable a nucleic acid to be linked
to the polypeptide it expresses. As used herein, a selection
display system is a system that permits the selection, by suitable
display means, of the individual members of the library by binding
the generic and/or target ligands.
[0192] Selection protocols for isolating desired members of large
libraries are known in the art, as typified by phage display
techniques. Such systems, in which diverse peptide sequences are
displayed on the surface of filamentous bacteriophage (Scott and
Smith (1990) Science, 249: 386), have proven useful for creating
libraries of antibody fragments (and the nucleotide sequences that
encoding them) for the in vitro selection and amplification of
specific antibody fragments that bind a target antigen (McCafferty
et al., WO 92/01047). The nucleotide sequences encoding the VH and
VL regions are linked to gene fragments which encode leader signals
that direct them to the periplasmic space of E. coli and as a
result the resultant antibody fragments are displayed on the
surface of the bacteriophage, typically as fusions to bacteriophage
coat proteins (e.g., pIII or pVIII). Alternatively, antibody
fragments are displayed externally on lambda phage capsids
(phagebodies). An advantage of phage-based display systems is that,
because they are biological systems, selected library members can
be amplified simply by growing the phage containing the selected
library member in bacterial cells. Furthermore, since the
nucleotide sequence that encode the polypeptide library member is
contained on a phage or phagemid vector, sequencing, expression and
subsequent genetic manipulation is relatively straightforward.
[0193] Methods for the construction of bacteriophage antibody
display libraries and lambda phage expression libraries are well
known in the art (McCafferty et al. (1990) Nature, 348: 552; Kang
et al. (1991) Proc. Natl. Acad. Sci. U.S.A., 88: 4363; Clackson et
al. (1991) Nature, 352: 624; Lowman et al. (1991) Biochemistry, 30:
10832; Burton et al. (1991) Proc. Natl. Acad. Sci. U.S.A., 88:
10134; Hoogenboom et al. (1991) Nucleic Acids Res., 19: 4133; Chang
et al. (1991) J. Immunol., 147: 3610; Breitling et al. (1991) Gene,
104: 147; Marks et al. (1991) supra; Barbas et al. (1992) supra;
Hawkins and Winter (1992) J. Immunol., 22: 867; Marks et al, 1992,
J. Biol. Chem., 267:16007; Lerner et al. (1992) Science, 258:1313,
incorporated herein by reference).
[0194] One particularly advantageous approach has been the use of
scFv phage-libraries (Huston et al., 1988, Proc. Natl. Acad. Sci.
U.S.A., 85: 5879-5883; Chaudhary et al. (1990) Proc. Natl. Acad.
Sci. U.S.A., 87:1066-1070; McCafferty et al. (1990) supra; Clackson
et al. (1991) Nature, 352: 624; Marks et al. (1991) J. Mol. Biol.,
222: 581; Chiswell et al. (1992) Trends Biotech., 10: 80; Marks et
at (1992) J. Biol. Chem., 267). Various embodiments of scFv
libraries displayed on bacteriophage coat proteins have been
described. Refinements of phage display approaches are also known,
for example as described in WO96/06213 and WO92/01047 (Medical
Research Council et al.) and WO97/08320 (Morphosys), which are
incorporated herein by reference.
[0195] Other systems for generating libraries of polypeptides
involve the use of cell-free enzymatic machinery for the in vitro
synthesis of the library members. In one method, RNA molecules are
selected by alternate rounds of selection against a target antibody
and PCR amplification (Tuerk and Gold (1990) Science, 249: 505;
Ellington and Szostak (1990) Nature, 346: 818). A similar technique
may be used to identify DNA sequences which bind a predetermined
human transcription factor (Thiesen and Bach (1990) Nucleic Acids
Res., 18: 3203; Beaudry and Joyce (1992) Science, 257: 635;
WO92/05258 and WO92/14843). In a similar way, in vitro translation
can be used to synthesize polypeptides as a method for generating
large libraries. These methods which generally comprise stabilized
polysome complexes, are described further in WO88/08453,
WO90/05785, WO90/07003, WO91/02076, WO91/05058, and WO92/02536.
Alternative display systems which are not phage-based, such as
those disclosed in WO95/22625 and WO95/11922 (Affymax) use the
polysomes to display polypeptides for selection.
[0196] A still further category of techniques involves the
selection of repertoires in artificial compartments, which allow
the linkage of a gene with its gene product. For example, a
selection system in which nucleic acids encoding desirable gene
products may be selected in microcapsules formed by water-in-oil
emulsions is described in WO99/02671, WO00/40712 and Tawfik &
Griffiths (1998) Nature Biotechnol 16(7), 652-6. Genetic elements
encoding a gene product having a desired activity are
compartmentalized into microcapsules and then transcribed and/or
translated to produce their respective gene products (RNA or
protein) within the microcapsules. Genetic elements which produce
gene product having desired activity are subsequently sorted. This
approach selects gene products of interest by detecting the desired
activity by a variety of means.
Library Construction
[0197] Libraries intended for selection, may be constructed using
techniques known in the art, for example as set forth above, or may
be purchased from commercial sources. Libraries which are useful in
the present invention are described, for example, in WO99/20749.
Once a vector system is chosen and one or more nucleic acid
sequences encoding polypeptides of interest are cloned into the
library vector, one may generate diversity within the cloned
molecules by undertaking mutagenesis prior to expression;
alternatively, the encoded proteins may be expressed and selected,
as described above, before mutagenesis and additional rounds of
selection are performed. Mutagenesis of nucleic acid sequences
encoding structurally optimized polypeptides is carried out by
standard molecular methods. Of particular use is the polymerase
chain reaction, or PCR, (Mullis and Faloona (1987) Methods
Enzymol., 155: 335, herein incorporated by reference). PCR, which
uses multiple cycles of DNA replication catalyzed by a
thermostable, DNA-dependent DNA polymerase to amplify the target
sequence of interest, is well known in the art. The construction of
various antibody libraries has been discussed in Winter et al.
(1994) Ann. Rev. Immunology 12, 433-55, and references cited
therein.
[0198] For example, PCR may be performed using template DNA (at
least 1 fg; more usefully, 1-1000 ng) and at least 25 pmol of
oligonucleotide primers; it may be advantageous to use a larger
amount of primer when the primer pool is heavily heterogeneous, as
each sequence is represented by only a small fraction of the
molecules of the pool, and amounts become limiting in the later
amplification cycles. A typical reaction mixture includes: 2 .mu.l
of DNA, 25 pmol of oligonucleotide primer, 2.5 .mu.l of
10.times.PCR buffer 1 (Perkin-Elmer, Foster City, Calif.), 0.4
.mu.l of 1.25 .mu.M dNTP, 0.15 .mu.l (or 2.5 units) of Taq DNA
polymerase (Perkin Elmer, Foster City, Calif.) and deionized water
to a total volume of 25 .mu.l. Mineral oil is overlaid and the PCR
is performed using a programmable thermal cycler. The length and
temperature of each step of a PCR cycle, as well as the number of
cycles, is adjusted in accordance to the stringency requirements in
effect. Annealing temperature and timing are determined both by the
efficiency with which a primer is expected to anneal to a template
and the degree of mismatch that is to be tolerated; obviously, when
nucleic acid molecules are simultaneously amplified and
mutagenised, mismatch is required, at least in the first round of
synthesis. The ability to optimize the stringency of primer
annealing conditions is well within the knowledge of one of
moderate skill in the art. An annealing temperature of between
30.degree. C. and 72.degree. C. is used. Initial denaturation of
the template molecules normally occurs at between 92.degree. C. and
99.degree. C. for 4 minutes, followed by 20-40 cycles consisting of
denaturation (94-99.degree. C. for 15 seconds to 1 minute),
annealing (temperature determined as discussed above; 1-2 minutes),
and extension (72.degree. C. for 1-5 minutes, depending on the
length of the amplified product). Final extension is generally for
4 minutes at 72.degree. C., and may be followed by an indefinite
(0-24 hour) step at 4.degree. C.
[0199] Vector constructs or libraries of vectors containing
polynucleotide molecules as described herein can be introduced to
selected host cells by any of a number of suitable methods known in
the art. For example, vector constructs may be introduced to
appropriate bacterial cells by infection, in the case of
bacteriophage vector particles such as lambda or M13, or any of a
number of transformation methods for plasmid vectors or for
bacteriophage DNA.
[0200] In one embodiment, a phage displayed repertoire of V.sub.H
or V.sub.L domains is screened by panning against either
IL-1.alpha. or IL-1.beta..
Combining Single Variable Domains
[0201] IL-1.alpha. or IL-1.beta. specific domains useful in the
invention, once selected, may be combined by a variety of methods
known in the art, including by covalent and non-covalent
methods.
[0202] Preferred methods include the use of polypeptide linkers, as
described, for example, in connection with scFv molecules (Bird et
al., (1988) Science 242:423-426). Discussion of suitable linkers is
provided in Bird et al. Science 242, 423-426; Hudson et al, Journal
Immunol Methods 231 (1999) 177-189; Hudson et al, Proc Nat Acad Sci
USA 85, 5879-5883. Linkers are preferably flexible, allowing the
two single domains to interact. One linker example is a (Gly.sub.4
Ser).sub.n linker, where n=1 to 8, e.g., 2, 3, 4, 5 or 7. The
linkers used in diabodies, which are less flexible, may also be
employed (Holliger et at, (1993) PNAS (USA) 90:6444-6448).
[0203] In one embodiment, the linker employed is not an
immunoglobulin hinge region.
[0204] Variable domains may be combined using methods other than
linkers. For example, the use of disulphide bridges, provided
through naturally-occurring or engineered cysteine residues, may be
exploited to stabilise VH-VH, VL-VL or VH-VL dimers (Reiter et al.,
(1994) Protein Eng. 7:697-704) or by remodelling the interface
between the variable domains to improve the "fit" and thus the
stability of interaction (Ridgeway et al., (1996) Protein Eng.
7:617-621; Zhu et. al., (1997) Protein Science 6:781-788).
Characterization of IL-1 Antibody
[0205] The binding of an antibody to its specific antigens or
epitopes can be tested by methods which will be familiar to those
skilled in the art and include ELISA.
[0206] In a one embodiment of the invention, binding of the single
domain of the invention is tested using monoclonal phage ELISA
according to standard methods.
[0207] Populations of phage produced at each round of selection can
be screened for binding by ELISA to the selected antigen or
epitope, to identify "polyclonal" phage antibodies. Phage from
single infected bacterial colonies from these populations can then
be screened by ELISA to identify "monoclonal" phage antibodies. It
is also desirable to screen soluble antibody fragments for binding
to antigen or epitope, and this can also be undertaken by ELISA
using reagents, for example, against a C- or N-terminal tag (see
for example Winter et al. (1994) Ann. Rev. Immunology 12, 433-55
and references cited therein.
[0208] The diversity of the selected phage monoclonal antibodies
may also be assessed by gel electrophoresis of PCR products (Marks
et al. 1991, supra; Nissim et al. 1994 supra), probing (Tomlinson
et al., 1992) J. Mol. Biol. 227, 776) or by sequencing of the
vector DNA.
[0209] Assays for detecting IL-1 are known in the art and may be
used to determine the ability of a dual-specific antibody for
neutralizing IL-1.alpha. and/or IL-113.
Structure of Dual-Specific Antibodies
[0210] As described above, an antibody is herein defined as an
antibody (for example IgG, IgM, IgA, IgA, IgE) or fragment (Fab,
Fv, disulphide linked Fv, scFv, diabody) which comprises at least
one heavy and a light chain variable domain, at least two heavy
chain variable domains or at least two light chain variable
domains. An antibody may be at least partly derived from any
species naturally producing an antibody, or created by recombinant
DNA technology; whether isolated from serum, B-cells, hybridomas,
transfectomas, yeast or bacteria).
[0211] In a preferred embodiment of the invention the dual-specific
antibody comprises at least one single heavy chain variable domain
of an antibody and one single light chain variable domain of an
antibody, or two single heavy or light chain variable domains. For
example, the antibody may comprise a VH/VL pair, a pair of VH
domains, or a pair of VL domains.
[0212] The first and the second variable domains of such an
antibody may be on the same polypeptide chain. Alternatively they
may be on separate polypeptide chains. In the case that they are on
the same polypeptide chain they may be linked by a linker, which is
preferentially a peptide sequence, as described above.
[0213] The first and second variable domains may be covalently or
non-covalently associated. In the case that they are covalently
associated, the covalent bonds may be disulphide bonds.
[0214] In the case that the variable domains are selected from
V-gene repertoires selected for instance using phage display
technology as herein described, then these variable domains
comprise a universal framework region, such that is they may be
recognised by a specific generic ligand as herein defined. The use
of universal frameworks, generic ligands and the like is described
in WO99/20749.
[0215] Where V-gene repertoires are used variation in polypeptide
sequence is preferably located within the structural loops of the
variable domains. The polypeptide sequences of either variable
domain may be altered by DNA shuffling or by mutation in order to
enhance the interaction of each variable domain with its
complementary pair. DNA shuffling is known in the art and taught,
for example, by Stemmer, 1994, Nature 370: 389-391 and U.S. Pat.
No. 6,297,053, both of which are incorporated herein by reference.
Other methods of mutagenesis are well known to those of skill in
the art.
[0216] According to another aspect of the invention,
advantageously, the epitope binding domains identified in the
screening process described above, are attached to a "protein
skeleton". Advantageously, a protein skeleton according to the
invention is an immunoglobulin skeleton.
[0217] According to the present invention, the term immunoglobulin
skeleton refers to a protein which comprises at least one
immunoglobulin fold and which acts as a nucleus for one or more
epitope binding domains, as defined herein.
[0218] Preferred immunoglobulin skeletons as herein defined
includes any one or more of those selected from the following: an
immunoglobulin molecule comprising at least (i) the CL (kappa or
lambda subclass) domain of an antibody; or (ii) the CH1 domain of
an antibody heavy chain; an immunoglobulin molecule comprising the
CH1 and CH2 domains of an antibody heavy chain; an immunoglobulin
molecule comprising the CH1, CH2 and CH3 domains of an antibody
heavy chain; or any of the subset (ii) in conjunction with the CL
(kappa or lambda subclass) domain of an antibody. A hinge region
domain may also be included. Such combinations of domains may, for
example, mimic natural antibodies, such as IgG or IgM, or fragments
thereof, such as Fv, scFv, Fab or F(ab').sub.2 molecules. Those
skilled in the art will be aware that this list is not intended to
be exhaustive.
[0219] Linking of the skeleton to the epitope binding domains, as
herein defined may be achieved at the polypeptide level, that is
after expression of the nucleic acid encoding the skeleton and/or
the epitope binding domains. Alternatively, the linking step may be
performed at the nucleic acid level. Methods of linking a protein
skeleton according to the present invention, to the one or more
epitope binding domains include the use of protein chemistry and/or
molecular biology techniques which will be familiar to those
skilled in the art and are described herein.
[0220] Skeletons may be based on immunoglobulin molecules or may be
non-immunoglobulin in origin as set forth above. Preferred
immunoglobulin skeletons as herein defined includes any one or more
of those selected from the following: an immunoglobulin molecule
comprising at least (i) the CL (kappa or lambda subclass) domain of
an antibody; or (ii) the CH1 domain of an antibody heavy chain; an
immunoglobulin molecule comprising the CH1 and CH2 domains of an
antibody heavy chain; an immunoglobulin molecule comprising the
CH1, CH2 and CH3 domains of an antibody heavy chain; or any of the
subset (ii) in conjunction with the CL (kappa or lambda subclass)
domain of an antibody. A hinge region domain may also be included.
Such combinations of domains may, for example, mimic natural
antibodies, such as IgG or IgM, or fragments thereof, such as Fv,
scFv, Fab or F(ab).sub.2 molecules. Those skilled in the art will
be aware that this list is not intended to be exhaustive.
[0221] In a one embodiment of the invention the dual-specific
antibody is a single chain Fv fragment. In an alternative
embodiment of the invention, the dual-specific antibody consists of
a Fab format.
[0222] In a further aspect, the present invention provides nucleic
acid encoding a dual-specific antibody as herein defined. In
another embodiment, the invention provides a nucleic acid encoding
a dAb identified herein.
[0223] One skilled in the art will appreciate that, depending on
the aspect of the invention, both antigens or epitopes may bind
simultaneously to the same antibody molecule. Alternatively, they
may compete for binding to the same antibody molecule. For example,
where both epitopes are bound simultaneously, both variable domains
of a dual-specific antibodies are able to independently bind their
target epitopes. Where the domains compete, the one variable domain
is capable of binding its target, but not at the same time as the
other variable domain binds its cognate target; or the first
variable domain is capable of binding its target, but not at the
same time as the second variable domain binds its cognate
target.
[0224] The variable regions may be derived from antibodies directed
against target antigens or epitopes. Alternatively they may be
derived from a repertoire of single antibody domains such as those
expressed on the surface of filamentous bacteriophage. Selection
may be performed as described above and in the Examples provided
herein.
[0225] In general, the nucleic acid molecules and vector constructs
required for the performance of the present invention may be
constructed and manipulated as set forth in standard laboratory
manuals, such as Sambrook et al. (1989) Molecular Cloning: A
Laboratory Manual, Cold Spring Harbor, USA.
[0226] The manipulation of nucleic acids useful in the present
invention is typically carried out in recombinant vectors.
[0227] Thus in a further aspect, the present invention provides a
vector comprising nucleic acid encoding at least a dual-specific
antibody as herein defined. In another embodiment, the invention
provides a vector comprising a nucleic acid encoding a dAb
identified herein.
[0228] Methods by which to select or construct and, subsequently,
use vectors are well known to one of ordinary skill in the art.
Numerous vectors are publicly available, including bacterial
plasmids, bacteriophage, artificial chromosomes and episomal
vectors. Such vectors may be used for simple cloning and
mutagenesis; alternatively gene expression vector is employed. A
vector of use according to the invention may be selected to
accommodate a polypeptide coding sequence of a desired size,
typically from 0.25 kilobase (kb) to 40 kb or more in length A
suitable host cell is transformed with the vector after in vitro
cloning manipulations. Each vector contains various functional
components, which generally include a cloning (or "polylinker")
site, an origin of replication and at least one selectable marker
gene. If given vector is an expression vector, it additionally
possesses one or more of the following: enhancer element, promoter,
transcription termination and signal sequences, each positioned in
the vicinity of the cloning site, such that they are operatively
linked to the gene encoding an antibody according to the
invention.
[0229] Both cloning and expression vectors generally contain
nucleic acid sequences that enable the vector to replicate in one
or more selected host cells. Typically in cloning vectors, this
sequence is one that enables the vector to replicate independently
of the host chromosomal DNA and includes origins of replication or
autonomously replicating sequences. Such sequences are well known
for a variety of bacteria, yeast and viruses. The origin of
replication from the plasmid pBR322 is suitable for most
Gram-negative bacteria, the 2 micron plasmid origin is suitable for
yeast, and various viral origins (e.g. SV 40, adenovirus) are
useful for cloning vectors in mammalian cells. Generally, the
origin of replication is not needed for mammalian expression
vectors unless these are used in mammalian cells able to replicate
high levels of DNA, such as COS cells.
[0230] Advantageously, a cloning or expression vector may contain a
selection gene also referred to as selectable marker. This gene
encodes a protein necessary for the survival or growth of
transformed host cells grown in a selective culture medium. Host
cells not transformed with the vector containing the selection gene
will therefore not survive in the culture medium. Typical selection
genes encode proteins that confer resistance to antibiotics and
other toxins, e.g. ampicillin, neomycin, methotrexate or
tetracycline, complement auxotrophic deficiencies, or supply
critical nutrients not available in the growth media.
[0231] Since the replication of vectors encoding an antibody
according to the present invention is most conveniently performed
in E. coli, an E. coli-selectable marker, for example, the
.beta.-lactamase gene that confers resistance to the antibiotic
ampicillin, is of use. These can be obtained from E. coli plasmids,
such as pBR322 or a pUC plasmid such as pUC18 or pUC19.
[0232] Expression vectors usually contain a promoter that is
recognised by the host organism and is operably linked to the
coding sequence of interest. Such a promoter may be inducible or
constitutive. 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.
[0233] Promoters suitable for use with prokaryotic hosts include,
for example, the .beta.-lactamase and lactose promoter systems,
alkaline phosphatase, the tryptophan (trp) promoter system and
hybrid promoters such as the tac promoter. Promoters for use in
bacterial systems will also generally contain a Shine-Delgarno
sequence operably linked to the coding sequence.
[0234] The preferred vectors are expression vectors that enable the
expression of a nucleotide sequence corresponding to a polypeptide
library member. Thus, selection with the first and/or second
antigen or epitope can be performed by separate propagation and
expression of a single clone expressing the polypeptide library
member or by use of any selection display system. As described
above, the preferred selection display system is bacteriophage
display. Thus, phage or phagemid vectors may be used, eg pIT1 or
pIT2. Leader sequences useful in the invention include pelB, stII,
ompA, phoA, bla and pelA. One example are phagemid vectors which
have an E. coli origin of replication (for double stranded
replication) and also a phage origin of replication (for production
of single-stranded DNA). The manipulation and expression of such
vectors is well known in the art (Hoogenboom and Winter (1992)
supra; Nissim et al. (1994) supra). Briefly, the vector contains a
.beta.-lactamase gene to confer selectivity on the phagemid and a
lac promoter upstream of a expression cassette that consists (N to
C terminal) of a pelB leader sequence (which directs the expressed
polypeptide to the periplasmic space), a multiple cloning site (for
cloning the nucleotide version of the library member), optionally,
one or more peptide tag (for detection), optionally, one or more
TAG stop codon and the phage protein pIII. Thus, using various
suppressor and non-suppressor strains of E. coli and with the
addition of glucose, iso-propyl thio-.beta.-D-galactoside (IPTG) or
a helper phage, such as VCS M13, the vector is able to replicate as
a plasmid with no expression, produce large quantities of the
polypeptide library member only or produce phage, some of which
contain at least one copy of the polypeptide-pIII fusion on their
surface.
[0235] The invention provides improved vectors for expressing an
antibody heavy or light chain, or an antigen-binding portion
thereof. The vectors of the invention may also be used for library
screening, as they provide efficient cloning for screening
purposes. In one embodiment, the invention provides an improved
recombinant expression vector comprising a shifter sequence, e.g.,
1 kb, which is in between an upstream signal sequence and a
downstream Ig constant region sequence. In order to make an
antibody heavy or light chain construct, the stuffer sequence in
the chosen master template (or vector) is removed by restriction
enzyme digestion followed by inserting the desired antibody V
domain sequence (e.g., VH, V.kappa., or V.gamma.).
[0236] The resulting plasmid construct is easily propogated in and
purified from E. coli and can be used to express antibody by
co-transfecting both a heavy chain and a light chain construct in
mammalian cells, such as COS cells. Examples of vector sequences
encompassed by the invention are provided in SEQ ID NO: 844, SEQ ID
NO: 845, SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848, SEQ ID NO:
849, SEQ ID NO: 850, SEQ ID NO: 851, SEQ ID NO: 852, SEQ ID NO:
853, SEQ ID NO: 854, and SEQ ID NO: 855. It should be noted that
nucleic acid sequences that are 80% identical, 90% identical, 95%
identical, 98% identical, and 99% identical to SEQ ID NO: 844, SEQ
ID NO: 845, SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848, SEQ ID
NO: 849, SEQ ID NO: 850, SEQ ID NO: 851, SEQ ID NO: 852, SEQ ID NO:
853, SEQ ID NO: 854, and SEQ ID NO: 855 are also contemplated as
part of the invention. An example of the vector of the invention is
also provided in FIG. 19.
[0237] The size of the stuffer sequence will vary depending on the
gene of interest being ligated or recombined into the vector.
Generally, the stuffer sequence cannot be too small, i.e., less
than 100 bp, as it should be large enough to visualize on an
analytical agarose gel to confirm the excision of the stuffer. In
theory, however, the theoretical lower limitation may be about 10
base pairs, as this size may provide enough distance between the
two sites for efficient enzyme cutting (without confirmation by gel
electrophresis). While there is no upper limit in size to the
stuffer sequence, it is not desirable to have a stutter sequence
which is larger than the vector itself, especially for
distinguishing between the stuffer and linear cut vector on an
agarose gel. In one embodiment, the stuffer sequence is less than 3
kilobases, but larger than 10 base pairs. In another embodiment,
the stuffer sequence is less than 3 kilobases, but larger than 50
base pairs. In another embodiment, the stuffer sequence is less
than 3 kilobases, but larger than 80 base pairs. In another
embodiment, the stuffer sequence is less than 3 kilobases, but
larger than 100 base pairs. In one embodiment, the stuffer sequence
is less than 1 kilobase, but larger than 10 base pairs. In another
embodiment, the stuffer sequence is less than 1 kilobase, but
larger than 50 base pairs. In another embodiment, the stuffer
sequence is less than 1 kilobase, but larger than 80 base pairs. In
another embodiment, the stuffer sequence is less than 1 kilobases,
but larger than 100 base pairs. Examples of stuffer sequences are
described in Table 36, and include nucleotides 124 to 1103 of SEQ
ID NO: 855, 124 to 1100 of SEQ ID NOs: 844 to 849, and 132 to 1100
of SEQ ID NO: 850. Also included within the invention are nucleic
acids having high homology to the stuffer sequences described
herein, i.e., nucleic acid sequences that are 80%, 90%, 95%, 98%,
or 99% identical to the stuffer sequences described herein.
[0238] In one embodiment, the stuffer sequence used in the vector
of the invention contains certain restriction enzyme sites at the
5', 3' or both the 5' and 3' ends. Examples of such sites include,
but are not limited to, NruI, FspAI, or a combination thereof at
the 5' end of the stuffer sequence and AfeI, SnaBI, BsiWI, HpaI,
SalI, or a combination thereof at the 3' end of the stuffer
sequence. Stuffer sequences having advantageous restriction sites
at the 5' and 3' ends are also described in FIG. 18.
[0239] Another important feature of the vector of the invention is
that it contains heavy or light chain constant region sequences
which may be operably linked to an insertion site for a nucleic
acid encoding a variable chain region or another protein (e.g, to
form an Fc fussion protein). The vector may contain any isotype of
antibody, e.g., IgG, IgA, IgE, IgM, and IgD. In one embodiment, the
constant region corresponds to a murine heavy or light chain
sequence, including murine lambda, murine IgG2b, and murine IgG3.
Alternatively, the constant region may be human, as described in
the vector of Example 4. Examples of constant regions that may be
used in the vector are described in Tables 36 and 37, as well as in
the vectors described in SEQ ID NO: 844, SEQ ID NO: 845, SEQ ID NO:
846, SEQ ID NO: 847, SEQ ID NO: 848, SEQ ID NO: 849, SEQ ID NO:
850, SEQ ID NO: 851, SEQ ID NO: 852, SEQ ID NO: 853, SEQ ID NO:
854, and SEQ ID NO: 855.
[0240] In one embodiment, the vector comprises an episomal origin
of replication which is an
[0241] SV40 origin of replication. The SV40 (Simian Virus 40)
origin of replication (described in FIG. 19) requires a single
viral protein, the large T-antigen, for initiation of replication
of the vector via this origin. The SV40 origin of replication may
be used in episomal vectors to replicate and maintain said vector
(see Cabs (1996) Trends Genetics 12: 462; Harrison et al. (1994) J
Virol 68:1913; Cooper et al. (1997) PNAS 94:6450; and Ascenziono et
al. (1997) Cancer Lett 118:135).
[0242] In one embodiment, the vector of the invention may be used
to express an Fc fusion protein. As used herein, the terms
"linked," "fused" or "fusion" are used interchangeably. These terms
refer to the joining together of two more elements or components,
by whatever means including chemical conjugation or recombinant
means. An "in-frame fusion" or "operably linked" refers to the
joining of two or more open reading frames (ORFs) to form a
continuous longer ORF, in a manner that maintains the correct
reading frame of the original ORFs. Thus, the resulting recombinant
fusion protein is a single protein containing two ore more segments
that correspond to polypeptides encoded by the original ORFs (which
segments are not normally so joined in nature.) Although the
reading frame is thus made continuous throughout the fused
segments, the segments may be physically or spatially separated by,
for example, an in-frame linker sequence.
[0243] As used herein, the term "Fc region" includes amino acid
sequences derived from the constant region of an antibody heavy
chain. In some embodiments, an Fc region includes a polypeptide
comprising the constant region of an antibody excluding the first
constant region immunoglobulin domain.
[0244] The terms "Fc fusion" or "Fc fusion protein", as used
herein, include a protein wherein one or more proteins,
polypeptides or small molecules is operably linked to an Fc region
or derivative thereof. The term "Fc fusion" as used herein is
intended to be synonymous with terms such as "Ig fusion", "Ig
chimera", and "receptor globulin" (sometimes with dashes) as used
in the prior art (Chamow et al., 1996, Trends Biotechnol 14:52-60;
Ashkenazi et al., 1997, Curr Opin Immunol 9:195-200). An Fc fusion
combines one or more Fc regions, or variant(s) thereof, of an
immunoglobulin with a fusion partner, which in general can be any
protein, polypeptide, peptide, or small molecule. In some
embodiments, the role of the non-Fc part of an Fc fusion, i.e., the
fusion partner, may be to mediate target binding, and thus it can
be functionally analogous to the variable regions of an
antibody.
[0245] Construction of vectors encoding antibodies according to the
invention employs conventional ligation techniques. Isolated
vectors or DNA fragments are cleaved, tailored, and religated in
the form desired to generate the required vector. If desired,
analysis to confirm that the correct sequences are present in the
constructed vector can be performed in a known fashion.
[0246] Suitable methods for constructing expression vectors,
preparing in vitro transcripts, introducing DNA into host cells,
and performing analyses for assessing expression and function are
known to those skilled in the art. The presence of a gene sequence
in a sample is detected, or its amplification and/or expression
quantified by conventional methods, such as Southern or Northern
analysis, Western blotting, dot blotting of DNA, RNA or protein, in
situ hybridisation, immunocytochemistry or sequence analysis of
nucleic acid or protein molecules. Those skilled in the art will
readily envisage how these methods may be modified, if desired.
[0247] Techniques for determining nucleic acid and amino acid
"sequence identity" also are known in the art. Typically, such
techniques include determining the nucleotide sequence of the mRNA
for a gene and/or determining the amino acid sequence encoded
thereby, and comparing these sequences to a second nucleotide or
amino acid sequence. In general, "identity" refers to an exact
nucleotide-to-nucleotide or amino acid-to-amino acid correspondence
of two polynucleotides or polypeptide sequences, respectively. Two
or more sequences (polynucleotide or amino acid) can be compared by
determining their "percent identity." The percent identity of two
sequences, whether nucleic acid or amino acid sequences, is the
number of exact matches between two aligned sequences divided by
the length of the shorter sequences and multiplied by 100. An
approximate alignment for nucleic acid sequences is provided by the
local homology algorithm of Smith and Waterman, Advances in Applied
Mathematics 2:482-489 (1981). This algorithm can be applied to
amino acid sequences by using the scoring matrix developed by
Dayhoff, Atlas of Protein Sequences and Structure, M. O. Dayhoff
ed., 5 suppl. 3:353-358, National Biomedical Research Foundation,
Washington, D.C., USA, and normalized by Gribskov, Nucl. Acids Res.
14(6):6745-6763 (1986). An exemplary implementation of this
algorithm to determine percent identity of a sequence is provided
by the Genetics Computer Group (Madison, Wis.) in the "BestFit"
utility application. The default parameters for this method are
described in the Wisconsin Sequence Analysis Package Program
Manual, Version 8 (1995) (available from Genetics Computer Group,
Madison, Wis.). A preferred method of establishing percent identity
in the context of the present invention is to use the MPSRCH
package of programs copyrighted by the University of Edinburgh,
developed by John F. Collins and Shane S. Sturrok, and distributed
by IntelliGenetics, Inc. (Mountain View, Calif.). From this suite
of packages the Smith-Wateman algorithm can be employed where
default parameters are used for the scoring table (for example, gap
open penalty of 12, gap extension penalty of one, and a gap of
six). From the data generated the "Match" value reflects "sequence
identity." Other suitable programs for calculating the percent
identity or similarity between sequences are generally known in the
art.
[0248] Two nucleic acid fragments are considered to "selectively
hybridize" as described herein. The degree of sequence identity
between two nucleic acid molecules affects the efficiency and
strength of hybridization events between such molecules. A
partially identical nucleic acid sequence will at least partially
inhibit a completely identical sequence from hybridizing to a
target molecule. Inhibition of hybridization of the completely
identical sequence can be assessed using hybridization assays that
are well known in the art (e.g., Southern blot, Northern blot,
solution hybridization, or the like, see Sambrook et al., 1989,
Molecular Cloning: A Laboratory Manual, Cold Spring Harbor
Laboratories, New York; or Ausubel et al. (Eds.), Current Protocols
In Molecular Biology, John Wiley & Sons, Inc., New York
(1997)). Such assays can be conducted using varying degrees of
selectivity, for example, using conditions varying from low to high
stringency. If conditions of low stringency are employed, the
absence of non-specific binding can be assessed using a secondary
probe that lacks even a partial degree of sequence identity (for
example, a probe having less than about 30% sequence identity with
the target molecule), such that, in the absence of non-specific
binding events, the secondary probe will not hybridize to the
target.
[0249] When utilizing a hybridization-based detection system, a
nucleic acid probe is chosen that is complementary to a target
nucleic acid sequence, and then by selection of appropriate
conditions the probe and the target sequence "selectively
hybridize," or bind, to each other to form a hybrid molecule. A
nucleic acid molecule that is capable of hybridizing selectively to
a target sequence under "moderately stringent" typically hybridizes
under conditions that allow detection of a target nucleic acid
sequence of at least about 10-14 nucleotides in length having at
least approximately 70% sequence identity with the sequence of the
selected nucleic acid probe. Stringent hybridization conditions
typically allow detection of target nucleic acid sequences of at
least about 10-14 nucleotides in length having a sequence identity
of greater than about 90-95% with the sequence of the selected
nucleic acid probe. Hybridization conditions useful for
probe/target hybridization where the probe and target have a
specific degree of sequence identity, can be determined as is known
in the art (see, for example, Nucleic Acid Hybridization: A
Practical Approach, editors B. D. Hames and S. J. Higgins, (1985)
Oxford; Washington, D.C.; IRL Press).
[0250] With respect to stringency conditions for hybridization, it
is well known in the art that numerous equivalent conditions can be
employed to establish a particular stringency by varying, for
example, the following factors: the length and nature of probe and
target sequences, base composition of the various sequences,
concentrations of salts and other hybridization solution
components, the presence or absence of blocking agents in the
hybridization solutions (e.g., formamide, dextran sulfate, and
polyethylene glycol), hybridization reaction temperature and time
parameters, as well as, varying wash conditions. The selection of a
particular set of hybridization conditions is selected following
standard methods in the art (see, for example, see Sambrook, et
al., supra or Ausubel et al., supra).
[0251] A first polynucleotide is "derived from" second
polynucleotide if it has the same or substantially the same base
pair sequence as a region of the second polynucleotide, its cDNA,
complements thereof, or if it displays sequence identity as
described above. A first polypeptide is "derived from" a second
polypeptide if it is (i) encoded by a first polynucleotide derived
from a second polynucleotide, or (ii) displays sequence identity to
the second polypeptides as described above.
[0252] The invention also provides a kit containing one or more
vectors of the invention in a suitable vessel such as a vial. The
expression vectors can contain at least one cloning site for
insertion of a selected sequence of interest, or can have a
specific gene of interest already present in the vector. The vector
an be provided in a dehydrated or lyophilized form, or in an
aqueous solution. The kit can include a buffer for reconstituting
the dehydrated polynucleotide. Other reagents can be included in
the kit, e.g., reaction buffers, positive and negative control
vectors for comparison. Generally, the kit will also include
instructions for use of the reagents therein.
Construction of IL-1 Dual-Specific Antibody: Selection of
Main-Chain Conformation
[0253] The members of the immunoglobulin superfamily all share a
similar fold for their polypeptide chain. For example, although
antibodies are highly diverse in terms of their primary sequence,
comparison of sequences and crystallographic structures has
revealed that, contrary to expectation, five of the six antigen
binding loops of antibodies (H1, H2, L1, L2, L3) adopt a limited
number of main-chain conformations, or canonical structures
(Chothia and Lesk (1987) J. Mol. Biol., 196: 901; Chothia et al.
(1989) Nature, 342: 877). Analysis of loop lengths and key residues
has therefore enabled prediction of the main-chain conformations of
H1, H2, L1, L2 and L3 found in the majority of human antibodies
(Chothia et al. (1992) J. Mol. Biol., 227: 799; Tomlinson et al.
(1995) EMBO J., 14: 4628; Williams et al. (1996) J. Mol. Biol.,
264: 220). Although the H3 region is much more diverse in terms of
sequence, length and structure (due to the use of D segments), it
also forms a limited number of main-chain conformations for short
loop lengths which depend on the length and the presence of
particular residues, or types of residue, at key positions in the
loop and the antibody framework (Martin et al. (1996) J. Mol.
Biol., 263: 800; Shirai et al. (1996) FEBS Letters, 399: 1).
[0254] The dual-specific antibodies of the present invention are
advantageously assembled from libraries of domains, such as
libraries of V.sub.H domains and/or libraries of V.sub.L domains.
Moreover, the dual-specific antibodies of the invention may
themselves be provided in the form of libraries. In one aspect of
the present invention, libraries of dual-specific antibodies and/or
domains are designed in which certain loop lengths and key residues
have been chosen to ensure that the main-chain conformation of the
members is known. Advantageously, these are real conformations of
immunoglobulin superfamily molecules found in nature, to minimise
the chances that they are non-functional, as discussed above.
Germline V gene segments serve as one suitable basic framework for
constructing antibody or T-cell receptor libraries; other sequences
are also of use. Variations may occur at a low frequency, such that
a small number of functional members may possess an altered
main-chain conformation, which does not affect its function.
[0255] Canonical structure theory is also of use to assess the
number of different main-chain conformations encoded by ligands, to
predict the main-chain conformation based on antibody sequences and
to choose residues for diversification which do not affect the
canonical structure. It is known that, in the human V.kappa.
domain, the L1 loop can adopt one of four canonical structures, the
L2 loop has a single canonical structure and that 90% of human
V.kappa. domains adopt one of four or five canonical structures for
the L3 loop (Tomlinson et al. (1995) supra); thus, in the V.kappa.
domain alone, different canonical structures can combine to create
a range of different main-chain conformations. Given that the
V.lamda. domain encodes a different range of canonical structures
for the L1, L2 and L3 loops and that V.kappa. and V.lamda. domains
can pair with any VH domain which can encode several canonical
structures for the H1 and H2 loops, the number of canonical
structure combinations observed for these five loops is very large.
This implies that the generation of diversity in the main-chain
conformation may be essential for the production of a wide range of
binding specificities. However, by constructing an antibody library
based on a single known main-chain conformation it has been found,
contrary to expectation, that diversity in the main-chain
conformation is not required to generate sufficient diversity to
target substantially all antigens. Even more surprisingly, the
single main-chain conformation need not be a consensus structure--a
single naturally occurring conformation can be used as the basis
for an entire library. Thus, in a preferred aspect, the
dual-specific ligands of the invention possess a single known
main-chain conformation.
[0256] The single main-chain conformation that is chosen is
preferably commonplace among molecules of the immunoglobulin
superfamily type in question. A conformation is commonplace when a
significant number of naturally occurring molecules are observed to
adopt it. Accordingly, in a preferred aspect of the invention, the
natural occurrence of the different main-chain conformations for
each binding loop of an immunoglobulin domain are considered
separately and then a naturally occurring variable domain is chosen
which possesses the desired combination of main-chain conformations
for the different loops. If none is available, the nearest
equivalent may be chosen. It is preferable that the desired
combination of main-chain conformations for the different loops is
created by selecting germline gene segments which encode the
desired main-chain conformations. It is more preferable, that the
selected germline gene segments are frequently expressed in nature,
and most preferable that they are the most frequently expressed of
all natural germline gene segments.
