U.S. patent application number 17/448730 was filed with the patent office on 2022-01-06 for anti-cea antibodies.
The applicant listed for this patent is Roche Glycart AG. Invention is credited to Thomas U. Hofer, Ralf Hosse, Ekkehard Moessner, Pablo Umana.
Application Number | 20220002441 17/448730 |
Document ID | / |
Family ID | 1000005853658 |
Filed Date | 2022-01-06 |
United States Patent
Application |
20220002441 |
Kind Code |
A1 |
Hofer; Thomas U. ; et
al. |
January 6, 2022 |
ANTI-CEA ANTIBODIES
Abstract
The present invention provides antigen binding molecules (ABMs)
which bind membrane-bound CEA, including ABMs with improved
therapeutic properties, and methods of using the same.
Inventors: |
Hofer; Thomas U.; (Zuerich,
CH) ; Hosse; Ralf; (Cham, CH) ; Moessner;
Ekkehard; (Kreuzlingen, CH) ; Umana; Pablo;
(Wollerau, CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Roche Glycart AG |
Schlieren |
|
CH |
|
|
Family ID: |
1000005853658 |
Appl. No.: |
17/448730 |
Filed: |
September 24, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16288587 |
Feb 28, 2019 |
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17448730 |
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15628854 |
Jun 21, 2017 |
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16288587 |
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14928819 |
Oct 30, 2015 |
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15628854 |
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14136299 |
Dec 20, 2013 |
9206260 |
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14928819 |
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13408171 |
Feb 29, 2012 |
8642742 |
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14136299 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 39/39566 20130101;
C07K 2317/92 20130101; C07K 16/3007 20130101; A61N 5/10 20130101;
C07K 2317/94 20130101; A61K 2039/505 20130101; C07K 2317/565
20130101; C07K 2317/24 20130101; C07K 2317/732 20130101; C07K
2317/41 20130101; C07K 16/42 20130101; A61K 45/06 20130101 |
International
Class: |
C07K 16/42 20060101
C07K016/42; C07K 16/30 20060101 C07K016/30; A61K 39/395 20060101
A61K039/395; A61K 45/06 20060101 A61K045/06; A61N 5/10 20060101
A61N005/10 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 2, 2011 |
EP |
11156665.9 |
Claims
1. An isolated antibody which binds membrane-bound CEA, wherein the
antibody comprises a heavy chain variable region comprising: a
heavy chain CDR1 selected from the group consisting of SEQ ID NO:1,
SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7,
SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:12, a heavy
chain CDR2 selected from the group consisting of SEQ ID NO:13, SEQ
ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18,
SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID
NO:23, and SEQ ID NO:24, a heavy chain CDR3 selected from the group
consisting of SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219 SEQ ID
NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:223, and SEQ ID
NO:224, and a light chain CDR1 selected from the group consisting
of SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID
NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, and
SEQ ID NO:45, and a light chain CDR2 selected from the group
consisting of SEQ ID NO:46, and SEQ ID NO:47, SEQ ID NO:48, SEQ ID
NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ
ID NO:54, and SEQ ID NO:55, and a light chain CDR3 of SEQ ID
NO:56.
2. The antibody of claim 1, wherein the heavy chain variable region
comprises: the heavy chain CDR1 of SEQ ID NO:1, the heavy chain
CDR2 of SEQ ID NO:13, a heavy chain CDR3 selected from the group
consisting SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219 SEQ ID
NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:223, and SEQ ID
NO:224; and wherein the light chain variable region comprises: the
light chain CDR1 of SEQ ID NO:39, the light chain CDR2 of SEQ ID
NO:49, and the light chain CDR3 of SEQ ID NO:56.
3. The antibody of claim 2, wherein the antibody comprises the
framework residues of CH1A1A (SEQ ID NO: 261) or CH1A1B (SEQ ID NO:
262).
4. The antibody of claim 1, wherein the antibody is stable at a
temperature that is at least 0.5, 1.0, 1.5, or 2.0 degree Celcius
higher than PR1A3 antibody or a humanized version of PR1A3
antibody.
5. The antibody of claim 4, wherein the PR1A3 antibody or humanized
version of PR1A3 antibody is a humanized version of PR1A3 antibody
which comprises the heavy chain variable region CH7A (SEQ ID
NO:101) and the light chain variable region 2F1 (SEQ ID NO:
209).
6. The antibody of claim 4, wherein the increase in stability is
measured using a dynamic light scattering assay.
7. An isolated antibody which binds membrane-bound CEA, wherein the
antibody comprises a heavy chain variable region comprising an
amino acid sequence that is at least 95% identical to a sequence
selected from the group consisting of SEQ ID NO: 233, SEQ ID NO:
234, SEQ ID NO: 235, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO:
242, SEQ ID NO: 243, and SEQ ID NO: 247 and a light chain variable
region comprising an amino acid sequence that is at least 95%
identical to the sequence of SEQ ID NO: 209, and wherein the
antibody has at least about 2-fold increase in stability as
compared to PR1A3 or a humanized version of PR1A3.
8. An isolated antibody which binds membrane-bound CEA, wherein the
antibody comprises a heavy chain variable region comprising an
amino acid sequence selected from the group consisting of SEQ ID
NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 239, SEQ ID NO:
241, SEQ ID NO: 242, SEQ ID NO: 243, and SEQ ID NO: 247 and a light
chain variable region comprising the amino acid sequence of SEQ ID
NO: 209.
9. The antibody of claim 1, wherein the antibody comprises an Fc
region that has been glycoengineered.
10. The antibody of claim 9, wherein at least about 20% to about
100% of the N-linked oligosaccharides in the Fc region are
nonfucosylated.
11. The antibody of claim 9, wherein at least about 20% to about
100% of the N-linked oligosaccharides in the Fc region are
bisected.
12. The antibody of claim 9, wherein at least about 20% to about
50% of the N-linked oligosaccharides in the Fc region are bisected,
nonfucosylated.
13. The antibody of claim 9, wherein the antibody has at least one
increased effector function.
14. The antibody of claim 13, wherein the at least one increased
effector function is selected from the group consisting of:
increased Fc receptor binding affinity, increased antibody-mediated
cellular cytotoxicity (ADCC), increased binding to NK cells,
increased binding to macrophages, increased binding to monocytes,
increased binding to polymorphonuclear cells, direct signaling
inducing apoptosis, increased dendritic cell maturation, and
increased T cell priming.
15. The antibody of claim 14, wherein the antibody has an increase
in ADCC of at least about 40% to about 100% as compared to the
non-glycoengineered parent antigen binding molecule.
16. An isolated polynucleotide encoding the antibody of claim
1.
17. A vector comprising the polynucleotide of claim 16.
18. A host cell comprising the vector of claim 17.
19. A composition comprising the antibody of claim 14 and a
pharmaceutically acceptable carrier.
20. A method of inducing cell lysis of a tumor cell, the method
comprising contacting the tumor cell with the antibody of any of
claim 1.
21. The method of claim 20, wherein the tumor cell is selected from
the group consisting of a colorectal cancer cell, NSCLC (non-small
cell lung cancer), gastric cancer cell, pancreatic cancer cell and
breast cancer cell.
22. The method of claim 20, wherein the cell lysis is induced by
antibody dependent cell cytotoxicity of the antibody.
23. A method of treating a subject having a cancer that abnormally
expresses CEA, the method comprising administering to the subject a
therapeutically effective amount of the antibody of claim 1.
24. The method of claim 24, wherein the antibody is administered in
combination with chemotherapy or radiation therapy.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. Ser. No.
16/288,587 filed Feb. 28, 2019 which is a continuation of U.S. Ser.
No. 15/628,854, filed on Jun. 21, 2017, which is a continuation of
U.S. Ser. No. 14/928,819, filed on Oct. 30, 2015, which is now
abandoned, which is a continuation of U.S. Ser. No. 14/136,299,
filed on Dec. 20, 2013, now U.S. Pat. No. 9,206,260, which is a
divisional of U.S. Ser. No. 13/408,171, filed on Feb. 29, 2012, now
U.S. Pat. No. 8,642,742, which claims the benefit of European
Patent Application No. 11156665.9, filed on Mar. 2, 2011, the
disclosures of which are incorporated herein by reference in their
entirety.
SEQUENCE LISTING
[0002] The present application contains a Sequence Listing which
has been submitted in ASCII format via EFS-Web and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Sep. 23, 2021, is named P27344_US_5_ST25_.TXT and is 165,910
bytes in size.
FIELD OF THE INVENTION
[0003] The present invention relates to antigen binding molecules
(ABMs). In particular embodiments, the present invention relates to
recombinant monoclonal antibodies, including chimeric, primatized
or humanized antibodies which bind to human carcinoembyronic
antigen (CEA).
BACKGROUND
Carcinoembryonic Antigen (CEA) and Anti-CEA Antibodies
[0004] Carcinoembryonic antigen (CEA, also known as CEACAM-5 or
CD66e) is a glycoprotein having a molecular weight of about 180
kDa. CEA is a member of the immunoglobulin superfamily and contains
seven domains that are linked to the cell membrane through a
glycosylphosphatidylinositol (GPI) anchor (Thompson J. A., J Clin
Lab Anal. 5:344-366, 1991) The seven domains include a single
N-terminal Ig variable domain and six domains (A1-B1-A2-B2-A3-B3)
homologous to the Ig constant domain (Hefta L J, et al., Cancer
Res. 52:5647-5655, 1992).
[0005] The human CEA family contains 29 genes, of which 18 are
expressed: 7 belonging to the CEA subgroup and 11 to the
pregnancy-specific glycoprotein subgroup. Several CEA subgroup
members are thought to possess cell adhesion properties. CEA is
thought to have a role in innate immunity (Hammarstrom S., Semin
Cancer Biol. 9(2):67-81 (1999)). Because of the existence of
proteins closely related to CEA, it can be challenging to raise
anti-CEA antibodies that are specific for CEA with minimal
cross-reactivity to the other closely related proteins.
[0006] CEA has long been identified as a tumor-associated antigen
(Gold and Freedman, J Exp Med., 121:439-462, 1965; Berinstein N.
L., J Clin Oncol., 20:2197-2207, 2002). Originally classified as a
protein expressed only in fetal tissue, CEA has now been identified
in several normal adult tissues. These tissues are primarily
epithelial in origin, including cells of the gastrointestinal,
respiratory, and urogential tracts, and cells of colon, cervix,
sweat glands, and prostate (Nap et al., Tumour Biol.,
9(2-3):145-53, 1988; Nap et al., Cancer Res., 52(8):2329-23339,
1992).
[0007] Tumors of epithelial origin, as well as their metastases,
contain CEA as a tumor associated antigen. While the presence of
CEA itself does not indicate transformation to a cancerous cell,
the distribution of CEA is indicative. In normal tissue, CEA is
generally expressed on the apical surface of the cell (Hammarstrom
S., Semin Cancer Biol. 9(2):67-81 (1999)), making it inaccessible
to antibody in the blood stream. In contrast to normal tissue, CEA
tends to be expressed over the entire surface of cancerous cells
(Hammarstrom S., Semin Cancer Biol. 9(2): 67-81 (1999)). This
change of expression pattern makes CEA accessible to antibody
binding in cancerous cells. In addition, CEA expression increases
in cancerous cells. Furthermore, increased CEA expression promotes
increased intercellular adhesions, which may lead to metastasis
(Marshall J., Semin Oncol., 30(a Suppl. 8):30-6, 2003).
[0008] CEA is readily cleaved from the cell surface and shed into
the blood stream from tumors, either directly or via the
lymphatics. Because of this property, the level of serum CEA has
been used as a clinical marker for diagnosis of cancers and
screening for recurrence of cancers, particularly colorectal cancer
(Goldenberg D M., The International Journal of Biological Markers,
7:183-188, 1992; Chau I., et al., J Clin Oncol., 22:1420-1429,
2004; Flamini et al., Clin Cancer Res; 12(23):6985-6988, 2006).
This property also presents one of the challenges for using CEA as
a target, since serum CEA binds most of the currently available
anti-CEA antibodies, hindering them from reaching their target on
the cell surface and limiting potential clinical effects.
[0009] Multiple monoclonal antibodies have been raised against CEA
for research purposes, as diagnostic tools, and for therapeutic
purposes (e.g., Nap et al., Cancer Res., 52(8):2329-23339, 1992;
Sheahan et al., Am. J. Clin. Path. 94:157-164, 1990; Sakurai et
al., J. Surg. Oncol., 42:39-46, 1989; Goldenberg D M., The
International Journal of Biological Markers, 7:183-188, 1992;
Ledermann J A, Br. J. Cancer, 58:654, 1988; Ledermann J A, Br. J.
Cancer, 68:69-73, 1993; Pedley R B, et al., Br. J. Cancer,
68:69-73, 1993; Boxer G M, et al., Br. J. Cancer, 65:825-831,
1992). Chester et al. have isolated a single chain anti-CEA
antibody from a phage display library to be used in
radioimmunodetection and radioimmunotherapy (U.S. Pat. No.
5,876,691), and the antibody was subsequently humanized (U.S. Pat.
No. 7,232,888). Anti-CEA antibodies have also been isolated from
human phage display libraries (U.S. Pat. No. 5,872,215).
[0010] The mouse monoclonal antibody PR1A3 was raised by fusion of
NS1 (P3/NS1/I-Ag-4-1) myeloma cells with spleen cells from mice
immunized with normal colorectal epithelium (Richman P. I. and
Bodmer W. F., Int. J. Cancer, 39:317-328, 1987). PR1A3 reacts
strongly to both well- and poorly-differentiated colorectal
carcinomas and has advantages over other colorectal
epithelium-reactive antibodies since its antigen appears to be
fixed to the tumor and does not appear in the lymphatics or normal
lymph nodes draining a tumor (Granowska M. et al., Eur. J. Nucl.
Med., 20:690-698, 1989). For example, PR1A3 reacted with 59/60
colorectal tumors (Richman P. I. and Bodmer W. F., Int. J. Cancer,
39:317-328, 1987), whereas the CEA reactive antibody B72.3 reacted
with only 75% of colorectal tumors (Mansi L., et al., Int J Rad
Appl Instrum B., 16(2): 127-35, 1989).
[0011] Epitope mapping of PR1A3 shows that the antibody targets the
B3 domain and the GPI anchor of the CEA molecule (Durbin H. et al.,
Proc. Natl. Scad. Sci. USA, 91:4313-4317, 1994). Consequently, the
PR1A3 antibody binds only to the membrane-bound CEA, and not the
soluble CEA form that can be found in the bloodstreams of cancer
patients. Because of this binding property, the PR1A3 antibody is
unlikely to be sequestered by the serum CEA; instead, it can target
CEA expressed on cancerous cells. The epitope bound by PR1A3 is a
conformational epitope, not a linear epitope, which is thought to
contribute to the loss of binding of PR1A3 to soluble CEA (Stewart
et al., Cancer Immunol Immunother, 47:299-06, 1999).