[0257] In designing dual-specific antibodies or libraries thereof
the incidence of the different main-chain conformations for each of
the six antigen binding loops may be considered separately. For H1,
H2, L1, L2 and L3, a given conformation that is adopted by between
20% and 100% of the antigen binding loops of naturally occurring
molecules is chosen. Typically, its observed incidence is above 35%
(i.e. between 35% and 100%) and, ideally, above 50% or even above
65%. Since the vast majority of H3 loops do not have canonical
structures, it is preferable to select a main-chain conformation
which is commonplace among those loops which do display canonical
structures. For each of the loops, the conformation which is
observed most often in the natural repertoire is therefore
selected. In human antibodies, the most popular canonical
structures (CS) for each loop are as follows: H1-CS1 (79% of the
expressed repertoire), H2-CS 3 (46%), L1-CS 2 of V.kappa. (39%),
L2-CS 1 (100%), L3-CS 1 of V.kappa. (36%) (calculation assumes a
.kappa.:.lamda. ratio of 70:30, Hood et al. (1967) Cold Spring
Harbor Symp. Quant. Biol., 48: 133). For H3 loops that have
canonical structures, a CDR3 length (Kabat et al. (1991) Sequences
of proteins of immunological interest, U.S. Department of Health
and Human Services) of seven residues with a salt-bridge from
residue 94 to residue 101 appears to be the most common. There are
at least 16 human antibody sequences in the EMBL data library with
the required H3 length and key residues to form this conformation
and at least two crystallographic structures in the protein data
bank which can be used as a basis for antibody modeling (2cgr and
1tet). The most frequently expressed germline gene segments that
this combination of canonical structures are the VH segment 3-23
(DP-47), the JH segment JH4b, the V.kappa. segment 012/02 (DPK9)
and the J.kappa. segment J.kappa.1. These segments can therefore be
used in combination as a basis to construct a library with the
desired single main-chain conformation.
[0258] Alternatively, instead of choosing the single main-chain
conformation based on the natural occurrence of the different
main-chain conformations for each of the binding loops in
isolation, the natural occurrence of combinations of main-chain
conformations is used as the basis for choosing the single
main-chain conformation. In the case of antibodies, for example,
the natural occurrence of canonical structure combinations for any
two, three, four, five or for all six of the antigen binding loops
can be determined. Here, it is preferable that the chosen
conformation is commonplace in naturally occurring antibodies and
most preferable that it observed most frequently in the natural
repertoire. Thus, in human antibodies, for example, when natural
combinations of the five antigen binding loops, H1, H2, L1, L2 and
L3, are considered, the most frequent combination of canonical
structures is determined and then combined with the most popular
conformation for the H3 loop, as a basis for choosing the single
main-chain conformation.
Construction of IL-1 Dual-Specific Antibody: Diversification of the
Canonical Sequence
[0259] Having selected several known main-chain conformations or,
preferably a single known main-chain conformation, dual-specific
antibodies according to the invention or libraries for use in the
invention may be constructed by varying the binding site of the
molecule in order to generate a repertoire with structural and/or
functional diversity. This means that variants are generated such
that they possess sufficient diversity in their structure and/or in
their function so that they are capable of providing a range of
activities.
[0260] The desired diversity is typically generated by varying the
selected molecule at one or more positions. The positions to be
changed can be chosen at random or are preferably selected. The
variation can then be achieved either by randomisation, during
which the resident amino acid is replaced by any amino acid or
analogue thereof, natural or synthetic, producing a very large
number of variants or by replacing the resident amino acid with one
or more of a defined subset of amino acids, producing a more
limited number of variants.
[0261] Various methods have been reported for introducing such
diversity. Error-prone PCR (Hawkins et al. (1992) J. Mol. Biol.,
226: 889), chemical mutagenesis (Deng et al. (1994) J. Biol. Chem.,
269: 9533) or bacterial mutator strains (Low et al. (1996) J. Mol.
Biol., 260: 359) can be used to introduce random mutations into the
genes that encode the molecule. Methods for mutating selected
positions are also well known in the art and include the use of
mismatched oligonucleotides or degenerate oligonucleotides, with or
without the use of PCR. For example, several synthetic antibody
libraries have been created by targeting mutations to the antigen
binding loops. The H3 region of a human tetanus toxoid-binding Fab
has been randomised to create a range of new binding specificities
(Barbas et al. (1992) Proc. Natl. Acad. Sci. USA, 89: 4457). Random
or semi-random H3 and L3 regions have been appended to germline V
gene segments to produce large libraries with unmutated framework
regions (Hoogenboom & Winter (1992) J. Mol. Biol., 227: 381;
Barbas et al. (1992) Proc. Natl. Acad. Sci. USA, 89: 4457; Nissim
et al. (1994) EMBO J., 13: 692; Griffiths et al. (1994) EMBO J.,
13: 3245; De Kruif et al. (1995) J. Mol. Biol., 248: 97). Such
diversification has been extended to include some or all of the
other antigen binding loops (Crameri et al. (1996) Nature Med., 2:
100; Riechmann et al. (1995) Bio/Technology, 13: 475; Morphosys,
WO97/08320, supra). Other methods for naive library diversification
are described in WO2005/093074 and WO2004/003019.
[0262] Since loop randomisation has the potential to create
approximately more than 10.sup.15 structures for H3 alone and a
similarly large number of variants for the other five loops, it is
not feasible using current transformation technology or even by
using cell free systems to produce a library representing all
possible combinations. For example, in one of the largest libraries
constructed to date, 6.times.10.sup.10 different antibodies, which
is only a fraction of the potential diversity for a library of this
design, were generated (Griffiths et al. (1994) supra).
[0263] In a preferred embodiment, only those residues which are
directly involved in creating or modifying the desired function of
the molecule are diversified. For many molecules, the function will
be to bind a target and therefore diversity should be concentrated
in the target binding site, while avoiding changing residues which
are crucial to the overall packing of the molecule or to
maintaining the chosen main-chain conformation.
Construction of IL-1 Dual-Specific Antibody: Diversification of the
Canonical Sequence as it Applies to Antibody Domains
[0264] In the case of dual-specific antibodies, the binding site
for the target is most often the antigen binding site. Thus, in a
highly preferred aspect, the invention provides libraries of or for
the assembly of dual-specific antibodies in which only those
residues in the antigen binding site are varied. These residues are
extremely diverse in the human antibody repertoire and are known to
make contacts in high-resolution antibody/antigen complexes. For
example, in L2 it is known that positions 50 and 53 are diverse in
naturally occurring antibodies and are observed to make contact
with the antigen. In contrast, the conventional approach would have
been to diversify all the residues in the corresponding
Complementarity Determining Region (CDR1) as defined by Kabat et
al. (1991, supra), some seven residues compared to the two
diversified in the library for use according to the invention. This
represents a significant improvement in terms of the functional
diversity required to create a range of antigen binding
specificities.
[0265] In nature, antibody diversity is the result of two
processes: somatic recombination of germline V, D and J gene
segments to create a naive primary repertoire (so called germline
and junctional diversity) and somatic hypermutation of the
resulting rearranged V genes. Analysis of human antibody sequences
has shown that diversity in the primary repertoire is focused at
the centre of the antigen binding site whereas somatic
hypermutation spreads diversity to regions at the periphery of the
antigen binding site that are highly conserved in the primary
repertoire (see Tomlinson et al. (1996) J. Mol. Biol., 256: 813).
This complementarity has probably evolved as an efficient strategy
for searching sequence space and, although apparently unique to
antibodies, it can easily be applied to other polypeptide
repertoires. The residues which are varied are a subset of those
that form the binding site for the target. Different (including
overlapping) subsets of residues in the target binding site are
diversified at different stages during selection, if desired.
[0266] In the case of an antibody repertoire, an initial `naive`
repertoire is created where some, but not all, of the residues in
the antigen binding site are diversified. As used herein in this
context, the term "naive" refers to antibody molecules that have no
pre-determined target. These molecules resemble those which are
encoded by the immunoglobulin genes of an individual who has not
undergone immune diversification, as is the case with fetal and
newborn individuals, whose immune systems have not yet been
challenged by a wide variety of antigenic stimuli. This repertoire
is then selected against a range of antigens or epitopes. If
required, further diversity can then be introduced outside the
region diversified in the initial repertoire. This matured
repertoire can be selected for modified function, specificity or
affinity.
[0267] Two different naive repertoires of binding domains may be
used for the construction of dual-specific ligands, or a naive
library of dual-specific antibodies, in which some or all of the
residues in the antigen binding site are varied. The "primary"
library mimics the natural primary repertoire, with diversity
restricted to residues at the centre of the antigen binding site
that are diverse in the germline V gene segments (germline
diversity) or diversified during the recombination process
(junctional diversity). Those residues which are diversified
include, but are not limited to, HS0, H52, H52a, H53, H55, H56,
H58, H95, H96, H97, H98, L50, L53, L91, L92, L93, L94 and L96. In
the "somatic" library, diversity is restricted to residues that are
diversified during the recombination process (junctional diversity)
or are highly somatically mutated). Those residues which are
diversified include, but are not limited to: H31, H33, H35, H95,
H96, H97, H98, L30, L31, L32, L34 and L96. All the residues listed
above as suitable for diversification in these libraries are known
to make contacts in one or more antibody-antigen complexes. Since
in both libraries, not all of the residues in the antigen binding
site are varied, additional diversity is incorporated during
selection by varying the remaining residues, if it is desired to do
so. It shall be apparent to one skilled in the art that any subset
of any of these residues (or additional residues which comprise the
antigen binding site) can be used for the initial and/or subsequent
diversification of the antigen binding site.
[0268] In the construction of libraries for use in the invention,
diversification of chosen positions is typically achieved at the
nucleic acid level, by altering the coding sequence which specifies
the sequence of the polypeptide such that a number of possible
amino acids (all 20 or a subset thereof) can be incorporated at
that position. Using the IUPAC nomenclature, the most versatile
codon is NNK, which encodes all amino acids as well as the TAG stop
codon. The NNK codon is preferably used in order to introduce the
required diversity. Other codons which achieve the same ends are
also of use, including the NNN codon, which leads to the production
of the additional stop codons TGA and TAA.
[0269] A feature of side-chain diversity in the antigen binding
site of human antibodies is a pronounced bias which favors certain
amino acid residues. If the amino acid composition of the ten most
diverse positions in each of the V.sub.H, V.kappa. and V.lamda.
regions are summed, more than 76% of the side-chain diversity comes
from only seven different residues, these being, serine (24%),
tyrosine (14%), asparagine (11%), glycine (9%), alanine (7%),
aspartate (6%) and threonine (6%). This bias towards hydrophilic
residues and small residues which can provide main-chain
flexibility probably reflects the evolution of surfaces which are
predisposed to binding a wide range of antigens or epitopes and may
help to explain the required promiscuity of antibodies in the
primary repertoire.
[0270] Since it is preferable to mimic this distribution of amino
acids, the distribution of amino acids at the positions to be
varied preferably mimics that seen in the antigen binding site of
antibodies. Such bias in the substitution of amino acids that
permits selection of certain polypeptides (not just antibody
polypeptides) against a range of target antigens is easily applied
to any polypeptide repertoire. There are various methods for
biasing the amino acid distribution at the position to be varied
(including the use of tri-nucleotide mutagenesis, see WO97/08320),
of which the preferred method, due to ease of synthesis, is the use
of conventional degenerate codons. By comparing the amino acid
profile encoded by all combinations of degenerate codons (with
single, double, triple and quadruple degeneracy in equal ratios at
each position) with the natural amino acid use it is possible to
calculate the most representative codon. The codons (AGT)(AGC)T,
(AGT)(AGC)C and (AGT)(AGC)(CT)--that is, DVT, DVC and DVY,
respectively using IUPAC nomenclature--are those closest to the
desired amino acid profile: they encode 22% serine and 11%
tyrosine, asparagine, glycine, alanine, aspartate, threonine and
cysteine. Preferably, therefore, libraries are constructed using
either the DVT, DVC or DVY codon at each of the diversified
positions.
[0271] Domains identified from library screening are then subjected
to mutagenesis and further screened in order to produce and select
variants with improved characteristics.
IV. Uses of IL-1 Dual-Specific Antibody
[0272] Given their ability to bind to both IL-1.alpha. and
IL-1.beta., the dual-specific antibodies, or portions thereof
(including single domain antibodies identified in the process of
making the dual-specific antibodies), of the invention can be used
to detect IL-1.alpha. and/or IL-1.beta. (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. The
invention provides a method for detecting IL-1.alpha. and/or
IL-1.beta. in a biological sample comprising contacting a
biological sample with an antibody, or antibody portion, of the
invention and detecting either the antibody (or antibody portion)
bound to IL-1.alpha. and/or IL-1.beta. or unbound antibody (or
antibody portion), to thereby detect IL-1.alpha. and/or IL-1.beta.
in the biological sample. The antibody 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 125I, 131I, 35S or 3H.
[0273] Alternative to labeling the antibody, IL-1.alpha. and/or
IL-1.beta. can be assayed in biological fluids by a competition
immunoassay utilizing IL-1.alpha. and/or IL-1.beta. standards
labeled with a detectable substance and an unlabeled
anti-IL-1.alpha. and/or anti-IL-1.beta. antibody. In this assay,
the biological sample, the labeled IL-1.alpha. and/or IL-1.beta.
standards and the anti-IL-1.alpha. and/or anti-IL-1.beta. antibody
are combined and the amount of labeled IL-1.alpha. and/or
IL-1.beta. standard bound to the unlabeled antibody is determined.
The amount of IL-1.alpha. and/or IL-1.beta. in the biological
sample is inversely proportional to the amount of labeled
IL-1.alpha. and/or IL-1.beta. standard bound to the
anti-IL-1.alpha. and/or anti-IL-1.beta. antibody.
[0274] The dual-specific antibodies and antibody portions of the
invention are capable of neutralizing IL-1.alpha. and IL-1.beta.
activity. Accordingly, the antibodies and antibody portions of the
invention can be used to inhibit IL-1.alpha. and/or IL-1.beta.
activity, e.g., in a cell culture containing IL-1.alpha. and/or
IL-1.beta., in human subjects. In one embodiment, the invention
provides a method for inhibiting IL-1.alpha. and IL-1.beta.
activity comprising contacting IL-1.alpha. and IL-1.beta. with an
antibody or antibody portion of the invention such that IL-1.alpha.
and IL-1.beta. activity is inhibited. Preferably, the IL-1.alpha.
and hIL-1.beta. is human IL-1.alpha. and IL-1.beta.. For example,
in a cell culture containing, or suspected of containing
hIL-1.alpha. and hIL-1.beta., an antibody or antibody portion of
the invention can be added to the culture medium to inhibit
hIL-1.alpha. and hIL-1.beta. activity in the culture.
[0275] In another embodiment, the invention provides a method for
inhibiting IL-1.alpha. and IL-Inactivity in a subject suffering
from a disorder in which that IL-1.alpha. and IL-1.beta. activity
is detrimental. The invention provides methods for inhibiting
IL-1.alpha. and IL-1.beta. activity in a subject suffering from
such a disorder, which method comprises administering to the
subject a dual-specificity antibody or antibody portion of the
invention such that IL-1.alpha. and IL-1.beta. activity in the
subject is inhibited. Preferably, the IL-1.alpha. and IL-1.beta. is
human IL-1.alpha. and IL-1.beta. and the subject is a human
subject. An antibody of the invention can be administered to a
human subject for therapeutic purposes. Moreover, an antibody of
the invention can be administered to a non-human mammal expressing
IL-1.alpha. and IL-1.beta. with which the antibody binds 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 of the invention
(e.g., testing of dosages and time courses of administration).
[0276] As used herein, the phrase "a disorder in which IL-1
activity is detrimental" or the phrase "a disorder in which
IL-1.alpha. and IL-1.beta. is detrimental," is intended to include
diseases and other disorders in which the presence of IL-1 (which
encompasses both IL-1.alpha. and IL-1.beta.) 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 IL-1 activity is detrimental is a
disorder in which inhibition of IL-1 activity (i.e., either or both
of IL-1.alpha. and IL-1.beta.) 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 IL-1
in a biological fluid of a subject suffering from the disorder
(e.g., an increase in the concentration of IL-1 in serum, plasma,
synovial fluid, etc. of the subject), which can be detected, for
example, using an anti-IL-1 antibody as described above.
[0277] Interleukin 1 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, rheumatoid arthritis, osteoarthritis, juvenile chronic
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
greata, seronegative arthopathy, arthropathy, Reiter's disease,
psoriatic arthropathy, ulcerative colitic arthropathy, enteropathic
synovitis, chlamydia, yersinia and salmonella associated
arthropathy, spondyloarthopathy, 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 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, 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 vasulitis 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,
diseases of the central nervous system (e.g., depression,
schizophrenia, Alzheimers, Parkinsons, etc.), acute and chronic
pain, and lipid imbalance. The human antibodies, and antibody
portions of the invention can be used to treat humans suffering
from autoimmune diseases, in particular those associated with
inflammation, including, rheumatoid spondylitis, allergy,
autoimmune diabetes, or autoimmune uveitis.
[0278] Preferably, the IL-1.alpha. and IL-1.beta. dual-specific
antibodies of the invention or antigen-binding portions thereof,
are used to treat rheumatoid arthritis, Crohn's disease, multiple
sclerosis, insulin dependent diabetes, mellitus and psoriasis.
[0279] An IL-1.alpha. and IL-1.beta. dual-specific antibody, or
antibody portion, of the invention also can be administered with
one or more additional therapeutic agents useful in the treatment
of autoimmune and inflammatory diseases.
[0280] It is understood that all of the above-mentioned IL1-related
disorders include both the adult and juvenile forms of the disease
where appropriate. It is also understood that all of the
above-mentioned disorders include both chronic and acute forms of
the disease.
[0281] Antibodies of the invention, or antigen binding portions
thereof may be used alone or in combination to treat such diseases.
It should be understood that the antibodies of the invention 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 of the present
invention. The additional agent also can be an agent which imparts
a beneficial attribute to the therapeutic composition e.g., an
agent which effects the viscosity of the composition.
[0282] It should further be understood that the combinations which
are to be included within this invention 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 which are part of this invention can be the antibodies
of the present invention 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.
[0283] Preferred combinations are non-steroidal anti-inflammatory
drug(s) also referred to as NSAIDS which include drugs like
ibuprofen and COX-2 inhibitors. Other preferred 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-IL-1 antibodies of this invention. Non-limiting examples
of therapeutic agents for rheumatoid arthritis with which an
antibody, or antibody portion, of the invention can be combined
include 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-2, IL-6,
IL-7, IL-8, IL-12, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF, and
PDGF. Antibodies of the invention, 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, or their ligands including CD154
(gp39 or CD40L).
[0284] Preferred combinations of therapeutic agents may interfere
at different points in the autoimmune and subsequent inflammatory
cascade; preferred examples include TNF antagonists like chimeric,
humanized or human TNF antibodies, D2E7 (adalimumab; PCT
Publication No. WO 97/29131), CA2 (Remicade.TM.), CDP 571, CDP 870,
Thalidomide 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 preferred
combinations include Interleukin 11. Yet another preferred
combination are other key players of the autoimmune response which
may act parallel to, dependent on or in concert with IL-1 function;
especially preferred are IL-12 and/or IL-18 antagonists including
IL-12 and/or IL-18 antibodies or soluble IL-12 and/or IL-18
receptors, or IL-12 and/or IL-18 binding proteins. It has been
shown that IL-12 and IL-18 have overlapping but distinct functions
and a combination of antagonists to both may be most effective. Yet
another preferred combination are non-depleting anti-CD4
inhibitors. Yet other preferred combinations include antagonists of
the co-stimulatory pathway CD80 (B7.1) or CD86 (B7.2) including
antibodies, soluble receptors or antagonistic ligands.
[0285] The antibodies of the invention, 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, cochicine, 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 signalling 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) and
antiinflammatory cytokines (e.g. IL-4, IL-10, IL-11, IL-13 and TGF
.beta.). Preferred combinations include methotrexate or leflunomide
and in moderate or severe rheumatoid arthritis cases,
cyclosporine.
[0286] Non-limiting examples of therapeutic agents for inflammatory
bowel disease with which an antibody, or antibody portion, of the
invention 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. monoclonal antibodies; anti-IL-6
monoclonal antibodies; growth factors; elastase inhibitors;
pyridinyl-imidazole compounds; antibodies to or antagonists of
other human cytokines or growth factors, for example, TNF, LT,
IL-2, IL-6, IL-7, IL-8, IL-12, IL-15, IL-16, IL-18, EMAP-II,
GM-CSF, FGF, and PDGF. Antibodies of the invention 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 of the invention, 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 signalling 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.beta.
converting enzyme inhibitors, T-cell signalling 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-6R) and
antiinflammatory cytokines (e.g. IL-4, IL-10, IL-11, IL-13 and
TGF.beta.).
[0287] Preferred examples of therapeutic agents for Crohn's disease
in which an antibody or an antigen binding portion can be combined
include the following: TNF antagonists, for example, anti-TNF
antibodies, D2E7 (adalimumab; PCT Publication No. WO 97/29131), CA2
(Remicade.TM.), CDP 571, TNFR-Ig constructs, (p75TNFRIgG
(Enbrel.TM. M) and p55TNFRIgG (Lenercept)) inhibitors and PDE4
inhibitors. Antibodies, or antigen binding portions thereof, of the
invention or antigen binding portions thereof, can be combined with
corticosteroids, for example, budenoside and dexamethasone,
Antibodies of the invention 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 of the invention or antigen binding portion thereof may
also be used with T cell signaling inhibitors, for example,
tyrosine kinase inhibitors 6-mercaptopurines. Antibodies of the
invention or antigen binding portions thereof, can be combined with
IL-11.
[0288] Non-limiting examples of therapeutic agents for multiple
sclerosis with which an antibody, or antibody portion, of the
invention 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); 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, for example, TNF, LT, IL-2,
IL-6, IL-7, IL-8, IL-12, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF,
and PDGF. Antibodies of the invention, 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, CD86, CD90 or their ligands. The antibodies of the invention,
or antigen binding portions thereof, may also be combined with
agents, such as methotrexate, cyclosporine, FK506, rapamycin,
mycophenolate mofetil, leflunomide, NSAIDs, for example, ibuprofen,
COX-2 inhibitors, corticosteroids such as prednisolone,
phosphodiesterase inhibitors, adensosine agonists, antithrombotic
agents, complement inhibitors, adrenergic agents, agents which
interfere with signalling 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 signalling 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-13 and
TGF.beta.).
[0289] Preferred examples of therapeutic agents for multiple
sclerosis in which the antibody or antigen binding portion thereof
can be combined to include interferon-.beta., for example, IFNP1a
and IFN.beta.1b; copaxone, corticosteroids, IL-1 inhibitors, TNF
inhibitors, and antibodies to CD40 ligand and CD80.
IV. Pharmaceutical Compositions and Pharmaceutical
Administration
[0290] The compositions of this invention 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. The preferred form depends on
the intended mode of administration and therapeutic application.
Typical preferred 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. The preferred
mode of administration is parenteral (e.g., intravenous,
subcutaneous, intraperitoneal, intramuscular). In a preferred
embodiment, the antibody is administered by intravenous infusion or
injection. In another preferred embodiment, the antibody is
administered by intramuscular or subcutaneous injection.
[0291] 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, the preferred 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.
[0292] 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. The antibodies and antibody-portions of the present
invention can be administered by a variety of methods known in the
art, although for many therapeutic applications, the preferred
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.
[0293] In certain embodiments, an antibody or antibody portion of
the invention 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 of the invention 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.
[0294] Supplementary active compounds can also be incorporated into
the compositions. In certain embodiments, an antibody or antibody
portion of the invention is coformulated with and/or coadministered
with one or more additional therapeutic agents that are useful for
treating disorders in which IL-1 activity is detrimental. For
example, an anti-hIL-1 antibody or antibody portion of the
invention 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 of the invention
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. It will be appreciated by the skilled
practitioner that when the antibodies of the invention are used as
part of a combination therapy, a lower dosage of antibody may be
desirable than when the antibody alone is administered to a subject
(e.g., a synergistic therapeutic effect may be achieved through the
use of combination therapy which, in turn, permits use of a lower
dose of the antibody to achieve the desired therapeutic
effect).
[0295] The pharmaceutical compositions of the invention may include
a "therapeutically effective amount" or a "prophylactically
effective amount" of an antibody or antibody portion of the
invention. 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 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.
[0296] 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. Dosage unit form as used
herein 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 of the invention 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.
[0297] An exemplary, non-limiting range for a therapeutically or
prophylactically effective amount of an antibody or antibody
portion of the invention is 0.1-20 mg/kg, more preferably 1-10
mg/kg. 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.
[0298] The contents of all references, patents and published patent
applications cited throughout this application are incorporated
herein by reference
[0299] This invention is further illustrated by the following
examples which should not be construed as limiting.
EXAMPLES
[0300] The following examples describes methods and compositions
for making a dual-specific antibody which is specific for and can
neutralize both IL-1.alpha. and IL-1.beta.. The following examples
detail how to select IL-1.alpha. and IL-1.beta. domain antibodies
(dAbs), how to affinity mature the identified dAbs to improve
desired characteristics (affinity and neutralization properties),
and how to construct a dual-specific antibody which retains the
affinity and neutralization properties of the parent antibody.
[0301] For each clone described herein, the first part of the name
relates to its specificity (IL-1.alpha. represented by ABT1; and
IL-1.beta. represented by ABT2), followed by the first number which
relates to the lineage from which that clone is derived and a
second number which is the unique identifier for that clone.
Example 1
Selection of IL-1.alpha. and IL-1.beta. Domain Antibodies
[0302] Human phage antibody libraries containing either variable
heavy or variable light single human frameworks were used in the
screen to identify specific IL-1.alpha. and IL-1.beta. domain
antibodies (dAbs).
[0303] The dAb libraries used in the screen were based on the DP47
or DPk9 germline sequences, i.e., a single human framework for VH
(V3-23 [locus] DP47 [V Base Entry] and JH4b) and VL (012/02 [locus]
DPk9 [V Base Entry] and J.LAMBDA.1) with side chain diversity
incorporated at positions in the antigen binding site that make
contacts with antigen in known molecular structures. Importantly,
these positions are also highly diverse in the mature repertoire.
The canonical structure (VH: 1-3, VL: 2-1-1) encoded by these
frameworks are the most common in the human antibody repertoire.
The CDR3 of the heavy chain was designed to be as short as possible
yet still able to form an antigen binding surface. The libraries
were selected and affinity matured without knowing the sequence of
selected clones.
[0304] The library format used was dAb displayed on phage (single
or multiple copies of a human variable VH or V.kappa. domain
displayed on a phage virus particle as gene III fusions). The VH
library size was 1.6.times.10.sup.10 and the V.kappa. library size
was 1.7.times.10.sup.10. The libraries were phage libraries;
therefore all the genes required to generate functional phage plus
the geneIII-dAb gene fusion were encoded on the phage vector (pDOM4
vector).
[0305] Three selection rounds were performed with clones screened
from the outputs of selections rounds 2 and 3. Selection type was
in solution using biotinylated antigen. Increase in phage output
titre was used as an indication of specific phage enrichment during
the selection rounds. Monoclonal phage were tested at rounds 2 and
3 for binding to antigen coated or captured by ELISA prior to
screening the soluble dAb from these selection outputs. There were
no antibody positive controls used during initial screening;
however MAb200 and MAb201 were used as positive inhibiting IgG in
the functional cell assay. Each single domain antibody was chosen
by binding of a phage repertoire to the antigen, t e., either human
IL-1.alpha. or human IL-1.beta.. Variable heavy and variable light
dAbs to both IL-1.alpha. and IL-1.beta. were identified.
[0306] DNA purification was by QIAprep Spin Miniprep Kit; QIAfilter
Plasmid Midi Kit or QIAfilter Plasmid Maxi Kit as appropriate for
the amount of DNA required (kits by QIAGEN Ltd., QIAGEN House,
Fleming Way, Crawley, West Sussex, RH10 9NQ), using the standard
protocols supplied with the kits.
[0307] Sequences of the single domain antibodies (dAbs) identified
in the screen are described in Table 60.
Example 2
Affinity Maturation of IL-1.alpha./IL-1.beta. Domain Antibodies
(dAbs)
[0308] Affinity maturation of the dAbs identified in the library
screen described in Example 1 was performed using different
affinity maturation processes, as described below.
2.1.1. Emulsion Affinity Maturation of IL-1.alpha. dAb ABT1-95
Overview.
[0309] One of the domain antibodies identified in the library
screen, i.e., ABT1-95, was affinity matured by Emulsion selection,
improving neutralization from an ND.sub.50 of 1 .mu.M to an
ND.sub.50 of 50 .mu.M for ABT1-95-15. This process of affinity
maturation was done in three incremental stages. In the first
stage, two ABT1-95 derived clones were isolated: ABT1-95-1 and
ABT1-95-2. By recombining these two mutants the clone ABT1-95-3 was
created and was determined to have an ND.sub.50 of 50 nM. Four
libraries, each mutating four positions at a time with NNS
diversification (wherein N=nucleotides A/T/C/G and S=nucleotides G
or C) targeting either CDR1 (2.times.), CDR2/3, and CDR3 were
created from ABT1-95-3. In addition to these libraries an error
prone library was made. After selection in emulsions nine clones
were identified with improved binding and ND.sub.50 values of about
1 nM. Analysis of the best clones identified positions 30, 31, 55,
and 56 as being highly relevant for high affinity binding.
Selections from an NNS library specifically targeting these
positions yielded three clones with ND.sub.50's in the 50-250 pM
range: ABT1-95-13, ABT1-95-14 and ABT1-95-15.
Detailed Description of Isolation Process for ABT1-95-15
[0310] Two ABT1-95 derived dAbs (Y88H named ABT1-95-1 (which
contained a Y88H mutation), and a mutant that contained the same
CDR2 as ABT1-141 named ABT1-95-2) were isolated from an ABT1-122
library selection. As shown in the table below, both clones had
moderate increases in affinity over the parental ABT1-95. However,
when these changes were combined by site directed mutagenesis, the
new clone, ABT1-95-3, showed a marked increase in affinity (7-12
nM). Below are the BIAcore kinetics using an IL-1.alpha. coated
chip with purified dAb flowed over the surface.
TABLE-US-00001 TABLE 1 BIAcore kinetics summary kon koff Kd (nM)
ABT1-122 2.9-6.2 .times. 10e.sup.5 0.025 40-86 ABT1-95-1 3.6-6.6
.times. 10e.sup.5 0.013-0.015 20-42 ABT1-95-2 2.3-2.4 .times.
10e.sup.5 7.6-9.6 .times. 10e-3 32-42 ABT1-95-3 4.7-6.4 .times.
10e.sup.5 4.7-5.7 .times. 10e-3 7-12
[0311] To create new affinity maturation libraries based on
ABT1-95-3, new `signature` templates were constructed. These
templates contained unique restriction sites, which would no longer
be present once libraries were constructed on the basis of these
templates. This made it possible to remove selectively parental
clones from the selection by restriction digest and focus on
library-derived dAbs.
[0312] Four signature templates were constructed and verified by
sequencing:
1) ABT1-95-3*: -BtsI+BamHI
2) ABT1-95-2*: -BtsI+BamHI
[0313] 3) ABT1-95-3*-k1: -BtsI+BamH1+KpnI after CDR1 4)
ABT1-95-3*-k3: -BtsI+BamHIH+KpnI after CDR3
[0314] These constructs were then used for construction of six
libraries.
#1) CDR1-1: Pro Ile NNS NNS NNS Leu NNS with template ABT1-95-3*-k1
#2) CDR1-2: NNS NNS Ile His NNS NNS Leu Arg with template
ABT1-95-3*-k1 #3) CDR2/3: NNS Ser Ser Ser NNS . . . NNS Tyr Arg Trp
Pro NNS: with template ABT1-95-3*-k3 #4) 88+CDR3: NNS Cys NNS Gln
NNS Tyr NNS Trp Pro Val with template ABT1-95-3*-k3 #5) Error-prone
1 with template ABT1-95-2* #6) Error-prone 2 with template
ABT1-95-3*
[0315] Diversity for libraries numbered 1-4 was complete, while for
the error-prone libraries diversity exceeded 1.times.10.sup.8,
which is higher than the diversity that can be assessed with
emulsion. Error rates were about 2.5 nucleotide mutations per dAb.
This was at the low end, but additional mutagenesis can be
performed after e.g. 3 rounds of selection.
[0316] The six ABT1-95-3 libraries were subjected to nine rounds of
emulsion selection. Libraries 1, 2, 3 and 4 went through 3 rounds
of selection at relatively high IL-1.alpha. concentrations (50, 20,
10 nM), were then mixed 1:1:1:1, and subjected to 6 rounds of
off-rate selection. During off-rate selection cold IL-1.alpha.
concentration was increased from 300 to 1000 nM, while `hot` IL-1a
was kept at 10 nM. All off-rate selections were performed for short
periods of time (i.e. 5-20 min.) and in the presence of free Ter
operator (3 nM, 2000-fold excess) to limit rebinding of Tus-dAb
complexes to non-corresponding DNA. Library 4 was also treated
separately, and went through 7 rounds of selection using the same
conditions as above. For the error-prone libraries, after each
three rounds of selection an error-prone amplification step was
introduced (using the GenemorphII mutagenesis kit). Error-prone
libraries that had gone through both two and three rounds of
mutagenesis were selected up to the ninth round. For additional
details on emulsions using the TUS DNA-binding protein, see WO
06/046042A2, incorporated by reference herein, which describes the
sequences of all oligonucleotides and vectors used herein.
[0317] After nine rounds, all dAbs were cloned into both pDOM-5 and
Tus vectors and screening of individual clones has been
initiated.
[0318] A total of 177 colonies isolated after nine round of
selection (using Tus) on IL-1.alpha. were analysed: 72 colonies
expressed dAbs (in pDOM5) with strong signals in ELISA on
IL-1.alpha.. Following BIAcore analysis for off-rates, 33 dAbs
showed a decreased off-rate when compared to ABT1-95-3: by
sequencing, it was shown that the vast majority of these clones had
unique sequences with mutations targeting primarily H30 and Q55.
Further detailed analysis of 8 clones revealed Kd between 0.8 and
2.9 nM (Kd ABT1-95-3 is 5.4 nM) and ND.sub.50 values between 0.3 nM
and 1 nM. ABT1 cell assay (100 pg/ml IL-1.alpha.) results are
described in Table 62.
[0319] Further off-rate selections (3 rounds) with the pooled
libraries did not yield improved clones: all the selected clones
were identical to those already isolated at round 9 (ABT1-95-10 and
ABT1-95-8).
[0320] Repeated prepping followed by cell assay reconfirmed that
all ABT-95-3 derived variants (i.e. 95-4 through-12) had increased
affinity for IL-1.alpha.. However, further affinity improvements
were needed. Therefore, a next generation of affinity maturation
was begun using ABT1-95-4, -8 and -12 as templates. Error-prone
libraries were made of all three, while of ABT1-95-4 also a NNS
library at positions 30, 31, 55, 56 was made. The latter was made
by 3 fragment SOE because the distance between positions was too
large to make a single oligonucleotide. All libraries were verified
by sequencing, with the error-prone libraries having an average of
2.4 by mutations/dAb. The NNS library was checked for parent by
digestion with FspI, which demonstrated no visible parent on gel.
In a first run, 6 rounds of selection were performed as noted
below, the first value for NNS library, the second for the ep
library.
round 1 20 nM, 10 nM round 2 10 nM+4/10 min, 500 nM cold round 3 10
nM+20/30 min, 750 nM cold round 4 10 nM+60/75 min, 1600 nM cold
[0321] Output at this stage was digested with BtsI to remove a
known contaminant. The NNS library was kept separate while the
error-prone libraries were pooled.
round 5n 10 nM+60 min 750 nM cold round 6n 10 nM+75 min 1 uM cold
round 7* 10 nM+105 min 1 uM cold round 8* 10 nM+105 min 1 uM cold
round 9 10 nM+105 min 1 uM cold
[0322] This time the yield of recovery of the dAb PCR was better
and maintained above 400 ng/well. Round 9 was recloned in pDOM5,
transformed to MACH1 cells, and 96 supernatants were screened on
BIAcore for off-rate. Only a small percentage (10%) of clones were
inactive, 15% had off-rates significantly faster than parent, while
75% had off-rates of at least parent. A subset of clones didn't
regenerate well, potentially indicating tighter binding. A total of
34 clones were taken further for purification and detailed
analysis. Sequence analysis indicated clones to be diverse but
focussed. Of these, 28 purified well and went into RBA (Receptor
Binding Assay described below).
[0323] IL-1 Receptor Binding Assay. The majority was at least as
good as ABT1-95-8 (1 nM), while 7 clones were better. These seven
were tested in the cell assay and 6 out of 7 were better than 95-8.
Summary of characterization is shown in Table 2 below. BIAcore was
also performed on purified clones, but values were not very
reliable, especially since the Koff values tend to be influenced by
drift of the baseline.
TABLE-US-00002 TABLE 2 After round 9, pools were cloned in pDOM5
and screened NNS ep Total Colonies picked 48 45 93 BIAcore screen
Inactive 5 3 8 (9%) faster off 14 5 19 (20%) off .ltoreq. 95-8 29
37 66 (71%) Purified 18 16 34 RBA 7/28 better than 95-8 MRC5 6/7
better than 95-8 Sequences, MRC5 data, and BIAcore for three best
clones ND50 kon koff KD ABT1-95-13 VR, ER (30, 120 pM 2.00E+06
1.90E-03 9.50E-10 31, 55, 56) ABT1-95-14 VK, ER 250 pM 1.60E+06
1.90E-03 1.20E-09 ABT1-95-15 VR, EP 45 pM 2.00E+06 1.30E-03
6.70E-10
Detailed Description on how Emulsions Described Above were
Performed
[0324] Library Construction.
[0325] Error-prone libraries: one pg of template DNA was PCR
amplified using GenemorphII for 35 cycles with the primer set
OA16/17n, followed by a restriction digest with SalI and NotI. The
clean fragments were ligated in the vector pIE7t.sup.3T using T4
DNA ligase. Small aliquots of the ligation product were amplified
in the presence of competitor DNA to establish that the number of
ligation events exceeded 10.sup.9. The libraries were PCR amplified
from the ligation reaction using Platinum pfx and primers AS12/18
(Tus libraries). For targeted diversification libraries,
oligonucleotides containing NNS codons at the indicated positions
were used and the dAb was assembled by overlap PCR. The assembly
reaction was PCR amplified with OA16/17n oligonucleotides using
PfuUltra DNA polymerase and cloned SalI/NotI in the respective IVT
vector.
In Vitro Expression and Emulsification.
[0326] For in vitro transcription/translation of the library of PCR
fragments the following reaction mixture was used: 1.5 .mu.l 100 mM
oxidized glutathione, 2 .mu.l 5 mM Methionine, 0.5 .mu.l DNA
(5.0.times.10.sup.8 molecules), 10 .mu.l H.sub.2O, biotinylated
antigen at varying concentrations, and 35 .mu.l of EcoPro T7
(Novagen) coupled transcription-translation extract. Immediately
after mixing, the extract was added to 0.7 ml of light white
mineral oil containing 4.5% (v/v) Span-80 and 0.5% (v/v) Triton
X-100. Emulsification was carried out by spinning a magnetic
stirrer for 5 min at 2000 rpm in a 5 ml glass vial. Microscopic
analysis of droplet formation confirmed that droplets were .about.2
.mu.m in diameter. The emulsion was incubated for 60 min at
25.degree. C. to allow expression and formation of the protein-DNA
complex to take place. Breaking of the emulsion was performed by
adding 0.25 ml of PBS/1% BSA/biotinylated antigen and 0.5 ml
hexane/20% (v/v) mineral oil, followed by brief vortex and
centrifugation for 1 min at 13 k rpm. After removal of the oil
phase, 1 ml of hexane/mineral oil was added and the procedure
repeated three times. The last extraction was performed using only
hexane.