[0012] The PR1A3 antibody was previously humanized by grafting the
CDRs of the murine parent antibody to the heavy chain framework
regions 1-3 of the human antibody RF-TS3'CL (retaining the murine
framework 4 of PR1A3) and the light chain framework regions of the
REI antibody. (Stewart et al., Cancer Immunol Immunother,
47:299-06, 1999). This humanized version of PR1A3 retained
specificity and for surface-expressed CEA with an affinity similar
to that of the murine antibody (Stewart et al., Cancer Immunol
Immunother, 47:299-06, 1999; U.S. Pat. No. 5,965,710). A humanized
PR1A3 (hPR1A3) antibody was shown to induce targeted killing of
colorectal cancer cell lines. (Conaghhan P. J., et al., Br. J.
Cancer, 98(7):1217-1225). However, the affinity of hPR1A3 for CEA
is relatively low.
[0013] Radio-labeled anti-CEA antibodies have been used in clinical
trials in patients with colorectal cancer. For example, an
.sup.1231-labeled chimeric minibody T84.66 (cT84.66) was used in a
pilot clinical study in patients with colorectal cancer. The
radio-labeled minibody was able to target cancer cells. (Wong J. Y.
et al., Clin Cancer Res. 10(15):5014-21, (2004)). In another
example, .sup.(131)I-labetuzumab, a radio-labeled humanized
anti-CEA antibody, was tested in adjuvant radioimmunotherapy in
patients with liver metastases of colorectal cancer, and was found
to provide a promising survival advantage. (Liersch T., et al.,
Ann. Surg. Oncol. 14(9):2577-90, (2007)).
[0014] Antibody Glycosylation
[0015] The oligosaccharide component can significantly affect
properties relevant to the efficacy of a therapeutic glycoprotein,
including physical stability, resistance to protease attack,
interactions with the immune system, pharmacokinetics, and specific
biological activity. Such properties may depend not only on the
presence or absence, but also on the specific structures, of
oligosaccharides. Some generalizations between oligosaccharide
structure and glycoprotein function can be made. For example,
certain oligosaccharide structures mediate rapid clearance of the
glycoprotein from the bloodstream through interactions with
specific carbohydrate binding proteins, while others can be bound
by antibodies and trigger undesired immune reactions. (Jenkins et
al., Nature Biotechnol. 14:975-81, 1996).
[0016] Mammalian cells have been the preferred hosts for production
of therapeutic glycoproteins due to their capability to glycosylate
proteins in the most compatible form for human application.
(Cumming et al., Glycobiology 1:115-30, 1991; Jenkins et al.,
Nature Biotechnol. 14:975-981, 1996). Bacteria very rarely
glycosylate proteins and, like other types of common hosts, such as
yeasts, filamentous fungi, insect and plant cells, yield
glycosylation patterns associated with rapid clearance from the
blood stream, undesirable immune interactions, and in some specific
cases, reduced biological activity. Among mammalian cells, Chinese
hamster ovary (CHO) cells have been most commonly used during the
last two decades. In addition to giving suitable glycosylation
patterns, these cells allow consistent generation of genetically
stable, highly productive clonal cell lines. They can be cultured
to high densities in simple bioreactors using serum-free media, and
permit the development of safe and reproducible bioprocesses. Other
commonly used animal cells include baby hamster kidney (BHK) cells,
NS0- and SP2/0-mouse myeloma cells. More recently, production from
transgenic animals has also been tested (Jenkins et al., Nature
Biotechnol. 14:975-81, 1996).
[0017] All antibodies contain carbohydrate structures at conserved
positions in the heavy chain constant regions, with each isotype
possessing a distinct array of N-linked carbohydrate structures,
which variably affect protein assembly, secretion or functional
activity. (Wright A. and Morrison S. L., Trends Biotech. 15:26-32,
1997). The structure of the attached N-linked carbohydrate varies
considerably, depending on the degree of processing, and can
include high-mannose, multiply-branched as well as biantennary
complex oligosaccharides. (Wright, A., and Morrison, S. L., Trends
Biotech. 15:26-32, 1997). Typically, there is heterogeneous
processing of the core oligosaccharide structures attached at a
particular glycosylation site such that even monoclonal antibodies
exist as a population of multiple glycoforms. Likewise, it has been
shown that major differences in antibody glycosylation occur
between cell lines, and even minor differences are seen for a given
cell line grown under different culture conditions. (Lifely, M. R.
et al., Glycobiology 5(8):813-22, 1995).
[0018] One way to obtain large increases in potency, while
maintaining a simple production process and potentially avoiding
significant, undesirable side effects, is to enhance the natural,
cell-mediated effector functions of monoclonal antibodies by
engineering their oligosaccharide component as described in Umana,
P. et al., Nature Biotechnol. 17:176-180 (1999) and U.S. Pat. No.
6,602,684, the entire contents of which are hereby incorporated by
reference in their entirety. IgG1-type antibodies, the most
commonly used antibodies in cancer immunotherapy, are glycoproteins
that have a conserved N-linked glycosylation site at Asn297 in each
CH2 domain. The two complex biantennary oligosaccharides attached
to Asn297 are buried between the CH2 domains, forming extensive
contacts with the polypeptide backbone, and their presence is
essential for the antibody to mediate effector functions such as
antibody dependent cellular cytotoxicity (ADCC) (Lifely, M. R., et
al., Glycobiology 5:813-822 (1995); Jefferis, R., et al., Immunol
Rev. 163:59-76 (1998); Wright, A. and Morrison, S. L., Trends
Biotechnol. 15:26-32 (1997)).
[0019] Umana et al. showed previously that overexpression of
.beta.(1,4)-N-acetylglucosaminyltransferase III ("GnTIII"), a
glycosyltransferase catalyzing the formation of bisected
oligosaccharides, in Chinese hamster ovary (CHO) cells
significantly increases the in vitro ADCC activity of an
anti-neuroblastoma chimeric monoclonal antibody (chCE7) produced by
the engineered CHO cells. (See Umana, P. et al., Nature Biotechnol.
17:176-180 (1999); and International Publication No. WO 99/54342,
the entire contents of which are hereby incorporated by reference).
The antibody chCE7 belongs to a large class of unconjugated mAbs
which have high tumor affinity and specificity, but have too little
potency to be clinically useful when produced in standard
industrial cell lines lacking the GnTIII enzyme (Umana, P., et al.,
Nature Biotechnol. 17:176-180 (1999)). That study was the first to
show that large increases of ADCC activity could be obtained by
engineering the antibody-producing cells to express GnTIII, which
also led to an increase in the proportion of constant region
(Fc)-associated, bisected oligosaccharides, including bisected,
nonfucosylated oligosaccharides, above the levels found in
naturally-occurring antibodies.
[0020] There remains a need for enhanced therapeutic approaches
targeting CEA, in particular, membrane-bound CEA for the treatment
of cancers.
BRIEF SUMMARY OF THE INVENTION
[0021] One aspect of the invention provides a variant antigen
binding molecule (ABM), such as an antibody, which binds
membrane-bound human carcinoembryonic antigen (CEA). In one
embodiment, the ABM has an increase in stability as compared to its
parent molecule. In one embodiment, the ABM has an increase in
stability and maintains, or has an improved, binding affinity for
membrane-bound CEA as compared to its parent molecule. In one
embodiment, ABM is stable at a temperature that is at least 0.5,
1.0, 1.5, or 2.0 degree Celcius higher than its parent molecule. In
one embodiment, the increase in stability is measured using a
dynamic light scattering assay. In some embodiments, the parent
comparator molecule is PR1A3 antibody or humanized version of PR1A3
antibody. In one embodiment, the parent comparator molecule is a
humanized version of PR1A3 antibody which comprises the heavy chain
variable region CH7A (SEQ ID NO:101) and the light chain variable
region 2F1 (SEQ ID NO: 209). In one embodiment, the variant antigen
binding molecule is stable at 67 degrees Celsius or higher, as
measured, for example by a dynamic light scattering assay. In one
embodiment, the variant antigen binding molecule binds
membrane-bound CEA at a Kd of 100 nM or lower. In one embodiment,
the variant antigen binding molecule binds membrane-bound CEA at a
Kd of 10 nM or lower.
[0022] In one embodiment, the ABM comprises a heavy chain variable
region comprising a heavy chain CDR1 selected from the group
consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5,
SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,
and SEQ ID NO:12, a heavy chain CDR2 selected from the group
consisting of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID
NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ
ID NO:21, SEQ ID NO:22, SEQ ID NO:23, and SEQ ID NO:24, and a heavy
chain CDR3 selected from the group consisting of SEQ ID NO:217, SEQ
ID NO:218, SEQ ID NO:219 SEQ ID NO:220, SEQ ID NO:221, SEQ ID
NO:222, SEQ ID NO:223, and SEQ ID NO:224. In one embodiment, the
ABM comprises a light chain variable region comprising a light
chain CDR1 selected from the group consisting of SEQ ID NO:36, SEQ
ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41,
SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, and SEQ ID NO:45, and a
light chain CDR2 selected from the group consisting of SEQ ID
NO:46, and SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50,
SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, and SEQ ID
NO:55, and a light chain CDR3 of SEQ ID NO:56. In another
embodiment, the heavy chain variable region of the ABM comprises
the heavy chain CDR1 of SEQ ID NO:1, the heavy chain CDR2 of SEQ ID
NO:13, a heavy chain CDR3 selected from the group consisting SEQ ID
NO:217, SEQ ID NO:218, SEQ ID NO:219 SEQ ID NO:220, SEQ ID NO:221,
SEQ ID NO:222, SEQ ID NO:223, and SEQ ID NO:224; and the light
chain variable region of the ABM comprises the light chain CDR1 of
SEQ ID NO:39, the light chain CDR2 of SEQ ID NO:49, and the light
chain CDR3 of SEQ ID NO:56. In a further embodiment, the ABM
comprises the framework residues of CH1A1A (SEQ ID NO: 261) or
CH1A1B (SEQ ID NO: 262).
[0023] In one embodiment, the heavy chain variable region of the
ABM comprises an amino acid sequence that is at least 95% identical
to a sequence selected from the group consisting of SEQ ID NO: 233,
SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 239, SEQ ID NO: 241, SEQ
ID NO: 242, SEQ ID NO: 243, and SEQ ID NO: 247 and the light chain
variable region of the ABM comprises an amino acid sequence that is
at least 95% identical to the sequence of SEQ ID NO: 209. In one
embodiment, the heavy chain variable region of the ABM comprises an
amino acid sequence selected from the group consisting of SEQ ID
NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 239, SEQ ID NO:
241, SEQ ID NO: 242, SEQ ID NO: 243, and SEQ ID NO: 247 and the
light chain variable region of the ABM comprises the amino acid
sequence of SEQ ID NO: 209. In some embodiments, the ABM comprises
an Fc region, for example, a human IgG Fc region. In certain
embodiments, the ABM or is an antibody or fragment thereof, such as
a whole antibody, an scFv fragment, an Fv fragment, an F(ab')2
fragment, a minibody, a diabody, a triabody, or a tetrabody.
[0024] Another aspect of the invention provides an isolated
antibody which binds membrane-bound CEA, wherein the antibody
comprises a heavy chain variable region comprising a heavy chain
CDR1 selected from the group consisting of SEQ ID NO:1, SEQ ID
NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID
NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:12, a heavy chain
CDR2 selected from the group consisting of SEQ ID NO:13, SEQ ID
NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ
ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23,
and SEQ ID NO:24, and a heavy chain CDR3 selected from the group
consisting of SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219 SEQ ID
NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:223, and SEQ ID
NO:224.
[0025] In one embodiment, the antibody has an increase in stability
as compared to its parent molecule. In one embodiment, the antibody
is stable at a temperature that is at at least 0.5, 1.0, 1.5, or
2.0 degree Celcius higher its parent molecule. In one embodiment,
the increase in stability is measured using a dynamic light
scattering assay. In some embodiments, the parent comparator
molecule is PR1A3 antibody or humanized version of PR1A3 antibody.
In one embodiment, the parent comparator molecule is a humanized
version of PR1A3 antibody which comprises the heavy chain variable
region CH7A (SEQ ID NO:101) and the light chain variable region 2F1
(SEQ ID NO: 209). In one embodiment, the antibody is stable at 67
degrees Celsius or higher, as measured, for example by a dynamic
light scattering assay. In one embodiment, the antibody binds
membrane-bound CEA at a Kd of 100 nM or lower. In one embodiment
the antibody binds membrane-bound CEA at a Kd of 10 nM or
lower.
[0026] In one embodiment, the antibody also comprises a light chain
variable region comprising a light chain CDR1 selected from the
group consisting of SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ
ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43,
SEQ ID NO:44, and SEQ ID NO:45, and a light chain CDR2 selected
from the group consisting of SEQ ID NO:46, and SEQ ID NO:47, SEQ ID
NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ
ID NO:53, SEQ ID NO:54, and SEQ ID NO:55, and a light chain CDR3 of
SEQ ID NO:56. In one embodiment, the heavy chain variable region of
the antibody comprises the heavy chain CDR1 of SEQ ID NO:1, the
heavy chain CDR2 of SEQ ID NO:13, a heavy chain CDR3 selected from
the group consisting SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219
SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:222, SEQ ID NO:223, and SEQ
ID NO:224; and the light chain variable region of the antibody
comprises the light chain CDR1 of SEQ ID NO:39, the light chain
CDR2 of SEQ ID NO:49, and the light chain CDR3 of SEQ ID NO:56. In
a further embodiment, the antibody comprises the framework residues
of CH1A1A (SEQ ID NO: 261) or CH1A1B (SEQ ID NO: 262). In one
embodiment, the heavy chain variable region of the antibody
comprises an amino acid sequence that is at least 95% identical to
a sequence selected from the group consisting of SEQ ID NO: 233,
SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 239, SEQ ID NO: 241, SEQ
ID NO: 242, SEQ ID NO: 243, and SEQ ID NO: 247 and the light chain
variable region of the antibody comprises an amino acid sequence
that is at least 95% identical to the sequence of SEQ ID NO: 209.
In one embodiment, the heavy chain variable region of the antibody
comprises an amino acid sequence selected from the group consisting
of SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID NO: 239,
SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, and SEQ ID NO: 247
and the light chain variable region of the antibody comprises the
amino acid sequence of SEQ ID NO: 209.
[0027] In certain embodiments, the ABM or antibody of the above
embodiments binds the same epitope as, or is capable of competing
for binding with, the murine monoclonal antibody PR1A3.
[0028] In one embodiment, the ABM or the antibody comprises an Fc
region that has been glycoengineered. In one embodiment, at least
about 20% to about 100% of the N-linked oligosaccharides in the Fc
region of the glycoengineered antibody are nonfucosylated. In one
embodiment, at least about 20% to about 100% of the N-linked
oligosaccharides in the glycoengineered Fc region are bisected. In
one embodiment, wherein at least about 20% to about 50% of the
N-linked oligosaccharides in the glycoengineered Fc region are
bisected, nonfucosylated. In one embodiment, the glycoengineered
ABM or antibody has at least one increased effector function. The
increased effector function is, for example, increased Fc receptor
binding affinity, increased antibody-mediated cellular cytotoxicity
(ADCC), increased binding to NK cells, increased binding to
macrophages, increased binding to monocytes, increased binding to
polymorphonuclear cells, direct signaling inducing apoptosis,
increased dendritic cell maturation, and increased T cell priming.
In one embodiment, the glycoengineered ABM or antibody has an
increase in ADCC of at least about 40% to about 100% as compared to
the non-glycoengineered parent antigen binding molecule.
[0029] Another aspect of the invention provides for an isolated
polynucleotide encoding the ABM or antibody of any of above
described embodiments. Another aspect of the invention provides for
a vector comprising the polynucleotide encoding the ABM or antibody
of any of above described embodiments. Another aspect of the
invention provides for host cell comprising this vector.
[0030] Another aspect of the invention provides a composition
comprising the ABM or antibody of any of above described
embodiments and a pharmaceutically acceptable carrier.
[0031] Another aspect of the invention provides a method of
inducing cell lysis of a tumor cell comprising contacting the tumor
cell with the ABM or antibody of any of above described
embodiments. In some embodiments, the tumor cell is a colorectal
cancer cell, NSCLC (non-small cell lung cancer), gastric cancer
cell, pancreatic cancer cell or breast cancer cell. In one
embodiment, the cell lysis is induced by antibody dependent cell
cytotoxicity of the ABM or antibody.
[0032] Another aspect of the invention provides a method of
treating a subject having a cancer that abnormally expresses CEA,
the method comprising administering to the subject a
therapeutically effective amount of the ABM or antibody of any of
above described embodiments
[0033] Another aspect of the invention provides a method of
increasing survival time in a subject having a cancer that
abnormally expresses CEA, said method comprising administering to
said subject a therapeutically effective amount of the ABM or
antibody of any of above described embodiments. In one embodiment,
the cancer is colorectal cancer, non-small cell lung cancer
(NSCLC), gastric cancer, pancreatic cancer or breast cancer.
[0034] In certain embodiments of these methods, the ABM, antibody,
or composition is administered in combination with chemotherapy or
radiation therapy. In one embodiment, the subject is a human.
[0035] Another aspect of the invention provides for use of the ABM
or antibody of any of above described embodiments in the
manufacture of a medicament for treating a subject having a cancer
that abnormally expresses CEA. In one embodiment, the cancer is
selected from the group consisting of colorectal cancer, non-small
cell lung cancer (NSCLC), gastric cancer, pancreatic cancer and
breast cancer.
BRIEF DESCRIPTION OF THE FIGURES
[0036] FIG. 1 shows a schematic diagram of the CEA (CEACAM-5,
CD66e) antigen. The PR1A3 antibody binds specifically to the B3
domain of the antigen when it is bound to the cell membrane.
[0037] FIG. 2 shows enhanced ADCC activity of a glycoengineered
chimeric PR1A3 antibody in comparison to non-glycoengineered
chimeric PR1A3 antibody with human PBMCs as effectors.
[0038] FIG. 3 shows antigen binding activity of a humanized PR1A3
antibody comprising a heavy chain variable region construct, CH7A,
and a light chain variable region construct, CL1A, in comparison to
chimeric PR1A3 antibody.
[0039] FIG. 4 shows randomization sites for generating an antibody
library for affinity maturation of the humanized PR1A3 antibody
light chain Positions marked with an X were randomized.
[0040] FIG. 5 shows randomization sites for generating an antibody
library for affinity maturation of the humanized PR1A3 antibody
heavy chain. Positions marked with an X were randomized.
[0041] FIG. 6 shows binding activity of affinity matured anti-CEA
antibodies derived from a humanized PR1A3 antibody comprising a
heavy chain variable region construct CH7ArF9 and a light chain
variable region construct CL1ArH11.
[0042] FIG. 7 shows the results of an efficacy study in SCID/bg
mice that were intrasplenically administered LS174T human
colorectal carcinoma cells in order to have an orthotopic tumor
model. Antibody therapy was started at seven days later by
injection of the antibodies at a dose of 25 mg/kg body weight,
followed by two additional weekly injections. "CH7A" represents a
humanized antibody comprising the CDRs of PR1A3 as described
herein. "SM3E" refers to a previously generated anti-CEA antibody.
"GA201" represents a humanized anti-EGF antibody used as a positive
control. "PBS" refers to phosphate buffered saline, which was used
as a negative control. Survival was measured according to the
termination criteria defined by the Swiss regulatory authority.
[0043] FIG. 8 shows the results of an efficacy study in SCID/bg
mice that were injected intravenously with A549 lung carcinoma
cells, where the tumor engrafts in the lung of the animals Antibody
therapy was started at seven days later by injection of the
antibodies at a dose of 25 mg/kg body weight, followed by two
additional weekly injections. "CH7A," "SM3E," and "GA201" are as
set forth for FIG. 7, above. The designation "CH7ArF9 CL1A rH11"
represents a CH7A antibody variant with affinity matured heavy and
light chains. The designation "ge" indicates that the antibody has
been glycoengineered to have reduced numbers of fucosylated
oligosaccharides in the Fc region. "Vehicle" refers to the negative
control. A549 lung carcinoma cells are strongly positive for EGFR
expression and weakly positive for CEA expression.
[0044] FIG. 9 shows the results of an efficacy study in SCID/bg
mice that were intrasplenically administered MKN45 gastric
carcinoma cells, which generates tumor metastasis in the liver of
the animals. The designations, "CH7ArF9 CL1A rH11," "SM3E," "ge,"
and "PBS" are as set forth for FIGS. 7 and 8, above.
[0045] FIG. 10 shows kinetic analysis of affinity matured clones:
(a) shows a sensorgram of anti-CEA Fabs with an affinity matured
heavy chain CH7A H4E9 (SEQ ID NO: 199) together with unmatured
light chain CL1A (SEQ ID NO:105); an affinity matured light chain
CL1A pAC18 (SEQ ID NO:209) combined with unmatured heavy chain
CH7A; and a combination thereof, CH7A H4E9 and CL1A pAC18 (SEQ ID
NOs:199 and 209); (b) summary of kinetic analysis of affinity
matured clones.
[0046] FIG. 11 shows a schematic overview of the PCR strategy for
the CDR1 and CDR2 randomization of the humanized CH7A anti-CEA
antibody heavy chain.
[0047] FIG. 12 shows a schematic overview of the PCR strategy for
the CDR1 and CDR2 randomization of the humanized CL1A anti-CEA
antibody light chain.
[0048] FIG. 13 shows a schematic overview of the PCR strategy for
the CDR3 randomization of the humanized CH7A anti-CEA antibody
heavy chain.
[0049] FIG. 14 shows a schematic overview of the PCR strategy for
the CDR3 randomization of the humanized CL1A anti-CEA antibody
light chain.
[0050] FIG. 15 shows binding affinity of anti-CEA antibodies for
membrane-bound CEA on MKN45 target cells. Humanized anti-CEA
antibodies with either an affinity matured light chain (Panel A,
CH7A,CL1ArH7 or CH7A,CL1ArH11) or affinity matured heavy and light
chains (Panel B, CH7A rB9, CL1A rH11 G2(1)) that have been
converted to IgG show improved binding as compared to the control
antibody (CH7A,CL1A).
[0051] FIG. 16 shows the results of an assay testing
antibody-dependent cellular cytotoxicity (ADCC) by affinity matured
antibodies (CH7ArB9, CL1A rH11G2(1), CH7Arf9, CL1A rH11G2(1), and
CH7A, CL1A rH11 G2(1)) compared to control antibodies (CH7A, CL1A
G2(R2).
[0052] FIG. 17 shows the results of a cell binding assay for
anti-CEA antibody with heavy chain CH1A as compared to the
mouse-human chimeric antibody chPR1A3.
[0053] FIG. 18 shows the results of a binding assay for anti-CEA
antibodies with heavy chain CH1A1, CH1A2, CH1A3, or CH1A4 and light
chain 2F1.
[0054] FIG. 19 shows the results of a stability assay for anti-CEA
antibodies with heavy chain CH1A1, CH1A2, CH1A3, or CH1A4.
[0055] FIG. 20 shows the result of Surface Plasmon Resonance (SPR)
analysis for anti-CEA antibodies generated from CH1A1.
[0056] FIG. 21 shows the result of cell binding assays for anti-CEA
antibodies generated from CH1A1.
[0057] FIG. 22 shows the Surface Plasmon Resonance (SPR)
measurements of the affinity (as measured in the bivalent form) of
stability engineered anti-CEA antibodies as compared to the parent
5HFF12 heavy chain.
[0058] FIGS. 23 and 24 show the results of a stability assay for
affinity-matured antibody 5HFF12 as compared to its parental heavy
chain CH7A with the individual point mutations introduced that were
selected in 5HFF12.
[0059] FIG. 25 shows Surface Plasmon Resonance analysis of the
combined framework and CDR-H3 variants.
[0060] FIG. 26 shows the ADCC activity of the CHA1A-based framework
variants.
[0061] FIG. 27 shows the ADCC activity of the CHA1A-based framework
variants.
[0062] FIG. 28 shows the ADCC activity of the combined framework
and CDR-H3 variants.
[0063] FIG. 29 shows the ADCC activity of the combined framework
and CDR-H3 variants.
[0064] FIG. 30 shows the efficacy of glycoengineered anti-CEA
antibody CH1A1A (Y98A/D99Y).times.2F1 in a colorectal carcinoma
xenograft model in SCID mice transgenic for human CD16.
[0065] FIG. 31 shows the efficacy of glycoengineered anti-CEA
antibody CH1A1A (Y98A/D99Y).times.2F1 in an A549 lung carcinoma
xenograft model in SCID mice transgenic for human CD16.
[0066] FIG. 32 shows the amino acid sequences of CDRs for various
anti-CEA ABMs.
[0067] FIG. 33 shows the amino acid sequences of the light chain
constructs for various anti-CEA ABMs.
[0068] FIG. 34A-B shows amino acid sequences of affinity matured
heavy and light chain CDRs and associated binding affinities
[0069] FIG. 35 shows affinity constants of the various affinity
matured antibody sequences.
[0070] FIG. 36 shows the amino acid sequences of CDR-H3 of various
anti-CEA ABMs.
[0071] FIG. 37A-C shows the amino acid sequences of VH regions of
various anti-CEA ABMs.
[0072] FIG. 38 shows the amino acid sequence alignments of VH
regions of various stability matured anti-CEA antibodies.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0073] Terms are used herein as generally used in the art, unless
otherwise defined as follows.
[0074] As used herein, the term "antigen binding molecule" refers
in its broadest sense to a molecule that specifically binds an
antigenic determinant A non-limiting example of an antigen binding
molecule is an antibody or fragment thereof that retains
antigen-specific binding. More specifically, as used herein, an
antigen binding molecule that binds membrane-bound human
carcinoembryonic antigen (CEA) is a ABM that specifically binds to
CEA, more particularly to cell surface or membrane-bound CEA. By
"specifically binds" is meant that the binding is selective for the
antigen and can be discriminated from unwanted or nonspecific
interactions.
[0075] As used herein, the term "antibody" is intended to include
whole antibody molecules, including monoclonal, polyclonal and
multispecific (e.g., bispecific) antibodies, as well as antibody
fragments having an Fc region and retaining binding specificity,
and fusion proteins that include a region equivalent to the Fc
region of an immunoglobulin and that retain binding specificity.
Also encompassed are antibody fragments that retain binding
specificity including, but not limited to, VH fragments, VL
fragments, Fab fragments, F(ab')2 fragments, scFv fragments, Fv
fragments, minibodies, diabodies, triabodies, and tetrabodies (see,
e.g., Hudson and Souriau, Nature Med. 9: 129-134 (2003)).
[0076] As used herein, the term "antigen binding domain" refers to
the part of an antigen binding molecule that comprises the area
which specifically binds to and is complementary to part or all of
an antigen. Where an antigen is large, an antigen binding molecule
may only bind to a particular part of the antigen, which part is
termed an epitope. An antigen binding domain may be provided by,
for example, one or more antibody variable domains. Preferably, an
antigen binding domain comprises an antibody light chain variable
region (VL) and an antibody heavy chain variable region (VH).
[0077] As used herein, the term "affinity matured" in the context
of antigen binding molecules (e.g., antibodies) refers to an
antigen binding molecule that is derived from a reference antigen
binding molecule, e.g., by mutation, binds to the same antigen,
preferably binds to the same epitope, as the reference antibody;
and has a higher affinity for the antigen than that of the
reference antigen binding molecule. Affinity maturation generally
involves modification of one or more amino acid residues in one or
more CDRs of the antigen binding molecule. Typically, the affinity
matured antigen binding molecule binds to the same epitope as the
initial reference antigen binding molecule.
[0078] As used herein "binding affinity" is generally expressed in
terms of equilibrium association or dissociation constants (K.sub.a
or K.sub.d, respectively), which are in turn reciprocal ratios of
dissociation and association rate constants (k.sub.d and k.sub.a,
respectively). Thus, equivalent affinities may comprise different
rate constants, so long as the ratio of the rate constants remains
the same.
[0079] As used herein, the term "Fc region" refers to a C-terminal
region of an IgG heavy chain Although the boundaries of the Fc
region of an IgG heavy chain might vary slightly, the human IgG
heavy chain Fc region is usually defined to stretch from the amino
acid residue at position Cys226 to the carboxyl-terminus.
[0080] As used herein, the term "region equivalent to the Fc region
of an immunoglobulin" is intended to include naturally occurring
allelic variants of the Fc region of an immunoglobulin as well as
variants having alterations which produce substitutions, additions,
or deletions but which do not decrease substantially the ability of
the immunoglobulin to mediate effector functions (such as
antibody-dependent cellular cytotoxicity). For example, one or more
amino acids can be deleted from the N-terminus or C-terminus of the
Fc region of an immunoglobulin without substantial loss of
biological function. Such variants can be selected according to
general rules known in the art so as to have minimal effect on
activity. (See, e.g., Bowie, J. U. et al., Science 247:1306-10
(1990).
[0081] As used herein, the term "membrane-bound human CEA" refers
to human carcinoembryonic antigen (CEA) that is bound to a
membrane-portion of a cell or to the surface of a cell, in
particular, the surface of a tumor cell. The term "membrane-bound
human CEA" may, in certain circumstances, refer to CEA which is not
bound to the membrane of a cell, but which has been constructed so
as to preserve the epitope to which the PR1A3 antibody binds. The
term "soluble CEA" refers to human carcinoembryonic antigen that is
not bound to or is cleaved from a cell membrane or cell surface
(e.g., a tumor cell surface) and/or which, typically, does not
preserve the conformation epitope that is bound by the PR1A3
antibody. Soluble CEA can, for example, be found in the blood
stream or lymphatics of a subject with cancer.
[0082] As used herein, the term "no substantial cross-reactivity
against soluble" CEA means that a molecule (e.g., an antigen
binding molecule) does not recognize or specifically bind to
soluble CEA, particularly when compared to membrane-bound CEA. For
example, an antigen binding molecule may bind less than about 10%
to less than about 5% soluble CEA, or may bind soluble CEA at an
amount selected from the group consisting of less than about 10%,
9%, 8% 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, or 0.1%, preferably
less than about 2%, 1%, or 0.5% soluble CEA, and most preferably
less than about 0.2% or 0.1% soluble CEA.