Selection and Amplification.
[0327] Selection for binders was performed by incubating the
extracted aqueous phase for 30 min in the presence 50-5 nM of
biotinylated antigen (See previous information for exact conditions
used per round). If off-rate selections were performed, this
incubation was followed by addition of an excess of unbiotinylated
antigen and incubation in the 5-105 min range. Each emulsion
reaction was then divided over 5 wells of a streptavidin coated PCR
plate (50 .mu.l/well), incubated for 15 min at 25.degree. C., and
washed 4 times with PBS/BSA. Fifty .mu.l of PCR mix containing
either OA16/17n primers, PfuUltra buffer, dNTPs, 2.5 u Pfu Ultra
DNA polymerase was added to each well. PCR was performed for 30
cycles. For Tus selections, the PCR product was cleaned and
digested with SalI/NotI. The fragment was then ligated in
pIE7t.sup.3T vector and amplified as described in library
construction.
[0328] For additional details on emulsions using the TUS
DNA-binding protein, see WO 06/046042A2, incorporated by reference
herein, which describes the sequences of all oligonucleotides and
vectors used.
Further Affinity Maturation of ABT-1-95-15
[0329] ABT1-95 was affinity matured by Emulsion selection from an
ND.sub.50 of 1 .mu.M to an ND.sub.50 of 50 pM for ABT1-95-15. This
was done in three incremental stages, as described above.
Generally, in the first stage, two ABT1-95 derived clones were
isolated: ABT1-95-1 and ABT1-95-2. By recombining these two mutants
the clone ABT1-95-3 was created and was determined to have an
ND.sub.50 of 50 nM.
[0330] Four libraries, each mutating four positions at a time with
NNS (wherein N is nucleotide A, T, C, or G and S is a nucleotide G,
or C) diversification targeting either CDR1 (2.times.), CDR2/3, and
CDR3 were created from ABT1-95-3. In addition to these libraries an
error prone library was made. After selection in emulsions nine
clones were identified with improved binding and ND.sub.50 values
of about 1 nM. Analysis of the best clones identified positions 30,
31, 55, and 56 as being highly relevant for high affinity binding.
Selections from an NNS library specifically targeting these
positions yielded three clones with ND.sub.50's in the 50-250 pM
range: ABT1-95-13, ABT1-95-15 and ABT1-95-15. ABT1-95-15 was then
further matured using the parallel strategies of NNS screening and
yeast display.
[0331] For the NNS screening, individual NNS libraries at every
diversified position in each CDR (Q27, P28, V30, R31, N32, R34,
Y49, S50, S51, Y53, E55, P56, Q89, G91, Y92, R93, W94, V96) were
created and screened, with mutations that led to activity
improvements combined to create ABT1-95-174 (ND.sub.50 of 2 .mu.M).
ABT1-95-15 was also matured using yeast display with each CDR being
individually diversified followed by sorting of the libraries using
FACS, then recombination of the outputs followed by further sorting
to create ABT1-95-A3, ABT1-95-A5, and ABT1-95-A6. Mutations from
the NNS screen identified as potentially detrimental to
neutralising activity were removed from ABT1-95-A3 and ABT1-95-A5
to create ABT1-95-181, ABT1-95-182 and ABT1-95-184. FIG. 21A and
Table 3 show the changes in both sequence, neutralising activity
and affinity that occurred as the maturation of ABT1-95
progressed.
TABLE-US-00003 TABLE 3 Summary of the activity improvements of
ABT1-95 dAb clones as the maturation program progressed Clone MRC5
ND.sub.50 K.sub.D ABT1-95 1 .mu.M 200 nM ABT1-95-1 40 nM ABT1-95-2
40 nM ABT1-95-3 50 nM 5-10 nM ABT1-95-4 0.9 nM 0.9 nM ABT1-95-8 0.9
nM 0.9 nM ABT1-95-13 110 pM ABT1-95-14 250 pM ABT1-95-15 50-150 pM
ABT1-95-A3 11 pM ABT1-95-A5 16 pM ABT1-95-A6 24 pM ABT1-95-174 *1-5
pM ABT1-95-181 14 pM ABT1-95-182 *1-5 pM ABT1-95-184 *1-5 pM
2.1.2 Affinity Maturation of ABT1-122
[0332] ABT1-122 was affinity matured by phage display using both
libraries diversified by using NNS codons in either CDR1, CDR2 or
CDR3 and error-prone PCR with selection taking place under both
equilibrium and off-rate conditions. Analyses of the outputs from
these selections lead to the identification of ABT1-122-18 which
has an ND.sub.50 of 5 nM compared to the parental ND.sub.50 of 50
nM.
[0333] Libraries, each diversified in blocks of three amino acids
scanning across all three CDRs were constructed and sorted by FACS.
This resulted in the lead clone ABT1-122-511 which has an ND.sub.50
of 80 nM when paired with ABT2-108 as an IgG compared to a parental
ND.sub.50 of 1 .mu.M for the same pairing. To mature this lead
clone further, each CDR was again individually diversified followed
by sorting of the libraries using FACS, then recombination of the
outputs followed by further sorting to create ABT1-122-750x which
has an ND.sub.50 in the MRC5 assay as a dAb of 934 pM and ND.sub.50
of 20nM in the MRC5 assay when paired as an IgG with ABT2-65-17.
FIG. 21B shows the sequences of clones ABT1-122 as maturation
progressed. In addition to the sequences described in FIG. 21B (as
well as Tables 60 and 64), clone ABT1-122-751x was also created
(CDR1 RASQHIWTELN/CDR2 GSASRQK/CDR3KQFAYYPNT). Table 4 below
describes the inhibitory activity of the clones.
TABLE-US-00004 TABLE 4 Summary of the activity improvements of
ABT1-122 dAb clones as maturation has progressed Clone MRC5
ND.sub.50 ABT1-122 50 nM ABT1-122-18 5 nM ABT1-122-511 ND
ABT1-122-750X 934 pM
2.1.3 Affinity Maturation of ABT1-141
[0334] ABT1-141 was affinity matured by phage display using both
libraries diversified using NNS codons in either CDR1, CDR2 or CDR3
and error-prone PCR with selection taking place under both
equilibrium and off-rate conditions (FIG. 21C shows the sequences
of the maturation process). Analyses of the outputs from these
selections identified ABT1-141-25, which has an ND.sub.50 of 5 nM
compared to the parental ND.sub.50 of 70 nM. An error-prone PCR
library based on ABT1-141-25 resulted in ABT1-141-47 which had an
improved ND.sub.50 of 1 nM and ABT1-141-76 which has an identical
ND.sub.50 to ABT1-141-47 but with the added benefit of no framework
mutations, compared to the germline DPK9/JK1 scaffold. Inhibitory
activity of the identified clones are described in Table 5.
TABLE-US-00005 TABLE 5 Summary of the activity improvements of
ABT1-141 dAb clones as maturation has progressed Clone MRC5
ND.sub.50 ABT1-141 70 nM ABT1-141-25 5 nM ABT1-141-47 1 nM
ABT1-141-76 1 nM
2.1.4 Affinity Maturation of ABT2-65
[0335] ABT2-65 was affinity matured by phage display using both
libraries diversified by using NNS codons in either CDR1, CDR2 or
CDR3 and error-prone PCR with selection taking place under both
equilibrium and off-rate conditions. A number of clones with
improved activity in the MRC5 assay were identified from the
library where the inner residues in CDR3 had been diversified, the
best of which was ABT2-65-17 which has an ND.sub.50 of 2 nM
compared to 266 nM for the parental clone. One framework mutation
and an N-linked glycosylation site were present in ABT2-65 and a
further framework mutation was present in ABT2-65-17. These were
all removed to create the clone ABT2-65-166. Clone sequences are
described in FIG. 21D, and inhibitory activity is described in
Table 6.
TABLE-US-00006 TABLE 6 Summary of the activity improvements of
ABT2-65 dAb clones. Clone MRC5 ND.sub.50 ABT2-65 266 nM ABT2-65-17
2 nM ABT2-65-166 4 nM
2.2.1. Affinity Maturation of IL-1.beta. dAb ABT2-108 by Yeast
Display
[0336] Previously ABT2-108 was identified as an anti-human
IL-1.beta. VH dAb (see Table 60 and Example 1). ABT2-108 has
.about.10-100 nM IL-1.beta. neutralizing potency as a dAb and
paired IgG. In order to improve the IL-1.beta. neutralization
potency to <200-pM, affinity maturation of ABT2-108 was
performed in the scFv format by yeast display, using the small
scanning library and CDR recombination methods.
[0337] The objective of the study was to affinity-mature ABT2-108
by yeast display in the scFv format by the use of small scanning
CDR libraries and CDR recombination in order to obtain <200 pM
IL-1.beta. neutralization potency when converted into the paired
IgG format.
Materials and Methods
[0338] Construction of the scFv Yeast Expression Vector.
[0339] ABT2-108/ABT1-122 scFv was subcloned into pYDs-TEV vector by
standard molecular cloning. The complete construct is shown in FIG.
1.
Construction of Small CDR Scanning Libraries
[0340] Gapped vectors for yeast recombination were generated by
long PCR using the pYDs-TEV-2-108/1-122 plasmid as a template and 7
sets of gapped primer pairs (Table 7).
TABLE-US-00007 TABLE 7 Gapped Vector Primers Gap Forward Reverse 1
2-108-Gap1Fwd 2-108-Gap1Rev TGG GTC CGC CAG GCT AAA GGT GAA TCC GGA
CCA G GGC TG 2 2-108-Gap2Fwd 2-108-Gap2Rev AAT ACA TAC TAC GCA GAC
TGA GAC CCA CTC GAG TCC GTG ACC 3 2-108-Gap3Fwd 2-108-Gap3Rev GAC
TCC GTG AAG GGC CGG ATC CTG CCC AAT ACG TGA G 4 2-108-Gap4Fwd
2-108-Gap4Rev CGG TTC ACC ATC TCC CGC GTA TGT ATT CTT ACC ATC CTG
CCC 5 2-108-Gap5Fwd 2-108-Gap5Rev CAT CAT CTT TTT GAC TAC TTT CGC
ACA GTA ATA TGG GGT C TAC CGC 6 2-108-Gap6Fwd 2-108-Gap6Rev GAC TAC
TGG GGT CAG AAC CCG ACC CGT ATA GGA AC TTT CG 7 2-108-Gap7Fwd
2-108-Gap7Rev TGG GGT CAG GGA ACC CTG ATG ATG AAC ACC AAC CCG
AC
[0341] 30 long oligos were ordered that span the gaps, 4 for HCDR1,
15 for HCDR2, and 11 for HCDR3. Each oligo contained a series of 9
random nucleotides (NNSNNSNNS, S.dbd.C or G), and each subsequent
oligo shifted the random sequence down by 3 nucleotides. Each
generated library would then randomize only 3 consecutive amino
acids, and this window of 3 amino acids would be walked across all
3 HDCRs. One oligo for HCDR1 randomized 3 non-consecutive amino
acids. FIG. 2 shows the randomization of HCDRs in the 30 small
libraries.
Yeast Libraries Generation.
[0342] 30 small yeast libraries were generated via homologous
recombination by co-transforming yeast (EBY100) with Gapped vectors
and their respective long oligos.
[0343] Libraries output sizes were estimated by plating aliquots of
transformed cells on SD(-UT) plates.
Yeast Libraries Labeling and Sorting.
[0344] Expression of scFv fragments on the surface of yeast was
induced by growing libraries in SRG(-UT) medium for 48 hours at
20.degree. C.
[0345] For library sorting 0.4 OD of yeast cells were labeled with
antigen (human IL-1.beta.) at a concentration of 6 nM followed by
staining with detection antibody (biotinylated goat polyclonal
anti-IL-1.beta. antibody, R&D Systems, Cat#BAF201, at
concentration 5 .mu.g/ml) and then Streptavidin-PE (Jackson
ImmunoResearch Laboratories, Inc, Cat #016-110-034). ScFv
expression was determined by staining libraries with anti-1/5 mAb
(Invitrogen, Cat#R96025) followed by anti-mouse IgG-FITC (Molecular
Probes, Cat #F11021).
[0346] Libraries were sorted on the Cytomation MoFlo for 2-3 rounds
individually, and then for 1-2 rounds pooled into 6 groups, H1
#1-4, H2 #1-5, H2 #6-10, H2 #11-15, H3 #1-5, and H3 #6-11. For each
round of sorting, the top .about.0.1% of cells were collected.
[0347] The outputs were analyzed for sequence diversity, and
individual clones were assayed for IL-1.beta. binding on the
surface of yeast. The best clones were converted into IgG and
tested for IL-1.beta. neutralization potency in an MRC-5 assay.
[0348] A 2-108 recombination library was generated by amplifying
the HCDR fragments from the best affinity matured clones as well as
2-108 parental for each CDR and then performing homologous
recombination in yeast with all the fragments (FIG. 3). One round
of sorting was performed on the recombination library using 2
different selection strategies, equilibrium sorting with 60 .mu.M.
IL-1.beta. and on-rate sorting with 600 .mu.M IL-1.beta. for 20
minutes labeling. Both outputs were analyzed for sequence
diversity, and individual clones were assayed for IL-1.beta.
binding on the surface of yeast. The best clones were converted
into IgG and tested for IL-1.beta. neutralization potency in a
MRC-5 assay.
MRC-5 Bioassay.
[0349] The MRC-5 cell line is a human lung fibroblast cell line
that produces IL-8 in response to human IL-1.alpha. and IL-1.beta.
in a dose-dependent manner. MRC-5 cells were originally obtained
from ATCC and subcultured in 10% FBS complete MEM and grown at 37 C
in a 5% CO.sub.2 incubator. To determine an antibody's neutralizing
potency against IL-1.alpha. or IL-1b, 4.times. concentrated Ab (50
ul) was added to a 96 well plate and pre-incubated with 50 ul of
4.times. concentrated IL-1a or IL-1b for 1 hr at 37 C, 5% CO.sub.2.
MRC-5 cells at a concentration of 1E5/ml were then added (100 ul)
to all wells and the plates were incubated overnight at 37 C in a
5% CO2 incubator. Antibody potency was determined by its ability to
inhibit IL-8 production. Human IL-8 production was measured by
ELISA.
Results
ABT2-108 Affinity Maturation by Small Scanning CDR Libraries
[0350] Improved clones were identified from the selections for all
3 HCDRs., and designated ABT2-108-501 through 532 see Table 8).
TABLE-US-00008 TABLE 8 CDR sequences of parental and affinity
matured 2-108 clones CDR1 CDR2 CDR3 Clone# ABT -2108 EYTMM
RIGQDGKNTYYADSVKG YTGRVGVHHLFDY WT -501 WEGMM Apr. 9, 2004 H1-1
scFV R2 6 nM output #1 -502 MESMM Apr. 30, 2004 H1-1~4 scFv pooled
R3 6 nM output #1 -503 EEKWM Apr. 30, 2004 H1-1~4 scFv pooled R3 6
nM output #2 -504 DEGMM Apr. 30, 2004 H1-1~4 scFv pooled R3 6 nM
output #3 -505 EYGLI Apr. 30, 2004 H1-1~4 scFv pooled R3 6 nM
output #9 -506 RCHEDGKNTYYADSVKG Apr. 30, 2004 H2-1~5 scFv pooled
R2 6 nM output #1 -507 RITWTGKNTYYADSVKG Apr. 30, 2004 H2-1~5 scFv
pooled R2 6 nM output #2 -508 RIGYMDKNTYYADSVKG Apr. 30, 2004
H2-1~5 scFv pooled R2 6 nM output #4 -509 RITYSGKNTYYADSVKG Apr.
30, 2004 H2-1~5 scFv pooled R2 6 nM output #5 -510
RCVWDGKNTYYADSVKG Apr. 30, 2004 H2-1~5 scFV pooled R2 6 nM output
#9 -511 RIGQDGKNTVIRDSVKG Apr. 30, 2004 H2-6~10 scFv pooled R2 6 nM
output #1 -512 RIGQDGKNTVLRDSVKG Apr. 30, 2004 H2-6~10 scFv pooled
R2 6 nM output #3 -513 RIGQDGKNTVDRDSVKG Apr. 30, 2004 H2-6~10 scFV
pooled R2 6 nM output #9 -514 RIGQDGKNTWTRDSVKG Apr. 30, 2004
H2-6~10 scFv pooled R2 6 nM output #10 -515 RIGQDGKNTYYRGYMKG May
3, 2004 H2-11~15 scFv pooled R2 6 nM output #1 -516
RIGQDGKNTYYRVDVKG May 3, 2004 H2-11~15 scFv pooled R2 6 nM output
#4 -517 RIGQDGKNTYYADRTDG May 3, 2004 H2-11~15 scFv pooled R2 6 nM
output #5 -518 RIGQDGKNTYYRMDVKG May 3, 2004 H2-11~15 scFv pooled
R2 6 nM output #6 -519 RIGQDGKNTYARMSVKG May 3, 2004 H2-11~15 scFv
pooled R2 6 nM output #8 -520 RIGQDGKNTYYADRSFG May 3, 2004
H2-11~15 scFv pooled R2 6 nM output #9 -521 YTGRILGHHLFDY Apr. 9,
2004 H3-5a scFv R2 6 nM output #1 -522 YTGRILHHHLFDY May 3, 2004
H3-1~5 scFv pooled R2 6 nM output #1 -523 YDGWVGVHHLFDY May 3, 2004
H3-1~5 scFV pooled R2 6 nM output #2 -524 YTGRVFNHHLFDY May 3, 2004
H3-1~5 scFv pooled R2 6 nM output #6 -525 YTGRILAHHLFDY May 3, 2004
H3-1~5 scFv pooled R2 6 nM output #9 -526 YTGRVLNHHLFDY May 3, 2004
H3-6~11 scFv pooled R2 6 nM output #1 -527 YTGRVFKHHLFDY May 3,
2004 H3-6~11 scFv pooled R2 6 nM output #3 -528 YTGRVLGHHLFDY May
3, 2004 H3-6~11 scFv pooled R2 6 nM output #6 -529 EEGMM May 14,
2004 H1-1~4 scFv pooled R4 2 nM output #1 -530 YTGRILEHHLFDY May
14, 2004 H3-1~5 scFv pooled R3 2 nM output #1 -531 YTGRIFSHHLFDY
May 14, 2004 H3-1~5 scFv pooled R3 2 nM output #3 -532
YTGRIFLHHLFDY May 14, 2004 H3-1~5 scFv pooled R3 2 nM output #5
-533x DEGMM RIGQDGKNTYYADSVKG YTGRILGHHLFDY Aug. 3, 2004 2-108 scFv
Recomb R1 S1 60 pM output #4 -534x DEGMM RITYSGKNTYYADSVKG
YTGRIFSHHLFDY Aug. 3, 2004 2-108 scFv Recomb R1 S1 60 pM output #9
-535x DEGMM RIGQDGKNTYYADSVKG YTGRIFSHHLFDY Aug. 3, 2004 2-108 scFv
Recomb R1 S1 60 pM output #10 -536x EEKWM RIGQDGKNTYYADSVKG
YTGRILGHHLFDY Aug. 3, 2004 2-108 scFv Recomb R1 S2 60 pM output #4
-537x DEGMM RIGQDGKNTYYRMDVKG YTGRILGHHLFDY Aug. 3, 2004 2-108 scFv
Recomb R1 20 min 600 pM output #2 -538x DEGMM RITYSGKNTYYADSVKG
YTGRILGHHLFDY Aug. 3, 2004 2-108 scFv Recomb R1 20 min 600 pM
output #6
[0351] K.sub.D curves on the surface of yeast for the clones in
relation to 2-108 parental were generated, with the best clones
showing 1-2 logs improvement in binding affinity (FIG. 4). EC50
values for the clones and wild type were as follows (clone name
(EC50 value)): 2-108/1-122 WT (69.29); 504 (1.054); 509 (5.318);
511 (1.848); 518 (4.404); 522 (2.963); 524 (3.103); and 527
(2.596).
[0352] The best clones when converted into IgG format paired with
ABT1-122 showed greater than 1 log improvement in IL-1.beta.
neutralization potency in a MRC-5 assay (FIG. 5).
ABT2-108 Affinity Maturation by CDR Recombination Library
[0353] Both the equilibrium and on rate sorting produced several
clones with improvement in IL-1.beta. affinity over the single CDR
affinity matured clones. Six of these clones were chosen for
further analysis, designated ABT2-108-533x through 538x (see Table
9).
TABLE-US-00009 TABLE 9 Dual-specificity pairings and neutralization
characteristics IL-1.alpha. IL-1.beta. Name Heavy Chain Light Chain
IC50 (nM) IC50 (nM) ABT2-108/ABT1-122 ABT2-108 ABT1-122 150, 500,
1500 15, 20, 40 hu IgG1/K wt ABT2-108-504/ABT1-122 ABT2-108-504
ABT1-122 1500 15 hu IgG1/K wt ABT2-108-518/ABT1-122 ABT2-108-518
ABT1-122 400 0.5 hu IgG1/K wt ABT2-108-521/ABT1-122 ABT2-108-521
ABT1-122 1000 0.2, 0.4 hu IgG1/K wt ABT2-108-533x/ABT1-122
ABT2-108-533x ABT1-122 1000 0.025, 0.15 hu IgG1/K wt
ABT2-108-534x/ABT1-122 ABT2-108-534x ABT1-122 1000 0.025, 0.035 hu
IgG1/K wt ABT2-108-537x/ABT1-122 ABT2-108-537x ABT1-122 1000 0.007,
0.02 hu IgG1/K wt ABT2-108-538x/ABT1-122 ABT2-108-538x ABT1-122
100, 1000 0.007, 0.02, 0.18 hu IgG1/K wt ABT2-108-603/ABT1-122
ABT2-108-603 ABT1-122 1500 10 hu IgG1/K wt ABT2-108-605/ABT1-122
ABT2-108-605 ABT1-122 20, 2000 1.5, 0.35 hu IgG1/K wt ABT2-108-612/
ABT1-122 ABT2-108-612 ABT1-122 100 1, 0.45 hu IgG1/K wt
ABT2-108-617/ABT1-122 ABT2-108-617 ABT1-122 150 0.05 hu IgG1/K wt
ABT2-108-620x/ABT1-122 ABT2-108-620x ABT1-122 150 0.05 hu IgG1/K wt
ABT2-65/ABT1-122 ABT2-65 ABT1-122 2000 2000 hu IgG1/K wt
ABT2-65-17/ABT1-122 ABT2-65-17 ABT1-122 70, 3000 0.1, 0.45 hu
IgG1/K wt ABT2-65-8/ABT1-122 ABT2-65-8 ABT1-122 1000 2.7 hu IgG1/K
wt ABT2-108/ABT1-122-505 ABT2-108 ABT1-122-505 2000 20 hu IgG1/K wt
ABT2-108/ABT1-122-508 ABT2-108 ABT1-122-508 400 20 hu IgG1/K wt
ABT2-65-17/ABT1-122-511 ABT2-108 ABT1-122-510 30, 200 0.1, 0.16 hu
IgG1/K wt ABT2-108/ABT1-122-511 ABT2-108 ABT1-122-511 40 8 hu
IgG1/K wt ABT2-108-521/ABT1-122-511 ABT2-108-521 ABT1-122-511 150
0.16 hu IgG1/K wt ABT2-108-538x/ABT1-122-511 ABT2-108-538x
ABT1-122-511 1, 10, 25, 200 0.006, 0.009, 0.01 hu IgG1/K wt
ABT2-108-620x/ABT1-122-511 ABT2-108-620x ABT1-122-511 10, 40, 200
0.01, 0.02, 0.03 hu IgG1/K wt ABT2-108-620x/ABT1-122-512
ABT2-108-620x ABT1-122-512 150 0.006 hu IgG1/K wt
ABT2-108-620x/ABT1-122-513 ABT2-108-620x ABT1-122-513 30, 100 0.01,
0.009 hu IgG1/K wt ABT2-108/ABT1-122-554 ABT2-108 ABT1-122-554 150
9 hu IgG1/K wt ABT2-108/ABT1-122-555 ABT2-108 ABT1-122-555 70 7 hu
IgG1/K wt ABT2-108-538x/ABT1-122-555 ABT2-108-538x ABT1-122-555 25
0.015 hu IgG1/K wt ABT2-108/ABT1-122-556 ABT2-108 ABT1-122-556 150
15 hu IgG1/K wt ABT2-108-620x/ABT1-122-750M ABT2-108-620x
ABT1-122-750M 18.46 0.00555 hu IgG1/K wt
ABT2-108-620x/ABT1-122-750MT ABT2-108-620x ABT1-122-750MT 10.14
0.00482 hu IgG1/K wt ABT2-108-538x/ABT1-122-750x ABT2-108-538x
ABT1-122-750x 4, 10, 100 0.005, 0.008, 0.01 hu IgG1/K wt
ABT2-108-620x/ABT1-122-750x ABT2-108-620x ABT1-122-750x 17.52 0.005
hu IgG1/K wt ABT2-108/ABT1-141 ABT2-108 ABT1-141 1000, 1500, 3000
15, 40, 50, 80 hu IgG1/K wt ABT2-65/ABT1-141 ABT2-65 ABT1-141 7000
100 hu IgG1/K wt ABT2-65-17/ABT1-141 ABT2-65-17 ABT1-141 70 0.4 hu
IgG1/K wt ABT2-65-8/ABT1-141 ABT2-65-8 ABT1-141 450 0.09 hu IgG1/K
wt ABT2-108/ABT1-141-25 ABT2-108 ABT1-141-25 N/A 150 hu IgG1/K wt
ABT2-108-521/ABT1-141-25 ABT2-108-521 ABT1-141-25 20 0.1 hu IgG1/K
wt ABT2-65-17/ABT1-141-25 ABT2-65-17 ABT1-141-25 400 1 hu IgG1/K wt
ABT2-65-17/ABT1-212 ABT2-65-17 ABT1-212 100 0.5, 0.09 hu IgG1/K wt
ABT2-65-8/ABT1-212 ABT2-65-8 ABT1-212 5000 3.5 hu IgG1/K wt
ABT2-108-538x/ABT1-221 ABT2-108-538x ABT1-221 200 0.01 hu IgG1/K wt
ABT2-108-538x/ABT1-221 ABT2-108-538x ABT1-222 200 0.01 hu IgG1/K wt
ABT2-108-538x/ABT1-95-15 ABT2-108-538x ABT1-95-15 0.9, 4, 10, 19
0.003, 0.01, 0.02, 0.03, 0.008 hu IgG1/K wt
ABT2-108-620x/ABT1-95-15 ABT2-108-620x ABT1-95-15 5 0.04 hu IgG1/K
wt ABT2-65-166/ABT1-95-15 ABT2-65-166 ABT1-95-15 1.89 0.13 hu
IgG1/K wt ABT2-65-17/ABT1-95-15 ABT2-65-17 ABT1-95-15 0.5, 1, 5,
5.35 0.01, 0.04, 0.08, 0.1 hu IgG1/K wt ABT2-65-201/ABT1-95-15
ABT2-65-201 ABT1-95-15 0.652 0.03 hu IgG1/K mut (234; 235)
ABT2-65-166/ABT1-95-184 ABT2-65-166 ABT1-95-184 0.068 0.15 hu IgG1
mut 234, 235/K ABT2-108/ABT1-95-3 ABT2-108 ABT1-95-3 1500 200 hu
IgG1/K wt ABT2-108-521/ABT1-95-A3 ABT2-108-521 ABT1-95-A3 0.027
0.098 hu IgG1/K wt ABT2-108-538x/ABT1-95-A3 ABT2-108-538x
ABT1-95-A3 0.095 0.035 hu IgG1 mut 234, 235/K
ABT2-108-620x/ABT1-95-A3 ABT2-108-620x ABT1-95-A3 0.002 0.0073 hu
IgG1/K mut (234; 235) ABT2-65-166/ABT1-95-A3 ABT2-65-166 ABT1-95-A3
0.026, 0.0178 0.17 hu IgG1/K wt ABT2-65-166/ABT1-95-A5 ABT2-65-166
ABT1-95-A5 0.015 0.145 hu IgG1 mut 234, 235/K
ABT2-65-166/ABT1-95-A6 ABT2-65-166 ABT1-95-A6 0.019, 0.35 0.32 hu
IgG1/K wt
[0354] All showed greater than 1 log improvement in affinity on the
surface of yeast in comparison to one of the best single CDR
affinity matured 2-108 clones (FIG. 6). EC50 values for the clones
and wild type were as follows (clone name (EC50 value)): 533x
(0.04322); 534x (0.2354); 535x (0.04475); 536x (0.07300); 537x
(0.03390); 538x (0.03956); and 521 (0.5222).
[0355] The best CDR recombination clones when converted into an IgG
format paired with 1-122 showed greater than 1 log improvement in
IL-1.beta. neutralization potency vs. one of the best single CDR
2-108 affinity matured clones in a MRC-5 assay. EC50 (nM) values
were determined as follows (clone name (EC50 value)):
2-108-533x/1-122 (0.03525); 2-108-534x/1-122 (0.02724);
2-108-537x/1-122 (0.006324); 2-108-538x/1-122 (0.005366);
2-108-521/1-122 (0.1878); and MAB201 (0.001218). The EC50 data
showed the IL-1.beta. neutralization potency of CDR recombination
vs. single CDR affinity matured 2-108 clones as IgG.
[0356] In conclusion, the above describes a successful experiment
which was able to affinity mature an IL-1.beta. binding dAb in a
scFv by yeast display. Through the use of the small scanning
libraries and subsequent CDR recombination, multiple clones were
identified that improved the affinity of 2-108 for IL-1.beta. by
greater than 3 logs. At least 4 of these clones, ABT2-108-533x,
534x, 537x, and 538x, were able to achieve the target goal of
better than 200 pM neutralization potency for IL-1.beta. in a MRC-5
assay when converted to an IgG format.
2.2.2 Affinity Maturation of Anti-IL-1.alpha. dAb Abt1-95-15 by
Yeast Display
[0357] ABT1-95 was previously identified as an anti-human
IL-1.alpha. Vic dAb (see Example 1 and Table 60). ABT1-95 had low
IL-1.alpha. neutralizing potency as a dAb and insignificant potency
when converted into IgG. Using an emulsion technology and CDR
mutagenesis, an improved ABT1-95-15 clone (dAb IC.sub.50 in the low
pM range) was generated (see above Example 2.1), with confirmed its
improved potency (IC.sub.50 .about.5 nM) in the IgG format with
pairing heavy chains ABT2-108 or ABT2-65 lineages (anti-IL-1.beta.
VH dAbs).
[0358] Since it is difficult to further improve the low pM affinity
of ABT1-95-15 in the dAb format, affinity maturation of ABT1-95-15
was performed in the scFv format by yeast display. The objective of
this study was to affinity-mature ABT1-95-15 by yeast display in
the scFv format by large CDR spiking libraries to sub-nM affinity
and potency in the IgG format.
Materials and Methods
[0359] Construction of the scFv Yeast Expression Vector.
[0360] ABT1-95-15 (VL to IL-1.alpha.) and ABT2-108-538x (VH to
IL-1.beta.) were subcloned into pYDs-TEV vector by standard
molecular cloning. The complete construct is shown in FIG. 11.
Construction of Large CDR Spiking Libraries.
[0361] The pYDs-TEV plasmid was linearized with Sfi I and Not I
restriction enzymes and purified.
[0362] ABT2-108-538x/ABT1-95-15 scFv fragment DNA was produced by
overlapping PCRs with ABT1-95-15 framework primers, pYD vector
specific primers, and long oligonucleotide primers containing
mutations in each of the three ABT1-95-15 CDRs. The mutations were
introduced by synthesizing the CDR region sequences at nucleotide
ratio 70-10-10-10 (70%-original nucleotide, the other three
nucleotides--10% each), (Table 10).
TABLE-US-00010 TABLE 10 Primers used for large CDR spiking
libraries construction Name Oligo Sequence 1-95-15-L1.sup.a
CATCCTCCCTGTCTGCATCTGTAGGAGACCGT GTCACCATCACTTGCCGGGCAAGTCAGCCG
ATTGTGCGGAACTTAAGGTGGTATCAGCAG AAACCAGG 1-95-15-L2.sup.a
TGGTATCAGCAGAAACCAGGGAAAGCCCCTAAG CTCCTGATCTATTCTTCGTCCTATTTGGAGC
CCGGGGTCCCATCACGTTTCAGTGGTAGTG GATCCG 1-95-15-L3.sup.a
ACTCTCACCATCAGCAGTCTGCAACCCGAAGAT TTTGCTACGTACCACTGTCAACAGGGGTA
TCGTTGGCCTGTTACGTTCGGCCAAGG GACCAAGGTGG 1-122 Gap 1 Rev
GCAAGTGATGGTGACACGGTCTCCTACAGATGC new 1-122 Gap 3 Rev
ATAGATCAGGAGCTTAGGGGCTTTCCCTGG new 1-95 Gap4 Rev
ACAGTGGTACGTAGCAAAATCTTCG pYD1 Fwd AGTAACGTTTGTCAGTAATTGC ABTVKSCFV
Rev TTAGGGATAGGCTTACCTTCGAAGGGCC CTCTAGACTCGAGGGCGGCCGCACGTT
TGATTTCCACCTTGG .sup.aSequences in bold are spiked
Yeast Libraries Generation and QC.
[0363] Large yeast libraries were generated via homologous
recombination by co-transforming yeast (EBY100) with linearized
vector and PCR fragments with spikes in each of ABT1-95-15 CDRs in
(FIG. 12).
[0364] Libraries output sizes were estimated by plating aliquots of
transformed cells on SD(-UT) plates. Libraries diversity was
verified by sequencing analysis of 24 individual colonies from each
library.
Yeast Libraries Labeling and Sorting.
[0365] Expression of scFv fragments on the surface of yeast was
induced by growing libraries in SRG(-UT) medium for 48 hours at
20.degree. C.
[0366] For equilibrium library sorting yeast cells
(5.times.10.sup.8 cells for each library at first round) were
labeled with antigen (human IL-1.alpha.) at a concentration of 660
.mu.M followed by staining with detection antibody (biotinylated
goat polyclonal anti-IL-1.alpha. antibody, R&D Systems,
Cat#BAF200, at concentration 5 .mu.g/ml) and then Streptavidin-PE
(Jackson ImmunoResearch Laboratories, Inc, Cat #016-110-034). ScFv
expression was determined by staining libraries with anti-V5 mAb
(Invitrogen, Cat#R96025) followed by anti-mouse IgG-FITC (Molecular
Probes, Cat #F11021).
[0367] For off-rate sorts aliquots of round 1 outputs
(9-25.times.10.sup.7 cells) were labeled with human IL-1.alpha. at
10 nM concentration followed by competition with IL-1.alpha.
specific IgG (ABT2-65-17/ABT1-95-15 A-897661.0) at concentration
100 .mu.g/ml for 60-80 minutes (3 to 4 half-lives). Antigen, still
bound to the yeast surface was detected as described above.
[0368] Round 2 outputs (8-20.times.10.sup.6 cells) were
subsequently sorted for round 3 as described above, except that
competition time was 100 min.
[0369] Fourteen to twenty-eight clones from each library Round 3
output were analyzed by sequencing. Three clones from each soiling
outputs were also analyzed on the surface of yeast for their
affinity improvement.
[0370] CDR1 fragments from ABT1-95-15 L1 round 3 output, CDR2
fragments from ABT1-95-15 L2 round 3 output, and CDR3 fragments
from ABT1-95-15 L3 round 3 output were mixed with linearized
pYDs-Tev vector and transformed into EBY100 yeast cells to generate
ABT1-95-15 CDR recombination library (FIG. 13)
[0371] Off-rate sort with competition for 3 hours (Round 1), 8
hours (Round 2) and 15 hours (Round 3) was used for CDR
recombination library sortings. Round 3 output of CDR recombination
library was analyzed by sequencing and 6 clones were selected for
analysis on the surface of yeast. Best clones were converted into
IgG paired with VH clones of 2-108 and 2-65 lineages.
Results
ABT1-95-15 Affinity Maturation by Large Single CDR Spiking
Libraries
[0372] Estimated libraries output sizes for ABT1-95-15 libraries
were 2-5.times.10.sup.7. Libraries QC analysis confirmed the
presence of designed mutations in all CDR spiking libraries.
Equilibrium sort was used for first round selection to remove
non-binders for IL-1.alpha. antigen. The libraries were labeled at
K.sub.D of ABT2-108-538x/ABT1-95-15 scFv with 660 pM human
IL-1.alpha. (see FIG. 14). Off-rate selection was used in
subsequent library selections. Half-life of ABT1-95-15/IL-1.alpha.
complex on the surface of yeast was determined to be 20 min (see
FIG. 15). Half life values were determined as follows (clone name
(half life)): 1-95-15 (19.77); L1-2B (162.7); L1-3B (52.56); L1-7
(31.56); L2-3 (38.26); L2-4 (39.47); L2-10 (21.43); L3-9 (46.10);
L3-12 (23.62); and L3-13 (32.08).
[0373] Based on three rounds of sorting results for the three
ABT1-95-15 individual CDR libraries (data not shown), all three
libraries showed some improvement over ABT1-95-15 after three
rounds of sorting.
[0374] Sequencing results of round 3 library selection output are
listed in Table 11.
TABLE-US-00011 TABLE 11 Sorting results for ABT1-95-15 affinity
maturation by large single CDR libraries Fre- quen- Clone CDR1 CDR2
CDR3 cy ABT1-95-15 RASQPIVRNLR SSSYLEP QQGYRWPVT a) RASQPIVRNLR 2
RASQSNLRNLR 1 RASRTPVRNLR 1 RANRTRVRNLR 2 L1-2B RKNNPFVRNLR 4
RKNHPNMRNLR 1 RKNHPHIRNLR 1 L1-7 RKTHPTVRNLR 2 RKAHPMVRNLR 1
RKLQPFVRNLR 1 RKSSPMVRNLR 1 LKRHPSVRNLR 1 LKRQPSVRNLR 1 L1-3B
RKSHPFVRNLR 2 RASQTRVQNLR 1 RARRAPVRNLR 1 RARNTPVRNLR 1 RASRPNVRNLR
1 RASRPIVRNLR 1 RGSRPGIRNLR 1 RRSGPNVRNLR 1 LKSHPHVRNLR 1 b)
SSSYLEP 1 L2-4 SVSYLEP 4 L2-3 SRSYLEP 2 L2-10 SKSYLEP 2 SVSWLEP 1
SISYLEP 1 STKNVER 1 SWSFLQP 1 SSNYLAP 1 ARSYLEP 1 c) QQGYRWPVT 10
RMGYRWPVT 1 L3-10 KAGYVWPVP 1 L3-12 KAGYRWPVT 1 L3-13 RSGYRWPVT a)
Clones from CDR1 library (L1) b) Clones from CDR2 library (L2) c)
Clones from CDR3 library (L3)
[0375] Sequencing results showed that round 3 selection outputs for
all 3 libraries were diverse, with most diversity observed in CDR1
library (L1). CDR3 library (L3) was least diverse and majority of
the clones had parental CDR3 sequence.
[0376] Three clones from each library were selected for the
analysis on the surface of yeast based on their frequency. Off-rate
analysis for these clones on yeast surface is presented in FIG.
15.
[0377] All analyzed clones showed improvement (2-5 fold) in their
off-rates over parental clone ABT1-95-15, and the most improvement
was observed in clones from the CDR1 library (L1).
ABT1-95-15 CDR Recombination Library
[0378] Round three library outputs for individual CDR libraries
were recombined in yeast to generate an ABT1-95-15 CDR
recombination library (data not shown).
[0379] Significant improvement in off-rate over parental clone
ABT1-95-15 was observed in the first round of library sorting.