[0083] As used herein, the terms "fusion" and "chimeric," when used
in reference to polypeptides such as ABMs, refer to polypeptides
comprising amino acid sequences derived from two or more
heterologous polypeptides, such as portions of antibodies from
different species. For chimeric ABMs, for example, the non-antigen
binding components may be derived from a wide variety of species,
including primates such as chimpanzees and humans. The constant
region of the chimeric ABM is generally substantially identical to
the constant region of a natural human antibody; the variable
region of the chimeric antibody generally comprises a sequence that
is derived from a recombinant anti-CEA antibody having the amino
acid sequence of the murine PR1A3 variable region. Humanized
antibodies are a particularly preferred form of fusion or chimeric
antibody.
[0084] As used herein, the term "humanized" is used to refer to an
antigen-binding molecule derived in part from a non-human
antigen-binding molecule, for example, a murine antibody, that
retains or substantially retains the antigen-binding properties of
the parent molecule but which is less immunogenic in humans. This
may be achieved by various methods (referred to herein as
"humanization") including, but not limited to (a) grafting the
entire non-human variable domains onto human constant regions to
generate chimeric antibodies, (b) grafting only the non-human
(e.g., donor antigen binding molecule) CDRs onto human (e.g.,
recipient antigen binding molecule) framework and constant regions
with or without retention of critical framework residues (e.g.,
those that are important for retaining good antigen binding
affinity or antibody functions), or (c) transplanting the entire
non-human variable domains, but "cloaking" them with a human-like
section by replacement of surface residues. Such methods are
disclosed in Jones et al., Morrison et al., Proc. Natl. Acad. Sci.,
81:6851-6855 (1984); Morrison and Oi, Adv. Immunol., 44:65-92
(1988); Verhoeyen et al., Science, 239:1534-1536 (1988); Padlan,
Molec. Immun, 28:489-498 (1991); Padlan, Molec. Immun,
31(3):169-217 (1994), all of which are incorporated by reference in
their entirety herein. There are generally 3 complementarity
determining regions, or CDRs, (CDR1, CDR2 and CDR3) in each of the
heavy and light chain variable domains of an antibody, which are
flanked by four framework subregions (i.e., FR1, FR2, FR3, and FR4)
in each of the heavy and light chain variable domains of an
antibody: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. A discussion of humanized
antibodies can be found, inter alia, in U.S. Pat. No. 6,632,927,
and in published U.S. Application No. 2003/0175269, both of which
are incorporated herein by reference in their entirety.
Humanization may also be achieved by transplanting truncated CDRs
that contain only the specificity-determining amino acid residues
for the given CDR onto a chosen framework. By
"specificity-determining residues" is meant those residues that are
directly involved in specific interaction with the antigen and/or
which are necessary for antigen-specific binding. In general, only
about one-fifth to one-third of the residues in a given CDR
participate in binding to antigen. The specificity-determining
residues in a particular CDR can be identified by, for example,
computation of interatomic contacts from three-dimensional modeling
and determination of the sequence variability at a given residue
position in accordance with the methods described in Padlan et al.,
FASEB J. 9(1):133-139 (1995), the contents of which are hereby
incorporated by reference in their entirety.
[0085] In some instances, framework region (FR) residues of the
human immunoglobulin are replaced by corresponding non-human
residues. Furthermore, humanized antigen binding molecules may
comprise residues which are not found in the recipient antibody or
in the donor antibody. These modifications are made to further
refine antigen binding molecule performance. In general, the
humanized antigen binding molecule will comprise substantially all
of at least one, and typically two, variable domains, in which at
least one, or substantially all, or all of the hypervariable
regions correspond to those of a non-human immunoglobulin and all
or substantially all of the FRs are those of a human immunoglobulin
sequence. The humanized antigen binding molecule optionally also
will comprise at least a portion of an immunoglobulin constant
region (Fc), typically that of a human immunoglobulin. See, e.g.,
Jones et al., Nature 321:522-525 (1986); Reichmann et al., Nature
332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596
(1992).
[0086] Similarly, as used herein, the term "primatized" is used to
refer to an antigen-binding molecule derived from a non-primate
antigen-binding molecule, for example, a murine antibody, that
retains or substantially retains the antigen-binding properties of
the parent molecule but which is less immunogenic in primates.
[0087] As used herein, the term "variant" (or analog)
polynucleotide or polypeptide refers to a polynucleotide or
polypeptide differing from a specifically recited polynucleotide or
polypeptide of the invention by insertions, deletions, and
substitutions, created using, e g., recombinant DNA techniques.
Specifically, recombinant variants encoding these same or similar
polypeptides may be synthesized or selected by making use of the
"redundancy" in the genetic code. Various codon substitutions, such
as the silent changes that produce various restriction sites, may
be introduced to optimize cloning into a plasmid or viral vector or
expression in a particular prokaryotic or eukaryotic system.
Mutations in the polynucleotide sequence may be reflected in the
polypeptide or domains of other peptides added to the polypeptide
to modify the properties of any part of the polypeptide, to change
characteristics such as ligand-binding affinities, interchain
affinities, or degradation/turnover rate.
[0088] As used herein, the term "variant anti-CEA antigen binding
molecule" refers to a molecule that differs in amino acid sequence
from a "parent" anti-CEA antigen binding molecule amino acid
sequence by virtue of addition, deletion and/or substitution of one
or more amino acid residue(s) in the parent antibody sequence. In a
specific embodiment, the variant comprises one or more amino acid
substitution(s) in one or more hypervariable region(s) or CDRs of
the heavy and/or light chain of the parent antigen binding
molecule. For example, the variant may comprise at least one, e.g.
from about one to about ten (i.e., about 1, 2, 3, 4, 5, 6, 7, 8, 9,
or 10), and preferably from about two to about five, substitutions
in one or more hypervariable regions or CDRs (i.e., 1, 2, 3, 4, 5,
or 6 hypervariable regions or CDRs) of the parent antigen binding
molecule. A variant anti-CEA antigen binding molecule may also
comprise one or more additions, deletions and/or substitutions in
one or more framework regions of either the heavy or the light
chain. Ordinarily, the variant will have an amino acid sequence
having at least about 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%,
or 100% amino acid sequence identity with the parent antigen
binding molecule heavy or light chain variable domain sequences,
typically at least about 80%, 90%, 95% or 99%. Identity with
respect to a sequence is defined herein as the percentage of amino
acid residues in the candidate sequence that are identical with the
parent antibody residues, after aligning the sequences and
introducing gaps, if necessary, to achieve the maximum percent
sequence identity. None of N-terminal, C-terminal, or internal
extensions, deletions, or insertions into the antibody sequence
shall be construed as affecting sequence identity or homology. The
variant antigen binding molecule retains the ability to bind
membrane-bound human CEA. In one embodiment, the anti-CEA ABM binds
the same epitope as that of the parent antigen binding molecule. In
one embodiment, the anti-CEA ABM competes for binding to
membrane-bound human CEA with the parent antigen binding molecule.
In one embodiment, the anti-CEA ABM binds to membrane-bound human
CEA and does not bind to soluble human CEA. The anti-CEA ABM has
properties which are superior to those of the parent antigen
binding molecule. For example, the variant may have a stronger
binding affinity, increased stability, and/or enhanced ability to
induce antibody-mediated cellular cytotoxicity in vitro and in
vivo. In one embodiment, the anti-CEA ABM has increased stability
and retains or has improved binding affinity for membrane-bound CEA
and retains or has an enhanced ability to induce antibody-mediated
cellular cytotoxicity in vitro and in vivo.
[0089] To analyze such properties, one should generally compare a
variant antigen binding molecule and the parent antigen binding
molecule in the same format; for example, an Fab form of the
variant antigen binding molecule to an Fab form of the parent
antigen binding molecule or a full length form of the variant
antigen binding molecule to a full length form of the parent
antigen binding molecule. In one embodiment, the variant antigen
binding molecule has at least about 2-fold, 3-fold, 4-fold, 5-fold,
6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 11-fold, 12-fold, 13-fold,
14-fold, 15-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold, or
20-fold enhancement in biological activity when compared to the
parent antigen binding molecule. In one embodiment, the variant
antigen binding molecule is a stability engineered variant that has
increased stability as compared to the parent antigen binding
molecule. Stability can be assayed by any method known in the art
and by methods described herein, specifically in Examples 3-6. In
specific embodiments, the variant antigen binding molecule has at
least about a 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold,
7-fold, 8-fold, 9-fold, 10-fold, 11-fold, 12-fold, 13-fold,
14-fold, 15-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold,
20-fold, 40-fold, 50-fold, 100-fold increase in stability as
compared to the parent antigen binding molecule.
[0090] In some embodiments, the variant antigen binding molecule
exhibits an increase in stability that is measured as a change in
stability parameter as compared to the parent antigen binding
molecule. In some embodiments, the variant antigen binding molecule
has at least about a 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold,
6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 11-fold, 12-fold, 13-fold,
14-fold, 15-fold, 15-fold, 16-fold, 17-fold, 18-fold, 19-fold,
20-fold, 40-fold, 50-fold, 100-fold change in a stability parameter
as compared to the parent antigen binding A stability parameter is
for example, temperature at which the variant antigen binding
molecule unfolds or denatures, the pressure at which the variant
antigen binding molecule unfolds or denature, or the time required
to denature or unfold the variant antigen binding molecule under
conditions designed to render the variant antigen binding molecule
unstable. In one embodiment, the increase in stability is
determined by a thermal denaturation assay, for example by
differential scanning calorimetry (DSC). In one embodiment, the
increase in stability is determined by a chemical denaturation
assay. In one embodiment, the increase in stability is determined
using a high pressure assay. In another embodiment, the stability
of the variant antigen binding molecule is determined using a
fluorescence polarization assay. In one embodiment, the stability
of the variant antigen binding molecule is determined using a
dynamic light scattering (DLS) assay. (See the Examples and, for
example, Nobbmann, U. et al., Biotech. Genetic Eng. Rev. 24:117-128
(2007). DLS monitors the integrity of a molecule, such as an
antibody, where, in general, an increase in light scattering
indicates protein unfolding or denaturation. The DLS of molecules
can be examined as a function of temperature or chemical
denaturants to compare relative stabilities. Those molecules that
remain in their native conformation (little or no increase in DLS
properties) are considered to be stable under the testing
conditions. In one embodiment, the variant antigen binding molecule
is stable at a temperature that is at least 0.25, 0.5, 1.0, 1.5,
2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 6.0, 7.0, 8.0, 9.0, or 10.0
degrees Celsius higher than the parent ABM, or other appropriate
reference molecule, when analyzed using a dynamic light scattering
assay. In one embodiment, the variant antigen binding molecule is
stable at 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80 degrees Celsius or higher. Thermal
stability can be measured, for example, using DLS, DSC, or
fluorescence polarization. In one embodiment, the thermal stability
of the variant antigen binding molecule is measured using DLS. In
one embodiment, the DLS assay is performed using 1 mg/ml of the ABM
or variant ABM in a buffer of 20 mM Histidine and 140 mM NaCl at pH
6.0. The DLS assay is conducted starting at 25.degree. C. with an
incremental temperature increase of 0.05.degree. C./min.
[0091] The term "parent" antigen binding molecule refers to an ABM
that is used as the starting point or basis for the preparation of
the variant. In a specific embodiment, the parent antigen binding
molecule has a human framework region and, if present, has human
antibody constant region(s). For example, the parent antibody may
be a humanized or human antibody.
[0092] Amino acid "substitutions" can result in replacing one amino
acid with another amino acid having similar structural and/or
chemical properties, e.g., conservative amino acid replacements.
"Conservative" amino acid substitutions may be made on the basis of
similarity in polarity, charge, solubility, hydrophobicity,
hydrophilicity, and/or the amphipathic nature of the residues
involved. For example, nonpolar (hydrophobic) amino acids include
alanine, leucine, isoleucine, valine, proline, phenylalanine,
tryptophan, and methionine; polar neutral amino acids include
glycine, serine, threonine, cysteine, tyrosine, asparagine, and
glutamine; positively charged (basic) amino acids include arginine,
lysine, and histidine; and negatively charged (acidic) amino acids
include aspartic acid and glutamic acid. "Insertions" or
"deletions" are generally in the range of about 1 to about 20 amino
acids, more specifically about 1 to about 10 amino acids, and even
more specifically, about 2 to about 5 amino acids. Non-conservative
substitutions will entail exchanging a member of one of these
classes for another class. For example, amino acid substitutions
can also result in replacing one amino acid with another amino acid
having different structural and/or chemical properties, for
example, replacing an amino acid from one group (e.g., polar) with
another amino acid from a different group (e.g., basic). The
variation allowed may be experimentally determined by
systematically making insertions, deletions, or substitutions of
amino acids in a polypeptide molecule using recombinant DNA
techniques and assaying the resulting recombinant variants for
activity.
[0093] As used herein, the term "single-chain Fv" or "scFv" refers
to an antibody fragment comprising a VH domain and a VL domain as a
single polypeptide chain Typically, the VH and VL domains are
joined by a linker sequence. See, e.g., Pluckthun, in: The
PHARMACOLOGY OF MONOCLONAL ANTIBODIES, vol. 113, Rosenburg and
Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).
[0094] As used herein, the term "minibody" refers to a bivalent,
homodimeric scFv derivative that contains a constant region,
typically the CH3 region of an immunoglobulin, preferably IgG, more
preferably IgG1, as the dimerisation region. Generally, the
constant region is connected to the scFv via a hinge region and/or
a linker region. Examples of minibody proteins can be found in Hu
et al. (1996), Cancer Res. 56: 3055-61.
[0095] As used herein, the term "diabody" refers to small antibody
fragments with two antigen-binding sites, which fragments comprise
a heavy chain variable domain (VH) connected to a light chain
variable domain (VL) in the same polypeptide chain (VH-VL). By
using a linker that is too short to allow pairing between the two
domains on the same chain, the domains are forced to pair with the
complementary domains of another chain and create two
antigen-binding sites. Diabodies are described more fully in, for
example, EP 404,097; WO 93/11161; and Hollinger et al., Proc. Natl.
Acad. Sci. USA 90:6444-6448 (1993). A triabody results from the
formation of a trivalent trimer of three scFvs, yielding three
binding sites, and a tetrabody is a tetravalent tetramer of four
scFvs, resulting in four binding sites.