Additional rounds of sorting with increasing competition time with
IL-1.alpha.-specific IgG ABT2-65-17/ABT1-95-15 did not lead to
better discrimination between the best clones and the rest of
population. The reason for this was probably the fact that off-rate
for IgG competitor was much faster then for affinity-matured clones
and therefore the competitor ABT2-65-17/ABT1-95-15 IgG had become
inefficient for antigen competition. As a result, round 3 sorting
output for CDR recombination library was very diverse (see Table
12).
[0380] Six clones from ABT1-95-15 CDR Recombined Library Round 3
output (ABT1-95-A1, -A2, -A3, A4, A5 and -A6) were selected for
further analysis on yeast surface based on their frequency and
frequencies of their individual CDRs.
[0381] On-rate, off-rate and K.sub.D analysis of these clones on
yeast surface is presented in FIGS. 16 and 17. A 10 to 30 fold
improvement was observed in KD on yeast surface for affinity
matured clones. On rate analysis half life was determined to be the
following (clone name (half life)): 1-95-A3 (2.817); 1-95-A5
(3.244); 1-95-A6 (4.485); 1-95-15 (6.924); amd 1-95-A1 (9.449). Off
rate analysis half life was determined to be (clone name (half
life)): 1-95-A3 (1.795); 1-95-A2 (2.235); 1-95-A4 (1.991); 1-95-A6
(1.795); 1-95-15 (0.3978); amd 1-95-A1 (3.132). EC50 values were
determined to be (clone name (EC50 value)): 1-95-A3 (0.01982);
1-95-A5 (0.02668); 1-95-A6 (0.03965); 1-95-A1 (0.7647); 1-95-15
(0.6731).
TABLE-US-00012 TABLE 12 1-95-15 CDR Recombined Library Round3
sorting outputs Clone CDR1 CDR2 CDR3 Frequency ABT1-95-15
RASQPIVRNLR SSSYLEP QQGYRWPVT ABT1-95-A3 RASKPGVRNMR SVSYLEP
LQGYRWPPT 11 RASKPGVRNMR SVSYLEP QQGYRWPVT 1 RASKPGVRNMR SVSYLEP
RQGYRWPVT 1 ABT1-95-A6 RASKPGVRNMR AKSYLEP KQGYRWPVQ 2 RASKPGVRNMR
AKSYLEP QQGYRWPVT 1 RASKPGVRNMR SKSYLEP QQGYRWPVT 1 RASKPGVRNMR
SSSYLNP QQGYRWPVT 2 RASKAGVRNLR SVSYLEP LQGYRWPPT 1 RASKAGVRNLR
SVSWLEP RQGYVWPVP 1 ABT1-95-A4 RASRPGVRNLR SRSFLEP LQGYRWPPT 2
RASRPGVRNLR SRSFLEP QQGYRWPVT 1 RASRPGVRNLR SVSFLEP RQGYVWPVP 1
RASRPGVRNLR SVSYLEP LQGYRWPPT 1 ABT1-95-A1 RASRPGVRNLR HVSDLEP
RQGYVWPVP 4 RGSRPGIRNLR SVSFLEP LQGYRWPPT 1 RGSRPGIRNLR ASSNLEP
QQGYRWPVT 1 ABT1-95-A5 RASRTPVRNLR SRSFLEP LQGYRWPPT 2 QASQPGVRNMR
SVSFLEP RQGYRWPVT 1 QASQPGVRNMR SVSYLEP QQGYRWPVT 1 QASQPGVRNMR
STSSLQP QQGYRWPVT 1 ABT1-95-A2 RILQPPGRNLR SKSFLEP QQGYRWPVT 6
RASHGGVRNLR SASYLEP QQGYRWPVT 1 RASHSPVRNLR SVSYLEP KAGYVWPVP 1
RAGHPRVRNLR SSSYLQP QQGYRWPVT 1 RASNQRVRNLR SVSYLEP QQGYRWPVT 2
RKNHPDVRNLR SSSLLEP QQGYRWPVT 1 RKSQPNMRNLR STSLLDR QQGYRWPVT 1
RASQPGVRNLR SSSYLEP QQGYRWPVT 2 RASQPSVRNLR SKSYLEP QQGYRWPVT 1
[0382] Based on the K.sub.D analysis on yeast surface, clones
ABT1-95-A3, -A5 and -A6 were selected for conversion into IgG
format with a pairing heavy chain containing VH of ABT2-108 and
ABT2-65 lineages. MRC-5 assays demonstrated up to 1000-fold
improvement in IL-1.alpha. neutralization potency for the
affinity-matured clones. MRC-5 IL-1.alpha. (50 pg/ml IL-1a)
inhibition assay values (EC50 nM) were determined to be 0.004457
for ABT2-108-620x/ABT1-95-A3 compared with 5.074
ABT2-108-620x/ABT1-95-15 (controls IL-1RA=0.6996 and
MAB200=0.07000).
[0383] In conclusion, the above example demonstrates that it is
possible to affinity mature an anti-IL-1.alpha. V.kappa. domain
antibody ABT1-95-15 to low pM potency by yeast display in the scFv
format. The three CDRs were mutated by spiking in 30% of mutating
nucleotides in the CDR regions, which resulted in three very large
yeast libraries. These libraries were selected for improved clones
through three rounds of sorting, followed by the generation of a
large CDR recombination library to combine all CDR mutations
present in the round 3 selection outputs. This approach maximizes
the chance to identify CDR mutations that are beneficial to the
antigen affinity without pre-determining the mutation positions
within the CDRs as they are determined by random and low frequency
mutagenesis. The subsequent CDR recombination library approach
seeks to identify individual CDR mutations that are compatible with
one another that as a whole further the affinity improvement of
these mutants. This method is time efficient.
2.3. Affinity Maturation of IL-1.alpha./.beta. Dual-Specific
Antibodies by a Yeast Surface Fab expression system
[0384] Two domain antibodies (anti-IL-1.beta. VH dAb ABT2-108 and
anti-IL-1.alpha. V.kappa. dAb ABT1-122), identified from single
domain antibody library selections (described above in Example 1)
were chosen for affinity maturation by yeast display. Both Fab and
scFv expression systems were employed to carry out the affinity
maturation works. This example focuses on the affinity maturation
works by the Fab expression system.
[0385] The objective of the study was to develop a yeast surface
Fab expression system for the affinity maturation of two IL-1 dAb
leads.
Materials and Methods
Construction of a Fab Expression Vector.
[0386] The pFab-B vector was based on the pESC-Trp vector
(Stratagene) for expressing Fab proteins on yeast surface. Both
ABT2-108 and ABT1-122 were subcloned into the pFab-B vector by
standard molecular cloning methods. The completed
pFab-B-2-108/1-122 vector is shown in FIG. 9.
Construction of Mini-Libraries.
[0387] Gapped Fab vectors were generated by PCR amplification using
Platinum HiFi DNA polymerase (Invitrogen) and `gap primers` that
flanks each CDRs. The PCR amplification produced `gapped vectors`
in which 15 to 21 nucleotides were deleted from pFabB-2-108/1-122,
leaving a gap across the CDR of interest. Seven gapped vectors were
generated for ABT 2-108 (1 for CDR1, 3 for CDR2, and 3 for CDR3)
and five gapped vectors for ABT 1-122 (2 for CDR1, 1 for CDR2, and
2 for CDR3). Primers used for gapped vectors generation are listed
below in Table 13.
[0388] Thirty homologous recombination oligos were made for
ABT2-108 and twenty-one oligos were made for ABT1-122. These oligos
contained the 15-21 nucleotides removed in the gapped vectors as
well as about 40 nucleotide-flanking sequences both up- and
downstream of the gap. Each homologous recombination oligos
randomizes three consecutive amino acids in the CDR gap.
TABLE-US-00013 TABLE 13 PCR primers used for gapped vectors
generation SEQ ID Name Oligo sequence NO: 2-108 Gap 1 Fwd new
TGGGTCCGCCAGGCTCCAGGGAAGGGT CTC 2-108 Gap 1 Rev new
AAAGGTGAATCCGGAGGCTGCACAGGA GAG 2-108 Gap 2 Fwd new
AATACATACTACGCAGACTCCGTGAAG GGC 2-108 Gap 2 Rev new
TGAGACCCACTCGAGACCCTTCCCTGG AGC 2-108 Gap 3 Fwd new
GACTCCGTGAAGGGCCGGTTCACCATC TCC 2-108 Gap 3 Rev new
ATCCTGCCCAATACGTGAGACCCACTC GAG 2-108 Gap 4 Fwd new
CGGTTCACCATCTCCCGCGACAATTCC AAG 2-108 Gap 4 Rev new
GTATGTATTCTTACCATCCTGCCCAAT ACG 2-108 Gap 5 Fwd new
CATCATCTTTTTGACTACTGGGGTCAG GGAAC 2-108 Gap 5 Rev new
TTTCGCACAGTAATATACCGCGGTGTC CTC 2-108 Gap 6 Fwd new
GACTACTGGGGTCAGGGAACCCTGGTC ACC 2-108 Gap 6 Rev new
AACCCGACCCGTATATTTCGCACAGTA ATATAC 2-108 Gap 7 Fwd
TGGGGTCAGGGAACCCTG 2-108 Gap 7 Rev ATGATGAACACCAACCCGAC 1-122 Gap 1
Fwd new ACTGAGTTAAATTGGTATCAGCAGAAA CCAGG 1-122 Gap 1 Rev new
GCAAGTGATGGTGACACGGTCTCCTAC AGATGC 1-122 Gap 2 Fwd new
TGGTATCAGCAGAAACCAGGGAAAGCC CCTAAG 1-122 Gap 2 Rev new
CGGCTGACTTGCCCGGCAAGTGATGG TG 1-122 Gap 3 Fwd new
GGGGTCCCATCACGTTTCAGTGGCAGT GG 1-122 Gap 3 Rev new
ATAGATCAGGAGCTTAGGGGCTTTCCC TGG 1-122 Gap 4 Fwd new
GCTACGTTCGGCCAAGGGACCAAGGTG GAAATC 1-122 Gap 4 Rev new
ACAGTAGTACGTAGCAAAATCTTCAGG TTGCAG 1-122 Gap 5 Fwd new
TTCGGCCAAGGGACCAAGGTGGAAATC AAACG 1-122 Gap 5 Rev new
AGCAAACTGTTGACAGTAGTACGTAGC AAAATC
Yeast Libraries Generation and QC.
[0389] Yeast mini-libraries were generated via homologous
recombination by co-transforming yeast (EBY100) with gapped vectors
and corresponding homologous recombination oligos.
[0390] Libraries output sizes were estimated by plating aliquots of
transformed cells on SD(-UT) plates. Libraries diversity was
verified by sequencing analysis of 5 individual colonies from each
library.
[0391] Yeast Libraries Labeling and Sorting.
[0392] Expression of Fab fragments on the surface of yeast was
induced by growing libraries in SRG(-UT) medium for 48 hours at
20.degree. C.
[0393] Aliquots of libraries (4.times.10.sup.6 cells for each
library) were labeled with antigen (IL-1.alpha. or IL-1.beta.) at
defined concentrations, followed by staining with detection
antibodies (biotinylated goat polyclonal anti-IL-1.alpha. antibody,
R&D Systems, Cat #BAF200, at concentration 5 .mu.g/ml, or
biotinylated goat polyclonal anti-IL-1.beta. antibody, R&D
Systems, Cat #BAF 201, at concentration 5 .mu.g/ml), and then
straptavidin-PE. Fab expression was determined by staining
libraries with either anti-HA niAb 12C5 (Roche) or anti-human Cx
(AbCam, Cat#Ab1050), followed by anti-mouse IgG-FITC (Molecular
Probes).
[0394] ABT2-108 libraries were individually sorted for Round 1 at
antigen (IL-1.beta.) concentration 220 pM. R1 outputs were sorted
individually and in pools at antigen concentration 220 pM (Round
2). Pooled R2 outputs were sorted at antigen concentration 220
.mu.M and 110 pM (Round 3). Ten clones from each pooled R3 output
were analyzed by sequencing
[0395] ABT1-122 libraries were individually sorted for Round 1 at
antigen (IL-1.alpha.) concentration 100 nM. R1 outputs were sorted
individually and in pools at antigen concentration 100 nM (Round
2). Pooled R2 outputs were sorted at antigen concentration 50 nM
(Round 3) Pooled R3 outputs were sorted at antigen concentration 20
nM (Round 4). Ten clones from each pooled R3 and R4 output were
analyzed sequencing.
[0396] Identified clones were analyzed for their KD on the surface
of yeast. Clones with improved KD were converted into full-length
IgG and expressed in COS cells. ABT2-108 CDR recombination
libraries generation and sorting
[0397] CDR1 fragments were PCR amplified from ABT2-108, -601, -602,
and -618 using primers 2-108-CDR1F (Table 14) and 2-108Gap2Rev-new.
CDR2 fragments were PCR amplified from ABT2-108, -603, -604, -605,
-606, -607, -607, -609, -610, -611, -615, and -619 using primers
2-108-CDR2F and 2-108-CDR2R. CDR3 fragments were amplified from
ABT2-108, -612, -613, -614, -616, and -617 using primers
2-108Gap4Fwd-new and 2-108-CDR3R. Overlapping CDR fragments were
transformed into and homologously recombined in EBY100 cells with
either ABT1-122 or ABT1-122-511 pairing light chain to generate CDR
recombination libraries. The estimated library output sizes are
2.2.times.10.sup.5 for ABT1-122-paired library and
3.4.times.10.sup.5 for ABT1-122-511-paired library. Twelve colonies
from each library were sequenced for QC.
TABLE-US-00014 TABLE 14 PCR primers used in CDR amplification for
recombination libraries Name Oligo Sequence 2-108-CDR1F
CAAGTATTTCGGAGTGCCTGAAC 2-108-CDR2F GTCCGCCAGGCTCCAGGGAAAGG
2-108-CDR2R TCGCACAGTAATATACCGCGGTG 2-108-CDR3R
GCATTGGTGACTATTGAGCACGTG
[0398] Two rounds of on-rate sorting were performed for these two
libraries. Round 1 was selected by labeling library with 500 pM
biotinylated IL-13 for 15 minutes incubation. Round 2 was selected
with both 250 pM and 500 pM biotinylated IL-1.beta. for 15 minutes
incubation. Eight clones from the 500 pM R2 sorts and 4 clones from
the 250 pM R2 sorts were sequenced from each library. An clone
ABT2-108-620x identified from the R2 selection was chosen for
conversion into IgG and tested by MRC-5 assay.
Results
ABT2-108 Affinity Maturation
[0399] Estimated libraries output sizes for ABT2-108 libraries were
6.times.10.degree. to 3.times.10.sup.5. Libraries QC analysis
confirmed the presence of designed mutations for most of the
libraries. Some colonies however showed mixed CDR sequences
presumably due to an initial uptake of more then one copy of the
plasmid by yeast cell. Sorting results for ABT2-108 affinity
maturation are shown in FIG. 8.
[0400] All 6 pooled libraries showed improved binding to IL-1.beta.
at concentration of 110 pM IL-1.beta. after 3 rounds of sorting.
Sequencing analysis of individual clones from round 3 outputs
(Table 15) revealed existence of hot spots for mutagenesis. The
same hot spots and similar or identical mutations were identified
during ABT2-108 affinity maturation in yeast display ScFv
format.
TABLE-US-00015 TABLE 15 Sorting outputs for ABT2-108 affinity
maturation Clone # CDR1 CDR2 CDR3 Frequency ABT2 -108 EYTMM
RIGQDGKNTYYADSVKG YTGRVGVHHLFDY -601 EESWM 28 -602 EEKYM 1 -603
RITDAGKNTYYADSVKG 14 -604 RVTYDGKNTYYADSVKG 2 -605
RIGQDGKNTYYREDVKG 8 -606 RIGQDGKNTYYRSSVKG 1 -607 RIGQDGKNTYYRSDVKG
1 -608 RIGQDGKNTYTRDSVKG 3 -609 RIGQDGKNTVYRDSVKG 3 -610
RIGQDGKNTVVRDSVKG 1 -611 RIGQDGKNTVLRDSVKG 2 -612 YTGRIMGHHLFDY 3
-613 YTGRVFEHHLFDY 6 -614 YTGRILRHHLFDY 6 -615 RIGQDGKNTVIRDSVKG 1
-616 YTGRIYNHHLFDY 2 -617 YTGRIFTHHLFDY 2 -618 EEYFM 3 -619
RHGWDGKK-YYADSVKG 7
[0401] KD analysis of identified clones on the surface of yeast
(FIG. 9) showed improvements in affinity from 5 fold (ABT2-108-604)
to 25 fold (ABT2-108-615, -617). EC50 values of the clones were as
follows (clone name (EC50 nM): 2-108 (5.778); 2-108-601 (0.5225);
2-108-602 (0.4741); 2-108-603 (1.349); 2-108-604 (1.348); 2-108-605
(0.2096); 2-108-606 (0.3432); 2-108-608 (0.3657); 2-108-611
(0.3508); 2-108-612 (0.2674); 2-108-613 (0.5273); 2-108-614
(1.010); 2-108-615 (0.2145); 2-108-616 (0.3237); and 2-108-617
(0.2236).
[0402] Six improved clones (ABT2-108-601, -603, -605, -612, -613
and -617) were converted into IgG constructs and expressed in COS
cells paired with ABT1-122 light chain.
[0403] MRC-5 analysis of these clones showed significant
improvement in neutralization potency (by at least 20-100 fold when
compared with the parental ABT2-108) for clones ABT2-108-605, -612,
-613 and -617.
ABT1-122 Affinity Maturation
[0404] Estimated libraries output sizes for ABT1-122 libraries were
8.times.10.sup.4 to 3.times.10.sup.5. Libraries QC analysis again
showed mixed CDR sequences in many libraries, presumably due to
initial uptake of more than one copy of the plasmid by yeast cell.
Furthermore, some of the clones were found to contain multiple
repeats of the recombination oligos and several clones showed
single base insertion or deletion in the coding region. In all,
only 25% of the clones analyzed had sequences with designed
mutations in appropriate CDRs. Three libraries covering the 3' half
of CDR3 were not sorted due to high background of parental vector
during library construction.
[0405] Sorting results for ABT1-122 affinity maturation are shown
in FIG. 10. None of the pooled ABT1-122 libraries showed
significant improvement in IL-1.alpha. binding after 3 rounds of
sorting. Many clones in these libraries showed lack of the staining
with anti-human C.kappa. antibody, presumably due to frame-shifting
mutations occurred during libraries construction.
[0406] Sequencing analysis of individual clones from Round 3
outputs is shown below in Table 16.
TABLE-US-00016 TABLE 16 Sorting outputs for 1-122 affinity
maturation Clone # CDR1 CDR2 CDR3 Frequency ABT1 -122 RASQPIWTELN
GSSSLQS QQFAYFPATF -501 RFSYPIWTELN 2 -502 RAVYLIWTELN 1 -503
EQFAYFPATF 2 -504 KQFAYFSATF 1 -505 KQFAYFPATF 9 -506 GSMHTQS 1
-507 RSSSLQS 1 -508 GSSSLQR 1 -509 GHAELQS 8
None of the identified clones, when analyzed on the surface of
yeast, showed improvements in KD, even though they were much
brighter than the parental clone ABT1-122. Similar results (i.e.
lack of improved IL-1.alpha.-binding clones) were found by
affinity-maturing ABT1-122 in the scFv format.
[0407] Three of the identified Fab clones (ABT1-122-505, -508 and
-510) were converted into IgG constructs and expressed in COS cells
with pairing ABT2-108 heavy chain. None of these clones showed
improvements in IL-1.alpha. neutralization potency. Mutations from
ABT1-122-505 and -508 were subsequently combined and produced the
ABT1-122-511 clone. This clone, when converted into IgG with
several of the improved anti-IL-1.beta. ABT2-108 heavy chains
showed improvements in its IL-1.alpha. neutralization potency by 4
to 5 folds. Representative IC50 data for ABT2-108/1-122-511 IgG is
described in Table 9.
ABT2-108 CDR Recombination
[0408] Analysis of the round 2 outputs of the 2-108 CDR
recombination clones is shown in Table 17. ABT2-108-601 was the
dominant CDR1 sequence seen, and ABT2-108-612 was the most frequent
CDR3 sequence observed. CDR2 sequences in the output showed more
diversity, with ABT2-108-605 being represented the most. Based on
these data, ABT2-108-620x(601-605-612) was converted in IgG with
ABT1-122 and ABT1-122-511 light chains and tested in MRC-5 assays.
Both pairs of ABT2-108-620x showed IL-1.beta. neutralization
potencies in the low pM range (see Table 18 below).
TABLE-US-00017 TABLE 17 ABT2-108 CDR recombination library R2
sorting outputs 1-122 Paired Library 1-122-511 Paired Library 0.5
nM Sort, R2 0.5 nM Sort, R2 CDR1-CDR2-CDR3 CDR1-CDR2-CDR3
601-WT-612 601-WT-612, 3x 601-605-612 601-605-612 601-606-612
601-607-612 601-608-612 601-605-616 601-609-612 601-605-617
601-605-613 601-605-WT 601-605-616 0.25 nM Sort, R2 601-607-616
601-WT-612, 2x 0.25 nM Sort, R2 601-605-612, 2x WT-605-612
601-609-612 601-606-612 601-605-616
TABLE-US-00018 TABLE 18 MRC-5 IL-1.beta. neutralization potencies
Clone numbers % Inhbition 2-108/1-122 ~15 nM 2-108-605/1-122 ~0.35
nM 2-108-612/1-122 ~0.45 2-108/1-122-511 ~8 nM 2-108-620x/1-122
~0.05 nM 2-108-620x/1-122-511 ~0.03 nM MAB201 ~0.004 nM
[0409] In conclusion, the above example demonstrates the
feasibility of affinity maturation of dual-specific antibody by a
newly developed Fab expression system on the surface of yeast
cells. ABT2-108 dAb was affinity matured by at least 20-100 fold in
this Fab expression system. This finding is consistent with that
observed by the well established scFv yeast display system
described above in Examples 2.2, as similarly improved clones were
identified by bath approaches.
[0410] The identification of improvement clones in the ABT1-122
individual CDR affinity maturation did not occur with the Fab
expression system, although this was also observed by a parallel
scFv approach (data not shown). The combination of two single amino
acid changes in CDR2 and CDR3, however, resulted in an ABT1-122-511
clone with 4-5 fold improved neutralization potency as compared
with parental ABT1-122 clone.
[0411] Thus, the results showed that domain antibodies can be
affinity matured, and, furthermore, that yeast Fab display system
can be used for their affinity maturation.
Example 3
Summary of dAbs
[0412] A summary of the domain antibodies with advantageous
neutralizing and affinity characteristics, i.e., dAbs with these
properties identified in the library screen and subjected to
affinity maturation processes described above, is provided in Table
61 and in Table 19 below. For each clone described herein, the
first part its name relates to its specificity (IL-1.alpha.
represented by ABT1; and IL-1.beta. represented by ABT2).
TABLE-US-00019 TABLE 19 Summary of lead dAb molecules Clone Name
Clone type Specificity MRC5 ND.sub.10 MRC5 ND.sub.50 ABT1-6-23 VH
(3.5G NH3) IL-1.alpha. 3 nM 30 nM ABT1-86 VH (4G H17) IL-1.alpha.
<200 nM 900 nM ABT1-95 VK (4G K) IL-1.alpha. 150 nM 700 nM
ABT1-96 VH (4G H15) IL-1.alpha. 3 nM 11 nM ABT1-98 VH (4G H11)
IL-1.alpha. 10 nM 30 nM ABT1-122 VK (4G K) IL-1.alpha. 9 nM 50 nM
ABT1-141 VK (4G K) IL-1.alpha. 10 nM 70 nM ABT2-13 VH (4G H11)
IL-1.beta. 100 nM 800 nM ABT2-42 VK (4G K) IL-1.beta. 100 nM 770 nM
ABT2-46 VK (4G K) IL-1.beta. 70 nM 386 nM ABT2-65 VH (4G H18)
IL-1.beta. 100 nM 266 nM ABT2-76 VK (4G K) IL-1.beta. 50 nM 456 nM
ABT2-108 VH (4G H17) IL-1.beta. 150 nM 250 nM
Detailed information regarding each of the identified IL-1.alpha.
and IL-1.beta. antibodies having advantageous neutralizing and
affinity characteristics is also provided below:
A. Domain Antibody: ABT1-6-23
Library: 3.5G VH3
[0413] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00020 EVQLLESGGGLVQPGGSLRLSCAASGFTFVRYDMAWVRQAPGKGLEWV
SSIYKSGALTSYDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA
KGWASFDYWGQGTLVTVSS
Clone ABT1-6-23 is an affinity matured variant; created by
combining CDR1 from ABT1-6-15 with CDR2 from ABT1-6-3 (these three
clones are all affinity matured derivatives of the parental clone
ABT1-6). The CDR3 for all three affinity matured clones is
unchanged from the parental clone ABT1-6. The CDR amino acid
sequences are detailed in Table 20.
TABLE-US-00021 TABLE 20 Summary of the CDR differences between the
parental ABT1-6 clone and the affinity matured progeny. Amino acid
differences from the parental clone are in bold and underlined
text. CDR1 CDR2 CDR3 MRC5 ND.sub.50 ABT1-6 GAYDMQ SINKSGALTSYDSVKG
GWASFDY 15 .mu.M ABT1-6-3 GAYDMQ SIYKSGALTSYDSVKG GWASFDY 0.08
.mu.M ABT1-6-15 VRYDMA SINKSGALTSYDSVKG GWASFDY 2 .mu.M ABT1-6-23
VRYDMA SIYKSGALTSYDSVKG GWASFDY 0.03 .mu.M
Calculated MW: 12565 Da
MRC5 ND.sub.50: 30 nM
Specificity: IL-1.alpha..
[0414] No significant inhibition by other human or murine IL-1
family members including IL-1.beta., IL-1 receptor antagonist,
IL-18, or human TNF. BIAcore Kinetic Analysis: The calculated
kinetic dissociation constant (K.sub.D), association rate constant
(k.sub.a) and dissociation rate constant (k.sub.d) are summarized
in Table 21.
TABLE-US-00022 TABLE 21 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 2.0 .times. 10.sup.5 1.2 .times. 10.sup.-2 6.1 .times.
10.sup.-8 1.94
B. Domain Antibody: ABT1-86
Library: 4G H17
[0415] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined. A single K to E framework 3 mutation
is highlighted in parentheses)
TABLE-US-00023 EVQLLESGGGLVQPGGSLRLSCAASGFTFDRYIMAWVRQAPGKGLEWVS
SITPSGAATYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCA (E)
EPADRYSTWTFDYWGQGTLVTVSS
Calculated MW: 13398 Da
MRC5 ND.sub.50: 900 nM
Specificity: IL-1.alpha..
[0416] No significant inhibition by other human or murine IL-1
family members including IL-1.beta., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0417] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarized in Table 22.
TABLE-US-00024 TABLE 22 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 8.2 .times. 10.sup.5 9 .times. 10.sup.-2 1.1 .times.
10.sup.-7 0.03
C. Domain Antibody: ABT1-95
Library: 4G K
[0418] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00025 DIQMTQSPSSLSASVGDRVTITCRASQPIHGNLRWYQQKPGKAPKLLIY
NISNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYRWPVT FGQGTKVEIKR
Calculated MW: 11930 Da
MRC5 ND.sub.50: 800 nM
Specificity: IL-1.alpha..
[0419] No significant inhibition by other human or murine IL-1
family members including IL-1.beta., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0420] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 23.
TABLE-US-00026 TABLE 23 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 4.9 .times. 10.sup.5 2.2 .times. 10.sup.-2 4.5 .times.
10.sup.-8 0.13
D. Domain Antibody: ABT1-96
Library: 4G H15
[0421] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00027 EVQLLESGGGLVQPGGSLRLSCAASGFTFNQYNMFWVRQAPGKGLEWVS
VISGSGRFTYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGW
WRRDPPFDYWGQGTLVTVSS
Calculated MW: 13408 Da
MRC5 ND.sub.50: 11 nM
Specificity: IL-1.alpha..
[0422] No significant inhibition by other human or murine IL-1
family members including IL-1.beta., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0423] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 24.
TABLE-US-00028 TABLE 24 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 3.7 .times. 10.sup.5 3.9 .times. 10.sup.-4 1.1 .times.
10.sup.-9 0.8
E. Domain Antibody: ABT1-98
Library: 4G H11
[0424] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00029 EVQLLESGGGLVQPGGSLRLSCAASGFTFDGYIMSWVRQAPGKGLEWVS
TISPLGSVTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK
KGPWFDYWGQGTLVTVSS
Calculated MW: 12645 Da
MRC5 ND.sub.50: 30 nM
Specificity: IL-1.alpha..
[0425] No significant inhibition by other human or murine IL-1
family members including IL-1.beta., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0426] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 25.
TABLE-US-00030 TABLE 25 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 2.0 .times. 10.sup.6 2.7 .times. 10.sup.-2 1.3 .times.
10.sup.-8 7.9
F. Domain Antibody: ABT1-122
Library: 4G K
[0427] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00031 DIQMTQSPSSLSASVGDRVTITCRASQPIWTELNWYQQKPGKAPKLLIY
GSSSLQSGVPRFSGSGSGTDFTLTISSLQPEDFATYYCQQFAYFPATFG QGTKVEIKR
Calculated MW: 11824 Da
MRC5 ND.sub.50: 50 nM
Specificity: IL-1.alpha..
[0428] No significant inhibition by other human or murine IL-1
family members including IL-1.beta., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0429] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 26.
TABLE-US-00032 TABLE 26 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 6.6 .times. 10.sup.6 0.28 4.2 .times. 10.sup.-8 2.4
G. Domain Antibody: ABT1-141
Library: 4G K
[0430] Amino Acid Sequence: (diversified residues are indicated in
bold the CDRs are underlined)
TABLE-US-00033 DIQMTQSPSSLSASVGDRVTITCRASQWIQKQLAWYQQKPGKAPKLLIY
SSSYLQSGVPRFSGSGSGTDFTLTISSLQPEDFATYYCQQHLRVPFTF GQGTKVEIKR
Calculated MW: 11998 Da
MRC5 ND.sub.50: 50 nM
Specificity: IL-1.alpha..
[0431] No significant inhibition by other human or murine IL-1
family members or human TNF.
BIAcore Kinetic Analysis:
[0432] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 27.
TABLE-US-00034 TABLE 27 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 1.3 .times. 10.sup.4 3.9 .times. 10.sup.-3 3.0 .times.
10.sup.-7 5.7
H. Domain Antibody: ABT2-13
Library: 4G H11
[0433] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined. A single V to A framework 2 mutation
is highlighted in parentheses)
TABLE-US-00035 EVQLLESGGGLVQPGGSLRLSCAASGFTFRDYVMYW(A)RQAPGKGLEW
VSRIDPMGSSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYY
CAKPEGNFDYWGQGTLVTVSS
Calculated MW: 12796 Da
MRC5 ND.sub.50: .about.1000 nM
Specificity: IL-1.beta..
[0434] No significant inhibition by other human or murine IL-1
family members including IL-1.alpha., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0435] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarise in Table 28.
TABLE-US-00036 TABLE 28 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 8.1 .times. 10.sup.4 7.5 .times. 10.sup.-2 9.3 .times.
10.sup.-7 0.03
I. Domain Antibody: ABT2-42
Library: 4G K
[0436] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined. A single K to T framework 2 mutation
is highlighted in parentheses)
TABLE-US-00037 DIQMTQSPSSLSASVGDRVTITCRASQYIEKWLTWYQQKPGKAP(T)LLI
YRGSLLQSGVPRFSGSGSGTDFTLTISSLQPEDFATYYCQQTEYWPFTFG QGTKVEIKR
Calculated MW: 12100 Da
MRC5 ND.sub.50: 770 nM
Specificity: IL-1.beta..
[0437] No significant inhibition by other human or murine IL-1
family members including IL-1.alpha., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0438] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 29.
TABLE-US-00038 TABLE 29 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 2.5 .times. 10.sup.3 0.13 5.4 .times. 10.sup.-5
0.02
J. Domain Antibody: ABT2-46
Library: 4G K
[0439] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00039 DIQMTQSPSSLSASVGDRVTITCRASQSIIEWLSWYQQKPGKAPKLLIYR
TSVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNEFWPFTFGQ GTKVEIKR
Calculated MW: 12049 Da
MRC5 ND.sub.50: 386 nM
Specificity:
[0440] No significant inhibition by other human or murine IL-1
family members including IL-1.alpha., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0441] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 30.
TABLE-US-00040 TABLE 30 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 2.3 .times. 10.sup.4 6.3 .times. 10.sup.-2 2.8 .times.
10.sup.-6 0.04
K. Domain Antibody: ABT2-65
Library: 4G H18
[0442] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined. A single N to D framework 3 mutation
is highlighted in parentheses).
TABLE-US-00041 EVQLLESGGGLVQPGGSLRLSCAASGFTFEDYQMGWVRQAPGKGLEWVSS
ISAMGNRTYYADSVKGRFTISRD(D)SKNTLYLQMNSLRAEDTAVYYCAK
NLVRTQSKMWMFDYWGQGTLVTVSS
Calculated MW: 13672 Da
MRC5 ND.sub.50: 266 nM
Specificity: IL-1.beta..
[0443] No significant inhibition by other human or murine IL-1
family members including IL-1.alpha., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0444] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarise in Table 31.
TABLE-US-00042 TABLE 31 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 2.5 .times. 10.sup.5 2.0 .times. 10.sup.-2 9.7 .times.
10.sup.-8 0.32
L. Domain Antibody: ABT2-76
Library: 4G K
[0445] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00043 DIQMTQSPSSLSASVGDRVTITCRASQSIDRWLAWYQQKPGKAPKLLIYR
GSILQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVAFWPPTFGQ GTKVEIKR
Calculated MW: 11908 Da
MRC5 ND.sub.50: 456 nM
Specificity: IL-1.beta..
[0446] No significant inhibition by other human or murine IL-1
family members including IL-1.alpha., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0447] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarise in Table 32.
TABLE-US-00044 TABLE 32 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) k.sub.d (s.sup.-1) K.sub.D (M)
.chi..sup.2 2.0 .times. 10.sup.4 2.7 .times. 10.sup.-2 1.3 .times.
10.sup.-6 0.99
M. Domain Antibody: ABT2-108
Library: 4G H17
[0448] Amino Acid Sequence: (diversified residues are indicated in
bold; the CDRs are underlined)
TABLE-US-00045 EVQLLESGGGLVQPGGSLRLSCAASGFTFAEYTMMWVRQAPGKGLEWVSR
IGQDGKNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKYT
GRVGVHHLFDYWGQGTLVTSS
Calculated MW: 13471 Da
MRC5 ND.sub.n: 200 nM
Specificity: IL-1.beta..
[0449] No significant inhibition by other human or murine IL-1
family members including IL-1.alpha., IL-1 receptor antagonist,
IL-18, or human TNF.
BIAcore Kinetic Analysis:
[0450] The calculated kinetic dissociation constant (K.sub.D),
association rate constant (k.sub.a) and dissociation rate constant
(k.sub.d) are summarised in Table 33.
TABLE-US-00046 TABLE 33 Kinetic constants determined by BIAcore
analysis k.sub.a (M s.sup.-1) kd (s.sup.-1) K.sub.D (M) .chi..sup.2
2.9 .times. 10.sup.4 3.1 .times. 10.sup.-3 1.1 .times. 10.sup.-7
0.06
[0451] Tables 62 and 63 provide an overview of the characteristics
of the IL-1.alpha. (ABT1) dAbs and IL-1.beta. dAbs, respectively.
In addition, the sequences of all of the dAb variable heavy and
light chains identified herein are provided in Table 64.
Example 4
Construction of Expression Vectors (P-Ef-Bos) for Recombinant
Antibody Expression
[0452] A panel of expression vectors, also referred to herein as
master template plasmids or master templates, for the expression of
recombinant human and mouse IgG were constructed from pre-existing
antibody constructs. The purpose of making these master templates
was to generate a panel of improved expression vectors, i.e.,
improved pEF-BOS--based cloning vectors, for mammalian expression
of recombinant human and mouse IgGs in transiently transfected COS
cells. The pEF-BOS vector was originally described in Mizushima and
Nagata, pEF-BOS, a powerful mammalian expression vector. Nucleic
Acids Res, 1990. 18(17): p. 5322.
[0453] The constructed mammalian expression vectors described
herein, all have a 1-kb stuffer sequence in between an upstream
signal peptide leader sequence and a downstream Ig constant region
sequence. In order to make an antibody heavy or light chain
construct, the stuffer sequence in the chosen master template is
removed by restriction enzyme digestion followed by inserting the
desired antibody V domain sequence (VH, Vx, or Vy). The resulting
plasmid constructs are easily proprigated in and purified from E.
coli and can be used to express antibody by co-transfecting both a
heavy chain and a light chain construct in COS cells.
Mouse Master Template Constructions
[0454] Master templates for mouse IgGs were constructed by first
introducing both FspA I and Afe I sites in between the signal
peptide leader sequences and the constant regions and subclone the
amplified products into the Srf I and Not I sites of pEF-BOS vector
backbone. The four primers used for pBOS-mC.kappa. overlapping PCR
were pEF-BOS-Fwd, mCkappa-Rev, mCkappa-Fwd, and pEF-BOS-Rev using
pA534 and pA246 DNA as templates. For pBOS-mC.gamma.1, the primers
were pEF-BOS-Fwd, mIgG1-Rev, mIgG1-Fwd, pEF-BOS-Rev and the
templates was pA245. Similarly, the primers for pBOS-mCy2a were
pEF-BOS-Fwd, nfIgG2a-Rev, mIgG2a-Fwd, and pEF-BOS-Rev using
templates pA245 and pA239. After the first step, a blunt-end
stuffer sequence, which was amplified by LambdaStuffer-Fwd and -Rev
primers from .lamda./Hind III digested DNA markers, was inserted
into these constructs linearized by FspA I and Afe Ito create the
final mouse master templates.
Human Master Template Constructions
[0455] Human heavy chain master templates were constructed by one
of two methods. For pBOS-hC.gamma.1,z,non-a and
pBOS-hC.gamma.1,z,non-a,mut(234,235) constructs, a heavy chain
leader sequence was linked to the stiffer sequence by overlapping
PCR using primers pEF-BOS-Fwd, SHS-Rev, SHS-Fwd, and hCGamma1-Rev
and the template vectors. The resulting PCR products were than
inserted into pBOS backbone vectors, via SrfI and Sal I sites. For
pBOS-hC.gamma.1,z,non-a,mut(234,237), a sniffer sequence was
amplified by SHS-Fwd and hCGamma1-Rev primers, digested by Sal I,
and subcloned into Nru I and Sal I-digested vector. The
pBOS-hC.gamma.1,z,a, pBOS-hC.gamma.4, and pBOS-hCy2(n-) were made
by subcloning the Sal I and Not I-digested constant regions from
constant chain containing vectors, and an RT-PCR product from
transiently transfected COS cells expressing IgG2 into
pBOS-hC.gamma.1,z,non-a,mut(234,237) linearized by Sal I and Not
I.
[0456] For the pBOS-hCy2, (n+) construct, a GVE to GME amino acid
change was introduced into the IgG2 Fc region in the pBOSAB198G2WT
template via overlapping PCR using primers 5'Sal198, 3'Sal198,
5'GME198, and 3'BSR198. Product from this overlapping PCR served as
template for the next round of overlapping PCR to introduce a PSS
to TSS amino acid change in the IgG2 Fc region using primers
5'Sal198, 3'TSS198, 5'BSR198, and 3'BSR198. This PCR product was
then inserted back into the pBOSAB198G2WT via BsrG I and Sal I
sites. Finally, the IgG2(n+) constant region from this vector was
excised and inserted into pBOS-hC.gamma.2(n-) via Sal I and Not I
sites to generate the pBOS-hC.gamma.2(n+) construct.