[0096] In the case where there are two or more definitions of a
term which is used and/or accepted within the art, the definition
of the term as used herein is intended to include all such meanings
unless explicitly stated to the contrary. A specific example is the
use of the term "complementarity determining region" ("CDR") to
describe the non-contiguous antigen combining sites (also known as
antigen binding regions) found within the variable region of both
heavy and light chain polypeptides. CDRs are also referred to as
"hypervariable regions" and that term is used interchangeably
herein with the term "CDR" in reference to the portions of the
variable region that form the antigen binding regions. This
particular region has been described by Kabat et al., U.S. Dept. of
Health and Human Services, "Sequences of Proteins of Immunological
Interest" (1983) and by Chothia et al., J. Mol. Biol. 196:901-917
(1987), which are incorporated herein by reference, where the
definitions include overlapping or subsets of amino acid residues
when compared against each other. Nevertheless, application of
either definition to refer to a CDR of an antibody or variants
thereof is intended to be within the scope of the term as defined
and used herein. The appropriate amino acid residues which
encompass the CDRs as defined by each of the above cited references
are set forth below in Table I as a comparison. The exact residue
numbers which encompass a particular CDR will vary depending on the
sequence and size of the CDR. Those skilled in the art can
routinely determine which residues comprise a particular CDR given
the variable region amino acid sequence of the antibody.
TABLE-US-00001 TABLE 1 CDR Definitions.sup.1 CDR Kabat Chothia
AbM.sup.2 V.sub.H CDR1 31-35 26-32 26-35 V.sub.H CDR2 50-65 52-58
50-58 V.sub.H CDR3 95-102 95-102 95-102 V.sub.L CDR1 24-34 26-32
24-34 V.sub.L CDR2 50-56 50-52 50-56 V.sub.L CDR3 89-97 91-96 89-97
.sup.1Numbering of all CDR definitions in Table 1 is according to
the numbering conventions set forth by Kabat et al. (see below).
.sup.2"AbM" with a lowercase "b" as used in Table 1 refers to the
CDRs as defined by Oxford Molecular's "AbM" antibody modeling
software.
[0097] Kabat et al. also defined a numbering system for variable
domain sequences that is applicable to any antibody. One of
ordinary skill in the art can unambiguously assign this system of
"Kabat numbering" to any variable domain sequence, without reliance
on any experimental data beyond the sequence itself. As used
herein, "Kabat numbering" refers to the numbering system set forth
by Kabat et al., U.S. Dept. of Health and Human Services, "Sequence
of Proteins of Immunological Interest" (1983). Unless otherwise
specified, references to the numbering of specific amino acid
residue positions in an ABM are according to the Kabat numbering
system. The sequences of the sequence listing (i.e., SEQ ID NO:1 to
SEQ ID NO:216) are not numbered according to the Kabat numbering
system. However, one of ordinary skill in the art is familiar with
how to convert the sequences in the Sequence Listing to Kabat
numbering.
[0098] By a nucleic acid or polynucleotide having a nucleotide
sequence at least, for example, 95% "identical" to a reference
nucleotide sequence of the present invention, it is intended that
the nucleotide sequence of the polynucleotide is identical to the
reference sequence except that the polynucleotide sequence may
include up to five point mutations per each 100 nucleotides of the
reference nucleotide sequence. In other words, to obtain a
polynucleotide having a nucleotide sequence at least 95% identical
to a reference nucleotide sequence, up to 5% of the nucleotides in
the reference sequence may be deleted or substituted with another
nucleotide, or a number of nucleotides up to 5% of the total
nucleotides in the reference sequence may be inserted into the
reference sequence.
[0099] As a practical matter, whether any particular nucleic acid
molecule or polypeptide is at least 80%, 85%, 90%, 95%, 96%, 97%,
98% or 99% identical to a nucleotide sequence or polypeptide
sequence of the present invention can be determined conventionally
using known computer programs. One method for determining the best
overall match between a query sequence (a sequence of the present
invention) and a subject sequence, also referred to as a global
sequence alignment, can be determined using the FASTDB computer
program based on the algorithm of Brutlag et al., Comp. App.
Biosci. 6:237-245 (1990). In a sequence alignment the query and
subject sequences are both DNA sequences. An RNA sequence can be
compared by converting U's to T's. The result of said global
sequence alignment is in percent identity. Preferred parameters
used in a FASTDB alignment of DNA sequences to calculate percent
identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1,
Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1,
Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length
of the subject nucleotide sequence, whichever is shorter.
[0100] If the subject sequence is shorter than the query sequence
because of 5' or 3' deletions, not because of internal deletions, a
manual correction must be made to the results. This is because the
FASTDB program does not account for 5' and 3' truncations of the
subject sequence when calculating percent identity. For subject
sequences truncated at the 5' or 3' ends, relative to the query
sequence, the percent identity is corrected by calculating the
number of bases of the query sequence that are 5' and 3' of the
subject sequence, which are not matched/aligned, as a percent of
the total bases of the query sequence. Whether a nucleotide is
matched/aligned is determined by results of the FASTDB sequence
alignment. This percentage is then subtracted from the percent
identity, calculated by the above FASTDB program using the
specified parameters, to arrive at a final percent identity score.
This corrected score is what is used for the purposes of the
present invention. Only bases outside the 5' and 3' bases of the
subject sequence, as displayed by the FASTDB alignment, which are
not matched/aligned with the query sequence, are calculated for the
purposes of manually adjusting the percent identity score.
[0101] For example, a 90 base subject sequence is aligned to a 100
base query sequence to determine percent identity. The deletions
occur at the 5' end of the subject sequence and therefore, the
FASTDB alignment does not show a matched/alignment of the first 10
bases at 5' end. The 10 unpaired bases represent 10% of the
sequence (number of bases at the 5' and 3' ends not matched/total
number of bases in the query sequence) so 10% is subtracted from
the percent identity score calculated by the FASTDB program. If the
remaining 90 bases were perfectly matched the final percent
identity would be 90%. In another example, a 90 base subject
sequence is compared with a 100 base query sequence. This time the
deletions are internal deletions so that there are no bases on the
5' or 3' of the subject sequence which are not matched/aligned with
the query. In this case the percent identity calculated by FASTDB
is not manually corrected. Once again, only bases 5' and 3' of the
subject sequence which are not matched/aligned with the query
sequence are manually corrected for. No other manual corrections
are made for the purposes of the present invention.
[0102] By a polypeptide having an amino acid sequence at least, for
example, 95% "identical" to a query amino acid sequence of the
present invention, it is intended that the amino acid sequence of
the subject polypeptide is identical to the query sequence except
that the subject polypeptide sequence may include up to five amino
acid alterations per each 100 amino acids of the query amino acid
sequence. In other words, to obtain a polypeptide having an amino
acid sequence at least 95% identical to a query amino acid
sequence, up to 5% of the amino acid residues in the subject
sequence may be inserted, deleted, or substituted with another
amino acid. These alterations of the reference sequence may occur
at the amino or carboxy terminal positions of the reference amino
acid sequence or anywhere between those terminal positions,
interspersed either individually among residues in the reference
sequence or in one or more contiguous groups within the reference
sequence.
[0103] As a practical matter, whether any particular polypeptide is
at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a
reference polypeptide can be determined conventionally using known
computer programs. One method for determining the best overall
match between a query sequence (a sequence of the present
invention) and a subject sequence, also referred to as a global
sequence alignment, can be determined using the FASTDB computer
program based on the algorithm of Brutlag et al., Comp. App.
Biosci. 6:237-245 (1990). In a sequence alignment the query and
subject sequences are either both nucleotide sequences or both
amino acid sequences. The result of said global sequence alignment
is in percent identity. Preferred parameters used in a FASTDB amino
acid alignment are: Matrix=PAM 0, k-tuple=2, Mismatch Penalty=1,
Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1,
Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05,
Window Size=500 or the length of the subject amino acid sequence,
whichever is shorter.
[0104] If the subject sequence is shorter than the query sequence
due to N- or C-terminal deletions, not because of internal
deletions, a manual correction must be made to the results. This is
because the FASTDB program does not account for N- and C-terminal
truncations of the subject sequence when calculating global percent
identity. For subject sequences truncated at the N- and C-termini,
relative to the query sequence, the percent identity is corrected
by calculating the number of residues of the query sequence that
are N- and C-terminal of the subject sequence, which are not
matched/aligned with a corresponding subject residue, as a percent
of the total bases of the query sequence. Whether a residue is
matched/aligned is determined by results of the FASTDB sequence
alignment. This percentage is then subtracted from the percent
identity, calculated by the above FASTDB program using the
specified parameters, to arrive at a final percent identity score.
This final percent identity score is what is used for the purposes
of the present invention. Only residues to the N- and C-termini of
the subject sequence, which are not matched/aligned with the query
sequence, are considered for the purposes of manually adjusting the
percent identity score. That is, only query residue positions
outside the farthest N- and C-terminal residues of the subject
sequence.
[0105] For example, a 90 amino acid residue subject sequence is
aligned with a 100 residue query sequence to determine percent
identity. The deletion occurs at the N-terminus of the subject
sequence and therefore, the FASTDB alignment does not show a
matching/alignment of the first 10 residues at the N-terminus. The
10 unpaired residues represent 10% of the sequence (number of
residues at the N- and C-termini not matched/total number of
residues in the query sequence) so 10% is subtracted from the
percent identity score calculated by the FASTDB program. If the
remaining 90 residues were perfectly matched the final percent
identity would be 90%. In another example, a 90 residue subject
sequence is compared with a 100 residue query sequence. This time
the deletions are internal deletions so there are no residues at
the N- or C-termini of the subject sequence which are not
matched/aligned with the query. In this case the percent identity
calculated by FASTDB is not manually corrected. Once again, only
residue positions outside the N- and C-terminal ends of the subject
sequence, as displayed in the FASTDB alignment, which are not
matched/aligned with the query sequence are manually corrected for.
No other manual corrections are to be made for the purposes of the
present invention.
[0106] Percent identity of polynucleotides and/or polypeptides can
also be determined using the BLAST programs available through the
National Center for Biotechnology Information (NCBI), with the
default parameters indicated in the programs.
[0107] As used herein, a nucleic acid that "hybridizes under
stringent conditions" to a nucleic acid sequence of the invention,
refers to a polynucleotide that hybridizes under specified
conditions, e.g., in an overnight incubation at 42.degree. C. in a
solution comprising 50% formamide, 5.times.SSC (750 mM NaCl, 75 mM
sodium citrate), 50 mM sodium phosphate (pH 7.6),
5.times.Denhardt's solution, 10% dextran sulfate, and 20 .mu.g/ml
denatured, sheared salmon sperm DNA, followed by washing the
filters in 0.1.times.SSC at about 65.degree. C.
[0108] As used herein, the term "polypeptide having GnTIII
activity" refers to polypeptides that are able to catalyze the
addition of a N-acetylglucosamine (GlcNAc) residue in .beta.-1-4
linkage to the .beta.-linked mannoside of the trimannosyl core of
N-linked oligosaccharides. This includes fusion polypeptides
exhibiting enzymatic activity similar to, but not necessarily
identical to, an activity of
.beta.(1,4)-N-acetylglucosaminyltransferase III, also known as
.beta.-1,4-mannosyl-glycoprotein
4-beta-N-acetylglucosaminyl-transferase (EC 2.4.1.144), according
to the Nomenclature Committee of the International Union of
Biochemistry and Molecular Biology (NC-IUBMB), as measured in a
particular biological assay, with or without dose dependency. In
the case where dose dependency does exist, it need not be identical
to that of GnTIII, but rather substantially similar to the
dose-dependence in a given activity as compared to the GnTIII
(i.e., the candidate polypeptide will exhibit greater activity or
not more than about 25-fold less and, preferably, not more than
about tenfold less activity, and most preferably, not more than
about three-fold less activity relative to the GnTIII).
[0109] As used herein, the term "Golgi localization domain" refers
to the amino acid sequence of a Golgi resident polypeptide which is
responsible for anchoring the polypeptide to a location within the
Golgi complex. Generally, localization domains comprise amino
terminal "tails" of an enzyme.
[0110] As used herein, the term "effector function" refers to those
biological activities attributable to the Fc region (a native
sequence Fc region or amino acid sequence variant Fc region) of an
antibody. Examples of antibody effector functions include, but are
not limited to, Fc receptor binding affinity, antibody-dependent
cellular cytotoxicity (ADCC), antibody-dependent cellular
phagocytosis (ADCP), cytokine secretion, immune-complex-mediated
antigen uptake by antigen-presenting cells, down-regulation of cell
surface receptors, etc.
[0111] As used herein, the terms "engineer, engineered,
engineering" particularly with the prefix "glyco-," as well as the
term "glycosylation engineering" are considered to include any
manipulation of the glycosylation pattern of a naturally occurring
or recombinant polypeptide or fragment thereof. Glycosylation
engineering includes metabolic engineering of the glycosylation
machinery of a cell, including genetic manipulations of the
oligosaccharide synthesis pathways to achieve altered glycosylation
of glycoproteins expressed in cells. Furthermore, glycosylation
engineering includes the effects of mutations and cell environment
on glycosylation. In one embodiment, the glycosylation engineering
is an alteration in glycosyltransferase activity. In a particular
embodiment, the engineering results in altered
glucosaminyltransferase activity and/or fucosyltransferase
activity.
[0112] As used herein, the term "host cell" covers any kind of
cellular system which can be engineered to generate the
polypeptides and antigen-binding molecules of the present
invention. In one embodiment, the host cell is engineered to allow
the production of an antigen binding molecule with modified
glycoforms. In a preferred embodiment, the antigen binding
molecule, or variant antigen binding molecule, is an antibody,
antibody fragment, or fusion protein. In certain embodiments, the
host cells have been further manipulated to express increased
levels of one or more polypeptides having GnTIII activity. Host
cells include cultured cells, e.g., mammalian cultured cells, such
as CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO myeloma cells,
P3X63 mouse myeloma cells, PER cells, PER.C6 cells or hybridoma
cells, yeast cells, insect cells, and plant cells, to name only a
few, but also cells comprised within a transgenic animal,
transgenic plant or cultured plant or animal tissue.
[0113] As used herein, the term "Fc-mediated cellular cytotoxicity"
includes antibody-dependent cellular cytotoxicity (ADCC) and
cellular cytotoxicity mediated by a soluble Fc-fusion protein
containing a human Fc-region. It is an immune mechanism leading to
the lysis of "targeted cells" by "human immune effector cells."
[0114] As used herein, the term "human immune effector cells"
refers to a population of leukocytes that display Fc receptors on
their surfaces, through which they bind to the Fc-region of antigen
binding molecules or of Fc-fusion proteins and perform effector
functions. Such a population may include, but is not limited to,
peripheral blood mononuclear cells (PBMC) and/or natural killer
(NK) cells.
[0115] As used herein, the term "targeted cells" refers to cells to
which antigen binding molecules comprising an Fc region (e.g.,
antibodies or fragments thereof comprising an Fc region) or
Fc-fusion proteins specifically bind. The antigen binding molecules
or Fc fusion-proteins bind to target cells via the protein part
that is N-terminal to the Fc region.
[0116] As used herein, the term "increased Fc-mediated cellular
cytotoxicity" is defined as either an increase in the number of
"targeted cells" that are lysed in a given time, at a given
concentration of antigen binding molecule or of Fc-fusion protein
in the medium surrounding the target cells, by the mechanism of
Fc-mediated cellular cytotoxicity defined above, and/or a reduction
in the concentration of antigen binding molecule or of Fc-fusion
protein, in the medium surrounding the target cells, required to
achieve the lysis of a given number of "targeted cells," in a given
time, by the mechanism of Fc-mediated cellular cytotoxicity. The
increase in Fc-mediated cellular cytotoxicity is relative to the
cellular cytotoxicity mediated by the same antigen binding molecule
or Fc-fusion protein produced by the same type of host cells, using
the same standard production, purification, formulation and storage
methods, (which are known to those skilled in the art) but that has
not been produced by host cells engineered to have an altered
pattern of glycosylation (e.g., to express the glycosyltransferase,
GnTIII, or other glycosyltransferases) by the methods described
herein.