[0457] Using pEF-BOS-Fwd, SLS-Rev, SLS-Fwd, hCkappa-Rev2,
hCkappa-Fwd2, and pEF-BOS-Rev primers and DNA templates, the human
x light chain master template was made by two overlapping PCRs
linking a x leader sequence to the stuffer sequence then to the
human .kappa. constant region before cloning the final product into
Srf I and Not I-lineared pEF-BOS-based backbone.
[0458] Similarly, for pBOS-hC.lamda. master template construction,
primers pEF-BOS-Fwd, LambdaSignal-Rev, LambdaSignal-Fwd,
hCLambda-Rev, hCLambda-fwd and pEF-BOS-Rev were used to amplify
.lamda. light chain leader sequence, the stuffer sequence, and
human X light chain constant region sequence together using
templates in two overlapping PCRs. The final product was cloned
into Srf I and Not I-lineared pEF-BOS-based backbone. PCR primers
used in the construction of the vectors are described in Table
34.
TABLE-US-00047 TABLE 34 PCR primers used in constructing pEF-BOS
master templates Name Oligo Sequence StufferLightSignal (SLS)-
TCGGCATACCATGCGCATCGCGAGCCGGGGAACCAC Rev StufferLightSignal (SLS)-
CGATGCGCATGGTATGCCGAAAGGGATGC Fwd StufferHeavySignal
GCACTGGACACCTTTTAAAATCGCG (SHS)-Rev StufferHeavySignal
CGATTTTAAAAGGTGTCCAGTGCGCATGGTATGCCGAAAGGG (SHS)-Fwd ATGC
pEF-BOS-Rev GGAGACCTGATACTCTCAAG pEF-BOS-Fwd TCAGGTGTCGTGAGGAAT
mIgG2a-Rev GTTTTAGCGCTACACTGGACACCTTTTAAAATCGCG mIgG2a-Fwd
GGTGTCCAGTGTAGCGCTAAAACAACAGCCCCATCG mIgG1-Rev
CGTTTTAGCGCTACACTGGACACCTTTTAAAATCG mIgG1-Fwd
GGTGTCCAGTGTAGCGCTAAAACGACACCCCCATC mCkappa-Rev
GCAGCATCAGCGCTGCATCGCGAGCCGGGGAACCAC mCkappa-Fwd
GGCTCGCGATGCAGCGCTGATGCTGCACCAACTG LambdaStuffer-Rev
GCTTGTCACCCAGGAACG LambdaStuffer-Fwd CGAAAGGGATGCTGAAATTGAG
LambdaSignal-Rev CATCCCTTTCGCGATAAGCTTCCTGTGCAG LambdaSignal-Fwd
GAAGCTTATCGCGAAAGGGATGCTGAAATTGAG hCLambda-Rev
GCCTTGGGTTAACGCTTGTCACCCAGGAACG hCLambda-Fwd
GGTGACAAGCGTTAACCCAAGGCTGCCCCCTCGGTC hCkappa-Rev2
CAGCCACCGTACGTAGCTTGTCACCCAGGAACG hCkappa-Fwd2
GTGACAAGCTACGTACGGTGGCTGCACCATCTG hCGamma1-Rev
CTTGGTCGACGCGCTTGTCACCCAGGAACG hCGamma1-Fwd
GGTGACAAGCGCGTCGACCAAGGGCCCATCGGTC 5'Sal198
TCTACGACTACGGTATGGACGTCTGG 3'Sal198 GCCGTCCACGTACCAGTTGAAC 5'BSR198
CCTCCAGCAACTTCGGCACCCAGACCTAC 3'BSR198 ACCTGGTTCTTGGTCATCTCCTCCC
5'GME198 GTTCAACTGGTACGTGGACGGCATGGAGGTG 3'TSS198
GTAGGTCTGGGTGCCGAAGTTGCTGGAGGTCACGGTCAC
Condition for DNA Amplification by PCR
[0459] All PCRs were carried out in an MJ Research DNA engine
tetrad 2 thermal cycler equipped with stainless steel heating
blocks. The proofreading Pfu Turbo Hotstart DNA polymerase
(Stratagene) and its accompanying buffer were used for DNA
amplification to minimize undesired sequence mutation and generate
blunt-ended products. Typical thermal cycling condition was an
initial DNA template denaturation at 94.degree. C. for 2 minutes;
followed by thermal cycles of 94.degree. C. for 20 seconds,
55.degree. C. for 20 seconds, and 72.degree. C. for 1 minute per
kb; finished up by an additional 5 minutes at 72.degree. C. and
kept at 4.degree. C. forever.
Sequencing Confirmation
[0460] All constructed master templates were sequenced and
confirmed in the region between the SrfI and Not I cloning sites
using primers pEF-BOS-Fwd, pEF-BOS-Rev, and Lambda844. Twenty two
additional primers as listed in Table 35 were used to obtain the
vector backbone sequence in both DNA strands of pBOS-mC.kappa..
This sequence was then applied to all master templates for their
electronic DNA sequence files by Vector NTI software package.
TABLE-US-00048 TABLE 35 Sequencing primers for pEF-BOS vector
backbone Name Oligo Sequence Lambda844 AAGCGTTTCACTAATGGGCG F1042
ATAAGCGGCCGCGACTCTAGAG F1541 TCACCCTCCACCTCTTCACC F2058
GGACTCCAACGTCAAAGGG F2538 AAACGCGCGAGACGAAAGG F3071
AAAGCATCTTACGGATGG F3553 ATGAACGAAATAGACAGATCGC F4068
TTACCGGATAAGGCGCAG F4557 ATACGCAAACCGCCTCTC F5060
TTGCAGCTAATGGACCTTCTAG F5551 AATCTGGTGGCACCTTCGCG F6068
TTCTCGAGCTTTTGGAGTACG R1527 GGGTGACAGTGGAGCTTCCT R2023
ACTCTTGTTCCAAACTGG R2530 TCTCGCGCGTTTCGGTGATG R3056
TGCTTTTCTGTGACTGGTGAG R3535 CATCCATAGTTGCCTGACTCC R4036
TCCAACCCGGTAAGACACG R4552 AGGCGGTTTGCGTATTGGGC R5051
CCATTAGCTGCAAAGATTCCTC R5548 AAGGTGCCACCAGATTCGC R6051
GAACTAATCGAGGTGCCTGG R6300 ATGGATCTCGAGGTCGAGGG
Results
Mouse Master Templates
[0461] Three mouse master templates were made: pBOS-mC.kappa.,
pBOS-mC.gamma.1, and pBOS-mC.gamma.2a. Sequences for the murine
vectors are described in SEQ ID NOs 844 to 855 and provided in
Table 36.
TABLE-US-00049 TABLE 36 Table of sequences and corresponding
vectors SEQ ID VECTOR NAME and CONSTANT REGION NO (constant region
and stuffer sequences indicated) 855 pBOS- hCg1, z, a- (Constant
region nt 1101 to 2093; stuffer sequence nt 124-1103) 844
pBOS-hCg1, z, non-a, mut (234, 235) (Constant region nt 1101 to
2093; stuffer sequence nt 124-1100) 845 pBOS-hCg1, z, non-a, mut
(234, 237) (Constant region nt 1101 to 2093; stuffer sequence nt
124-1100) 846 pBOS-hCg1, z, non-a (Constant region nt 1101 to 2093;
stuffer sequence nt 124-1100) 847 pBOS-hCg2, (n-) (Constant region
nt 1101 to 2081; stuffer sequence nt 124-1100) 848 pBOS-hCg2, (n+)
(Constant region nt 1101 to 2081; staffer sequence nt 124-1100) 849
pBOS-hCg4 (Constant region nt 1101 to 2084; stuffer sequence nt
124-1100) 850 pBOS-hCk (Constant region nt 1114 to 1434; stuffer
sequence nt 132-1100) 851 pBOS-hCl (Constant region nt 1095-1412;
stuffer sequence nt 127-1095) 852 pBOS-mCg1 (Constant region nt
1101 to 2075; stuffer sequence nt 124-1100) 853 pBOS-mCg2a
(Constant region nt 1101 to 2093; stuffer sequence nt 124-1100) 854
pBOS-mCk (Constant region nt 1109 to 1429; stuffer sequence nt
132-1108)
[0462] The pBOS-mC.kappa. may be used for cloning mouse V.kappa.
region sequence for expressing a mouse Ig light chain, while
pBOS-mC.gamma.1 and pBOS-mC.gamma.2a may be used for cloning mouse
V.sub.H sequence for expressing a heavy chain with a .gamma.1 and
.gamma.2a constant region, respectively. The 1-kb .lamda. stuffer
sequence in all three master templates can be released from the
master template DNA by FspA I and Afe I (or Eco47 III) restriction
enzymes. There is an Nru I site upstream of the FspA I site that
can be used also. The resulting linearized vectors will have blunt
ends at both 5' and 3' ends and are ready for ligation with
blunt-ended V region sequences. If PCR amplification method is used
to prepare the V region for subcloning, Pfu DNA polymerase or other
DNA polymerases that produce blunt-end products should be used and
the products need to be phosphorylated prior to ligating to the
linearized and dephosphorylated pBOS vectors. Alternatively,
bacterial homologous recombination can be used to subclone the
desired V region into the pBOS master templates easily. For this
approach, the V region amplification forward primer should have
additional 30 nucleotides at the 5' end that overlaps with the 3'
end of leader sequence in the vector, and the reverse primer should
have additional 30 nucleotides at the 3' end that overlaps with the
5' end of the constant region sequence. After PCR amplification,
the amplified V region products are gel-purified and mixed with the
linearized vector DNA at greater than 5 to 1 molar ratio and
co-transformed into a recombination-capable competent cells. The
DH5.alpha. competent cells sold by Invitrogen work well for this
purpose. A schematic showing the design and use of mouse pBOS
templates is described in FIG. 18.
Human Master Templates
[0463] Table 37 lists the eight human pBOS master templates. The
pBOS-hC.kappa. template can be linearized by FspA I and BsiW I
enzymes to remove the A stuffer sequence and ligated to a V.kappa.
sequence for kappa light chain expression (see FIG. 18B).
Alternative 5' Nru 1 and 3' SnaB I restriction enzyme sites are
available for cloning purpose if FapA I or BsiW I site is present
in the V.kappa. sequence and thus prevents their use in cloning
work. The pBOS-hC.lamda. should be linearized by Nru I and Hpa I
and ligated to V.lamda. sequence (FIG. 18B). Alternatively, the
linearized vector can be homologously recombined with the V.lamda.
sequence in bacteria as described in 5.1 to avoid the use of less
efficient blunt-end ligation process or the presence of either Nru
I or Hpa I site in the V.lamda. sequence. It should be noted that
the creation of the Hpa I site at the 5' end of C.lamda. requires
the creation of a C to T point mutation and this mutation must be
corrected back to a C in the V.lamda. sequence to be ligated to the
vector backbone. Keeping this mutation in the constructed plasmid
will result in truncated open reading frame as the C to T mutation
created a termination codon at the 5' end of C.lamda..
[0464] All six human Ig heavy chain pBOS master templates can be
used by the same cloning strategy. They all can be lineared by FspA
I and Sal I restriction enzyme digestions and ligated to a VH
sequence that has compatible ends (FIG. 18B). Again, there is an
available Nru I site upstream of the FspA I site as an alternative
cloning site.
TABLE-US-00050 TABLE 37 List of human pBOS master templates
Template Name Isotype Allotype Mutations pBOS-hC.kappa. Kappa
pBOS-hC.lamda. Lambda pBOS-hC.gamma.1, z, a Gamma1 z, a
pBOS-hC.gamma.1, z, non-a Gamma1 z, non-a pBOS-hC.gamma.1, z,
non-a, Gamma1 z, non-a a.a. 234 -> Ala, mut(234, 235) a.a. 235
-> Ala pBOS-hC.gamma.1, z, non-a, Gamma1 z, non-a a.a. 234 ->
Ala, mut(234, 237) a.a. 237 -> Ala pBOS-hC.gamma.2 (n-) Gamma2
n- pBOS-hC.gamma.2 (n+) Gamma2 n+ pBOS-hC.gamma.4 Gamma4
pEF-BOS Vector Backbone
[0465] The pBOS-mC.kappa. was submitted for complete double-strand
sequencing using primers listed in the methods section. A sequence
contig was assembled and used as the template to generate all pBOS
templates' electronic sequence file in Vector NTI program. Other
than several single base-pair differences and difference in the
length of a poly-A region, there was no major difference from the
old pEF-BOS vector sequence. The new vector backbone sequence is a
perfect match to the pUC119 plasmid sequence from which the pEF-BOS
was derived (Mizushima and Nagata,pEF-BOS, a powerful mammalian
expression vector. Nucleic Acids Res, 1990. 18(17): p. 5322).
[0466] The master templates described herein provide a 1-kb stuffer
sequence unrelated to any Ig sequence in the vector. The 1-kb
staffer facilitates restriction enzyme digestion, easy removal by
gel purification, and identifying correct clones from background
colonies after transformation by either colony PCR or restriction
digestion of miniprep DNAs.
Example 5
Expression of Paired Single Domain Antibodies as a Functional
IL-1.alpha. and IL-10 Dual-Specific Molecule
[0467] The overall objective of the study was to generate a
dual-specific antibody that can bind and neutralize both
IL-1.alpha. and .beta.. Previous studies combined separate VH or VL
dAbs with affinities for either IL-1.alpha. or IL-1.beta. into a
dual-specific IgG or IgG-like molecule that binds and neutralizes
both IL-1.alpha. and 1l. Initial attempts to pair VH/VH or VL/VL to
generate IgG-like proteins had poor yields and unpaired heavy/light
chains using a COS cell expression system. This example provides an
alternative format for expressing VH/VH or VL/VL single chain-Fe
fusion proteins by directly pairing two dAbs with an inflexible
linker and fused to a human IgG1 Fc region.
[0468] The purpose of the study was to determine if scFv-Fc fusion
constructs could be made by linking an IL-1.alpha. and an
IL-1.beta.-specific VH dAb together, and if these constructs could
maintain high neutralization potencies for both IL-1.alpha. and
IL-1.beta. in a bioassay. Constructs pairing ABT2-108-538x (an
IL-1.beta. specific VH dAb) with ABT1-207 or ABT1-98 (IL-1.alpha.
specific VH dAbs) were generated and analyzed to test this
approach.
Materials and Methods
[0469] VH dAbs ABT2-108-538, ABT1-207, ABT1-98 were amplified from
their corresponding pBOS plasmid constructs via PCR using primers
SCFCVH 5' and GSBAMVH 3', resulting in the addition of a 3'
(Gly.sub.4Ser).sub.3 linker, and gel purified. The Fc region was
amplified from pBOS-hCgl,z,non-a via PCR using primers FCBAMB 5'
and 1926 3', resulting in the addition of an overlapping 5'
(Gly.sub.4Ser).sub.3 linker, and gel purified. Overlapping PCR
using the purified VH-(Gly.sub.4Ser).sub.3 and
(Gly.sub.4Ser).sub.3-FC PCR products and primers SVFVH 5' and 1926
3' was performed, and the resulting products gel purified. The
VH-FC fusions were inserted into vectors pBOS-ABT1-98,
pBOS-ABT1-207, and pBOS-ABT2-108-538x via Not I and Sal I sites to
generate constructs pBOS-1-98/2-108-538x-scFc,
pBOS-1-207/2-108-538x-scFc, pBOS-2-108-538x/1-98-scFc, and
pBOS-2-108-538x/1-207-scFc (FIG. 19). ScFc protein (scFc=ScFv. Fc
fusion construct) for each construct was generated by transfecting
the constructed plasmids into COS cells and purifying proteins
using protein A column chromatography. The purified proteins were
quantified by BCA assay using BSA as standard and also separated on
reducing and non-reducing SDS-PAGE gels. Purified scFc proteins
were tested for neutralization potencies against both IL-1.alpha.
and IL-1.beta. in MRC-5 assays.
TABLE-US-00051 TABLE 38 Primers used in PCR to generate scFc
constructs Primer name Sequence SCFCVH 5'
Gactgcgtcgacgcgtacggaggtgcagctgttggagt ctggg GSBAMVH 3'
tccacctccaccggatccaccacctccgctcgagac ggtgaccagggttccctg FCBAMB 5'
ggaggtggtggatccggtggaggtggatctggtggtggt
ggatcacccaaatcttgtgacaaaactcacacatgccca 1926 3'
Ggagacctgatactctcaag
Results
[0470] The purified scFc protein for all 4 constructs produced well
in COS cells, and gave one uniform band of slightly more than 100
kDa unreduced and 50 kDa reduced by SDS-PAGE. Protein yields are
described below in Table 39.
TABLE-US-00052 TABLE 39 Protein yields of scFc constructs ScFc
Construct Protein yield (per L supernatant) ABT1-98/2-108-538 6.2
mg ABT1-207/2-108-538 4.6 mg ABT2-108-538/1-98 4.4 mg
ABT2-108-538/1-207 1 mg
Table 40 show the results of the MRC-5 assays, using MAB200 and
MAB201 as reference antibodies, to measure IL-1.alpha. and .beta.
neutralization potencies of the scFc proteins
TABLE-US-00053 TABLE 40 Estimated EC50 for IL-1.alpha. and
IL-.beta. neutralization from MRC-5 assays Protein IL-1.alpha. (nM)
IL-.beta. (nM) ABT1-98/ABT2-108-538x 30 1.3 ABT1-207/ABT2-108-538x
25 1.1 ABT2-108-538x/ABT1-98 3 <0.001 ABT2-108-538x/ABT1-207 50
0.002 MAB200 0.17 -- MAB201 -- 0.005
[0471] All scFc constructs generated showed neutralization for both
IL-1.alpha. and 3. The order of the VH dAbs in the scFc construct
seemed to impact their neutralization potencies, however. For
instance, ABT2-108-538x lost about 3 logs of potency against
IL-1.beta. when it was placed in between an N-terminal IL-1.alpha.
dAb and a C-terminal Fc in the scFc construct than if it was placed
N-terminal to both the IL-1.alpha. dAb and the Fc. The placement of
the IL-1.alpha. dAb in the scFc construct seemed to have less
impact on the neutralization potency. In the case of ABT1-98, it
lost about one log of IL-1.alpha. neutralization potency going from
the C-terminal to the N-terminal VH position in the scFc, whereas
ABT1-207 lost about 2-fold potency going from the N-terminal to the
C-terminal VH in the scFc construct with the IL-1.beta. VH
2-108-538x. The scFv construct that showed the best neutralization
potency for both IL-1.alpha. and p was ABT2-108-538x/ABT1-98, with
EC50s of .about.3 nM against IL-1.alpha. and less than 1 .mu.M
against IL-1.beta.
SUMMARY of Variable Domain Pairing and Neutralization Data
[0472] Table 41 describes 10 Fab molecules that were specific to
IL-1.alpha./IL-1.beta. and were also able neutralizing in a
standard MRC5 in vitro bioassay. In addition, Fab molecule
ABT1-6-23/ABT2-13 was effective at neutralizing with respect to
IL-1.alpha., however the molecule was not been tested at a
sufficient concentration to determine a value for IL-1.beta.
neutralisation. For each clone described herein, the first part its
name relates to its specificity (IL-1.alpha. represented by ABT1;
and IL-1.beta. represented by ABT2).
TABLE-US-00054 TABLE 41 Summary of the most potent IL-1
neutralising dAb pairing as determined in the MRC5 bioassay. Clone
ABT1-6-23 is an affinity matured clone derived from the parental
ABT1-6. Fab pairing dAb type Fab IL-1.alpha. ND.sub.10 Fab
IL-1.beta. ND.sub.10 ABT1-96/ABT2-42 VH/VK 3 nM 10 nM
ABT1-122/ABT2-108 VK/VH 3 nM 50 nM ABT1-141/ABT2-108 VK/VH 10 nM 15
nM ABT1-96/ABT2-13 VH/VH 0.9 nM 80 nM ABT1-6-23/ABT2-46 VH/VK 4 nM
150 nM ABT1-141/ABT2-65 VK/VH 80 nM 300 nM ABT1-96/ABT2-46 VH/VK 6
nM 400 nM ABT1-95/ABT2-13 VK/VH 200 nM 400 nM ABT1-122/ABT2-65
VK/VH 40 nM 1500 nM ABT1-98/ABT2-76 VH/VK 100 nM 1500 nM
ABT1-6-23/ABT2-13 VH/VH 40 nM >200 nM
A summary of the IL-1.alpha. and IL-1.beta. pairs is described in
more detail below and is also described above in Table 9. Pairing
ABT1-122 with ABT2-108 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 42.
TABLE-US-00055 TABLE 42 ND.sub.50 and ND.sub.10 values for the dAb
and Fab molecules. Clone Name ND.sub.10 IL-1.alpha. ND.sub.50
IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb ABT1-122
9 nM 50 nM -- -- dAb ABT2-108 -- -- 150 nM 250 nM Fab 3 nM 60 nM 50
nM 150 nM
Pairing ABT1-141 with ABT2-108 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 43.
TABLE-US-00056 TABLE 43 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-141 10 nM 70 nM -- -- dAb ABT2-108 -- -- 150 nM 250 nM Fab 10
nM 150 nM 15 nM 40 nM
Pairing ABT1-96 with ABT2-42 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 44.
TABLE-US-00057 TABLE 44 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-96 3 nM 11 nM -- -- dAb ABT2-42 -- -- 100 nM 770 nM Fab ~3 nM
30 nM ~10 nM 150 nM
Pairing ABT1-141 with ABT2-65 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 45.
TABLE-US-00058 TABLE 45 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-141 10 nM 70 nM -- -- dAb ABT2-65 -- -- 100 nM 266 nM Fab 80
nM 300 nM 300 nM 1000 nM
Pairing ABT1-122 with ABT2-65 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 46.
TABLE-US-00059 TABLE 46 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-122 9 nM 50 nM -- -- dAb ABT2-65 -- -- 100 nM 266 nM Fab 40 nM
800 nM 1500 nM 5000 nM
Pairing ABT1-96 with ABT2-13 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 47.
TABLE-US-00060 TABLE 47 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-96 3 nM 11 nM -- -- dAb ABT2-13 -- -- 100 nM 800 nM Fab 0.9 nM
7 nM 80 nM N.D.
Pairing ABT1-95 with ABT2-13 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay are summarised in Table 48.
TABLE-US-00061 TABLE 48 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-95 150 nM 700 nM -- -- dAb ABT2-13 -- -- 100 nM ~800 nM Fab
200 nM 600 nM 400 nM N.D.
Pairing ABT1-96 with ABT2-46 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 49.
TABLE-US-00062 TABLE 49 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-96 3 nM 11 nM -- -- dAb ABT2-46 -- -- 70 nM 386 nM Fab 6 nM 40
nM 400 nM N.D.
Pairing ABT1-6-23 with ABT2-46 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 50.
TABLE-US-00063 TABLE 50 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-6-23 3 nM 30 nM -- -- dAb ABT2-46 -- -- 70 nM 386 nM Fab 4 nM
100 nM 150 nM 5000 nM
Pairing ABT1-98 with ABT2-76 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 51.
TABLE-US-00064 TABLE 51 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-98 <100 nM <100 nM -- -- dAb ABT2-46 -- -- 50 nM 456 nM
Fab 100 nM 300 nM 1500 nM N.D.
Pairing ABT1-6-23 with ABT2-13 The ND.sub.10 and ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 52.
TABLE-US-00065 TABLE 52 ND.sub.50 and estimated ND.sub.10 values
for the dAb and Fab molecules. Clone Name ND.sub.10 IL-1.alpha.
ND.sub.50 IL-1.alpha. ND.sub.10 IL-1.beta. ND.sub.50 IL-1.beta. dAb
ABT1-6-23 3 nM 30 nM -- -- dAb ABT2-46 -- -- 100 nM 800 nM Fab 40
nM 200 nM >500 nM >500 nM
[0473] In conclusion, these results demonstrate that it is possible
to use a scFv-Fc fusion construct to express two VH dAbs to two
different antigens in one recombinant protein and maintain
dual-specificity and high potency.
Example 6
Dual Specific IL-1.alpha./IL-1.beta. IgGs
[0474] As described above, phage display and emulsion technologies
and yeast display, together with rational mutagenesis, produced a
range of improved dAbs to IL-1.alpha. and IL-1.beta. (some of which
appeared to exceed the potency of the cell based assay, indicating
that they have monomeric potencies in the very low picomolar or
high femtomolar range). These dAbs were then used these to create a
panel of dual targeting IgGs.
[0475] Examples 6.1-6.2 provide experiments that show that using
dAbs identified and affinity matured, IgGs could be constructed
that have sub-200 pM potencies to BOTH IL-1.alpha. and IL-1.beta.,
thus demonstrating that a fully human IgG can be generated that
neutralises two separate targets as well as the best murine
antibodies that are only able to neutralise one or the other
target. Furthermore, in vitro these molecules inhibited IL-1
signalling better than the natural antagonist (IL-1ra) and yet did
not bind to it, suggesting that in vivo the dual targeting IL-1 IgG
should act in concert with IL-1ra.
[0476] The goal of the experiments in Example 6 was to provide a
panel of IgGs that met the following criteria: [0477] Two
high-affinity Fab or Fab-like molecules, each with the following
properties: [0478] K.sub.d for IL-1.alpha.: 5.times.10.sup.-10M or
lower [0479] K.sub.d for IL-1.beta.: 5.times.10.sup.-10 M or lower
k.sub.off for IL-1.alpha.: 10.sup.-4s.sup.-1 or lower [0480]
k.sub.off for IL-1.beta.: 10.sup.-4 s.sup.-1 or lower [0481] Two
IgG molecules corresponding to re-cloned versions of the two Fab or
Fab-like molecules described above, each with the following
properties: [0482] IC.sub.50 in RB assay for IL-1.alpha.:
9.times.10.sup.-10 M or lower [0483] IC.sub.50 in RB assay for
IL-1.beta.: 9.times.10.sup.-10 M or lower [0484] ND.sub.50 in MRC5
assay for IL-1.alpha.: 2.times.10.sup.-10 M or lower [0485]
ND.sub.50 in MRC5 assay for IL-1.beta.: 2.times.10.sup.-10 M or
lower
[0486] IgGs or Fabs containing the lead clones have also been
tested in the Receptor Binding Assay (RBA) and the kinetics of
their interaction with IL-1.alpha. and IL-1.beta. were analysed
using BIAcore.
[0487] The below experiments shows that multiple clones from the
ABT1-95, ABT2-65 and ABT2-108 lineages formatted into IgGs meet the
above criteria in the MRC5 and Receptor Binding Assays.
Example 6.1
MRC5 and Receptor Ligand Binding (RBA) Assay Data for IgG
Pairings
[0488] Five clones ABT1-95, ABT1-122, ABT1-141, ABT2-65 and
ABT2-108 were paired as IgGs, and their inhibitory activity was
tested in the MRC5 assay. Table 53 contains a summary of the IgG
pairings that were analysed in the MRC5 assay as the affinity
maturation of each dAb progressed. This culminated in the
identification of dual specific pairings that met the above
criteria (ND.sub.50 of <200 pM)--these are highlighted in bold
in Table 53.
TABLE-US-00066 TABLE 53 Summary of the neutralisation of the dAb
pairings as determined in the MRC5 bioassay over the course of the
maturation program. ABT1 clone ABT2 clone MRC5 IL-1.alpha. (nM)
MRC5 IL-1.beta. (nM) 1-95-A3 2-108-538X 0.032 0.023 1-95-A3
2-65-166 0.017 0.15 1-95-A2 2-65-166 0.08 0.121 1-95-A5 2-65-166
0.014 0.197 1-95-A6 2-65-166 0.019 0.207 1-95-15 2-65-17 2.7 0.094
1-95-15 2-108-538X 8 0.042 1-122-750X 2-65-17 20 1 1-122-750X
2-108-538X 75 0.0094 1-122-511 2-108 80 8 1-122-511 2-108-538X 259
0.009 1-95-3 2-108 300 1-141-25 2-65-17 400 0.975 1-122 2-108-521
1000 1 1-141 2-108 1500 22
The IgG pairings were also tested in a standard Receptor Binding
Assay (RBA). Table 54 summarises the activity of the lead IgG pairs
in the RBA. Only the lead pairings were tested in this assay and
all pairings tested exceeded the above neutralisation criteria of
900 pM.
TABLE-US-00067 TABLE 54 Summary of the activity of IgG pairs in the
Receptor Binding Assay. Receptor Binding Assay ABT1 clone MRC5
IL-1.beta. ABT2 clone MRC5 IL-1.alpha. 1-95-15 540 pM 2-108-538X 71
pM 1-95-A2 67 pM 2-65-166 73 pM 1-95-A3 83 pM 2-65-166 709 pM
1-95-A6 108 pM 2-65-166 537 pM
Example 6.2
Comparison of dAb Monomers and IgG Pairs
[0489] Pairing ABT1-95-A2 with ABT2-65-166
[0490] The ND.sub.50 Values Determined from the MRC5 IL-1.alpha.
and IL-1.beta. Neutralisation Assay Data from 24 are summarised in
Table 55, as well as FIG. 20.
TABLE-US-00068 TABLE 55 ND.sub.50 values for the dAb and IgG
molecules. Clone Name ND.sub.50 IL-1.alpha. ND.sub.50 IL-1.beta.
dAb ABT1-95-A2 133 pM -- dAb ABT2-65-166 -- 4 nM IgG 80 pM 121
pM
Pairing ABT1-95-A3 with ABT2-65-166
[0491] The ND.sub.50 values determined from the MRC5 IL-1.alpha.
and IL-1.beta. neutralisation assay data is summarised in Table
56.
TABLE-US-00069 TABLE 56 ND.sub.50 values for the dAb and IgG
molecules. Clone Name ND.sub.50 IL-1.alpha. ND.sub.50 IL-1.beta.
dAb ABT1-95-A3 11 pM -- dAb ABT2-65-166 -- 4 nM IgG 17 pM 150
pM
Pairing ABT1-95-A5 with ABT2-65-166
[0492] The ND.sub.50 values determined from the MRC5 IL-1.alpha.
and IL-1.beta. neutralisation assay data is summarised in Table
57.
TABLE-US-00070 TABLE 57 ND.sub.50 values for the dAb and IgG
molecules. Clone Name ND.sub.50 IL-1.alpha. ND.sub.50 IL-1.beta.
dAb ABT1-95-A5 16 pM -- dAb ABT2-65-166 -- 4 nM IgG 14 pM 197
pM
Pairing ABT1-95-A3 with ABT2-108-5 38X The ND.sub.50 values
determined from the MRC5 IL-1.alpha. and IL-1.beta. neutralisation
assay data are summarised in Table 58.
TABLE-US-00071 TABLE 58 ND.sub.50 values for the dAb and IgG
molecules. Clone Name ND.sub.50 IL-1.alpha. ND.sub.50 IL-1.beta.
dAb ABT1-95-A3 11 pM -- dAb ABT2-65-166 -- 10 nM IgG* 32 pM 23 pM
*There was significant precipitation in this IgG sample.
Example 7
Potency of Dual Specific 12G in MRC-5 Neutralization Assay
[0493] The potency of a dual specific IgG, i.e.,
ABT2-108-620x/ABT1-95-A3, was determined using the MRC-5
neutralization assay.
MRC5 Assay
[0494] The following provides how the MRC5 neutralization assays
described herein were performed. The MRC5 neutralization assay is
based on the amount of IL-8 in the supernatant of human lung
fibroblast cells having activated IL-1 receptor. First, antibody is
pre-incubated with IL-1.alpha. or IL-1.beta.. Following the
incubation, the antibody (bound to IL-1.alpha., or IL-1.beta. is
added to human lung fibroblast cells, which are trypsinized and
plated in a 96 well plate. The plate is then incubated for about 20
hours at 37.degree. C. with 5% CO.sub.2. Following the incubation,
supernatant from the cells is taken and the IL-8 level is measured
using standard ELISA. The binding of IL-1.alpha. or IL-1.beta. to
IL-1 receptor on cells produces IL-8.
Results
[0495] With respect to IL-1.alpha., the results of the MRC-5
neutralization assay showed that the potency (IC50) of
ABT2-108-620x/ABT1-95-A3 was better than 1 .mu.M antibody
concentration (versus an EC50 of 0.1447 for control MAD200). For
IL-1.beta., the results also showed that the dual specific IgG was
able to inhibit IL-8 production, where ABT2-108-620x/ABT1-95-A3 had
an IC50 value of 0.005585 nM and the control antibody (MAB201) had
an IC50 value of 0.003586.
Example 8
Biacore Analysis of ABT2-108-620x/ABT1-95-A3
[0496] Biacore analysis was performed on ABT2-108-620x/ABT1-95-A3
to determine the binding affinity of the dual specific antibody.
This analysis was done according to the following:
Simultaneous and Non Interferring Independent Binding
[0497] The antibody (1-5 ug/ml) was first captured on a goat anti
human IgG FC surface. Then individual binding of the antigens at
100 nM for recombinant human IL-1 alpha (rhIL-1 alpha) and 200 nM
for recombinant human IL-1 beta (rhIL-1 beta) was determined.
rhIL-1 alpha (100 nM) was injected followed by rhIL-1 beta(200 nM).
The analysis of this injection showed that the stoichiometry for
rhIL-1 alpha was 2 and for rhIL-1 beta was 0.9 (see FIG. 22). In
the next cycle, rhIL-1 beta (200 nM) was injected first followed by
rhIL-1 alpha, and, in this case, the stoichiometry for the antigens
remained the same as before.
[0498] The analysis showed that the antibody has a higher affinity
for rhIL-1 alpha than rhIL-1 beta. The difference in affinity is in
the off rate. In addition, the stoichiometry was different for
these two analyte. Simultaneous binding of rhIL-1 alpha followed by
rhIL-1 beta to the dual specific antibody showed non interfering
independent binding (see FIG. 22). The reverse was also found,
i.e., simultaneous binding of rhIL-1 beta followed by rhIL-1 alpha
to ABT2-108-620x/ABT1-95-A3 showed non interfering independent
binding (stoichiometry rhIL-1 alpha 2.0 and stoichiometry rhIL-1
beta 0.98).
Kinetic Analysis
[0499] Following the above, the kinetic analysis was preformed
individually for rhIL-1 alpha and rhIL-1 beta. In case on rhIL-1
alpha the concentrations used were between 100-0.78 nM and for
rhIL-1 beta the concentration range used was between 200-1.56 nM.
Various concentrations of rhIL-1 alpha or rhIL-1 beta were injected
over the captured antibody (1-5 ug/ml) using goat anti hu IgG FC
capture. The surface was regenerated using 10 mM Glycine pH
1.5.
[0500] Results from the kinetic assay (bound IL-1 alpha or IL-1
beta to captured antibody) are provided in Table 59.
TABLE-US-00072 TABLE 59 Summary of Kinetic rate parameters by
Biacore analysis: Captured antibody: ABT2-108-620x/ABT1-95-A3
rhIL-1 alpha rhIL-1 beta On Rate-ka Expt 1: 4.79 .times. 10.sup.5
Expt 1: 3.15 .times. 10.sup.5 (1/Ms) Expt 2: 4.92 .times. 10.sup.5
Expt 2: 2.83 .times. 10.sup.5 Average : 4.855 .times. 10.sup.5
Average : 2.99 .times. 10.sup.5 Off Rate-kd Expt 1: 4.25 .times.
10.sup.-5 Expt 1: 1.38 .times. 10.sup.-4 (1/s) Expt 2: 4.11 .times.
10.sup.-5 Expt 2: 1.02 .times. 10.sup.-4 Average : 4.18 .times.
10.sup.-5 Average : 1.2 .times. 10.sup.-4 KD (pM) Expt 1: 88.7 Expt
1: 446 Expt 2: 83.5 Expt 2: 361 Average : 86.1 Average : 403.5
Thus, the binding to a first antigen does not interfere with the
binding to a second antigen.
Example 9
In Vivo Inhibition of IL-1.alpha. or IL-1.beta. Using Dual Specific
IgG
[0501] The following example provides an in vivo experiment using
the dual specific antibody, ABT2-108-620x/ABT1-95-A3, to inhibit
IL-1.alpha. or IL-1.beta..
[0502] At days 7, 4, or 1 prior to cytokine challenge, C57131/6N
mice at 8 weeks of age were dosed with 200 .mu.l of
ABT2-108-620x/ABT1-95-A3 via intra-peritoneal injection (200 .mu.g
for IL-1.alpha. challenged animals and 50 .mu.g for IL-1.beta.
challenged animals).
[0503] Animals were then challenged with 40 ng of rhIL-1.alpha.
cytokine (Roche) or 60 ng of rhIL-1.beta. cytokine (R&D
Systems) in 100 .mu.l by subcutaneous injection in the scruff of
the neck. At 2 hours post cytokine challenge, animals were
euthanized and terminally bled via cardiac puncture for plasma.
[0504] As shown in FIG. 23, treatment of animals with 50 .mu.g of
the anti-IL-1.alpha./.beta. antibody ABT2-108-620x/ABT1-95-A3
significantly inhibited IL-6 production in response to rhIL-1.beta.
up 10 to 7 days prior to challenge. IL-6 production in response to
rhIL-1.alpha. was significantly inhibited by treatment with 200
.mu.g of antibody 1 day prior to challenge, but not 4 or 7 days
prior to challenge.
[0505] The following Tables (Tables 60 to 64) relate to the above
examples.