[0117] By "antigen binding molecule having increased antibody
dependent cellular cytotoxicity (ADCC)" is meant an antigen binding
molecule, as that term is defined herein, having increased ADCC as
determined by any suitable method known to those of ordinary skill
in the art. One accepted in vitro ADCC assay is as follows: [0118]
1) the assay uses target cells that are known to express the target
antigen recognized by the antigen-binding region of the antibody;
[0119] 2) the assay uses human peripheral blood mononuclear cells
(PBMCs), isolated from blood of a randomly chosen healthy donor, as
effector cells; [0120] 3) the assay is carried out according to
following protocol: [0121] i) the PBMCs are isolated using standard
density centrifugation procedures and are suspended at 5.times.106
cells/ml in RPMI cell culture medium; [0122] ii) the target cells
are grown by standard tissue culture methods, harvested from the
exponential growth phase with a viability higher than 90%, washed
in RPMI cell culture medium, labeled with 100 micro-Curies of 51Cr,
washed twice with cell culture medium, and resuspended in cell
culture medium at a density of 105 cells/ml; [0123] iii) 100
microliters of the final target cell suspension above are
transferred to each well of a 96-well microtiter plate; [0124] iv)
the antibody is serially-diluted from 4000 ng/ml to 0.04 ng/ml in
cell culture medium and 50 microliters of the resulting antibody
solutions are added to the target cells in the 96-well microtiter
plate, testing in triplicate various antibody concentrations
covering the whole concentration range above; [0125] v) for the
maximum release (MR) controls, 3 additional wells in the plate
containing the labeled target cells, receive 50 microliters of a 2%
(V/V) aqueous solution of non-ionic detergent (Nonidet, Sigma, St.
Louis), instead of the antibody solution (point iv above); [0126]
vi) for the spontaneous release (SR) controls, 3 additional wells
in the plate containing the labeled target cells, receive 50
microliters of RPMI cell culture medium instead of the antibody
solution (point iv above); [0127] vii) the 96-well microtiter plate
is then centrifuged at 50.times.g for 1 minute and incubated for 1
hour at 4.degree. C.; [0128] viii) 50 microliters of the PBMC
suspension (point i above) are added to each well to yield an
effector:target cell ratio of 25:1 and the plates are placed in an
incubator under 5% CO2 atmosphere at 37.degree. C. for 4 hours;
[0129] ix) the cell-free supernatant from each well is harvested
and the experimentally released radioactivity (ER) is quantified
using a gamma counter; [0130] x) the percentage of specific lysis
is calculated for each antibody concentration according to the
formula (ER-MR)/(MR-SR).times.100, where ER is the average
radioactivity quantified (see point ix above) for that antibody
concentration, MR is the average radioactivity quantified (see
point ix above) for the MR controls (see point v above), and SR is
the average radioactivity quantified (see point ix above) for the
SR controls (see point vi above); [0131] 4) "increased ADCC" is
defined as either an increase in the maximum percentage of specific
lysis observed within the antibody concentration range tested
above, and/or a reduction in the concentration of antibody required
to achieve one half of the maximum percentage of specific lysis
observed within the antibody concentration range tested above. The
increase in ADCC is relative to the ADCC, measured with the above
assay, mediated by the same antibody, produced by the same type of
host cells, using the same standard production, purification,
formulation and storage methods, which are known to those skilled
in the art, but that has not been produced by host cells engineered
to overexpress GnTIII.
Anti-CEA Antigen Binding Molecules, Polypeptides, and
Polynucleotides
[0132] CEA has long been used as a cancer marker for diagnostic
purposes. It is abnormally expressed (e.g., overexpressed and/or
distributed in a different pattern in the cell) in many tumor
tissues compared to non-tumor tissues of the same cell type.
However, because CEA is generally cleaved from the tumor cell
surface and most of the available anti-CEA antibodies also bind
soluble CEA, unconjugated antibodies to CEA are generally not used
for therapeutic purposes. For example, the anti-CEA antibodies that
are currently in pilot trials are administered as radioconjugates
(Wong et al., 2004; Liersch et al., 2007). Several mechanisms are
involved in the therapeutic efficacy of anti-CEA antibodies,
including antibody dependent cellular cytotoxicity (ADCC) and
complement-dependent cytotoxicity (CDC). Increased CEA expression
promotes increased intercellular adhesion, which may lead to
metastasis of cancerous cells (Marshall J., Semin Oncol. 30(3)
Suppl. 8:30-36). Thus, anti-CEA antigen binding molecules may also
play a role in inhibiting CEA-mediated cell adhesion and metastasis
of cancerous cells.
[0133] In one aspect, the invention is directed to a variant
antigen binding molecule (e.g., an antibody or fragment thereof)
comprising one or more (e.g., one, two, three, four, five, or six)
CDRs of the murine PR1A3 antibody, wherein at least one of the CDRs
has substitution of at least one amino acid residue compared to the
corresponding CDR of PR1A3, and wherein the variant antigen binding
molecule has improved affinity for CEA, preferably membrane-bound
CEA compared to a parent PR1A3 antigen binding molecule.
International Patent Application WO2011023787 describes anti-CEA
antigen binding molecules with improved affinity for CEA as
compared to a parent PR1A3 antigen binding molecule.
[0134] In another aspect, the invention is directed to a variant
antigen binding molecule comprising one or more (e.g., one, two,
three, four, five, or six) CDRs of the murine PR1A3 antibody,
wherein at least one of the CDRs has a substitution of at least one
amino acid residue compared to the corresponding CDR of PR1A3, and
wherein the variant antigen binding molecule has increased
stability compared to a parent PR1A3 antigen binding molecule. In
one embodiment, the parent PR1A3 antigen binding molecule is a
humanized PR1A3 antigen binding molecule. In one embodiment, the
parent PR1A3 antigen binding molecule comprises the heavy chain
variable region of CH7A (SEQ ID NO:101). In one embodiment, the
parent PR1A3 antigen binding molecule comprises the heavy chain
variable regions CH7A (SEQ ID NO:101) and the light chain variable
region 2F1 (SEQ ID NO: 209). Such one or more CDRs can be truncated
CDRs and will contain, at a minimum, the specificity-determining
residues (SDRs), as that term is defined herein, for a given CDR.
In one embodiment, the variant antigen binding molecule comprises
at least one (e.g., one, two, three, four, five or six) of the CDRs
selected from SEQ ID NOs: 1-3, 5-10, 12-56 and 217-224 (FIG. 32 and
FIG. 36), comprising the residues of the CDRs that will retain
specific binding. In another embodiment, the variant antigen
binding molecule comprises at least one (e.g., one, two, three,
four, five, or six) CDR selected from SEQ ID NOs: 1-3, 5-10, 12-56
and 217-224, or a variant or truncated form thereof containing at
least the specificity-determining residues for said CDR, and
comprising a sequence derived from a heterologous polypeptide. In a
specific embodiment, where the variant antigen binding molecule
comprises a heavy chain CDR1 variant of PR1A3, the HCDR1 has a
glutamate substituted for a valine at Kabat position 31. In a
specific embodiment, where the variant antigen binding molecule
comprises a heavy chain CDR3 variant of PR1A3, the HCDR3 has an
alanine substituted for a tyrosine at Kabat position 98 or a
tyrosine substituted for an aspartate at Kabat position 99. In a
specific embodiment, where the variant antigen binding molecule
comprises a heavy chain CDR3 variant of PR1A3, the HCDR3 has an
alanine substituted for a tyrosine at Kabat position 98 and a
tyrosine substituted for an aspartate at Kabat position 99.
[0135] In one embodiment, the variant antigen binding molecule
comprises one heavy chain CDR3 selected from SEQ ID NOs: 217-224
(FIG. 36) and two heavy chain CDRs (e.g., HCDR1 and HCDR2) selected
from SEQ ID NOs: 1-3, 5-10, and 12-24 and/or three light chain CDRs
(e.g. LCDR1, LCDR2, and LCDR3) selected from SEQ ID NOs: 36-56, or
variants or truncated forms thereof containing at least the
specificity-determining residues for each of the CDRs. In a more
specific embodiment, the variant antigen binding molecule comprises
one heavy chain CDR3 selected from SEQ ID NOs: 217-224 and two
heavy chain CDRs (e.g., HCDR1 and HCDR2) selected from SEQ ID NOs:
1-3, 5-10, and 12-24 and three light chain CDRs (e.g. LCDR1, LCDR2,
and LCDR3) selected from SEQ ID NOs: 36-56. In another embodiment,
the variant antigen binding molecule comprises the variable
region(s) of an antibody light and/or heavy chain, preferably both
a heavy and light chain variable region. In a more particular
embodiment, the heavy chain and/or light chain variable region is
selected from the heavy and/or light chain variable region selected
from SEQ ID NOs: 99-108, SEQ ID NOs: 188-216, and SEQ ID NOs:
225-248 (FIG. 33 and FIG. 37A-C) or a combination thereof, wherein,
the heavy and light chain variable region is not a combination of
SEQ ID NO:99 and SEQ ID NO:103 or SEQ ID NO:100 and SEQ ID NO:104.
In some embodiments, the heavy chain comprises the framework
residues of CH1A1 A (SEQ ID NO: 261) or CH1A1B (SEQ ID NO: 262)
(FIG. 37C). In one embodiment, the variant antigen binding molecule
comprises a heavy chain variable region selected from SEQ ID NO:
225, SEQ ID NO: 226, SEQ ID NO: 231, SEQ ID NO: 234, SEQ ID NO:
235, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO:
243, or SEQ ID NO: 247.
[0136] In one embodiment, the variant antigen binding molecule is a
chimeric antibody, more specifically, a humanized antibody. In
another embodiment, the variant antigen binding molecule comprises
an Fc region. In another embodiment, the variant antigen binding
molecule is affinity matured. In another embodiment, the variant
antigen binding molecule is engineered to have increased stability
(stability matured). In another embodiment, the variant antigen
binding molecule has increased ADCC activity compared to PR1A3. In
one embodiment, the increased ADCC of the variant antigen binding
molecule is due to an increase in affinity of the variant antigen
binding molecule for membrane-bound CEA, for example by affinity
maturation or other methods of improving affinity (see Tang et al.,
J. Immunol. 2007, 179:2815-2823, the entire contents of which is
herein incorporated by reference). In another embodiment, the
variant antigen binding molecule comprises an Fc region that is
glycoengineered. In another aspect, the invention is also directed
to methods of making such variant antigen binding molecules and
their use in the treatment of disease, particularly cell
proliferation disorders wherein CEA is expressed, particularly
wherein CEA is abnormally expressed (e.g., overexpressed or
expressed in a different pattern in the cell) compared to normal
tissue of the same cell type. Such disorders include, but are not
limited to colorectal cancer, NSCLC (non-small cell lung cancer),
gastric cancer, pancreatic cancer and breast cancer. CEA expression
levels may be determined by methods known in the art and those
described herein (e.g., via immunohistochemistry assay,
immunofluorescence assay, immunoenzyme assay, ELISA, flow
cytometry, radioimmunoassay, Western blot, ligand binding, kinase
activity, etc.).
[0137] In another aspect, the invention is also directed to an
isolated polynucleotide comprising a sequence that encodes a
polypeptide comprising one or more (e.g., one, two, three, four,
five, or six) complementarity determining regions of the murine
PR1A3 antibody, or variants or truncated forms thereof containing
at least the specificity-determining residues for said
complementarity determining regions. Typically, such isolated
polynucleotides encode one or more fusion polypeptides that form an
antigen binding molecule. In one embodiment, the polynucleotide
comprises a sequence encoding one or more (e.g., one, two, three,
four, five or six) of the CDRs selected from SEQ ID NOs: 1-3, 5-10,
12-56 and 217-224, comprising the residues of the CDRs that will
retain specific binding. In one embodiment, the polynucleotide
comprises a sequence that encodes at least three heavy chain CDRs
(e.g., HCDR1, HCDR2, and HCDR3) and/or three light chain CDRs (e.g.
LCDR1, LCDR2, and LCDR3 selected from SEQ ID NOs: 1-3, 5-10, 12-56
and 217-224, or variants or truncated forms thereof containing at
least the specificity-determining residues (SDRs) for each of said
three complementarity determining regions. In a more specific
embodiment, the polynucleotide encodes a polypeptide comprising one
heavy CDR3 selected from SEQ ID NOs: 217-224 and two heavy chain
CDRs (e.g., HCDR1 and HCDR2) selected from SEQ ID NOs: 1-3, 5-10,
and 12-24 and three light chain CDRs (e.g. LCDR1, LCDR2, and LCDR3)
selected from SEQ ID NOs: 36-56. In another embodiment, the
polynucleotide encodes a polypeptide comprising the variable
region(s) of an antibody light and/or heavy chain. The
polynucleotides encoding the heavy and light chain variable region
polypeptides can be expressed in one or more expression vectors. In
a more particular embodiment, the polynucleotide encoding a heavy
chain and/or light chain variable region selected from SEQ ID NOs:
99-108, SEQ ID NOs: 188-216, and SEQ ID NOs: 225-248 is selected
from the group of polynucleotides of SEQ ID NOs: 159-187 and SEQ ID
NOs: 249-256 or a combination thereof, wherein, the heavy and light
chain variable regions are not encoded by a combination of SEQ ID
NO:11 and SEQ ID NO:115 or SEQ ID NO:112 and SEQ ID NO:116. In one
embodiment, the heavy and light chain variable region polypeptides
encoded by the polynucleotides combine to form a chimeric antibody,
more specifically, a humanized antibody. In a specific embodiment,
where the polynucleotide comprises a sequence that encodes heavy
chain CDR1 of PR1A3 or a variant thereof, said polynucleotide
encodes a glutamate substituted for a valine at Kabat position 31.
In a specific embodiment, where the polynucleotide comprises a
sequence that encodes heavy chain CDR3 of PR1A3 or a variant
thereof, said polynucleotide encodes an alanine substituted for a
tyrosine at Kabat position 98 or a tyrosine substituted for an
aspartate at Kabat position 99. In a specific embodiment, where the
polynucleotide comprises a sequence that encodes heavy chain CDR3
of PR1A3 or a variant thereof, said polynucleotide encodes an
alanine substituted for a tyrosine at Kabat position 98 and a
tyrosine substituted for an aspartate at Kabat position 99. In one
embodiment, the polynucleotide encodes an alanine substituted for a
tyrosine at Kabat position 98 or a tyrosine substituted for an
aspartate at Kabat position 99 in the framework of CH1A1A (SEQ ID
NO: 261) or CH1A1B (SEQ ID NO: 262). In one embodiment, the
polynucleotide comprises a sequence encodes the heavy chain of SEQ
ID NO: 225, SEQ ID NO: 226, SEQ ID NO:231, SEQ ID NO:234, SEQ ID
NO:235, SEQ ID NO:239, SEQ ID NO:241, SEQ ID NO: 242, SEQ ID NO:
243, or SEQ ID NO: 247. In another embodiment, the polynucleotide
comprises a sequence that encodes an Fc region. The invention is
further directed to the polypeptides encoded by such
polynucleotides.