TABLE-US-00073 TABLE 60 Domain Antibody Sequences Seq ID Sequences
VH 53 VH Dummy evqllesggglvqpggslrlscaasgftfs wvrqapgkglewv s
rftisrdnskntlylqmnslraedtavyyc ak wgqgtlvtvss VH to IL-1alpha 54
ABT1-207 evqllesgggliqpggslrlscaasgftfgryymswvrqapgkglewv
ssidylgtntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akltrirppnfdywgqgtlvtvss 55 ABT1-6
evqllesggglvqpggslrlscaasgftfgaydmqwvrqapgkglewv
ssinksgaltsyadsvkgrftisrdnskntlylqmnslraedtavyyc
akgwasfdywgqgtlvtvss 56 ABT1-6-15
evqllesggglvqpggslrlscaasgftfvrydmawvrqapgkglewv
ssinksgaltsyadsvkgrftisrdnskntlylqmnslraedtavyyc
akgwasfdywgqgtlvtvss 4 ABT1-6-23
evqllesggglvqpggslrlscaasgftfvrydmawvrqapgkglewv
ssiyksgaltsyadsvkgrftisrdnskntlylqmnslraedtavyyc
akgwasfdywgqgtlvtvss 57 ABT1-6-3
evqllesggglvqpggslrlscaasgftfgaydmqwvrqapgkglewv
ssiyksgaltsyadsvkgrftisrdnskntlylqmnslraedtavyyc
akgwasfdywgqgtlvtvss 8 ABT1-86
evqllesggglvqpggslrlscaasgftfdryimawvrqapgkglewv
ssitpsgaatyyadsvkgrftisrdnskntlylqmnslraedtavyyc
aeepadrystwtfdywgqgtlvtvss 16 ABT1-96
evqllesggglvqpggslrlscaasgftfnqynmfwvrqapgkglewv
svisgsgrftyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akgwwrrdppfdywgqgtlvtvss 20 ABT1-98
evqllesggglvqpggslrlscaasgftfdgyimswvrqapgkglewv
stisplgsvtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akkgpwfdywgqgtlvtvss VH to IL-1beta 58 ABT2-10
evqllesggglvqpggslrlsctasgftfnrynmawarqapgkglewv
seidlkgsqtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akvsisayhmfdywgqgtlvtvss 52 ABT2-108
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 59 ABT2-108-504
evqllesggglvqpggslrlscaasgftfadegmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 60 ABT2-108-509
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
sritysgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 61 ABT2-108-511
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntvirdsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 62 ABT2-108-
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv 512 (=611)
srigqdgkntyvlrsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 63 ABT2-108-518
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyrmdvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 64 ABT2-108-521
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrilghhlfdywgqgtlvtvss 65 ABT2-108-524
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvfnhhlfdywgqgtlvtvss 66 ABT2-108-527
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvfkhhlfdywgqgtlvtvss 67 ABT2-108-533x
evqllesggglvqpggslrlscaasgftfadegmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrilghhlfdywgqgtlvtvss 68 ABT2-108-534x
evqllesggglvqpggslrlscaasgftfadegmmwvrqapgkglewv
sritysgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrifshhlfdywgqgtlvtvss 69 ABT2-108-537x
evqllesggglvqpggslrlscaasgftfadegmmwvrqapgkglewv
srigqdgkntyyrmdvkgrftisrdnskntlylqmnslraedtavyyc
akytgrilghhlfdywgqgtlvtvss 70 ABT2-108-538x
evqllesggglvqpggslrlscaasgftfadegmmwvrqapgkglewv
sritysgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrilghhlfdywgqgtlvtvss 71 ABT2-108-601
evqllesggglvqpggslrlscaasgftfaeeswmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 72 ABT2-108-602
evqllesggglvqpggslrlscaasgftfaeekymwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 73 ABT2-108-603
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
sritdagkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 74 ABT2-108-604
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srvtydgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 75 ABT2-108-605
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyredvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 76 ABT2-108-606
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyrssvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 77 ABT2-108-607
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyrsdvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvgvhhlfdywgqgtlvtvss 78 ABT2-108-612
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrimghhlfdywgqgtlvtvss 79 ABT2-108-613
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrvfehhlfdywgqgtlvtvss 80 ABT2-108-617
evqllesggglvqpggslrlscaasgftfaeytmmwvrqapgkglewv
srigqdgkntyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akytgrifthhlfdywgqgtlvtvss 81 ABT2-108-620S
evqllesggglvqpggslrlscaasgftfseeswmwvrqapgkglewv
srigqdgkntyyredvkgrftisrdnskntlylqmnslraedtavyyc
akytgrimghhlfdywgqgtlvtvss 82 ABT2-108-620x
evqllesggglvqpggslrlscaasgftfaeeswmwvrqapgkglewv
srigqdgkntyyredvkgrftisrdnskntlylqmnslraedtavyyc
akytgrimghhlfdywgqgtlvtvss 32 ABT2-13
evqllesggglvqpggslrlscaasgftfrdyvmywarqapgkglewv
sridpmgsstyyadsvkgrftisrdnskntlylqmnslraedtavyyc
akpegnfdywgqgtlvtvss 44 ABT2-65
evqllesggglvqpggslrlscaasgftfedyqmgwvrqapgkglewv
ssisamgnrtyyadsvkgrftisrddskntlylqmnslraedtavyyc
aknlvrtqskmwmfdywgqgtlvtvss 83 ABT2-65-166
evqllesggglvqpggslrlscaasgftfedyqmgwvrqapgkglewv
ssisamggrtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 84 ABT2-65-166S
evqllesggglvqpggslrlscaasgftfsdyqmgwvrqapgkglewv
ssisamggrtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 85 ABT2-65-166SK
evqllesggglvqpggslrlscaasgftfsdyqmgwvrqapgkglewv
ssisamggrtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
aknlvrlgrsrwmfdywgqgtlvtvss 86 ABT2-65-17
evqllesggglvqpggslrlscvasgftfedyqmgwvrqapgkglewv
ssisamgnrtyyadsvkgrftisrddskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 87 ABT2-65-201
evqllesggglvqpggslrlscvasgftfedyqmgwvrqapgkglewv
ssisamgrrtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 88 ABT2-65-203
evqllesggglvqpggslrlscvasgftfedyqmgwvrqapgkglewv
ssisamgfrtyyadsvkgrftisrdnskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 89 ABT2-65-8
evqllesggglvqpggslrlscaasgftfedyqmgwvrqapgkglewv
ssisamgnrtyyadsvkgrftisrddskntlylqmnslraedtavyyc
aqnlvrmdsrrwmfdywgqgtlvtvss 90 ABT2-65-B1
evqllesggglvqpggslrlscvasgftfedyqmgwvrqapgkglewv
ssisamggrtyyadsvkgrftisrddskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 91 ABT2-65-B2
evqllesggglvqpggslrlscvasgftfedyqmgwvrqapgkglewv
ssisamgrrtyyadsvkgrftisrddskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss 92 ABT2-65-B3
evqllesggglvqpggslrlscvasgftfedyqmgwvrqapgkglewv
ssisamgnrayyadsvkgrftisrddskntlylqmnslraedtavyyc
aqnlvrlgrsrwmfdywgqgtlvtvss VL 93 Vk Dummy diqmtqspsslsasvgdrvtitc
wyqqkpgkapkll iy gvpsrfsgsgsgtdftltisslqpedfatyyc fgqgtkveikr VL to
IL-1alpha 24 ABT1-122
diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr 94
ABT1-122-505 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf patfgqgtkveikr 95
ABT1-122-508 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqrgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr 96
ABT1-122-510 diqmtqspsslsasvgdrvtitcrfsypiwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr 97
ABT1-122-511 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqrgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf patfgqgtkveikr 98
ABT1-122-512 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygsakrqrgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf patfgqgtkveikr 99
ABT1-122-513 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygsgsrqrgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf patfgqgtkveikr 100
ABT1-122-551 diqmtqspsslsasvgdrvtitcrakfniwtelnwyqqkpgktpkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr 101
ABT1-122-552 diqmtqspsslsasvgdrvtitcraslgvwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr 102
ABT1-122-553 diqmtqspsslsasvgdrvtitcrasqpiwtelkwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr 103
ABT1-122-554 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqetkveikr 104
ABT1-122-555 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgktpkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patfgqgtkveikr
105 ABT1-122-556 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyhcqqfayf patfgqgtkveikr 106
ABT1-122-557 diqmtqspsslsasvgdrvtitcrasqpiwtelnwyqqkpgkapkll
iygssslqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf patlgqgtkveikr 107
ABT1-122-750M diqmtqspsslsasvgdrvtitcsasqhiwtemswyqqkpgkapkll
iygsasrqkgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf pntfgqgtkveikr 108
ABT1-122-750MH diqmtqspsslsasvgdrvtitcsasqhiwtemswyqqkpgkapkll
iygsasrqkgvpsrfsgsgsgtdftltisslqpedfatyhckqfayf pntfgqgtkveikr 109
ABT1-122-750MT diqmtqspsslsasvgdrvtitcsasqhiwtemswyqqkpgktpkll
iygsasrqkgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf pntfgqgtkveikr 110
ABT1-122-750T diqmtqspsslsasvgdrvtitcsasqhiwteiswyqqkpgkapkll
iygsasrqkgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf pntfgqgtkveikr 111
ABT1-122-750x diqmtqspsslsasvgdrvaitcsasqhiwteiswyqqkpgkapkll
iygsasrqkgvpsrfsgsgsgtdftltisslqpedfatyyckqfayf pntfgqgtkveikr 112
ABT1-122-750xH diqmtqspsslsasvgdrvaitcsasqhiwteiswyqqkpgkapkll
iygsasrqkgvpsrfsgsgsgtdftltisslqpedfatyhckqfayf pntfgqgtkveikr 28
ABT1-141 diqmtqspsslsasvgdrvtitcrasqwiqkqlawyqqkpgkapkll
iysssylqsgvpsrfsgsgsgtdftltisslqpedfatyycqqhlrv pftfgqgtkveikr 113
ABT1-141-25 diqmtqspsslsasvgdrvtitcrasqwiqkqlawyqqkpgkapkll
iysssylqsgvpsrfsgsgsgtdftltisslqpedfatyycqqhlrv pmtfgqgtkveikr 114
ABT1-141-29 diqvtqspsslsasvgdrvtitcrasqwiqkqlawyqlkpgkapkll
iysssylqsgvpsrfsgsgsgtdftltisslqpedfatyycqqhlrv pftfgqgtkveikr 115
ABT1-18 diqmtqspsslsasvgdrvtitcrasqsiqrwlawyqqkpgkapkll
iyfasqlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqllrl pktfgqgtkveikr 116
ABT1-212 diqmtqspsslsasvgdrvtitcrasqringnlrwyqqkpgkapkll
iysvsqlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqgydw pptfgqgtkvetkr 117
ABT1-221 diqmtqspsslsasvgdrvtitcrasqdiwpylmwyqqkpgkapkll
iyyssmlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfrrw pytfgqgtkveikr 118
ABT1-222 diqmtqspsslsasvgdrvtitcrasqpitrnlrwyqqkpgkapkll
iyhssvlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqgyrw pvtfgqgtkveikr 119
ABT1-3 diqmtqspsslsasvgdrvtitcrasqsiwtelkwyqqkpgkapkll
iygasllqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfayf pftfgqgtkveikr 120
ABT1-9 diqmtqspsslsasvgdrvtitcrasqsigssllwyqqkpgkapkll
iylasrlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqfrst pntfgqgtkveikr 12
ABT1-95 diqmtqspsslsasvgdrvtitcrasqpihgnlrwyqqkpgkapkll
iynisnlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqgyrw pvtfgqgtkveikr 121
ABT1-95-15 diqmtqspsslsasvgdrvtitcrasqpivrnlrwyqqkpgkapkll
iysssylepgvpsrfsgsgsgtdftltisslqpedfatyhcqqgyrw pvtfgqgtkveikr 122
ABT1-95-174 diqmtqspsslsasvgdrvtitcrasrpivrnlrwyqqkpgkapkll
iysvsylepgvpsrfsgsgsgtdftltisslqpedfatyhcrqgyrw pvtfgqgtkveikr 123
ABT1-95-177 diqmtqspsslsasvgdrvtitcrasqpivrnlrwyqqkpgkapkll
iyassylepgvpsrfsgsgsgtdftltisslqpedfatyhcrqgyrw pvtfgqgtkveikr 124
ABT1-95-181 diqmtqspsslsasvgdrvtitcraskpivrnmrwyqqkpgkapkll
iysvsylepgvpsrfsgsgsgtdftltisslqpedfatyhclqgyrw pptfgqgtkveikr 125
ABT1-95-182 diqmtqspsslsasvgdrvtitcraskpgvrnlrwyqqkpgkapkll
iysvsylepgvpsrfsgsgsgtdftltisslqpedfatyhclqgyrw pptfgqgtkveikr 126
ABT1-95-184 diqmtqspsslsasvgdrvtitcrasrtpvrnlrwyqqkpgkapkll
iysrsylepgvpsrfsgsgsgtdftltisslqpedfatyhclqgyrw pptfgqgtkveikr 127
ABT1-95-3 diqmtqspsslsasvgdrvtitcrasqpihgnlrwyqqkpgkapkll
iysssylqsgvpsrfsgsgsgtdftltisslqpedfatyhcqqgyrw pvtfgqgtkveikr 128
ABT1-95-A1 diqmtqspsslsasvgdrvtitcrasrpgvrnlrwyqqkpgkapkll
iyhvsdlepgvpsrfsgsgsgtdftltisslqpedfatyhcrqgyvw pvpfdqgtkveikr 129
ABT1-95-A2 diqmtqspsslsasvgdrvtitcrilqppgrnlrwyqqkpgkapkll
iysksflepgvpsrfsgsgsgtdftltisslqpedfatyhcqqgyrw pvtfgqgtkveikr 130
ABT1-95-A3 diqmtqspsslsasvgdrvtitcraskpgvrnmrwyqqkpgkapkll
iysvsylepgvpsrfsgsgsgtdftltisslqpedfatyhclqgyrw pptfgqgtkveikr 131
ABT1-95-A4 diqmtqspsslsasvgdrvtitcrasrpgvrnlrwyqqkpgkapkll
iysrsflepgvpsrfsgsgsgtdftltisslqpedfatyhclqgyrw pptfgqgtkveikr 132
ABT1-95-A5 diqmtqspsslsasvgdrvtitcrasrtpvrnlrwyqqkpgkapkll
iysrsflepgvpsrfsgsgsgtdftltisslqpedfatyhclqgyrw pptfgqgtkveikr 133
ABT1-95-A6 diqmtqspsslsasvgdrvtitcraskpgvrnmrwyqqkpgkapkll
iyaksylepgvpsrfsgsgsgtdftltisslqpedfatyhckqgyrw pvqfgqgtkveikr VL
to IL-1beta 36 ABT2-42
diqmtqspsslsasvgdrvtitcrasqyiekwltwyqqkpgkaptll
iyrgsllqsgvpsrfsgsgsgtdftltisslqpedfatyycqqteyw pftfgqgtkveikr 40
ABT2-46 diqmtqspsslsasvgdrvtitcrasqsiiewlswyqqkpgkapkll
iyrtsvlqsgvpsrfsgsgsgtdftltisslqpedfatyycqqnefw pftfgqgtkveikr 48
ABT2-76 diqmtqspsslsasvgdrvtitcrasqsidrwlawyqqkpgkapkll
iyrgsilqsgvpsrfsgsgsgtdftltisslqpedfatyycqqvafw pptfgqgtkveikr
TABLE-US-00074 TABLE 61 Summary of identified single domain
antibodies IL1.alpha. Clone Name MRC5 AMINO ACID SEQUENCE (Clone
type) CDR1 CDR2 CDR3 ND.sub.50 K.sub.D (M) (diversified residues
are indicated in bold; CDRs are underlined) ABT1-6-23 VRYDMA
SIYKSGALTSYDS GWASFDY 0.03 .mu.M 6.1 .times. 10.sup.-8
EVQLLESGGGLVQPGGSLRLSCAASGFTFVRYDMAWVRQAPGKGLEWVS (VH) (SEQ ID NO:
1) VKG (SEQ ID SIYKSGALTSYDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKG
(SEQ ID NO: 2) NO: 3) WASFDYWGQGTLVTVSS (SEQ ID NO: 4) ABT1-86
RYIMA SITPSGAATYYAD EPADRYSTW 900 nM 1.1 .times. 10.sup.-7
EVQLLESGGGLVQPGGSLRLSCAASGFTFDRYIMAWVRQAPGKGLEWVS (VH) (SEQ ID NO:
5) SVKG TFDY SITPSGAATYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAE (SEQ
ID NO: 6) (SEQ ID EPADRYSTWTFDYWGQGTLVTVSS (SEQ ID NO: 8) NO: 7)
ABT1-95 RASQPIHGNLR NISNLQS QQGYRWPVT 800 nM 4.5 .times. 10.sup.-8
DIQMTQSPSSLSASVGDRVTITCRASQPIHGNLRWYQQKPGKAPKLLIY (VK) (SEQ ID NO:
9) (SEQ ID (SEQ ID
NISNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYRWPVTF NO: 10) NO: 11)
GQGTKVEIKR (SEQ ID NO: 12) ABT1-96 QYNMF VISGSGRFTYADS GWWRRDPPF 11
nM 1.1 .times. 10.sup.-9
EVQLLESGGGLVQPGGSLRLSCAASGFTFNQYNMFWVRQAPGKGLEWVS (VH) (SEQ ID VKG
DY VISGSGRFTYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKG NO: 13) (SEQ
ID (SEQ ID WWRRDPPFDYWGQGTLVTVSS (SEQ ID NO: 16) NO: 14) NO: 15)
ABT1-98 GYIMS TISPLGSVTYYAD KGPWFDY 30 nM 1.3 .times. 10.sup.-8
EVQLLESGGGLVQPGGSLRLSCAASGFTFDGYIMSWVRQAPGKGLEWVS (VH) (SEQ ID SVKG
(SEQ ID TISPLGSVTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK NO: 17)
(SEQ ID NO: 19) KGPWFDYWGQGTLVTVSS (SEQ ID NO: 20) NO: 18) ABT1-122
RASQPIWTELN GSSSLQS QQFAYFPAT 50 nM 4.2 .times. 10.sup.-8
DIQMTQSPSSLSASVGDRVTITCRASQPIWTELNWYQQKPGKAPKLLIY (VK) (SEQ ID (SEQ
ID (SEQ ID GSSSLQSGVPRFSGSGSGTDFTLTISSLQPEDFATYYCQQFAYFPATFG NO:
21) NO: 22) NO: 23) QGTKVEIKR (SEQ ID NO: 24) ABT1-141 RASQWIQKQLA
SSSYLQS QQHLRVPFT 50 nM 3.0 .times. 10.sup.-7
DIQMTQSPSSLSASVGDRVTITCRASQWIQKQLAWYQQKPGKAPKLLIY (VK) (SEQ ID (SEQ
ID (SEQ ID SSSYLQSGVPRFSGSGSGTDFTLTISSLQPEDFATYYCQQHLRVPFTFG NO:
25) NO: 26) NO: 27) QGTKVEIKR (SEQ ID NO: 28) IL1.beta. Clone Name
ABT2-13 DYVMY RIDPMGSSTYYAD PEGNFDY ~1000 nM 9.3 .times. 10.sup.-7
EVQLLESGGGLVQPGGSLRLSCAASGFTFRDYVMY+EEWARQAPGKGLEWVS (VH) (SEQ ID
SVKG (SEQ ID RIDPMGSSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK NO:
29) (SEQ ID NO: 31) PEGNFDYWGQGTLVTVSS (SEQ ID NO: 32) NO: 30)
ABT2-42 RASQYIEKWLT RGSLLQS QQTEYWPFT 770 nM 5.4 .times. 10.sup.-5
DIQMTQSPSSLSASVGDRVTITCRASQYIEKWLT+EEWYQQKPGKAPTLLIY (VK) (SEQ ID
(SEQ ID (SEQ ID RGSLLQSGVPRFSGSGSGTDFTLTISSLQPEDFATYYCQQTEYWPFTFG
NO: 33) NO: 34) NO: 35) QGTKVEIKR (SEQ ID NO: 36) ABT2-46
RASQSIIEWLS RTSVLQS QQNEFWPFT 386 nM 2.8 .times. 10.sup.-6
DIQMTQSPSSLSASVGDRVTITCRASQSIIEWLSWYQQKPGKAPKLLIY (VK) (SEQ ID (SEQ
ID (SEQ ID RTSVLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNEFWPFTF NO:
37) NO: 38) NO: 39) GQGTKVEIKR (SEQ ID NO: 40) ABT2-65 DYQMG
SISAMGNRTYYAD NLVRTQSKM 266 nM 2.8 .times. 10.sup.-8
EVQLLESGGGLVQPGGSLRLSCAASGFTFEDYQMGWVRQAPGKGLEWVS (VH) (SEQ ID SVKG
WMFDY SISAMGNRTYYADSVKGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCAK NO: 41)
(SEQ ID (SEQ ID NLVRTQSKMWMFDYWGQGTLVTVSS (SEQ ID NO: 44) NO: 42)
NO: 43) ABT2-76 RASQSIDRWLA RGSILQS QQVAFWPPT 456 nM 1.3 .times.
10.sup.-6 DIQMTQSPSSLSASVGDRVTITCRASQSIDRWLAWYQQKPGKAPKLLIY (VK)
(SEQ ID (SEQ ID (SEQ ID
RGSILQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVAFWPPTF NO: 45) NO: 46)
NO: 47) GQGTKVEIKR (SEQ ID NO: 48) ABT2-108 EYTMM RIGQDGKNTYYAD
YTGRVGVHH 200 nM 1.1 .times. 10.sup.-7
EVQLLESGGGLVQPGGSLRLSCAASGFTFAEYTMMWVRQAPGKGLEWVS (VH) (SEQ ID SVKG
LFDY RIGQDGKNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK NO: 49) (SEQ
ID (SEQ ID YTGRVGVHHLFDYWGQGTLVTSS (SEQ ID NO: 52) NO: 50) NO:
51)
TABLE-US-00075 TABLE 62 ABT 1 dAbs Clone Name dAb IC.sub.50 [nM]
ND.sub.50 [nM] K.sub.D [nM] ABT1-3 VK 1000 ABT1-6 VH 3000 3000
ABT1-6-3 VH 80 60 ABT1-6-15 VH 200 500 ABT1-6-23 VH 30 30 6 ABT1-7
VH ABT1-8 VH ABT1-9 VK 4000 ABT1-9-500 VK ABT1-10 VK >20000
ABT1-11 VK >20000 ABT1-12 VK ABT1-13 VK ABT1-14 VK ABT1-15 VH
ABT1-17 VH ABT1-18 VK >10000 ABT1-19 VH ABT1-20 VH 1000 ABT1-21
VH ABT1-22 VH 7000 ABT1-23 VH 5000 ABT1-24 VK ABT1-25 VK ABT1-26 VK
ABT1-27 VK ABT1-28 VK ABT1-29 VK ABT1-30 VK ABT1-31 VH ABT1-32 VH
ABT1-33 VH ABT1-34 VH ABT1-35 VH ABT1-36 VH ABT1-37 VH ABT1-38 VH
ABT1-40 VH ABT1-41 VH ABT1-42 VH ABT1-43 VH ABT1-45 VH ABT1-46 VH
ABT1-47 VH 5000 ABT1-49 VH ABT1-50 VH ABT1-51 VH ABT1-52 VH ABT1-53
VK ABT1-54 VK ABT1-56 VK ABT1-57 VK ABT1-59 VK ABT1-60 VK ABT1-61
VK ABT1-62 VK ABT1-63 VK ABT1-64 VK ABT1-65 VK ABT1-66 VK ABT1-67
VK 0.044 ABT1-68 VK ABT1-75 VH ABT1-76 VH ABT1-77 VH ABT1-78 VH
ABT1-79 VH ABT1-81 VH ABT1-82 VK ABT1-84 VH >10000 3000 ABT1-85
VH >10000 30000 ABT1-86 VH 1000 900 ABT1-87 VH >10000
>10000 ABT1-88 VH >10000 >10000 ABT1-89 VH >10000
>10000 ABT1-90 VH >10000 >10000 ABT1-91 VH >10000
>10000 ABT1-92 VH >10000 >10000 ABT1-93 VH >10000 5000
ABT1-94 VH ABT1-95 VK 500 800 ABT1-95-3 VK 4 80 7 ABT1-95-4 VK 0.7
2 1.5 ABT1-95-6 VK 0.8 1 2.1 ABT1-95-8 VK 0.8 1 2.1 ABT1-95-9 VK
0.8 -- 1 ABT1-95-10 VK 0.8 -- 0.9 ABT1-95-11 VK 0.9 0.300 1.1
ABT1-95-12 VK 0.8 -- 0.9 ABT1-95-13 VK 0.3 0.120 0.9 ABT1-95-14 VK
0.3 0.250 1.2 ABT1-95-15 VK 0.3 0.045 0.4 ABT1-95-21 VK 0.5 0.61
ABT1-95-22 VK 0.5 1 ABT1-95-23 VK 0.5 0.5 ABT1-95-24 VK 0.5 0.27
ABT1-95-25 VK 0.5 0.43 ABT1-95-26 VK 0.5 0.71 ABT1-95-27 VK 0.5 1
ABT1-95-28 VK >0.5 2 ABT1-95-29 VK >0.5 1 ABT1-95-30 VK >1
>10 ABT1-95-31 VK >0.5 3.3 ABT1-95-32 VK >0.5 1.2
ABT1-95-33 VK 0.5 1 ABT1-95-34 VK 0.1 1 ABT1-95-35 VK 0.2
ABT1-95-36 VK <0.1 ABT1-95-37 VK 0.1 ABT1-95-38 VK <0.1 0.119
ABT1-95-39 VK 0.2 ABT1-95-40 VK ABT1-95-41 VK <0.1 0.150
ABT1-95-42 VK 0.1 ABT1-95-43 VK ABT1-95-44 VK <0.1 0.150
ABT1-95-45 VK ABT1-95-46 VK 0.8 ABT1-95-47 VK 0.1 0.139 ABT1-95-48
VK ABT1-95-49 VK 274 ABT1-95-50 VK 0.5 ABT1-95-51 VK 615 ABT1-95-52
VK 1 ABT1-95-53 VK 0.3 ABT1-95-54 VK 1 ABT1-95-55 VK 0.6 ABT1-95-56
VK 0.3 ABT1-95-57 VK 0.6 ABT1-95-58 VK 0.6 ABT1-95-59 VK 0.3
ABT1-95-60 VK 0.7 ABT1-95-61 VK 0.3 ABT1-95-62 VK 0.1 ABT1-95-63 VK
0.6 ABT1-95-64 VK 3 0.438 ABT1-95-65 VK 0.2 ABT1-95-67 VK 0.1 0.044
ABT1-95-68 VK 0.2 ABT1-95-69 VK 1 ABT1-95-70 VK 0.1 ABT1-95-71 VK 1
ABT1-95-72 VK 1 ABT1-95-73 VK 3 ABT1-95-74 VK 0.3 ABT1-95-75 VK 0.3
ABT1-95-76 VK 0.3 ABT1-95-77 VK 0.3 ABT1-95-78 VK 0.3 ABT1-95-79 VK
0.3 ABT1-95-80 VK 0.3 0.008 ABT1-95-81 VK 0.3 ABT1-95-82 VK 0.4
ABT1-95-83 VK 0.2 ABT1-95-84 VK ABT1-95-85 VK 0.2 0.713 ABT1-95-86
VK 0.1 0.2 ABT1-95-87 VK 0.2 ABT1-95-88 VK 0.2 0.714 ABT1-95-89 VK
0.3 ABT1-95-90 VK 0.5 ABT1-95-91 VK 0.3 ABT1-95-92 VK <0.1 0.5
ABT1-95-93 VK 0.1 ABT1-95-94 VK 0.1 ABT1-95-95 VK 0.3 ABT1-95-96 VK
0.1 ABT1-95-97 VK ABT1-95-98 VK 0.8 ABT1-95-99 VK ABT1-95-100 VK
ABT1-95-101 VK 0.5 ABT1-95-102 VK 0.1 0.184 ABT1-95-103 VK
ABT1-95-104 VK ABT1-95-105 VK 0.1 ABT1-95-106 VK 0.1 ABT1-95-107 VK
0.1 0.5 ABT1-95-108 VK 0.5 ABT1-95-109 VK ABT1-95-110 VK 0.1
ABT1-95-111 VK <0.1 ABT1-95-112 VK 0.1 ABT1-95-113 VK <0.1
ABT1-95-114 VK 0.2 ABT1-95-115 VK 0.2 0.230 ABT1-95-116 VK 0.2
0.168 ABT1-95-117 VK 0.2 0.1 ABT1-95-118 VK 0.2 0.122 ABT1-95-119
VK ABT1-95-120 VK 0.1 0.1 ABT1-95-121 VK 0.2 0.1 ABT1-95-122 VK 0.1
0.116 ABT1-95-123 VK 0.1 ABT1-95-124 VK 0.2 ABT1-95-125 VK 0.2
ABT1-95-126 VK 0.8 ABT1-95-127 VK 0.1 ABT1-95-128 VK >1
ABT1-95-129 VK >10 ABT1-95-130 VK >1 ABT1-95-131 VK 0.1 0.085
ABT1-95-132 VK 0.1 2 ABT1-95-133 VK 0.050 ABT1-95-134 VK 0.075
ABT1-95-135 VK 0.1 ABT1-95-136 VK 0.1 ABT1-95-137 VK 0.09
ABT1-95-139 VK 0.2 0.1 ABT1-95-140 VK 0.2 ABT1-95-141 VK 1
ABT1-95-142 VK 0.5 ABT1-95-143 VK 10 ABT1-95-144 VK 0.5 ABT1-95-145
VK 0.8 ABT1-95-146 VK 0.8 ABT1-95-147 VK 0.2 ABT1-95-148 VK 0.2
0.190 ABT1-95-149 VK 0.2 0.064 ABT1-95-150 VK 0.2 ABT1-95-151 VK
0.2 ABT1-95-152 VK 0.8 ABT1-95-153 VK 0.5 ABT1-95-154 VK >10
ABT1-95-155 VK 10 ABT1-95-156 VK 10 ABT1-95-157 VK 10 ABT1-95-158
VK 0.5 ABT1-95-159 VK 0.1 0.127 ABT1-95-160 VK 0.1 0.238
ABT1-95-161 VK 0.1 0.5 ABT1-95-162 VK 0.186 ABT1-95-163 VK 0.05
ABT1-95-164 VK 0.150 ABT1-95-165 VK 0.391 ABT1-95-166 VK 0.035
ABT1-95-167 VK 0.469 ABT1-95-168 VK 2.3 ABT1-95-169 VK 0.093
ABT1-95-170 VK 0.2 ABT1-95-171 VK 0.059 ABT1-95-172 VK 0.171
ABT1-95-173 VK 0.151 ABT1-95-174 VK 0.01 ABT1-95-175 VK 0.017
ABT1-95-176 VK 0.027 ABT1-95-177 VK 0.008 ABT1-95-178 VK 0.004
ABT1-95-179 VK 0.022 ABT1-95-181 VK 0.014 ABT1-95-182 VK 0.005
ABT1-95-183 VK ABT1-95-184 VK 0.006 ABT1-95-500 VK ABT1-95-501 VK
ABT1-95-502 VK ABT1-95-503 VK ABT1-95-A1 VK ABT1-95-A2 VK 0.133
ABT1-95-A3 VK 0.011 ABT1-95-A5 VK 0.016 ABT1-95-A6 VK 0.024 ABT1-96
VH 17 ABT1-97 VH 3000 ABT1-98 VH <100 ABT1-99 VH 500 ABT1-101 VH
ABT1-102 VH ABT1-103 VH ABT1-104 VH ABT1-105 VH ABT1-106 VH
ABT1-109 VH >500 ABT1-110 VH ABT1-111 VH ABT1-113 VH >500
ABT1-117 VH >500 ABT1-118 VH ABT1-119 VK ABT1-120 VK ABT1-121 VK
ABT1-122 VK 80 100 ABT1-122-18 VK 5 7 ABT1-122-21 VK 80 100
ABT1-122-511 VK 10 18 ABT1-122-750X VK 0.713 ABT1-123 VK >500
ABT1-124 VK ABT1-125 VK ABT1-126 VK ABT1-127 VK ABT1-128 VK
ABT1-129 VK ABT1-130 VK 300 ABT1-131 VK 80 ABT1-132 VK 300 ABT1-133
VK ABT1-134 VK ABT1-135 VK ABT1-136 VK ABT1-137 VH 200 ABT1-138 VH
>500 ABT1-139 VH 80 ABT1-140 VH ABT1-141 VK 50 142 ABT1-141-1 VK
>100 ABT1-141-2 VK 100 ABT1-141-3 VK 80 ABT1-141-6 VK 20
ABT1-141-7 VK 10 ABT1-141-8 VK 10 ABT1-141-9 VK 10 ABT1-141-10 VK
30 ABT1-141-11 VK ABT1-141-12 VK ABT1-141-13 VK ABT1-141-14 VK 10
ABT1-141-15 VK 50 ABT1-141-16 VK 20 ABT1-141-17 VK 30 ABT1-141-18
VK 80 ABT1-141-19 VK 50 ABT1-141-20 VK 10 ABT1-141-21 VK 60
ABT1-141-22 VK 30 ABT1-141-23 VK 80 ABT1-141-24 VK ABT1-141-25 VK 3
5 ABT1-141-27 VK 20 ABT1-141-28 VK 200 ABT1-141-29 VK 20
ABT1-141-30 VK 40 ABT1-141-31 VK 2 ABT1-141-32 VK 10 ABT1-141-33 VK
3 ABT1-141-34 VK 30 ABT1-141-35 VK 10 ABT1-141-42 VK 3 5
ABT1-141-43 VK 3 5 ABT1-141-44 VK 3 5 ABT1-141-45 VK 100 100
ABT1-141-46 VK 3 1 ABT1-141-47 VK 1 1 ABT1-141-48 VK 3 1
ABT1-141-49 VK 3 5 ABT1-141-50 VK 10 20 ABT1-141-51 VK 10 20
ABT1-141-52 VK 100 100 ABT1-141-53 VK 50 10 ABT1-141-54 VK 3 5
ABT1-141-70 VK 1 1 ABT1-141-75 VK 3 5 ABT1-141-76 VK 1 1
ABT1-141-77 VK 3 1 ABT1-141-78 VK 3 0.7 ABT1-141-79 VK 3 2.4
ABT1-141-80 VK 3 1.7 ABT1-141-500 VK ABT1-141-501 VK ABT1-141-502
VK ABT1-141-503 VK ABT1-141-504 VK ABT1-141-505 VK ABT1-141-506 VK
ABT1-141-507 VK ABT1-141-508 VK ABT1-141-509 VK ABT1-141-510 VK
ABT1-141-511 VK ABT1-141-512 VK ABT1-141-513 VK ABT1-141-514 VK
ABT1-141-516 VK ABT1-141-519 VK ABT1-141-520 VK ABT1-141-521 VK
ABT1-141-522 VK ABT1-141-523 VK ABT1-141-524 VK ABT1-141-525 VK
ABT1-141-526 VK ABT1-141-527 VK ABT1-141-528 VK ABT1-141-529 VK
ABT1-141-530 VK ABT1-141-531 VK 1 ABT1-141-532 VK 2 ABT1-141-533 VK
ABT1-141-534 VK ABT1-141-535 VK ABT1-141-536 VK ABT1-141-537 VK
ABT1-141-538 VK ABT1-141-539 VK ABT1-141-541 VK ABT1-141-542 VK
ABT1-141-543 VK ABT1-141-544 VK ABT1-141-545 VK ABT1-141-546 VK
ABT1-141-547 VK ABT1-141-548 VK ABT1-141-549 VK ABT1-141-550 VK
ABT1-141-551 VK ABT1-141-552 VK ABT1-141-553 VK ABT1-141-554 VK
ABT1-141-555 VK ABT1-141-556 VK ABT1-141-557 VK ABT1-141-558 VK
ABT1-141-559 VK ABT1-141-561 VK ABT1-141-562 VK >10 ABT1-141-563
VK ABT1-141-564 VK ABT1-141-565 VK ABT1-141-567 VK ABT1-141-568 VK
ABT1-141-569 VK ABT1-141-572 VK ABT1-141-573 VK ABT1-141-574 VK
ABT1-142 VK 200 ABT1-143 VK >500 ABT1-144 VK 100 ABT1-145 VK
>500 ABT1-146 VK >500 ABT1-147 VH >500 300 ABT1-148 VK 400
100 ABT1-149 VK 100 100 ABT1-150 VK 150 ABT1-151 VK 200 ABT1-152 VK
300 ABT1-153 VK 300 ABT1-154 VK >1000 ABT1-155 VK 20 20 ABT1-156
VK ABT1-157 VK 300 ABT1-158 VK 2 <100 ABT1-159 VK 200 ABT1-160
VK 300 ABT1-161 VK >1000 8 ABT1-162 VK 1000 6.2 ABT1-163 VK 1000
105 ABT1-164 VK >1000 14 ABT1-165 VK >1000 1000 ABT1-166 VK
>1000 ABT1-167 VK >1000 ABT1-168 VK >1000 ABT1-169 VK 1000
ABT1-170 VK >1000 ABT1-171 VK >1000 ABT1-172 VK >1000
ABT1-221 VK 100
TABLE-US-00076 TABLE 63 ABT2 dAbs Clone Name dAb IC.sub.50 [nM]
ND.sub.50 [nM] K.sub.D [nM] ABT2-6 VK >20000 ABT2-7 VK >20000
ABT2-8 VK >20000 ABT2-10 VH 6000 ABT2-11 VH >10000 ABT2-12 VH
ABT2-13 VH 175 1000 ABT2-14 VH ABT2-15 VH ABT2-16 VH ABT2-17 VH
ABT2-19 VH ABT2-20 VH ABT2-21 VH ABT2-22 VH ABT2-23 VH 5000 ABT2-24
VH 2000 ABT2-25 VH ABT2-26 VH ABT2-27 VH ABT2-28 VH ABT2-29 VH
ABT2-30 VH ABT2-31 VH ABT2-32 VH 1000 ABT2-33 VH ABT2-35 VH
>10000 ABT2-36 VH ABT2-37 VH ABT2-38 VH ABT2-39 VH >10000
ABT2-40 VH ABT2-41 VH ABT2-42 VK 770 ABT2-43 VK >10000 ABT2-44
VK ABT2-46 VK 390 ABT2-53 VK ABT2-54 VK ABT2-55 VK ABT2-56 VK
ABT2-57 VK ABT2-58 VK ABT2-59 VK 700 >1000 ABT2-60 VK 1000
>1000 ABT2-61 VK 1000 >1000 ABT2-62 VK 1000 >1000 ABT2-63
VK 1000 ABT2-64 VH >10000 ABT2-65 VH 100 505 ABT2-65-1 VH 100
ABT2-65-2 VH 20 ABT2-65-7 VH 30 200 ABT2-65-8 VH 10 3 ABT2-65-9 VH
100 ABT2-65-10 VH 20 ABT2-65-11 VH 50 ABT2-65-12 VH 20 ABT2-65-13
VH 20 ABT2-65-14 VH 1000 ABT2-65-15 VH 100 ABT2-65-16 VH 200
ABT2-65-17 VH 2.5 5 ABT2-65-18 VH 20 ABT2-65-19 VH >1000
ABT2-65-20 VH >1000 ABT2-65-21 VH >1000 ABT2-65-22 VH 100
ABT2-65-23 VH 800 ABT2-65-24 VH 50 ABT2-65-25 VH 4 ABT2-65-26 VH 20
ABT2-65-27 VH 3.5 ABT2-65-28 VH 5 ABT2-65-29 VH 20 ABT2-65-30 VH 20
ABT2-65-31 VH 3 ABT2-65-32 VH 4 ABT2-65-33 VH 20 ABT2-65-34 VH 20
ABT2-65-35 VH 4 ABT2-65-36 VH >1000 ABT2-65-37 VH 100 ABT2-65-38
VH 30 ABT2-65-39 VH 300 ABT2-65-40 VH ABT2-65-41 VH >1000
ABT2-65-42 VH >1000 ABT2-65-43 VH ABT2-65-44 VH >1000
ABT2-65-45 VH >1000 ABT2-65-46 VH 30 ABT2-65-47 VH 20 80
ABT2-65-48 VH 20 80 ABT2-65-49 VH 20 ABT2-65-50 VH ABT2-65-51 VH
ABT2-65-52 VH ABT2-65-53 VH ABT2-65-54 VH ABT2-65-55 VH ABT2-65-56
VH ABT2-65-57 VH ABT2-65-58 VH ABT2-65-59 VH ABT2-65-6 VH 30
>1000 ABT2-65-60 VH ABT2-65-61 VH ABT2-65-62 VH ABT2-65-63 VH
ABT2-65-64 VH ABT2-65-65 VH ABT2-65-66 VH ABT2-65-67 VH ABT2-65-68
VH ABT2-65-69 VH ABT2-65-70 VH ABT2-65-71 VH ABT2-65-72 VH
ABT2-65-73 VH ABT2-65-74 VH >1000 ABT2-65-75 VH 30 ABT2-65-76 VH
8 ABT2-65-77 VH 50 ABT2-65-78 VH 30 ABT2-65-79 VH 6 2 ABT2-65-80 VH
7 0.5 ABT2-65-81 VH 10 2 ABT2-65-82 VH 8 2 ABT2-65-83 VH 300
ABT2-65-84 VH 60 ABT2-65-85 VH 7 2 ABT2-65-86 VH >1000
ABT2-65-87 VH 50 ABT2-65-88 VH 20 ABT2-65-89 VH 10 2 ABT2-65-90 VH
300 ABT2-65-113 VH 3.3 3 ABT2-65-114 VH 4 ABT2-65-115 VH 4
ABT2-65-116 VH ABT2-65-117 VH 10 ABT2-65-118 VH 4.6 ABT2-65-119 VH
4 ABT2-65-120 VH 4.2 ABT2-65-121 VH 3.9 ABT2-65-122 VH 3.8
ABT2-65-123 VH ABT2-65-124 VH 3.8 ABT2-65-125 VH 4 ABT2-65-126 VH
4.3 ABT2-65-127 VH 5.5 ABT2-65-128 VH ABT2-65-129 VH 3.4
ABT2-65-130 VH 4.3 ABT2-65-131 VH 13.2 ABT2-65-132 VH 5 ABT2-65-133
VH 30 ABT2-65-134 VH ABT2-65-135 VH 5 ABT2-65-136 VH ABT2-65-137 VH
5 ABT2-65-138 VH 5 ABT2-65-139 VH 4 ABT2-65-140 VH 2 1 ABT2-65-141
VH 5 ABT2-65-142 VH ABT2-65-143 VH 5 ABT2-65-144 VH 6 ABT2-65-145
VH 5 ABT2-65-146 VH 6 ABT2-65-147 VH ABT2-65-148 VH 4 ABT2-65-149
VH 4 ABT2-65-150 VH ABT2-65-151 VH 6 ABT2-65-152 VH 5 ABT2-65-154
VH 3 1.5 ABT2-65-155 VH 5 ABT2-65-156 VH 5 ABT2-65-157 VH
ABT2-65-158 VH 5 ABT2-65-159 VH 5 ABT2-65-160 VH 2 ABT2-65-163 VH 2
ABT2-65-166 VH 4 4 ABT2-65-167 VH 30 ABT2-65-168 VH 30 ABT2-65-169
VH 9 ABT2-65-170 VH 10 ABT2-65-171 VH 7.5 ABT2-65-172 VH 6
ABT2-65-173 VH 30 ABT2-65-174 VH 10 ABT2-65-175 VH 10 ABT2-65-176
VH 5 ABT2-65-177 VH 20 ABT2-65-500 VH ABT2-65-501 VH ABT2-65-502 VH
2 ABT2-65-503 VH ABT2-65-504 VH 4 ABT2-65-505 VH ABT2-65-506 VH
ABT2-65-507 VH ABT2-65-508 VH ABT2-65-509 VH ABT2-65-510 VH 1
ABT2-65-511 VH ABT2-65-512 VH 3 ABT2-65-513 VH ABT2-65-514 VH
ABT2-65-515 VH ABT2-66 VH >10000 ABT2-67 VK >10000 ABT2-68 VK
>10000 ABT2-69 VH >10000 ABT2-70 VH ABT2-71 VH ABT2-72 VH
>10000 ABT2-73 VH ABT2-74 VH ABT2-75 VH ABT2-76 VK 500 ABT2-77
VK ABT2-79 VK ABT2-80 VK >500 ABT2-81 VK ABT2-83 VK ABT2-84 VK
ABT2-85 VK ABT2-86 VK >500 ABT2-87 VK ABT2-88 VK ABT2-89 VK
ABT2-90 VK ABT2-91 VK >500 ABT2-93 VK >500 ABT2-94 VK >500
ABT2-95 VK >500 ABT2-96 VK >500 ABT2-97 VK >500 ABT2-98 VK
>500 ABT2-99 VH >500 ABT2-101 VH >500
ABT2-104 VH >500 ABT2-105 VH >500 ABT2-106 VH >500
ABT2-107 VH >500 ABT2-108 VH 70 ABT2-108-533X VH 5 ABT2-108-534X
VH ABT2-108-537X VH 206 ABT2-108-538X VH 5 ABT2-108-620X VH
TABLE-US-00077 TABLE 64 DNA sequences for ABT1 and ABT2 dAbs dAb
Name DNA sequence ABT1 VH dAbs ABT1-6
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGGCTTATGATATGCAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTAATAAGTCTGGTGCTTTGACATCTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTTGGGCGTCTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 134) ABT1-6-3
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGGCTTATGATATGCAGTGGGTCCGCCAGGCTCCAG
GCAAGGGTCTAGAGTGGGTCTCAAGTATTTATAAGTCTGGTGCTTTGACATCTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTTGGGCGTCTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 135) ABT1-6-23
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGTACGCTATGACATGGCATGGGTCCGCCAGGCTCCAG
GgAAGGGTCTAGAGTGGGTCTCAAGTATTTATAAGTCTGGTGCTTTGACATCTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTTGGGCGTCTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 136) ABT1-7
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGCGGTATCGGATGCAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTAGTTGGGAGGGTACGAGGACACTTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTGACTCAGAAGGGTT
TGTTGAATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 137)
ABT1-8
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCCTTAAGGCGTATAATATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACTGATTAGTAGTACGGGTATGTTTACAGATTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGGGGGTTAGGCGGG
GGCTTTATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 138)
ABT1-15
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGCGGTATCGGATGCAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTAATAAGTCTGGTGCTTTGACATCTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTTGGGCGTCTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 139) ABT1-17
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGCGGTATCGGATGCCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTAGTTGGGAGGGTACGAGGACACTTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTGACTCAAAAGGGTT
TGCTGAATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 140)
ABT1-19
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCCTGTTTATAGTATGCCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACTGATTCCGTGGCCTGGTTTGAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGAGACGTCTAATTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 141) ABT1-20
GAGGTGCAGCTGTTGGAGACTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGCCTTATCGGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACATATTGGGATTTGGGGTGCTAATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGCGCCGAGGACACCGCGGTATATTACTGTGCGAAACATCCGAGGCCTTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 142) ABT1-21
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGCCGTATGCGATGACTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTGGGCGGACGGGTACTCGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGGCGGCGGAGGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 143) ABT1-22
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCCTTAAGGCGTATAATATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACTGATTAGTAGTACGGGTATGTTTACAGATTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGGGGGTTAGGCGGG
GGCTTTATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 144)
ABT1-23
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCGTCGTATCCTATGGAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCGATTGCGTGGCCGGGTAGTATGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACGCATCGGCTTTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 145) ABT1-31
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGGTATTAATCGGTCTGGTACGCGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGGATTCGTCATTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 146) ABT1-32
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCCTGATTATGATATGAAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAACTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCTGTATATTACTGTGCGAAAATTGTTGGTTTTGCGT
GGGATTATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 147)
ABT1-33
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATACACCTTTAGGAGTTATAGTATGAATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCGATTAGTCCGCATGGTACGTATACAAAGTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACGGCGTATTCGTTCTT
CGGGTTTTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 148)
ABT1-34
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGCTAGGTATTCGATGAGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAATGATTGCTCCTGCGGGTAGGATGACATTGTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACGGGTTTGACTCGGA
GGACGCGGTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 149)
ABT1-35
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAAGTTGTATGAGATGGAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACGGATTGATAGTATGGGTGGGATTACAGAGTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACTGAGCTTCCTTATC
CTTCGGCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 150)
ABT1-36
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTTCGTCGGGTGGTAGTGCTACAGCGTACGCAGAC
TCCGTGATGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACGCCGCGGTATATTACTGTGCGAAAAGGTCTTATCTGTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 151) ABT1-37
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCATGATTATGATATGAAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATGGATTTCGTTTGATGGTGCTCGGACATTTTACGCAGAC
TCCGTGCAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGAGGAGACGGGTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 152) ABT1-38
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGCAGTATCGGATGGGGTGGGTCCGCCAGGCTCCAG
GGAGGGGTCTAGAGTGGGTCTCACGGATTGAGAGTGATGGTGCGGAGACATCTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCTGATTGGATTTTTG
ACTACAGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 153) ABT1-40
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCCTATTTATTTTATGGTTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACTGATTAGTAGTACGGGTATGTTTACAGATTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGGGGGTTAGGCGGG
GGCTTTATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 154)
ABT1-41
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCAGTATTATGTTATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATGGATTGCTACGTATGGTAGTCATACATGGTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACAGGGTAATCATCGTT
TGGGTCATTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 155)
ABT1-42
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGAAGTATGAGATGGCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTGGGCCTATGGGTTTTGGTACAAATTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATTTGGGAAGCATCCTC
CGGGTACTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 156)
ABT1-43
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGAAGTATGAGATGGCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTGGGCCTATGGGTTTTGGTACAAATTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATTTGGGAAGCATCCTC
AGGGTACTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 157)
ABT1-45
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGCCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCATAATTATAGGATGTTGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATCGGCGCCTATTCCGA
TGAGTATGTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 158)
ABT1-46
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGAATTATATGATGGATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACTGGGGCATCCGATGG
GTAATGGTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 159)
ABT1-47
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGGCTTATGATATGCACTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATGTATTAATAAGTCTGGTGCTTTGACATCTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTTGTTAGTCTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 160) ABT1-49
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAATTTGTATGCGATGAGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTTCGCCGATGGGTAAGGGTACATATTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTGGGGTATATTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 161) ABT1-50
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCT
CCTGTGCAGCCACCGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCC
AGGGAAGGGTCTAGAGTGGGTCTCATCTATTTCGTCGGGTGGTAGTGCTACAGCGTACGCA
GACTCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGC
AAATGAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGGTCTTATCT
GTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 162)
ABT1-51
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGTGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGTTATGGTGCTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 163) ABT1-52
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGAGTTATCGGATGAGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTTCTGCTATGGGTCGTCGTACACTTTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTTGGGGCGCGTTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 164) ABT1-75
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTACTCCTGGTGGTTCTGGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGGAAGTTTGCTTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 165) ABT1-76
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCCTCTTTATGATATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGGATTCGTGCGAATGGTGGTCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACGGCGGCTTGGGTTA
AGGGTAATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no:
166) ABT1-77
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGTGCGTTATTCTATGCTGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAACCTTCTAGGATGAGTC
TTAAGAAGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 167)
ABT1-78
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGCATTATACGATGACTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCCAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGTATTGAGCTGTCGG
GTGGGGCGTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 168)
ABT1-79
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAAGGCTTCTGGTCGTC
GTTCGATGTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 169)
ABT1-81
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGCTCGGGTGGCGCCGG
CTGTTCTGTTTGACTACCGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 170)
ABT1-84
GAGGTGCAGCTGTCGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAATGATTATAGGATGGTGTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTAGTAAGTCTGGTCGGACTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAATATACGATTCCGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 171) ABT1-85
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGACTCACCTTTAGTTTGTATGGGATGGCGTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTTCGTCTAGTGGTAGTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACAGCGTTGGTCTCATG
GTAGTGTGCTTGGTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id
no: 172) ABT1-86
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGATCGTTATATTATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTGGAGTGGGTCTCATCGATTACTCCTAGTGGTGCTGCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAGAGCCTGCTGATAGGT
ATAGTACGTGGACTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id
no: 173) ABT1-87
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGGCGTATAATATGAATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTTCTGCTTCGGGTCGTTATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTTCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAGGGTTTATTTCTGTGG
ATGAGTATGTGCCGTATCATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq
id no: 174) ABT1-88
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGTTGTATAAGATGTCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTAGTTCTAATGGTGAGGGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACTTTGTATCTGTGGG
GGTCGAGGCAGGCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id
no: 175) ABT1-89
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCGGTTGTATGGGATGAATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTTCTTCGGATGGTCAGAGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGGTGGTTGAGGGGTC
ATAATCTTATGCGGACGGAGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq
id no: 176) ABT1-90
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCT
CCTGTGCAGCCTCCGGATTCACCTTTCATGATTATTGGATGACGTGGGTCCGCCAGGCTCC
AGGGAAGGGTCTGGAGTGGGTCTCATCTATTTCTGCTTTGGGTGGTGCTACATACTACGCA
GACTCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGC
AAATGAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGGCTTCTTCG
GGAGCCGCAGTGGCGGCAGGGTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCG AGC
(SEQ ID NO: 177) ABT1-91
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGGATTATACTATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTGAGCGGSATGGTCGGCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAGGGCAGGCGAAGTTTT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 178) ABT1-92
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTACGTATACTATGACTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTTTTCCTGGGGGTTCGACGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATGTTGATTTTGTGA
AGTTGTCTTTTGACTACTGGGGTCAGAGAACCCTGGTCACCGTCTCGAGC (seq id no: 179)
ABT1-93
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAATGAGTATAATATGGTGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTTCTGGGGGTGGTGGGTTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAACCTCTGAGGGGGTTTG
TGTGTTCGCAGCATTGTTCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq
id no: 180) ABT1-94
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCATGTGTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATTTATTGATCGGATGGGTCGTCGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAACCTGGGTATCCGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 181) ABT1-96
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAATCAGTATAATATGTTTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTAGTGGTTCGGGTCGGTTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAGGGTGGTGGAGGCGTG
ATCCTCCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 182)
ABT1-97
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTGGGAGTATGATATGTATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACGGATTTCTGGTTCGGGTCGTTATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGGTCGCTTACTCGTC
CGAGTCAGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 183)
ABT1-98
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGATGGTTATATTATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTAGTCCGTTGGGTTCTGTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAAGGGGCCGTGGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 184) ABT1-99
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGTAAGTATTCTATGACGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTGATCCGTGGGGTCATTATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATCGGTTGATGCTACGC
TGTTGCGTAGGTATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id
no: 185) ABT1-101
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCGAAGTATTGGATGAGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTGGAGTGGGTCTCATCTATTTCTCCTGATGGTAAGACTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTGCGTTTGCGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 186) ABT1-102
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCGGAATTATGCGATGTCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTACTCCGCTGGGTTCTAGTACATACTATGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGGGCGTGGGCTTTTG
ACTACTGGGGTCAGGGAA-CCTGGTCACCGTCTCGAGC (seq id no: 187) ABT1-103
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACGGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGCGGTATTGGATGACTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATATATTTCGAGTAGTGGTACGGCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGGGCTCCGAATTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 188) ABT1-104
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAAGTCTTATCCTATGCGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTAGTCCTTTGGGTTCTACGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGCTTCTTATTCTTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 189) ABT1-105
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCTGGATTCACCTTTTCGCAGTATCGTATGTTTTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTAGTACTTCGGGTAATAAGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTTTCTAAGCCGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 190) ABT1-106
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGAATTATACTATGAAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTTCGCCTGTTGGTTCGGTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAACCGTCGCCTTTTTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id no: 191) ABT1-109
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGCTGGGTATAATATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTGCGTATGATGGTTATCGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATATGCAGATTACTA
GGCCGCGTCCGACGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id
no: 192) ABT1-110
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGTCGGTATAATATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTTCGTATGATGGTTTTCGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCATTCTACTCAGG
GTAAGGCGAATGTGTCTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (seq id
no: 193) ABT1-111
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTAATTATGGTATGCAGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATATATTAGTGGTTCTGGTTTGCTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAATCTGATGTGGGTTTGC
GGCTGCCTGCTTCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 194) ABT1-113
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCAGAATTATGATATGACGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTTCTAGTGGTGGTTCGTTGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACTTAGTTATCGTTGGT
CGTATACGTTTTCGGATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 195) ABT1-117
GAGGTGCAGCTGTTGGAGTCCGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGCGGGGTATGATATGCTGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAATGATTCTGGGGACTGGTAGTGGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACGG (SEQ ID NO: 196)
ABT1-118
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGGCGTATACTATGTATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACATATTTCGCCGGTGGGTTCTGATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATTTGTTTTTTTGGCGG
GGCGTTTGCCTACTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 197) ABT1-137
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGAGGTATTGGATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAATATTAATCTTACTGGTAGTGCGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAGGGGCTTTTCAGTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 198) ABT1-138
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAATGAGTATCTTATGTATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTGGAGTGGGTCTCATCTATTAGTCCGCTTGGTTATCATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACGTGCTCGGTGGTATT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 843) ABT1-139
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTTACCTTTTCTGATTATACGATGATTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTGGAGTGGGTCTCATCTATTTCGGCTCGTGGTTGGGGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTAGTCCTCAGCTTG
TTCTGCCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 199)
ABT1-140
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATGAGATGGATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTTCTCCTCATGGTCTGCTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACGTTTGTGGCCTCAGA
GGAAGTGGACTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO:
200) ABT1-147
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTACTTATAATATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTACGCATGAGGGTACGATGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATGGTCGTATTAGTC
AGAATCGGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 201)
ABT1 VK dAbs ABT1-3
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTTGGACGGAGTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTGCATCCCTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCGTATTTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 202) ABT1-9
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGTTCGTCGTTACTTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTTGGCATCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTCGGTCTACGCCTAATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 203) ABT1-9-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 500
TACCTGCCGTGCGAGCCATCGTATTCAGGTGAGCCTGAACTGGTATCAGCGTAAACCGGGCA
AAGCGCCGAAACTGCTGGTGTATCTGGCGAGCCGTCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCAGCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGGCGATTT
TGCGACCTATTATTGCCAGCAGTTTCGTAGCACCCCGAACACCTTTGGCCAGGGCACCAAAG
TGGAAATTCGTCGT (SEQ ID NO: 204) ABT1-10
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGTGCCGAATTTAAGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTGCATCCACGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGTTATGTTTATCCTGGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 205) ABT1-11
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCCGTATAATTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGTGCATCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATAGTTGGCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 206) ABT1-12
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTTGGACGGAGTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTGCATCCCTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCGTATTTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 207) ABT1-13
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCGTACTTTTTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTGGGCATCCCCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTAGTCGGGCTCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (seq id no: 208) ABT1-14
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTTGGACGGAGTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTGCATCCCTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCGTATTTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 209) ABT1-18
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCAGCGTTGGTTAGCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTTTGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTGCTGAGGTTGCCTAAGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 210) ABT1-24
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGGGCTAAGTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGGCTCGGCATCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 211) ABT1-25
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGCCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGGGCTAAGTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGGCTCGGCATCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 212) ABT1-26
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCCGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGTAATGTTTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTGCAGCCATGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGCGGACTTATCCTTGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 213) ABT1-27
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGCAGAAGTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTGGGCATCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGCATCTGAAGCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 214) ABT1-28
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGAATCGGTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTGCATCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGTATCATATTCCTAGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 215) ABT1-29
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCGTTCTCGTTTATCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGTTTTATTTGGCCTAGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 216) ABT1-30
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAATAGGGGTTTAAGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTGCATCCGTTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGTGAGTCGTCGTCCTCGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 217) ABT1-53
GACATCCAGATGACCCAGTCTCCATCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGGGCTAAGTTACGTTGGTACCAGCAGAAACCAGGGA
AGGCCCCTAAGCTCCTGATCTATAAGGCATCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGTGGGCTTCTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TAGAAATCAAACGG (SEQ ID NO: 218) ABT1-54
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGTAGGAAGTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATGCATCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGGTTCGTATGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 219) ABT1-56
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGACTCGTTTACAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATGCATCCGTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTACAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGAAGATGCGTCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 220) ABT1-57
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGCGTGAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGAAGTCGCATCCTCGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 221) ABT1-59
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGGCGGCGTTTAGCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTGGGCATCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGAATGTGTGGCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (seq id no: 222) ABT1-60
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGTCGTCGTTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCAGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATAGTCATCATCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 223) ABT1-61
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGGCATCGTTTAAGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCCTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGTGCGTGCGTATCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 224) ABT1-62
GACATCCAGATGACCCAGTCTCCATCCCCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCCGTTTTATTTAGGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGGCATCCATGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGATTCTTCAGGCTCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (seq id no: 225) ABT1-63
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGTGCGTTTTTAGGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGGCATCCCCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCGGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCAGCGTGTGGTTCCTGGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (seq id no: 226) ABT1-64
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTTCGAAGGTGTTAGGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCATTTTGCAAAGTGGGGTCCCATCACGTCTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGCGCTTGATTTTCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 227) ABT1-65
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGATTATGATTTTCCTATTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 228) ABT1-66
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTATGCGGAAGTTAAGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGTCATCGGAGGCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 229) ABT1-67
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCATCGTTCTTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATGCATCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTATAAGCGGCGTCCTTCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 230) ABT1-68
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCATAAGCAGTTAACGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGGCATCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTATCATCATAGGCCTTCAACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 231) ABT1-82
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGGAAGTCGTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTGCATCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGTTATGTTTGGGCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 232) ABT1-95
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATATTTCCAATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 233)
ABT1-95-3
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 234) ABT1-95-4
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTGTGCGCAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 235) ABT1-95-6
GACATCCAGATGACCCAGTCACCATCCTCTCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTCAGGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AGGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGTGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 236) ABT1-95-8
GACATCCAGATGACCCAGTCACCATCCTCTCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTCAGGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGAGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 237) ABT1-95-9
GACATCCAGATGACCCAGTCACCATCCTCTCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTCAGGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGAGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCATCCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 238) ABT1-95-
GACATCCAGATGACCCAGTCACCATCCTCTCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 10
CACTTGCCGGGCAAGTCAGCCGATTCAGGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGAGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCATCCTGAAGATCT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 239) ABT1-95-
GACATCCAGATGACCCAGTCACCATCCTCTCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 11
CACTTGCCGGGCAAGTCAGCCGATTCAGGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
TAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGAGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCATCCTGAAGATCT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 240) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 12
CACTTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGTTGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 241) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 13
CACTTGCCGGGCAAGTCAGCCGATTGTGCGCAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGAGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 242) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 14
CACTTGCCGGGCAAGTCAGCCGATTGTGAAGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAAAGAGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 243) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 15
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 244) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 21
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 245) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 22
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCACTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 246) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 23
CACTTGCCGGGCAAGTCAGCCGACTGTGCGGAACCTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTACCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 247) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 24
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTTGCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 248) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 25
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTGACCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 249) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 26
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTGGCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 250) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 27
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAAGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTGCGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 251) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 28
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATGGCTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 252) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 29
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCAGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 253) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 30
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGTCCTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 254) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 31
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCTGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 255) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 32
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATTCCTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 256) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 33
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCGCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 257) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 34
CACTTGCCGGGCAAGTGCGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 258) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 35
CACTTGCCGGGCAAGTGGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGAA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 259) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCGCCAT 36
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 260) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 37
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 261) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGCCTGCATCTGTAGGAGACCGTGTCACCAT 38
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCCAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 262) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 39
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCGGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 263) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 40
CACTTGCCGGGCAAGTCCCCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 264) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 41
CACTTGCCGGGCAAGTTACCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 265) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 42
CACTTGCCGGGCAAGTTTCCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCCACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 266) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 43
CACTTGCCGGGCAAGTCTCCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 267) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 44
TACTTGCCGGGCAAGTTCCCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 268) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 45
CACTTGCCGGGCAAGTCCCCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAAATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 269)
ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 46
CACTTGCCGGGCAAGTTCGCCGATTGTGCAGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 270) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 47
CACTTGCCGGGCAAGTATGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 271) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 48
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 272) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTATCTGCATCTGTAGGAGACCGTGTCACCAT 49
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 273) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 50
CACTTGCCGGGCAAGTCAGCCGATTCCGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 274) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 51
CACTTGCCGGGCAAGTCAGCCGATTATCCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 275) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 52
CACTTGCCGGGCAAGTCAGCCGATTGTGTCGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 276) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 53
CACTTGCCGGGCAAGTCAGCCGATTGTGGCCAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 277) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 54
CACTTGCCGGGCAAGTCAGCCGATTGTGACCAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 278) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 55
CACTTGCCGGGCAAGTCAGCCGATTGTGCACAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 279) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 56
CACTTGCCGGGCAAGTCAGCCGATTGTGCGCAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 280) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 57
CACTTGCCGGGCAAGTCAGCCGATTGTGGAGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 281) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 58
CACTTGCCGGGCACGTCAGCCGATTGTGTCGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 282) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 59
CACTTGCCGGGCAAGTCAGCCGATTGTGAAGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 283) ABT1-95-
GACATCCAGATGACCCAGTCTCCAGCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 60
CACTTGCCGGGCAAGTCAGCCGATTGTGGCCAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 284) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 61
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTACGCTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 285) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 62
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTACTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 286) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 63
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACTAGGGA
AAGCCCCTAAGCTCCTGATCTTCTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 287) ABT1-95-
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 64
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGGTCCACTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAGTCAAACGG (SEQ ID NO: 288) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 65
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGCTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 289) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 66
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGGTCTATTTCTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 290) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 67
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 291) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 68
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 292) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 69
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 293) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 70
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGGTCTATGGCTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 294) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 71
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAACTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 295) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 72
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCACTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 296) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 73
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCAGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 297) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 74
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAGACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGGGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 298) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 75
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGCCTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 299) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 76
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGGCTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 300) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 77
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTGAGCTCCTGATCTATTCTGGCTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 301) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 78
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGAGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 302) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 79
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTAGCTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 303) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 80
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 304) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTTCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 81
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTAGCTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 305) ABT1-95-
GACATCCCGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACTAT 82
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTACCTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 306) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 83
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCACCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 307) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 84
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTGGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 308) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 85
CACTTGCCGGGCAGGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGGGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 309) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 86
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTACGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 310) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 87
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCGGTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 311) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 88
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTACTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 312) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 89
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCTCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 313) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 90
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCAGTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 314) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 91
CACTTGCCGGGCGAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCAATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 315) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 92
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCACCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 316) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 93
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCATCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTGGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 317) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 94
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTCCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 318) ABT1-95-
GACATCCAGATGATCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 95
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCAGTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 319) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 96
CGCTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCAGGTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 320) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 97
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTTCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 321) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 98
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCGCGTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 322) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCTCCAT 99
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCACGTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 323) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 100
CACTTGCCGGGCAAGTCGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCTCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 324) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCCGTAGGAGACCGTGTCACCAT 101
CACTTGCCGGGCGAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGGTCTATTCTTCGTCCCTCTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 325) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 102
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 326) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 103
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGAAGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 327) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 104
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGTCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 328) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 105
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCGCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 329) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 106
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGAGGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTGCGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 330) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 107
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCACGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG(SEQ ID NO: 331) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 108
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGGAGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 332) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 109
CACTTGCCGGGCAAATCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGAACGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 333) ABT1-95-
GACATCCAGATGACCCAGTCACCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 110
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGGCGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 334) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 111
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGGACGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 335) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 112
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGAACGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 336) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 113
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 337) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 114
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGGTGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 338) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 115
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTTTGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 339) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 116
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTGTGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 340) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 117
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTATGTACCACTGTGTCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 341) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 118
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTTCGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 342) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 119
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTGAGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 343) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 120
CACTTGCCGGGCAAGTCAGCCGATTGCGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGTGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 344) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 121
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATGAGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 345) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 122
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCATTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 346) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 123
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATGCGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 347) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 124
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATATGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 348) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 125
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTCTGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 349) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 126
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTTCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 350) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 127
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCGGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTATTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 351) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 128
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGGCACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAGCGG (SEQ ID NO: 352) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 129
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCACCCCGAAGATCT
AGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 353) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 130
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCTGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 354) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 131
CACTTGCCGGGCAAGTCAGCCGACTGTGCGGAACCTAAGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 355) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 132
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
TAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCCGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 356) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 133
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 357) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 134
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTGCCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 358) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 135
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATTCCTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 359) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 136
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCACCCCGAAGATCT
AGCTACGTACCACTGTCAACAGGGGTATGGCTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 360) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 137
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCTGTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 361) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 139
CACTTGCCGGGCAAGTCAGCAGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 362) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 140
CACTTGCCGGGCAAGTCAGGGCATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 363) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 141
CACTTGCCGGGCAAGTCAGCCGATTTTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 364) ABT1-95-
GACATTCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 142
CACTTGCCGGGCAAGTCAGCCGATTACCCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 365) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 143
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCACGTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 366) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 144
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCTGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGAGCCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 367) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCATGTCACCAT 145
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGTCCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 368) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 146
CACTTGCCGGGCAAGTCAGAACATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 369) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 147
CACTTGCTGGGCAAGTCAGGGGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 370) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 148
CACTTGCCGGGCAAGTCAGACCATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 371) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 149
CACTTGCCGGGCAAGTCAGGCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 372) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 150
CACTTGCCGGGCAAGTCAGCACATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 373) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 151
CACTTGCCGGGCAGGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGACCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 374) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 152
CACTTGCCGGGCAGGTCAGCCGATTCCGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 375) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 153
CACTTGCCGGGCAAGTCAGCCGATTATCCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGGTTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 376) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 154
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTCCTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 377) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 155
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCACCTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTC
CGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 378) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 156
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGCCTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 379) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 157
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCACGTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGGCAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 380) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 158
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTTCTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 381) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCA 159
TCACTTGCCGGGCAAGTCAGCCGATTGCGCGGAACTTAAGGTGGTATCAGCAGAAACCAGG
GAAAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGT
TTCAGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAG
ATTTTGCTACGTACCACTGTCAACAGGTGTATCGTTGGCCTGTTACGTTCGGCCAAGGGAC
CAAGGTGGAAATCAAACGG (SEQ ID NO: 382) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCA 160
TCACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGG
GAAAGCCCCTAAGCTCCTGATCTATTCTTCATCCTATTTGGAGCCCGGGGTCCCATCACGT
TTCAGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAG
ATTTTGCTACGTACCACTGTCAACAGGGGTATGTGTGGCCTGTTACGTTCGGCCAAGGGAC
CAAGGTGGAAATCAAACGG (SEQ ID NO: 383) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 161
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 384) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 162
CACTTGCCGGGCAAGTAAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 385) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 163
CACTTGCCGGGCAAGTAGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAGATCAAACGG (SEQ ID NO: 386) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 164
CACTTGTCGGGCAAGTCAGCCGGGAGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 387) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 165
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACATGAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 388) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 166
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTAAGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 389) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 167
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTCGGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGGCAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 390) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 168
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTTTTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 391) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 169
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCTCCTAAGCTCCTGATCTATGCAGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 392) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 170
CACTTGCCGGGCAAGTAGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 393) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 171
CACTTGCCGGGCAAGTAGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 394) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 172
CACTTGCCGGGCAAGTAGGCCGATTGTGCGGAATTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCTCCTAAGCTCCTGATCTATGCAGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 395) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 173
CTCTTGCCGGGCAAGTAGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGCCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 396) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTATCTGCATCTGTAGGAGACCGTGTCACCAT 174
CACTTGCCGGGCAAGTAGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 397) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 175
CACTTGCCGGGCAAGTAGGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 398) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 176
CACTTGCCGGGCAAGTAGGCCAATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCTCCTAAGCTCCTGATCTATGCAGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 399) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 177
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCGTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 400) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 178
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGTGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGGCAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 401) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 179
CACTTGCCGGGCAAGTCAGCCGATTGTGCGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGGCAGGGGTATCGTTGGCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 402) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 181
CACTTGCCGGGCTAGTAAGCCGATCGTGCGTAACATGAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGGTGTCCTATTTGGAACCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTTCAGGGGTATCGTTGGCCTCCTACATTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 403) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 182
CACTTGCCGGGCTAGTAAGCCGGGTGTGCGTAACTTGAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGGTGTCCTATTTGGAACCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTTCAGGGGTATCGTTGGCCTCCTACATTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 404) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 183
CACTTGCCGGGCAAGTCGGACGATAGTGCGGAACTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCAAGGAGCTTTTTGGAGCCGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTACAGGGGTATCGTTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 405) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 184
CACTTGCCGGGCAAGTCGGACGCCTGTGCGGAACTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCAAGGAGCTATTTGGAGCCGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTACAGGGGTATCGTTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 406) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 500
CACTTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATATCTCCAATTTGCAAAGTGGGGTCCCATCGCGTTTC
AGTGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 407) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 501
CACTTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTATAGTATTTCCAATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 408) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGTATCTGTAGGAGACCGTGTCACCAT 502
CACCTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGTGTCCAGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCGTTGGCCTGTTACATTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 409) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 503
CACTTGCCGGGCAAGTCAGCCGATTCATGGGAATTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGTGTCCAGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCGAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 410) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT A1
CACTTGCCGAGCCAGTCGGCCGGGTGTGAGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATGTGAGCGATTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCGACAGGGGTATGTTTGGCCTGTTCCCTTCGACCAAGGGACCAAGG
TGGAAATCAAACGT (SEQ ID NO: 411) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT A2
CACTTGCCGGATACTTCAGCCCCCTGGCAGGAACTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCAAAGAGCTTTTTGGAGCCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCAACAGGGGTATCGTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGT (SEQ ID NO: 412) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT A3
CACTTGCCGGGCTAGTAAGCCGGGTGTGCGTAACATGAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGGTGTCCTATTTGGAACCCGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTTCAGGGGTATCGTTGGCCTCCTACATTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 413) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT A5
CACTTGCCGGGCAAGTCGGACGCCTGTGCGGAACTTACGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCAAGGAGCTTTTTGGAGCCGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTCTACAGGGGTATCGTTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 414) ABT1-95-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT A6
CACTTGCCGGGCTAGTAAGCCGGGTGTGCGTAACATGAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCTAAATCCTATTTGGAGCCGGGGGTCCCATCACGTTTC
AGTGGTAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCCGAAGATTT
TGCTACGTACCACTGTAAACAGGGGTATCGTTGGCCAGTTCAGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 415) ABT1-119
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGCGATTGGTGAGTGGTTAGGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGACGTCCGTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTTCTTTTTTTCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 416) ABT1-120
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTTCTGTGCTTTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCGTCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTGGCGGTTTAGTCCCCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 417) ABT1-121
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTTATACTGAGTTATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTTCGTCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCGTATCATCCTGTGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 418) ABT1-122
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTTGGACTGAGTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGGTCGTCCTCGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCTTATTTTCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 419) ABT1-122-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 18
CACTTGCCGGGCAAGTCAGCCGATTTGGACTGAGTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGGTCTATGGGTCGTCCTCGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCTTATTTTCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 420) ABT1-122-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 21
CACATGCCGGGCAAGTCAGCCGATTTGGACTGAGTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGGTCGTCCTCGTTGCAAAGTGGGGTCCCAACACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCTTATTTTCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 421) ABT1-122-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 511
CACTTGCCGGGCAAGTCAGCCGATTTGGACTGAGTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGGTCGTCCTCGTTGCAAAGAGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTAAACAGTTTGCTTATTTTCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 422) ABT1-122-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCGCCAT 750X
CACTTGCTCGGCATCTCAGCATATTTGGACTGAGATATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGCTCGGCCTCGCGGCAAAAAGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTAAACAGTTTGCTTATTTTCCTAATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGGG (SEQ ID NO: 423) ABT1-123
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTTGGACGGAGTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTGCATCCCTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCTTATCATCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 424) ABT1-124
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTGAGGGTAATTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGTGTCCCTTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCCGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGGTATCGTTATCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 425) ABT1-125
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTCAGAAGTTTTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGGGTTCCGGGTTGCAAAGTGGGGTCCCAACACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAGGATTT
TGCTACGTACTACTGTCAACAGGAGTATCGGATGCCTCTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 426) ABT1-126
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCTATTTATAAGTATTTAAGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTATGCTCCTGATCTATAGGGGGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTATGCGTTTTCGCCTTTTACGTTCGGCCAAGGGACCAAGG
TAGAAATCAAACGG (SEQ ID NO: 427) ABT1-127
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGTATTAGGTATTATTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTGGGATTCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTAGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTGCGTCGGCTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 428) ABT1-128
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCTATTAGGGAGTTTTTACATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGTCTTCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGAGTATAGGTATCCTCTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 429) ABT1-129
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGGGACAGTGTCGCCAT
CACTTGCCGGGCAAGTCAGCCGATTGGGGCGTTTTTATCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCGATTTCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCAGATGATTTCGCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 430) ABT1-130
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGAGATTGGTTTGTGGTTATCTTGGTACCAGCAGAAATCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGCATTCCAGTTCGCAAAGTGGGGTTCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTTATAAGGCGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 431) ABT1-131
GACATCCAGATGACCCAGTCTCCATCTTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGGATTAATGGGAATTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGTTTCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGTTATGATTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAACCAAACGG (SEQ ID NO: 432) ABT1-132
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTGCTACGGCGTTACTTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAGGCTCCTGATCTATGATTCGTCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGATGTATTGGGTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 433) ABT1-133
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACGATGACTGCTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 434) ABT1-134
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCATATTTATGATATGTTATCGTGGTACCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTACGTCCCTTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTTACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATCATCGGAGGCCTCATAGGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 435) ABT1-135
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTATATTGGTCGGAAGTTAAGGTGGTATCAGCAGAAACCAGGGA
AAGCCCTTAAGCTCCTGATCTATCGGACGTCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACGGGGCAGCATCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 436) ABT1-136
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGATATTGCGAATAATTTAGTGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGGCATTCCTTTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGATGCAGCAGGCTCCTTTTACGTCCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 437) ABT1-141
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 438) ABT1-141-1
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGTGGTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGCTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 439) ABT1-141-2
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATACAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGTGCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGCTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 440) ABT1-141-3
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAACCCCTAAGCTCCTGATCTATAGTCAGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 441) ABT1-141-6
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCCGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCTACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 442) ABT1-141-7
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTATCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 443) ABT1-141-8
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGTTCTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 444) ABT1-141-9
GACATCCAGGTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 445) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 10
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTCGCTTCGGGTGCCCTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 446) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 11
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTGGATGAGTCCTCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 447) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 12
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGATCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 448) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
13 CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATTTGAGGACTCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 449) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 14
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGCGCCGCGTCTGCTGATTTATAGCAGCAGCTATCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGAACCTGAGCGTGCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 450) ABT1-141-
GACATCCAGATGACCCAGTCTCCACCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 15
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 451) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCGCCAT 16
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACATCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 452) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 17
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 453) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 18
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGTCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 454) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 19
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGAGTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 455) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 20
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGGCTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 456) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 21
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCAGATGTCGTCTCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 457) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 22
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCAGATGGCTCCTCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 458) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 23
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCAGATGCAGCCTCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 459) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 24
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGACTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 460) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 25
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 461) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 27
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGGTCTTAGGCAGCCTATGACTTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 462) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 28
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGCTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 463) ABT1-141-
GACATCCAGGTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 29
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 464) ABT1-141-
GACATCCAGGTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 30
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 465) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 31
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 466) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTATCACCAT 32
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 467) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 33
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGTTCTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 468) ABT1-141-
GACATCCAGACGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 34
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 469) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 35
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTGCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 470) ABT1-141-
GACCTCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 42
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 471) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 43
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCACCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 472) ABT1-141-
GACATCCAGATTACCCAGTCTCCATCATCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 44
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGATTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 473) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 45
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGCGGATCTGGGACAGATTTCACTCTCACCATCAGTAGTCTGCAACTTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGCTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 474) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 46
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGTAATCAAACGG (SEQ ID NO: 475) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 47
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTCAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 476) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 48
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTTTTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTTCGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 477) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCA 49
TCACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCTGAAACCAGG
GAAAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTCGCAAAGTGGGGTCCCATCACGT
TTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAG
ATTTTGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGAC
CAAGGTGGAAATCAAACGG (SEQ ID NO: 478) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 50
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAACCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAATG
TGGAAATCAAACGG (SEQ ID NO: 479) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 51
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGTCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGTAATCAAACGG (SEQ ID NO: 480) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 52
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGCTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 481) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCA 53
TCACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCTGAAACCAGG
GAAAACCCCTAAGCTCCTGATCTTTTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGT
TTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAG
ATTTTGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGAC
CAAGGTGGAAATCAAACGG (SEQ ID NO: 482) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 54
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTTGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 483) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 70
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATATGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCAGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 484)
ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 75
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 485) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 76
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 486) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 77
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTTCTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTCAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 487) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 78
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATACGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTCAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 488) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 79
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTTCTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 489) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 80
CACTTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATACGGGACAGATTTCACTCTCACCATTAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 490) ABT1-141-
GACATCCAGATGACCCAGTATCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 500
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGCTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCGGCCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 491) ABT1-141-
GACATCTTGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 501
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTACTCCGCCTCCACGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTGGCTGTCTATTCCTCATACGTTCGGCCAAGGGACCAAGG
TGGGAATCAAACGG (SEQ ID NO: 492) ABT1-141-
GACATCCAGGTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 502
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCCAGGCTCCTGATCTATGCTGCGTCCGCTTTGCGAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCCACGTACTACTGTCAACAGACTTTGAGGAGCCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 493) ABT1-141-
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCATCTGTAGGAGGCCGTGTCACCAT 503
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCCAAGCTCCTGGTCTATGCGGCTTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGTCTGAGGGCGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 494) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 504
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGG
GAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATCTGTCGGTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 495) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 505
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATCTGTCGGTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 496) ABT1-141-
GACATCCAGATGACCCAGTCCCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 506
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAGCCAGGGA
GAGCCCCTAGGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGCGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATCTGTCGGTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 497) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCACCAGCGTGGGCGATCGTGTGACCAT 507
TACCTGCCGTGCGAGCCTGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGCGCCGAAACTGCTGATTTATGCGGGCAGCTTTCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTGCGCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGACCCTGTTTTATCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 498) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 508
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCC
GTGCGCCGAAACTGCTGATTTATAGCAGCAGCTATCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGGCGTATTATTGCCAGCAGCATCTGCGTGTGCCGAACACCTTTAGCCAGGGCACCAAAG
TGGAAGTGAAACGT (SEQ ID NO: 499) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 509
TACCTGCCGTGCGAACCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGCGCTGAAACTGCTGGTGTATAGCGCGAGCTATCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGCATCTGCGTGTGCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 500) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 510
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGCGCCGAAACTGCTGGTGTATGCGGCGAGCTGGCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGAGCCTGCGTGCGCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 501) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 511
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAACCCCGAAACTGCTGATTTATAGCAGCAGCTATCTGCAGAGCGGCATTCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGCATCTGCGTGTGCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 502) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 512
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGTGCCGAAACTGCTGATTTATAGCAGCAGCTATCTGCAGAGCGGCGTGCCGAGCCGTTTT
AACGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGACCCTGACCGTGCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 503) ABT1-141-
GATATTCAGGTGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 513
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGCGCCGCGTCTGCTGATTTATGCGGCGAGCGCGCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCGGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCGGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGACCCTGCGTAGCCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 504) ABT1-141-
GATATTCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCGAGCGTGGGCGATCGTGTGACCAT 514
TACCTGCCGTGCGAGCCAGTGGATTCAGAAACAGCTGGCGTGGTATCAGCAGAAACCGGGCA
AAGCGCCGCGTCTGCTGATTTATAGCAGCAGCTATCTGCAGAGCGGCGTGCCGAGCCGTTTT
AGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGCCGGAAGATTT
TGCGACCTATTATTGCCAGCAGAACCTGAGCGTGCCGTTTACCTTTGGCCAGGGCACCAAAG
TGGAAATTAAACGT (SEQ ID NO: 505) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 516
CAATTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AGGCCCTTAAGCTCCTGGTCTATAGTGCGTCCTATCTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGTCAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTGAGGGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 506) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCTT 519
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGTCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTTCTGTCAACAGACTTTGACTGTGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 507) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 520
CAATTGCCGGGCAAATCAGTGGATTCAGAAGCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AGGCCCTTAAGCTCCTGGTCTATAGTGCGTCCTATCTGCAAAGTGGGGTCCCATCAAGGTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 508) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 521
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 509) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 522
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCTGGTGGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 510) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 523
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 511) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 524
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCTGGTTGCAAAGGGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 512) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 525
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCTGGTTGCAAAGTGGGGCCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 513) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCGCCAT 526
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 514) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 527
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTCTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 515) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 528
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGATGCATAAGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 516) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 529
CACTTGCCGGGCAAACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCACGTTGCAAAGTGGGACCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGCTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 517) ABT1-141-
AACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 530
CACTTGCCGGGCATCGGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 518) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 531
CACTTGCCGGGCAGACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 519) ABT1-141-
GACATCCAGGTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 532
CACTTGCCGGGCAGACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 520)
ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 533
CACTTGCCGGACGAACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATAACCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 521) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 534
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGAACCATCCGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAGTCAAACGG (SEQ ID NO: 522) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 535
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGGCGCATGCGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 523) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 536
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGGCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 524) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 537
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGATGCATACCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 525) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 538
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGAACCATTCCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 526) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 539
CACTTGCCGGGCGAACGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGATGCATCCCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 527) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 541
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGATGCATCCGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 528) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 542
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGATGCATTCCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 529) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 543
CACTTGCCGGTCGAACGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGACCCATCGGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 530) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTTTCTGCATCTGTAGGAGACCGTGTCACCAT 544
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCTGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCATATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 531) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 545
CACTTGCCGGGCAAACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 532) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 546
CACTTGCCGGGCATCGGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 533) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 547
CACTTGCCGGGCAAACTCGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTCTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 534) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 548
CACTTGCCGGGCAGACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 535) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 549
CACTTGCCGGGCAAACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 536) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 550
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCTGGTTGCAAAGTGGGATCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 537) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 551
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 538) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 552
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 539) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 553
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCTGGTTGCAAAGTGGGGCCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 540) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 554
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 541) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 555
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCCACTCGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 542) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 556
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 543) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 557
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGATGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 544) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 558
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATATGTCGTCCGAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 545) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 559
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCTGGTCGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 546) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 561
CACTTGCCAGGCACACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 547) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 562
CACTTGCCGGGCAAACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAGCTCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 548) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 563
CACTTGCCGGGCAGACAGCTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGAGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTTCGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 549) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 564
CACTTGCCGGAGCAACGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 550) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 565
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGATGCATCTCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 551) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 567
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTACTCTTCGAACCATCCCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 552) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 568
CACTTGCCGGGCAAGTCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGGGCCATCCCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 553) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 569
CACTTGCCGGGCCAACCAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATCCCCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 554) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 572
CACTTGCCGGAGCAACGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGTCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 555) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 573
CACTTGCCGGACCAACGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGGCCCATAGGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGGTTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 556) ABT1-141-
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT 574
CACTTGCCGGATCAACGAGTGGATTCAGAAGCAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCTTCGACACATCCGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATCTTCGTGTTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 557) ABT1-142
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTAATGATGTGTTAGCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAGGCTCCTGATCTATTCGGCTTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTATATTAGTCTTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 558) ABT1-143
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGGATTGATCGTCATTTATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGACTTCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACACTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAGCAGCTTCATGCTAAGCCTTTTACGTTCGGTCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 559) ABT1-144
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACTAT
CACTTGCCGGGCAAGTCAGCGGATTAAGAAGTATTTAGCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAGGCTCCTGATCTATCGTTCTTCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTCGTTTAAGAGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 560) ABT1-145
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGATATTGATGATGGTTTAGCGTGGTATCAGCAGAAACCCGGGA
AAGCCCCTAAGCTCCTGATCTATCGTACGTCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACCGACTTGGCTGCCTCCTTGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 561) ABT1-146
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTAAGACGTTTTTACATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTGTTTCCCATTTGCAAAGTGGGGTTCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGAGTATCGTATTCCTCTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 562) ABT1-148
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGCTCTGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTATCGTCATCCTTCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 563) ABT1-149
GACATCCAGATGACCCAGTCTCCATCCTCTCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGTGGGCTGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTGGCGGTGGCCTTCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAGCGGGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGAT (SEQ ID NO: 564)
ABT1-150
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGTGTTTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTGGCGGTGGCCTTCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGGGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGAT (SEQ ID NO: 565)
ABT1-151
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGTCGTTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGGGCAGTGGTATCGGCATCCTGCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGGGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGAT (SEQ ID NO: 566)
ABT1-152
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGGAGGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCGGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGCATGCAGTGGTATAGGGCGCCTAGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGGGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGAT (SEQ ID NO: 567)
ABT1-153
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTAGTCCTATTGTGGCTAATTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTACTCAGGGGTATGTGTGGCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGGGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGAT (SEQ ID NO: 568)
ABT1-154
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTACTGAGATTGTGCGTAATTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTCGGGTGTTTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGTTCAGGGTTATTCTTGGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 569) ABT1-155
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTAGTCCTATTGTGGCTAATTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTACTCAGGGGTATGTGTGGCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 570) ABT1-156
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTTCGACTATTTTGGATAAGTTAGAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTCTGGTGCGTCCTGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCGGCAGGCGTTGTGGGATCCTCCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 571) ABT1-157
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGTGTGTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTCTCGGCCGCCTCGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 572) ABT1-158
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTAGGTCTATTGGGCGTGGGTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGGTGGTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGCTCAGGCGTCGCCGCTTCCGACGTTCGGCCAAGGGACCAAGGTGG
AAATCAAACGG (SEQ ID NO: 573) ABT1-159
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTAGTCCTATTGGTATGGATTTATTTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAGGCTCCTGATCGATGGTGTGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTTCTCAGCATTGGCCTGCGCCTCTGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 574) ABT1-160
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGTCGGGCAAGTACTGAGATTGTGCGTAATTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAATCTCCTGATCTGGGGGGGTTCCACTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGTTCAGGGTTATTCTTTGCCTGTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 575) ABT1-161
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCATGCTATTACTGGTAGTTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCAGTGGTGGGTCCGTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGCTCAGTGGTTGGGGGGGCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 576) ABT1-162
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGGCCGCGTCACCAT
CACTTGCCGGGCAAGTCATTATATTAGGAATCGGTTACATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGCGCGTGGGTCCGCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGTGCAGGATGGTTATCTGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 577) ABT1-163
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTGGGAATATTGTGCATAATTTACGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGCGCAGGGGTATGAGTGGCCTCTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACAG (SEQ ID NO: 578) ABT1-164
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCGTCCGATTGCTGGTAATTTACGGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTGGGGGGCGTCCTCGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGTGCAGGGGTGGCAGTGGCCTATTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 579) ABT1-165
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTTCTTCGATTAGGGCGGGTTTAGCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGCCTGAGTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGGGCAGGGGATTGATGGTCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 580) ABT1-166
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGTGGTCTGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGGTCAGTGGGCGCGTGCTCCTATGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 581) ABT1-167
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGTGGGCTGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTGGCGGTATCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 582) ABT1-168
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGGCTGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCTGCAGTGGTTTTCTGAGCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 583) ABT1-169
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGTGGGTTGTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTGGCGGTGGCCTTCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 584) ABT1-170
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGTCGGTCCGGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTAAGCAGTGGTATAGGTATCCTTCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 585) ABT1-171
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCGGGGGGCGGTCCGGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTATGCAGTGGTTTCGTCCGCCTAGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 586) ABT1-172
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCGTATTCGTAATGGTTTAGAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTGGGGGTCGTCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTGCGCAGACGGTTTGGGGTCCTGCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 587) ABT1-221
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGGGACCGTGTTACCAT
CACTTGCCGGGCAAGTCAGGATATTTGGCCTTATTTAATGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTAATCTATTATTCTTCCATGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTAGGAGGTGGCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 588) ABT2 VH dAbs ABT2-10
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTACAGCCTCCGGATTCACCTTTAATAGGTATAATATGGCGTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGAGATTGATCTTAAGGGTAGTCAGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTGAGTATTAGTGCGT
ATCATATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 589)
ABT2-11
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGATAATTATTCTATGACGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAATGTATTGATCGTTTGC
CTTGTCTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 590)
ABT2-12
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGGGTTATGATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTGATTTATGCTTATGGTGAGTCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAATGACTTCTGCTTCTT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 591) ABT2-13
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCGGGATTATGTTATGTATTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGGATTGATCCTATGGGTAGTTCGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCTGAGGGTAATTTTG
ACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 592) ABT2-14
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCCTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGCGGATTATGATATGTCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTTATGGTTTTGGTTTTGCGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACGTTGGCTGGGGGGT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 593) ABT2-15
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAATGGCTAATGCTAAGT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 594) ABT2-16
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATAAGGGGCATGGGC
AGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 595)
ABT2-17
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAATGAAGGGGAGTACTT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 596) ABT2-19
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACGTCTCCGGAGTCTT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 597) ABT2-20
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACTCCGCGGTATATTACTGTGCGAAACCTAGTATTGATGGTT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 598) ABT2-21
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGTTAGTAGTGATACTT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 599) ABT2-22
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCCTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGGCCTTTGTCTACGC
ATGATAGTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 600)
ABT2-23
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTGTATTACTGTGCGAAAGGGAATTCGAGTTTGG
ATCCGAATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 601)
ABT2-24
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAACTCAGCCTAGGTCTC
TGGATAATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 602)
ABT2-25
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTAAGTATAAGATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTAGTAGTTCTGGTTCTACTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GGACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCTCTGTTGTTGGATG
TGGAGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 603)
ABT2-26
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTAATTATAAGATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTAATACTTCTGGTTCTACTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GGACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCTCTGTTGTTGGATG
TGGAGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 604)
ABT2-27
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGCGCATTATTCGATGTGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCACGTATTGGGTCTCCGGGTAATGATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATTTGGGTTTAGGTCTG
CGGAGTCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 605)
ABT2-28
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGGGTTATGATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTGATTTATGCTTATGGTGAGTCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAACCTGTGCAGGGGTCGT
TGTTGGGGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 606)
ABT2-29
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGCGACGTTTGGGAATC
TGGAGGAGTCGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO:
607) ABT2-30
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATTCGGCTGGGCCTT
TTGGGCAGACGCTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 608) ABT2-31
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGGGTTATGATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTGATTTATGCTTATGGTGAGTCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAAGAGTAAGGGGCCTT
CTGGTCTTCGTACGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 609) ABT2-32
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGGGTTATGATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTGATTTATGCTTATGGTGAGTCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACTACGCCGAAGTCTA
ATCTGCGTGATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO:
610) ABT2-33
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATGCTATGGCTTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTTTGGCGGATGGTCATTCGACACACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCTTATGCGGCGGTTT
TTAATCGTGTGTATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 611) ABT2-35
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGTGAGTATGATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAATGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAATGGGTGGTGGGCTTG
TTTCTCTTCCTCAGTCTAAGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ
ID NO: 612) ABT2-36
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCAGTCGTATGATATGTGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATGTATTTATGCTTATGGTACGAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCTGCTGTTGGTACGC
ATCGTCAGCGGGCTGTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 613) ABT2-37
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTCGGTATTCGATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGGGATTGATTCTACGGGTGTTCATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGGTAGTTATGCTCTTT
GGGGTCGTGAGCCGACTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 614) ABT2-38
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGATCGGTATGCGATGGGGTGGGTCCGCCAGGTTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTAATGTTCCGGGTACGTTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAGGGCATTTTCCGCGTG
GTGGGGCGATTGTTCCTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 615) ABT2-39
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAATTGATCGGGATGCGA
GTCTGCCGACGGGGGAGGGGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ
ID NO: 616) ABT2-40
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAACCGTGTGCGATTTGTG
ATACGGATTCGTTGGGTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 617) ABT2-41
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACTACTGATATTAATA
AGAGGTGGCCTACTGGGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 618) ABT2-64
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGATGATTATGGGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTTGAGTGGGTCTCACTTATTTATCCGTTTGGTGGGGGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAGTTACGCAGACTGGTA
AGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 619)
ABT2-65
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 620) ABT2-65-1
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAAGAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 621) ABT2-65-2
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTACTTATAATATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAATAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 622) ABT2-65-6
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 623)
ABT2-65-7
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTACGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGACGA
GGATTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 624) ABT2-65-8
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTTTATTACTGTGCGCAAAATCTGGTTCGTATGG
ATAGTAGGCGTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 625) ABT2-65-9
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACATCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCGTTGGTATCCGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 626) ABT2-65-
AGGTGCAGCTGTCGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTCC 10
TGTGCAGCCTCTGGATTCACCTTTACGGAGTATAATATGGGTTGGGTCCGCCAGGCTCCAGG
GAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGACT
CCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAATG
AACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAATAATCTGGTTCGTACTCA
GTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 627) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 11
CTGTGCAGCCTCCGGATTCACCTTTAGGGCGTATCCTATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAATAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 628) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 12
CTGTGCAGCCTCCGGATTCACCTTTCGGGAGTATTGGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAACAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 629) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 13
CTGTGCAGCCTCCGGATTCACCTTTAGGGAGTATCAGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAATAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 630) ABT2-65-
GAGGTGCAGCTGTTGGAGTCCGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 14
CTGTGCAGCCCCCGGATTCACCTTTCCGTATTATCCGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 631) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 15
CTGTGCAGCCTCCGGATTCACCTTTGGTATTTATCAGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 632) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 16
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAAGTGGA
GGAGTGCGGAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 633) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 17
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 634 ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 18
CTGTGCAGCCTCCGGATTCACCTTTGGGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCAGTGGA
GGATTGGTGCGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 635) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 19
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCAAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTTTTTATG
ATGCGATGAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 636) ABT2-65-
GAGGCGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 20
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTAGTCAGTTGGGTTGGAATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 637) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 21
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTTCGCAGCTGGGTTGGCATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 638) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 22
CTGTGCAGCCTCCGGATTCACCTTTCTTGGGTATCCGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCTCCCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 639) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGAGTCCCTGCGTCTCTC 23
CTGTGCAGCCTCCGGATTCACCTTTAAGGTGTATCCTATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 640) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 24
CTGTGCAGCCTCCGGATTCACCTTTGCGGCTTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 641) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 25
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTTATC
GTAGTTATTATTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTTGAGC (SEQ ID
NO: 642) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 26
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCTGGTGC
GTTCGGGTTATTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 643) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 27
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTCTGT
GGAAGGGTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 644) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 28
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCAGTTTC
GTCGTTCTCAGTGGATGTTTGATTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 645) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 29
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTTTGGTTC
GGGTTGGGTGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 646) ABT2-65-
GAGGTGCGGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 30
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTCTGA
ATAGGAGGGATTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 647) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 31
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAAGATTA
TGAGGTTGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 648) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 32
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTGCGGTGT
CGCGGGGTCGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 649) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 33
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGTATA
TGAAGTCTCAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 650) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 34
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTA
GGTCTAAGAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 651) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 35
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGA
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 652) ABT2-65-
GAGGTGCGGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 36
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 653) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTAGTACAGCCTGGGGGGTCCCTGCGTCTCTC 37
CTGTGCAGCCTCCGGATTCACCTTTGCGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 654) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 38
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTACCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 655) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 39
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGGAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 656) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 40
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAAGACGT
TTAAGAGTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 657) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 41
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGGTGACTCGTACTC
AGTCTAAGCTGGGGCTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 658) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 42
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGGTGACTCGTACTC
AGTCTAAGTTGGGTCGGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 659) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 43
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGAGGTATCGTATTC
AGTCTAAGCTGGGGCTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 660) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 44
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGTTGGGTCGTATTC
AGTCTAAGTTGGGTCTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 661) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 45
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGCGTCGGCGTACTC
AGTCTAAGCTGGGGTATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 662) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 46
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTTTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGTTGACGCGTACTC
AGTCTAAGTTGGGGTATTTTGACTACTGGGGTCAAGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 663) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 47
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGAGGAAGCGTACTC
AGTCTAAGCTTGGTTATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 664) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 48
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATGGCAGATGCGTACTC
AGTCTAAGCTTGGTTATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 665) ABT2-65-
GGGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 49
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 666) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 50
CTGTGCAGCCTCCGGATTCACCTTTCTGACGTATCCGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCGGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 667) ABT2-65-
GAGGTGCAGGTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 52
CTGTGCAGCCTCCGGATTCACCTTTTATAGTTATCCTATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 668) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 53
CTGTGCAGCCTCCGGATTCACCTTTTATGTTTATCCTATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCTGAGGACACCGCGGTATATTACTGTGCGATAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 669) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 54
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTAGTCAGTTGGGTTGGATGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 670) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGCACAGCCTGGGGGGTCCCTGCGTCTCTC 55
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCGATTACTCAGCTTGGTTGGTATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTATGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 671) ABT2-65-
GAGGTGCAGTTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 56
CTGTGCAGCCTCCGGATTCACCTTTCCTGAGTATTATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 672) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 57
CTGTGCAGCCTCCGGATTCACCTTTAGGGAGTATGGTATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 673) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 58
CTGTGCAGCCTCCGGATTCACCTTTGTTATTTATCCGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 674) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 59
CTGTGCAGCCTCCGGATTCACCTTTGATACTTATTGGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 675) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 60
CTGTGCAGCCTCCGGATTCACCTTTAGTATGTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 676) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 61
CTGTGCAGCCTCCGGATTCACCTTTAGGGAGTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 677) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 62
CTGTGCAGCCTCCGGATTCACCTTTCGGGAGTATTTTATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 678) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 63
CTGTGCAGCCTCCGGATTCACCTTTGATGGTTATAATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 679) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 64
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTAGGACTCCTGGTAAGTTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 680) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 65
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTAGTGTGGCTGGTAAGCCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 681) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 66
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTGCGCTGCTGGGTGGGCCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 682) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 67
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTCAGGTTTTTGGTGGTAAGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 683) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 68
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAAGTATTCGGCCGACTGGTCCGTTTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 684) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 69
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTTATAGTCGTGGTACGCCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 685) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 70
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTCCGAGGGTGGGTATGCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC
(SEQ ID NO: 686) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 71
CTGTATAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTCGGAATGTTGGTAGGGCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 687) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 72
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTGATTGGTTTTCCTGGTAGGCTGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 688) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 73
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTCGGGCGGTTGGTGTTGGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGGACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 689) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 74
CTGTGCAGCCTCCGGATTCACCTTTCAGCATTATTATATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGTACTC
AGTCTAAGATGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 690) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCATGGGGGGTCCCTGCGTCTCTC 75
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAAGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 691) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 76
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCGG
GGAAGGGTCTAGAGTGGGTCACATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTACCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 692) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCATGGGGGGTCCCTGCGTCTCTC 77
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCTTGGTCACTGTCTCGAGC (SEQ ID
NO: 693) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCATGGGGGGTCCCTGCGTCTCTC 75
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAAGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 691) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 76
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCGG
GGAAGGGTCTAGAGTGGGTCACATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTACCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 692) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCATGGGGGGTCCCTGCGTCTCTC 77
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCTTGGTCACTGTCTCGAGC (SEQ ID
NO: 693) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 78
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGACGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAGCCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 694) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 79
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGGGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCATGGTCACAGTCTCGAGC (SEQ ID
NO: 695) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCGC 80
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGTCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 696) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 81
CTGTGTAGCCTCCGGATTCACCTTTGAGGAATATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACTATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 697) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACGGCCTGGGGGGTCCCTGCGTCTCTC 82
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATCCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 698) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 83
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCAGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 699) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 84
CTGTGTAGCCTCCGGATTCTCCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAC
GGAAGGGTCTAGAGTGGATCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 700) ABT2-65-
GAGGTGCAGCTGATGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 85
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAACCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCGCCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCAAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 701) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACGGCCTGGGGGGTCCCAGCGTCTCTC 86
CTGTGTAGCCTCCGGATTCACCTCTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 702) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 87
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGATCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 703) ABT2-65-
GAGGTGCAGTTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTA 88
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACAATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 704) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 89
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGAGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATATGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 705) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 90
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCAAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCACGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 706) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 113
CTGTGCAGCCTCCGGATTCACCTTTGTGCAGTATCAGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 707) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 114
CTGTGCAGCCTCCGGATTCACCTTTAGGGAGTATATGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 708) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 115
CTGTGCAGCCTCCGGATTCACCTTTATTGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 709) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 116
CTGTGCAGCCTCCGGATTCACCTTTGCGGGTTATTATATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 710) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACGGCCTGGGGGGTCCCTGCGTCTCTC 117
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGATCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 711) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 118
CTGTGCAGCCTCCGGATTCACCTTTGCGACTTATACGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 712) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 119
CTGTGCAGCCTCCGGATTCACCTTTATTGCGTATCCGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 713) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCCTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 120
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCTGCTATGGGTATGCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 714) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
121 CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTGCGGCTCATGGTAATAGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 715) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACGGCCTGGGGGGTCCCTGCGTCTCTC 122
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 716) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 123
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGAGATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACATCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGGTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 717) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 124
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTGTGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACATCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 718) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 125
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGATCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 719) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTATCTC 126
CTGTGTAACCTCCGGGTTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGAATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGGTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 720) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 127
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCACATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 721) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGAGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 128
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGATATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 722) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 129
CTGTGTAGCCTCCGGTTTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGGTTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCATGAAGGGCCGGTTCACCATCTCTCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCAGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 723) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 130
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGTTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCAGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 724) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTCGGGGGTCCCTGCGTCTCTC 131
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GCAAGGGTCTAGAGTGGGTCTCATCGATTTCGACTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCAGAGGACACCGCGGTATATCACTGTGCGCAAAATCTGGTTAGGTTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 725) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 132
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTCTTG
CTTCGAGGAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 726) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 133
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCATTGGA
AGAGGGGGGGTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 727) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 134
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAAGTTGT
CTAAGAGTCGTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 728) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 135
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAGGATTA
AGAAGATGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 729) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 136
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAAGCTGT
CGCGTACTGAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 730) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 137
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTTACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGGACGG
CTAAGCGTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 731) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 138
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGGGTTA
CGGCGCGGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 732) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 139
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGGTGC
GTGTGCGTCGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 733) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 140
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAAGATTT
CGAGGCGTCATTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 734) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 141
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAGGATTA
AGCAGACGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 735) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 142
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAAGATTT
CTGCGAGGGAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 736) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 143
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTCTGC
AGAGGGATAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 737) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 144
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTGCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTATTT
CGAAGAAGCAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 738) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 145
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAGGATTC
AGAAGGTTAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 739) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 146
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAGGACTA
TGAAGCGGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 740) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGTGGCTTGGTGCAGCCTGGGGGGTCCCTGCGTCTCTC 147
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGGCTTG
GGCGGATTCGTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 741) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 148
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGGCTTG
GGTCGAAGAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 742) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 149
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GAAAGGGTCTAGAGTGGGTCTCGTCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTGTTG
ATAGGAGGCGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 743) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 150
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCTGATTG
CGCGTGGTAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 744) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 151
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTTGGA
GGAAGCATGCGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 745) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 152
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGTTGGA
AGAGTTCGGCGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 746) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 154
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACACACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGATTC
GTAGGACGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 747) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 155
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTTTGGTTC
AGAGGAGGAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 748) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 156
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGGATTG
GGCGGAATAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 749) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 157
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACATGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGGCTTA
GTCTGCATCGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 750) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 158
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAAGTGGA
AGAGGGATTTTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 751) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 159
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAGGGTGA
GTAAGGCGAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 752) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 160
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTCCCCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 753) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 163
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTGGGCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 754) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 166
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTGGACGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACAGTCTCGAGC (SEQ ID
NO: 755) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 167
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGGTGGA
AGAAGAATGAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 756) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 168
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGTGGA
AGTTGGAGTCTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 757) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 169
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAATTGGC
GGCGTGGGTCTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 758) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 170
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGGGTTC
TTCTTAATAAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 759) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 171
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTCGGTTGA
ATAGGTCGCAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 760) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 172
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTCGGTGGA
AGAAGGGTTCTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 761) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 173
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGTTTC
GGGCGAAGGAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 762) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 174
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGTTTA
AGAAGACGCAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 763) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 175
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAGGATTG
CGGGGGATCGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 764) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 176
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAGTCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGGAAAATCTGGTTAAGGTGG
CGCGGGGTCGTTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 765) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 177
CTGTGCAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCTTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGTTTA
AGGTGCTGCAGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 766) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 500
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGGTTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGGGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTTGAGC (SEQ ID
NO: 767) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 501
CTGTGTAGCCTCCGGATTCGCCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAGGGGCCGGTTCACCATCTCCCGCGACGGTTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGCATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGGTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 768) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 502
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGGTTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATTTCCCGCGACGGTTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 769) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 503
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGCGGGTCTCATCGGTTTCGGCTATGGGTAATCGGACACACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGGGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 770) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 504
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGCGGGTCTCATCGGTTTCGGCTATGGGTAATCGGACACACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGGGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 771) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 505
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCGGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGGTTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGGTTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAAGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 772) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACGGCCTGGGGGGTCCCTGCGTCTCTC 506
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTGCTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCAAGC (SEQ ID
NO: 773) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCCC 507
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTGTCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATTTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 774) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 508
CTGTGTAGCCTCCGGATTCCCCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGGAGGGTCTAGAGTGGGCCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCAACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGGGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 775) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 509
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACGCCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCTAGC (SEQ ID
NO: 776) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 510
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCGAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 777)
ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCCCTC 511
CTGTGTAGCCTCCGGATTCACCTTTGGGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAGTCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACCACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 778) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 512
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCACCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 779) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 513
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTGTGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAGAATCTGGTTAGGCTGG
GGAGGTCTAGGCGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 780) ABT2-65-
GAGGTGCAGCTGTTGGAGTCTGGGGGGGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 514
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAAATGGGGTGGGCCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 781) ABT2-65-
GAGTGCAGCTGTTGGAGGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 515
CTGTGTAGCCTCCGGATTCACCTTTGAGGATTATCAGATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCGATTTCGGCTATGGGTAATCGGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACGATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAAATCTGGTTAGGCTGG
GGAGGTCTAGGTGGATGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 782) ABT2-66
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTCTCATTATACTATGGCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTGGTTCTTCTGGTAATAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAAATGCGGGGGCCGTATT
ATACTTTTGACTACCGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 783)
ABT2-69
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTACGGGTTATGATATGGCTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTGATTTATGCTTATGGTGAGTCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAGTGTATTCGAATCAGT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 784) ABT2-70
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCGGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAGGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACGTCTCCTAATGCGA
GTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 785)
ABT2-71
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAAGGGGTCGTCAGTCGT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 786) ABT2-72
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACTTCTTCTGATGGGT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 787) ABT2-73
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTTTATTACTGTGCGAAATTTTCTCCGCTTAGGG
CTCCTGTGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 788)
ABT2-74
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGATCTGTATGATATGCATTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACGATTTATGCTTATGGTTATGCGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAAAGCATGGTATGTATG
ATCATTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 789)
ABT2-75
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGGGCAGTATCCGATGGCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAACTATTTCTCCGAAGGGTGATCGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAGCGATGTATTCGTATC
AGTATAAGGAGAAGGGTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 790) ABT2-99
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTACAGCCTCCGGATTCACCTTTCGTGATTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAACTTCGTCTGTTAATT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 791) ABT2-101
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCTTATGATATGGGGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGTTATTTATGCGTGGGGTACTAGTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGTGAAACGGGGGGAGGGTAAGT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 792) ABT2-104
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGAGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGGGCGTATCGGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATGGATTTGGCCTAATGGTTCTCATGCATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGCAATTTCGTGATAATCAGC
GGTTTGACTACCGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 793)
ABT2-105
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTTTTGCTTATGAGATGTCGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCATCTATTTCTTATAATGGTACTCAGACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGATACCGCGGTATATTACTGTGCGAAAAGGCCGGATCTTTCGA
GTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 794)
ABT2-106
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTAGTGATTATGGGATGAGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTAGAGTGGGTCTCAGCTATTGCTACTGATGGCGTGTCTACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATTTTTATCCGAATT
TTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 795) ABT2-107
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTCCGCCTTATGCGATGGGTTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTGGAGTGGGTCTCATTGATTTATGAGAATGGTTTTAATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAAACTTTGGGTTCTAATT
TGTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID NO: 796)
ABT2-108
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC
CTGTGCAGCCTCCGGATTCACCTTTGCTGAGTATACTATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTCGAGTGGGTCTCACGTATTGGGCAGGATGGTAAGAATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATACGGGTCGGGTTG
GTGTTCATCATCTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 797) ABT2-108-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 533X
CTGTGCAGCCTCCGGATTCACCTTTGCTGACGAGGGGATGATGTGGGTCCGCCAGGCTCCAG
GGAAAGGTCTCGAGTGGGTCTCACGTATTGGGCAGGATGGTAAGAATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATACGGGTCGGATCC
TCGGGCATCATCTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 798) ABT2-108-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 534X
CTGTGCAGCCTCCGGATTCACCTTTGCTGACGAGGGGATGATGTGGGTCCGCCAGGCTCCAG
GGAAAGGTCTCGAGTGGGTCTCACGTATTACCTACAGCGGTAAGAATACATACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATACGGGTCGGATCT
TCTCCCATCATCTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 799) ABT2-108-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 537X
CTGTGCAGCCTCCGGATTCACCTTTGCTGACGAGGGGATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTCGAGTGGGTCTCACGTATTGGGCAGGATGGTAAGAATACATACTACCGGATG
GACGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATACGGGTCGGATCC
TCGGGCATCATCTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 800) ABT2-108-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 538X
CTGTGCAGCCTCCGGATTCACCTTTGCTGACGAGGGGATGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTCGAGTGGGTCTCACGTATTACCTACAGCGGTAAGAATACGTACTACGCAGAC
TCCGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTGTATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATACGGGTCGGATCC
TCGGGCATCATCTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 801) ABT2-108-
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGCGTCTCTC 620X
CTGTGCAGCCTCCGGATTCACCTTTGCTGAGGAGTCGTGGATGTGGGTCCGCCAGGCTCCAG
GGAAGGGTCTCGAGTGGGTCTCACGTATTGGGCAGGATGGTAAGAATACATACTACCGCGAG
GACGTGAAGGGCCGGTTCACCATCTCCCGCGACAATTCCAAGAACACGCTATATCTGCAAAT
GAACAGCCTGCGTGCCGAGGACACCGCGGTATATTACTGTGCGAAATATACGGGTCGGATCA
TGGGCCATCATCTTTTTGACTACTGGGGTCAGGGAACCCTGGTCACCGTCTCGAGC (SEQ ID
NO: 802) ABT2 VK dAbs: ABT2-6
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGGGCTACTGTTAACTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATATGGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTGGCATCGTCCTCCTAGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 803) ABT2-7
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGTCTGGGTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATATGGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTGGCATCGTCCTCCTAGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 804) ABT2-8
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGTCTATGTTAGCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATATGGCATCCAATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTATTGTTAAGCCTTCTACGTTCGGCCAAGGGACTAAGG
TGGAAATCAAACGG (SEQ ID NO: 805) ABT2-42
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTATATTGAGAAGTGGTTAACTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTACGCTCCTGATCTATCGTGGGTCCTTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTGAGTATTGGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 806) ABT2-43
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTGATACGTATTTAACTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAGGCTCCTGATCTATGGGGCTTCCACGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGTGCTTATTATCCTACGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 807) ABT2-44
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGATATTGGTGAGTGGTTAGAGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGGCGTCCACTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATGCGTATTATCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 808) ABT2-46
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGTATTATTGAGTGGTTAAGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTACTTCCGTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATGAGTTTTGGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 809) ABT2-53
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGAAGCATTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCATCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTAAGCGGGAGCCTAAGACGTTTGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 810) ABT2-54
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGTCTGGGTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATATGGCATCCCAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTGGCATCGTCCTCCTAGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 811) ABT2-55
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAAGACTAGGTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCTGGCATCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGTGTGGCGTCTTCCTAGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 812) ABT2-56
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACCTGCCGGGCAAGTCAGAGCATTGGGAGGCGGTTAAGTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGTGTTAGTGTTCCTCGGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 813) ABT2-57
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTGCGCGTCAGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGCATCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTATCGGATGCGGCCTAAGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 814) ABT2-58
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTTCTAAGAAGTTAGATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGGCATCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGCGGCGTAAGCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 815) ABT2-59
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCTAGTCAGAGCATTAATGTATTTTTATCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCATCCAATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGGCTCTTTATCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 816) ABT2-60
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGGGTTTTTTTATCTTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCATCCAATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGGCTCTTTATCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 817) ABT2-61
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGGGTTTTTTTATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAACGCATCCCATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGGCTCTTTATCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 818) ABT2-62
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTCAGACTTATTTAAGTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCATCCAATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGGCTCTTTATCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 819) ABT2-63
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGCATTAGGGTTTTTTTATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCATCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCGGGCTCTTTATCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 820) ABT2-67
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGAGTCACCAT
CACTTGCCGGGCAAGTCAGTATATTGGGGAGCATTTAGCTTGGTACCAGCAGAAACCGGGGA
AAGCCCCTAAGCTCCTGATCTATCATAATTCCGCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATGCGTTTCTTCCTAATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 821) ABT2-68
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTTGGTATCATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCTTGGTTCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
CGCTACGTACTACTGTCAACAGATTCATCATGATCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 822) ABT2-76
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTGATCGGTGGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGGGTTCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGTTGCGTTTTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 823) ABT2-77
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTGGGGCGCATTTAAAGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTACTTCCCGGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTTACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATCATACTCGTCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 824) ABT2-79
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCAGATTGGTGAGTGGTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGACGTCCCTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTAATTTTTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 825) ABT2-80
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGCTATTTATCATAATTTAAAGTGGTACCAGCAGAAACCAGGGA
AGGCCCCTAAGCTCCTGATCTATCATACGTCCTATTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACGTGGGCTTATCCTTATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 826) ABT2-81
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCGATTTATACTGAGTTATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGGTTCGTCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTGCGTATCATCCTGTGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 827) ABT2-83
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCCTATTCAGACTAAGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATAATTCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGTTTCGTAAGCATCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 828) ABT2-84
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGATTATTGATACGTGGTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCTTAAGCTCCTGATCTATCGTACTTCCTCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGTAATTATTGGCCTGCTACGTTTGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 829) ABT2-85
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGGATTCGTAAGCGTTTAAAGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATTCGTCGTCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATTTTTCTAAGCCTTCGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 830) ABT2-86
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAGTATTAAGAAGTATTTAGCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCTGGCTTCCACGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTAGCAGTCTGCAACCTGAAGATTT
CGCTACGTACTACTGTCAACAGACGCTGACGTATCCTTCTACGTTCGGCCAAGGGACTAAGG
TGGAAATCAAACGG (SEQ ID NO: 831) ABT2-87
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTAGTGTTTATTTATCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAATGCTTCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAGGGCTCATTATCCTACTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 832) ABT2-88
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCTATTGATGAGTGGTTAGAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGTGCGTCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGCGGCTTCGTGGCCTCGTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 833) ABT2-89
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTGATCGGTGGTTAGCGTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGGGTTCCATTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGTTGCGTTTTGGCCTCCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 834) ABT2-90
GACATCCAGATGACTCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGAATATTGATCAGTGGTTAGCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGACGTCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCCGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
CGCTACGTACTACTGTCAACAGGTTGCTGCGTTTCCTGCTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 835) ABT2-91
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCTTATTGGGAAGCATTTAAATTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATGAGTCCGCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATGCGTTTTCTCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 836) ABT2-93
GACATCCAGATGACCCAGGCTCCATCCTCCCTGTCTGCATCAGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTCGATTGGTCCGCATTTAAATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCATATTTCCACTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCCGCAACCTGAAGATTT
TGCTACGTACTACTGTCAGCAGCATGCGTTTTCTCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 837) ABT2-94
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGCGATTGGTATTCATTTAGATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATGATACTTCCTCTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGCATGCGCTTTTGCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 838) ABT2-95
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGTTTATTGGTCATTATTTAGCTTGGTATCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCCGGCGTCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGACTTATCTGAATCCTACGACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 839) ABT2-96
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGGGTATTAAGAGGAAGTTAAAGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCAGGCTTCCCGTTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGAATGCGCAGCGTCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 840) ABT2-97
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTCACCAT
CACTTGCCGGGCAAGTCAGCAGATTGATCAGTGGTTATCGTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATCGGACTTCCTTGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAACCTGAAGATTT
TGCTACGTACTACTGTCAACAGGCTGAGTATTGGCCTTTTACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 841) ABT2-98
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACCGTGTTACCAT
CACTTGCCGGGCAAGTCAGTATATTGGTAATAATTTACATTGGTACCAGCAGAAACCAGGGA
AAGCCCCTAAGCTCCTGATCTATAAGGGGTCCAAGTTGCAAAGTGGGGTCCCATCACGTTTC
AGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTT
CGCTACGTACTACTGTCAACAGAATTCTGCTCGTCCTCATACGTTCGGCCAAGGGACCAAGG
TGGAAATCAAACGG (SEQ ID NO: 842)
[0506] Forming part of the present disclosure is the appended
Sequence Listing.
EQUIVALENTS
[0507] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents to the specific embodiments of the invention described
herein. Such equivalents are intended to be encompassed by the
following claims. The contents of all references, patents and
published patent applications cited throughout this application are
incorporated herein by reference
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=US20130171146A1).
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=US20130171146A1).
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