[0138] In one aspect, the present invention is related to antigen
binding molecules or variant antigen binding molecules (e.g., an
antibody or fragment thereof) and polypeptides having the same
binding specificity of the murine PR1A3 antibody (e.g., binding to
the same epitope of membrane-bound CEA), and having comparable or
improved biological activities (e.g., improved affinity for
membrane-bound CEA, increased stability, and/or enhanced ADCC). In
one embodiment, the variant antigen binding molecule binds the same
epitope as that of the parent antigen binding molecule. In one
embodiment, the variant antigen binding molecule competes for
binding to membrane-bound human CEA with the parent antigen binding
molecule. In one embodiment, the variant antigen binding molecule
binds to membrane-bound human CEA and does not bind to soluble
human CEA. In one aspect, the present invention is related to
antigen binding molecules and the variant antigen binding molecule
and polypeptides having increased stability as compared to murine
PR1A3 antibody, or humanized variant thereof. In one aspect, the
present invention is related to antigen binding molecules and
variant antigen binding molecules (e.g., an antibody or fragment
thereof) and polypeptides which bind membrane bound CEA and have an
increased stability as compared to a humanized PR1A3 antibody which
comprises the heavy chain variable region of CH7A (SEQ ID NO:101).
In one aspect, the present invention is related to antigen binding
molecules and variant antigen binding molecules (e.g., an antibody
or fragment thereof) and polypeptides which bind membrane bound CEA
and have increased stability as compared to a humanized PR1A3
antibody which comprises the heavy chain variable region CH7A (SEQ
ID NO:101) and the light chain variable region 2F1 (SEQ ID NO:
209)
[0139] In one embodiment, the variant antigen binding molecule or
polypeptide comprises at least one (e.g., one, two, three, four,
five or six) of the CDRs selected from SEQ ID NOs: 1-3, 5-10, 12-56
and 217-224 (FIG. 32 and FIG. 36). In one embodiment, the variant
antigen binding molecule or polypeptide comprises: (a) a heavy
chain CDR1 sequence selected from the group consisting of: SEQ ID
NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ ID
NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:12; (b)
a heavy chain CDR2 sequence selected from the group consisting of:
SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID
NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ
ID NO:22, SEQ ID NO:23, and SEQ ID NO:24: and (c) a heavy chain
CDR3 sequence selected from the group consisting of: SEQ ID NO:217,
SEQ ID NO:218, SEQ ID NO:219, SEQ ID NO:220, SEQ ID NO:221, SEQ ID
NO:222, SEQ ID NO:223, and SEQ ID NO:224. In another embodiment,
the variant antigen binding molecule or polypeptide comprises: (a)
a light chain CDR1 sequence selected from the group consisting of:
SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID
NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, and
SEQ ID NO:45; (b) a light chain CDR sequence selected from the
group consisting of: SEQ ID NO:46, and SEQ ID NO:47, SEQ ID NO:48,
SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID
NO:53, SEQ ID NO:54, and SEQ ID NO:55; and (c) a light chain CDR3
of SEQ ID NO:56. In some embodiments, the variant antigen binding
molecule or polypeptide comprising the CDRs also comprise the
framework residues of CH1A1 A (SEQ ID NO: 261) or CH1A1B (SEQ ID
NO: 262).
[0140] In one aspect, the invention is directed to a variant
antigen binding molecule or polypeptide which binds membrane-bound
human CEA comprising a heavy chain variable region and/or a light
chain variable region. In one embodiment, the heavy chain and/or
light chain variable region is selected from the heavy and/or light
chain variable region selected from SEQ ID NOs: 99-108, SEQ ID NOs:
188-216, and SEQ ID NOs: 225-248 (FIG. 33 and FIG. 37A-C). In one
embodiment, the heavy chain variable region comprises a polypeptide
having the sequence of any one of SEQ ID NOs: 225-248. In another
specific embodiment, the heavy chain variable region comprises a
polypeptide having a sequence that is at least about 80%, 85%, 90%,
95%, 96%, 97%, 98% or 99% identical to the sequence of any one of
SEQ ID NOs: 225-248.
[0141] In one embodiment, the heavy chain variable region comprises
a polypeptide having the sequence of SEQ ID NO: 233; SEQ ID NO:
234, SEQ ID NO: 235, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO:
242, SEQ ID NO: 243, and SEQ ID NO: 247. In another embodiment, the
heavy chain variable region comprises a polypeptide having at least
about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity to the
sequence of SEQ ID NO: 233, SEQ ID NO: 234, SEQ ID NO: 235, SEQ ID
NO: 239, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, and SEQ ID
NO: 247. In one embodiment, the light chain variable region
comprises a polypeptide having the sequence of SEQ ID NO: 209. In
another embodiment, the heavy chain variable region comprises a
polypeptide having at least about 80%, 85%, 90%, 95%, 96%, 97%,
98%, or 99% identity to the sequence of SEQ ID NO: 209.
[0142] In one embodiment, the antigen binding molecule, variant
antigen binding molecule, or polypeptide which binds membrane-bound
human CEA comprises a heavy chain variable region and a light chain
variable region. In a specific embodiment, the heavy chain variable
region comprises a polypeptide having the sequence of SEQ ID NO:4
as follows:
TABLE-US-00002
QVQLVQSGSELKKPGASVKVSCKASGYTFTEX.sup.1X.sup.2MX.sup.3WVRQAPGQ
GLEWMGX.sup.4INTKX.sup.5GEAX.sup.6YX.sup.7EEFKGRFVFSLDTSVSTAYLQISSLK
AEDTAVYYCARWDX.sup.8X.sup.9X.sup.10YX.sup.11X.sup.12X.sup.13X.sup.14DYWGQG-
TTVTVSS
wherein X.sup.1 is Y or F; X.sup.2 is S or G; X.sup.3 is N or S;
X.sup.4 is W or Y; X.sup.5 is N, T or S; X.sup.6 is T or N; X.sup.7
is V or I; X.sup.8 is F or A; X.sup.9 is Y, A, V, F or S; X.sup.10
is D, H, W, E, or Y; X.sup.11 is V, L or F; X.sup.12 is E, K or Q;
X.sup.13 is A or T; and X.sup.14 is M or L.
[0143] In a specific embodiment, the light chain variable region
comprises a polypeptide having the sequence of SEQ ID NO:11 as
follows:
TABLE-US-00003 DIQMTQSPSSLSASVGDRVTITCKASX15X16X17X18X19X20VAWYQQ
KPGKAPKX21LIYX22ASX23X24X25X26GVPSRFSGSGSGTDFTLTIS
SLQPEDFATYYCHQYYTYPLFTFGQGTKLEIK
[0144] wherein X15 is Q, A, K, or H; X16 is N, A, Y, I, K, T, or F;
X17 is V, A, G, or M; X18 is G, S, T, or L; X19 is T, N, P, or A;
X20 is N or Y; X21 is P or L; X22 is S, L, or W; X23 is Y, N, or H;
X24 is R, L, P, or H; X25 is Y, S, Q, K, E, F, or P; and X26 is S,
G, I, or R.
[0145] In another aspect, the invention is further directed to
isolated polynucleotides encoding the antigen binding molecules,
variant antigen binding molecules, or polypeptides which bind
membrane-bound CEA. In one embodiment, the polynucleotide comprises
a sequence that encodes at least three heavy chain CDRs (e.g.,
HCDR1, HCDR2, and HCDR3) and/or three light chain CDRs (e.g. LCDR1,
LCDR2, and LCDR3 selected from SEQ ID NOs: 1-3, 5-10, 12-56 and
217-224, or variants or truncated forms thereof containing at least
the specificity-determining residues (SDRs) for each of said three
complementarity determining regions. In a more specific embodiment,
the polynucleotide encodes a polypeptide comprising one heavy CDR3
selected from SEQ ID NOs: 217-224 and two heavy chain CDRs (e.g.,
HCDR1 and HCDR2) selected from SEQ ID NOs: 1-3, 5-10, and 12-24 and
three light chain CDRs (e.g. LCDR1, LCDR2, and LCDR3) selected from
SEQ ID NOs: 36-56. In another embodiment, the polynucleotide
encodes a polypeptide comprising the variable region(s) of an
antibody light and/or heavy chain. The polynucleotides encoding the
heavy and light chain variable region polypeptides can be expressed
in one or more expression vectors. In a more particular embodiment,
the polynucleotide encoding a heavy chain and/or light chain
variable region selected from SEQ ID NOs: 99-108, SEQ ID NOs:
188-216, and SEQ ID NOs: 225-248 is selected from the group of
polynucleotides selected from SEQ ID NOs: 159-187 and SEQ ID NOs:
249-256 or a combination thereof, wherein, the heavy and light
chain variable regions are not encoded by a combination of SEQ ID
NO:11 and SEQ ID NO:115 or SEQ ID NO:112 and SEQ ID NO:116. In one
embodiment, the heavy and light chain variable region polypeptides
encoded by the polynucleotides combine to form a chimeric antibody,
more specifically, a humanized antibody.
[0146] In one embodiment, the antigen binding molecule, variant
antigen binding molecule, or polypeptide binds membrane-bound CEA
comprises an Fc region. In a more specific embodiment, the antigen
binding molecule, variant antigen binding molecule, or polypeptide
is glycoengineered to have an altered pattern of glycosylation in
the Fc region. In a particular embodiment, the affinity for
membrane-bound CEA of the variant antigen binding molecule or
polypeptide is increased compared to the parent PR1A3 antibody. In
another embodiment, the stability of the variant antigen binding
molecule or polypeptide is increased compared to the parent PR1A3
antibody. In another embodiment, the variant antigen binding
molecule or polypeptide has increased ADCC activity. In one
embodiment, the increased ADCC of the variant antigen binding
molecule or polypeptide is due to an increase in affinity of the
polypeptide for membrane-bound CEA, for example by affinity
maturation or other methods of improving affinity.
[0147] In another aspect, the invention is also directed to use of
the antigen binding molecule, variant antigen binding molecule, or
polypeptide in the treatment of disease, particularly cell
proliferation disorders wherein CEA is expressed, particularly
wherein CEA is abnormally expressed (e.g., overexpressed or
expressed in a different pattern in the cell) compared to normal
tissue of the same cell type. Such disorders include, but are not
limited to colorectal cancer, NSCLC (non-small cell lung cancer),
gastric cancer, pancreatic cancer and breast cancer. CEA expression
levels may be determined by methods known in the art and those
described herein (e.g., via immunohistochemistry assay,
immunofluorescence assay, immunoenzyme assay, ELISA, flow
cytometry, radioimmunoassay, Western blot, ligand binding, kinase
activity, etc.).
[0148] In a particular embodiment, the invention is directed to a
humanized antigen binding molecule or a portion or fragment thereof
that binds membrane-bound CEA comprising a heavy chain variable
region comprising the sequence of any one of SEQ ID NOs: 225-248.
In another embodiment, the invention is directed to a humanized
antigen binding molecule or a portion or fragment thereof that
binds membrane-bound CEA comprising a light chain variable region
comprising the sequence of any one of SEQ ID NOs: 105, 108, or
207-216. In a particular embodiment, the humanized antigen binding
molecule or a portion or fragment thereof that binds membrane-bound
CEA comprises a heavy chain variable region comprising the sequence
of any one of SEQ ID NOs: 225-248 and a light chain variable region
comprising the sequence of any one of SEQ ID NOs:105, 108, or
207-216. In one embodiment, the humanized antigen binding molecule
further comprises a human heavy chain constant region and/or a
human light chain constant region. Such constant regions are
described herein and are known in the art. In a more particular
embodiment, the humanized antigen binding molecule comprises an Fc
region, more particularly, an Fc region that has been
glycoengineered.
[0149] Methods for humanizing non-human antibodies are known in the
art. For example, humanized ABMs of the present invention can be
prepared according to the methods of U.S. Pat. No. 5,225,539 to
Winter, U.S. Pat. No. 6,180,370 to Queen et al., or U.S. Pat. No.
6,632,927 to Adair et al., the entire contents of each of which is
hereby incorporated by reference. Preferably, a humanized antibody
has one or more amino acid residues introduced into it from a
source which is non-human. These non-human amino acid residues are
often referred to as "import" residues, which are typically taken
from an "import" variable domain. Humanization can be essentially
performed following the method of Winter and co-workers (Jones et
al., Nature, 321:522-525 (1986); Riechmann et al., Nature,
332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536
(1988)), by substituting hypervariable region sequences for the
corresponding sequences of a human antibody. Accordingly, such
"humanized" antibodies are chimeric antibodies (U.S. Pat. No.
4,816,567) wherein substantially less than an intact human variable
domain has been substituted by the corresponding sequence from a
non-human species. Typically, humanized antibodies are human
antibodies in which some hypervariable region residues and possibly
some FR residues are substituted by residues from analogous sites
in a non-human (e.g., rodent) antibodies. The subject humanized
anti-CEA antibodies will optionally comprise constant regions from
a human immunoglobulin.
[0150] The choice of light and heavy chain human variable domains
for making the humanized antibodies is very important to reduce
antigenicity. According to the so-called "best-fit" method, the
sequence of the variable domain of a donor (e.g., rodent) antibody
is screened against the entire library of known human
variable-domain sequences. The human sequence which is closest to
that of the donor (e.g., rodent) is then accepted as the human
framework region (FR) for the humanized antibody (Sims et al., J.
Immunol., 151:2296 (1993); Chothia et al., J. Mol. Biol., 196:901
(1987)). Another method of selecting the human framework sequence
is to compare the sequence of each individual subregion of the full
donor (e.g., rodent) framework (i.e., FR1, FR2, FR3, and FR4) or
some combination of the individual subregions (e.g., FR1 and FR2)
against a library of known human variable region sequences that
correspond to that framework subregion (e.g., as determined by
Kabat numbering), and choose the human sequence for each subregion
or combination that is the closest to that of the rodent (Leung
U.S. Patent Application Publication No. 2003/0040606A1, published
Feb. 27, 2003). Another method uses a particular framework region
derived from the consensus sequence of all human antibodies of a
particular subgroup of light or heavy chains (Carter et al., Proc.
Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immunol.,
151:2623 (1993)). In one embodiment, the human framework regions
are selected from a collection of human germline sequences. Such
collections of human germline sequences can be found in databases
such as IMGT or VBase. Framework regions can be selected
individually (e.g., the FR1-3 selected for the acceptor for the
heavy and/or light chain variable regions of the humanized anti-CEA
ABMs can be encoded by different germline genes) or as part of the
same germline gene. In a more specific embodiment, heavy chain
FR1-3 are encoded by the IGHV7_4_1*02 human immunoglobulin germline
gene sequence (Accession No. X62110, SEQ ID NO:114). In another
specific embodiment, light chain FR1-3 are encoded by the
IMGT_hVK_1_39 human immunoglobulin germline gene sequence
(Accession No. X59315, SEQ ID NO:118). In another specific
embodiment, heavy chain FR4 is encoded by the JH6 germline gene
sequence (See GenBank Accession No. M63030). In another specific
embodiment, light chain FR4 is encoded by the JK2 germline gene
sequence (See Genbank Accession No. X61584).
[0151] It is generally desirable that antigen binding molecules,
such as antibodies and fragments thereof, be humanized with
retention of high affinity for the antigen and other favorable
biological properties. Accordingly, in one embodiment, humanized
antibodies are prepared by analyzing the parental sequences and
various conceptual humanized products using three-dimensional
models of the parental and humanized sequences. Three-dimensional
immunoglobulin models are commonly available and are familiar to
those skilled in the art. Computer programs are available which
illustrate and display probable three-dimensional conformational
structures of selected candidate immunoglobulin sequences. Analysis
of these displays helps to elucidate the likely role of the
residues in the functioning of the candidate immunoglobulin
sequence, e.g., the analysis of residues that influence the ability
of the candidate immunoglobulin to bind its antigen. In this way,
FR residues can be selected and combined from the recipient and
import sequences so that the desired antibody characteristic, such
as increased affinity for the target antigen(s), is achieved. In
general, the hypervariable region residues are directly and most
substantially involved in influencing antigen binding.
[0152] In one aspect, the invention is directed to humanized,
affinity matured and/or variant anti-CEA antigen binding molecules
with desirable properties and characteristics that include, but are
not limited to: strong binding affinity for the CEA antigen--in
particular, membrane-bound CEA--while having substantially no
cross-reactivity against soluble CEA; an ability to induce cell
lysis of CEA-expressing cells in vitro and ex vivo, preferably in a
dose-dependent manner; an ability to inhibit CEA-mediated cell
adhesion in vitro; an ability to inhibit tumor tissue growth and/or
induce tumor tissue regression (for example, as demonstrated in
tumor models (e.g., xenograft mouse)).
[0153] As described herein, in some embodiments, variant antigen
binding molecules of the invention have increased binding affinity,
for example, due to affinity maturation of a parent antibody
comprising one or more CDRs of the PR1A3 antibody. Affinity of the
antigen binding molecules and variant antigen binding molecules of
the invention can be determined by methods known in the art and as
described herein. In a specific embodiment, humanized or variant
antigen binding molecules of the invention bind to human CEA,
preferably membrane-bound CEA, with a monovalent affinity constant
(K.sub.D) value of no more than about 1 .mu.M to about 0.001 nM,
more specifically no more than about 800 nM to about 1 nM, and even
more specifically no more than about 550 nM to about 10 nM. In a
specific embodiment, the variant antigen binding molecule is an
affinity matured antibody or fragment thereof that binds to
membrane-bound CEA with a monovalent affinity constant (K.sub.D)
value of no more than about 100 nM to about 10 nM. In one
embodiment, the variant antigen binding molecule is an affinity
matured antibody or fragment thereof that binds to membrane-bound
CEA with a monovalent affinity constant (K.sub.D) value of no more
than about 100 nM to about 0.01 nM. In one embodiment, the variant
antigen binding molecule is an affinity matured antibody or
fragment thereof that binds to membrane-bound CEA with a monovalent
affinity constant (K.sub.D) value of no more than about 10 nM to
about 0.1 nM. In one embodiment, the variant antigen binding
molecule is an affinity matured antibody or fragment thereof that
binds to membrane-bound CEA with a monovalent affinity constant
(K.sub.D) value of 100 nM, 10 nM, 1 nM, 0.1 nM, or lower. In some
embodiments, the variant antigen binding molecule is a stability
matured (engineered to have increased stability) antibody or
fragment thereof that retains the increased binding affinity of its
affinity matured parent. In one embodiment, the variant antigen
binding molecule binds to the membrane-bound CEA with a higher
affinity than it binds to shed CEA. In one embodiment, the variant
antigen binding molecule binds to the membrane-bound CEA with 1.5
fold, 2 fold, 3 fold, 4 fold, 5 fold, 10 fold, 20 fold, or higher
affinity than it binds to shed CEA.
[0154] In one embodiment, the variant antigen binding molecule of
the invention typically binds the same epitope as recognized by the
mouse antibody PR1A3, or is capable of competing with the PR1A
antibody for binding to membrane-bound CEA. To screen for
antibodies that bind to the epitope on human CEA bound by an
antibody of interest (e.g., those that block binding of the PR1A3
antibody to human CEA), a routine cross-blocking assay such as that
described in ANTIBODIES, A LABORATORY MANUAL, Cold Spring Harbor
Laboratory, Harlow and Lane eds. (1988), can be performed.
Alternatively, epitope mapping, e.g. as described in Champe et al.,
J. Biol. Chem. 270:1388-1394 (1995), can be performed to determine
whether the antibody binds an epitope of interest.
[0155] In one embodiment, variant antigen bind molecules specific
for human CEA are made from a parent anti-CEA antigen binding
molecule comprising at least one CDR of the monoclonal antibody
PR1A3, wherein the parent anti-CEA antibody binds the same epitope
as the PR1A3 antibody and is capable of competing with PR1A3 for
antigen binding. In one embodiment, the parent antigen binding
molecule comprises at least one, two, or typically three, heavy
chain CDRs of the PR1A3 antibody; in another embodiment, the parent
antigen binding molecule comprises at least one, two, or typically
three, light chain CDRs of the PR1A3 antibody; in another
embodiment, the parent antigen binding molecule comprises the three
heavy chain CDRs and the three light chain CDRs of the PR1A3
antibody. Preferably, where the variant antigen binding molecule
comprises HCDR1 of PR1A3, said HCDR1 comprises a substitution of
glutamate for valine at Kabat position 31. The variant ABMs
typically have a greater affinity for CEA than the parent. In one
embodiment, the variant ABMS have increased stability as compared
to the parent. In one embodiment, the variant ABM comprises an Fc
region. In one embodiment, the variant ABM is glycoengineered. In
one embodiment the variant ABM has increased ADCC activity compared
to the parent ABM. In a particular embodiment, the increased ADCC
is result of the increased affinity, achieved, for example, by
affinity maturation of the parent ABM to generate the variant ABM.
In a more particular embodiment, the increase in ADCC is at least
about 40% to about 100% as compared to said parent antigen binding
molecule. In another particular embodiment, the variant ABM
increases ADCC by at least about 10% to about 100% in an in vitro
cytotoxicity assay. In a more particular embodiment, the variant
ABM is at least from about 10-fold to about 1000-fold more potent
at inducing ADCC at a given concentration compared to the murine
PR1A3 antibody. In another particular embodiment, the increased
ADCC activity is a result of glycoengineering of the Fc region. In
another particular embodiment, the increased ADCC activity is a
result of a combination of increased affinity and
glycoengineering.
[0156] In one embodiment, the variant antigen binding molecules of
the invention comprise one or more amino acid substitutions in at
least one CDR. The number of amino acid substitution(s) can range
from one to ten (e.g, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10), preferably
from two to five (e.g., 2, 3, 4, or 5). In one embodiment, at least
one heavy chain CDR comprises one or more amino acid
substitution(s). In another embodiment, at least one light chain
CDR comprises one or more amino acid substitution(s). In another
embodiment, at least one heavy chain CDR comprises one or more
substitutions, and at least one light chain CDR comprises one or
more substitutions. Preferably, where the variant antigen binding
molecule comprises HCDR1 of PR1A3, said HCDR1 comprises a
substitution of glutamate for valine at Kabat position 31.
[0157] Substantial modifications in the biological properties of
the antigen binding molecules are accomplished by selecting
substitutions that differ significantly in their effect on
maintaining (a) the structure of the polypeptide backbone in the
area of the substitution, for example, as a sheet or helical
conformation, (b) the charge or hydrophobicity of the molecule at
the target site, or (c) the bulk of the side chain Variant antigen
binding molecules comprising amino acid substitutions may have
improved biological activities, for example, improved antigen
binding affinity and enhanced ADCC, compared to the parent antigen
binding molecule Amino acid substitutions can be introduced by
various techniques known in the art including, but not limited to,
site directed mutagenesis and/or affinity maturation of the parent
antigen binding molecule e.g., by phage display.
[0158] In order to identify candidate sites, e.g., hypervariable
region residues, for modification, alanine scanning mutagenesis can
be performed to find residues that contribute significantly to
antigen binding. Alternatively, or in addition, it may be
beneficial to analyze a crystal structure of the antigen-antibody
complex to identify contact points between the antibody and human
CEA. Such contact residues and neighboring residues are candidates
for substitution according by methods known in the art and/or
described herein. Once such variants are generated, the panel of
variants can be screened by methods known in the art and/or
described herein and antibodies with superior properties in one or
more relevant assays can be selected for further development.
[0159] Phage display can be used to generate a repertoire of
hypervariable region sequences from a parent antigen binding
molecule that containing random amino acid mutation(s). For
example, several hypervariable region sites (e.g. 6-7 sites) are
mutated to generate all possible amino substitutions at each site.
Alternatively, random mutagenesis can be performed on heavy and/or
light chain variable regions. Mutations can be generated by
techniques known in the art, including but not limited to using
mutagenesis primers, controlling the number of cycles and using
mutagenic nucleotide analogues 8-oxo-dGTP and dPTP during PCR
amplification. The antibody variants thus generated are displayed
in a monovalent fashion from filamentous phage particles as fusions
to the gene III product of M13 packaged within each particle. The
phage-displayed variants are then screened for their biological
activities (e.g. binding affinity) as herein disclosed and
candidates that have one or more improved activities will be used
for further development. Methods for making phage display libraries
can be found in Huse et al., Science, 246:1275-1281 (1989); Proc.
Nat'l Acad. Sci., USA, 88:4363-4366 (1991), the entire contents of
each of which are hereby incorporated by reference. An alternative
method for identifying affinity matured antigen binding molecules
can be found in, for example, U.S. Pat. No. 7,432,063 to Balint et
al., the entire contents of which are hereby incorporated by
reference.
[0160] In some embodiments, the antigen binding molecules and
variant antigen binding molecules of the present invention comprise
a Fc region, preferably a human Fc region. The sequences and
structures of Fc regions are known in the art and have been
characterized. In a specific embodiment, the human constant region
is IgG1, as set forth in SEQ ID NOs 121 and 122.
[0161] However, variants and isoforms of the human Fc region are
also encompassed by the present invention. For example, variant Fc
regions suitable for use in the present invention can be produced
according to the methods taught in U.S. Pat. No. 6,737,056 to
Presta (Fc region variants with altered effector function due to
one or more amino acid modifications); or in U.S. Pat. Appl. Nos.
60/439,498; 60/456,041; 60/514,549; or WO 2004/063351 (variant Fc
regions with increased binding affinity due to amino acid
modification); or in U.S. patent application Ser. No. 10/672,280 or
WO 2004/099249 (Fc variants with altered binding to Fc.gamma.R due
to amino acid modification), the contents of each of which is
herein incorporated by reference in its entirety. In a particular
embodiment, the anti-CEA ABMs and variant ABMs comprise an Fc
region that has been glycoengineered to alter the effector function
activity of the ABM (e.g., decrease fucosylation, improve Fc
receptor binding affinity, increase ADCC, etc.). Methods of
glycoengineering that can be used are described in detail herein
below and are known in the art.
[0162] In one embodiment, the antigen binding molecule or a variant
antigen binding molecule of the present invention is conjugated to
an additional moiety, such as a radiolabel or a toxin. Such
conjugated antigen binding molecules can be produced by numerous
methods that are well known in the art. Anti-CEA ABM conjugates of
the invention are described in detail herein below in the section
entitled "Anti-CEA Antigen Binding Molecule Conjugates."
Expression Vectors and Host Cells
[0163] In one aspect, the present invention is directed to an
expression vector and/or a host cell that comprises one or more
isolated polynucleotides of the present invention. For example, the
host cell or expression vector comprises any one or more of the
polynucleotides or polynucleotides encoding the polypeptides, ABMs
and/or variant ABMs described herein. In another aspect, the
present invention is directed to a method of producing an ABM that
binds membrane-bound human CEA, the method comprising: culturing a
host cell comprising one or more isolated polynucleotides of the
present invention or an expression vector comprising one or more
isolated polynucleotides of the present invention in a medium under
conditions allowing the expression of said one or more
polynucleotide, wherein said one or more polynucleotides encodes
one or more polypeptides that form part of the ABM; and recovering
said ABM, wherein said ABM or a portion thereof binds
membrane-bound CEA.
[0164] Generally, any type of cultured cell line can be used to
express the ABM of the present invention. In a preferred
embodiment, CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO
myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells
or hybridoma cells, other mammalian cells, yeast cells, insect
cells, or plant cells are used as the background cell line to
generate the engineered host cells of the invention.
[0165] In a specific embodiment, the host cell or expression vector
comprises one or more polynucleotides encoding an anti-CEA ABM as
provided herein. In one embodiment, the antibody is affinity
matured. The affinity matured antibody generally has improved
binding affinity than that of the reference antibody from which the
affinity matured antibody is derived. In another embodiment, the
antibody has desirable therapeutic properties including but not
limited to: strong binding affinity for the CEA antigen, in
particular, membrane-bound CEA, while having substantially no
cross-reactivity against soluble CEA; an ability to induce cell
lysis of CEA-expression cells in vitro and ex-vivo, preferably in a
dose-dependent manner; an ability to inhibit CEA mediated cell
adhesion in vitro; an ability to inhibit tumor tissue growth and/or
induce tumor tissue regression in tumor models in mice (e.g.,
xenograft mouse). In another embodiment, the antibody has increased
stability as compared to the parent antibody from which the more
stable antibody is derived. In another embodiment, the variant
antibody or fragment thereof comprises a human Fc.
[0166] In one embodiment, one or several polynucleotides encoding
an ABM of the present invention may be expressed under the control
of a constitutive promoter or, alternately, a regulated expression
system. Suitable regulated expression systems include, but are not
limited to, a tetracycline-regulated expression system, an
ecdysone-inducible expression system, a lac-switch expression
system, a glucocorticoid-inducible expression system, a
temperature-inducible promoter system, and a metallothionein
metal-inducible expression system. If several different nucleic
acids encoding an ABM of the present invention are comprised within
the host cell system, some of them may be expressed under the
control of a constitutive promoter, while others are expressed
under the control of a regulated promoter. The maximal expression
level is considered to be the highest possible level of stable
polypeptide expression that does not have a significant adverse
effect on cell growth rate, and will be determined using routine
experimentation. Expression levels are determined by methods
generally known in the art, including Western blot analysis using
an antibody specific for the ABM or an antibody specific for a
peptide tag fused to the ABM; and Northern blot analysis. In a
further alternative, the polynucleotide may be operatively linked
to a reporter gene; the expression levels of an ABM disclosed
herein are determined by measuring a signal correlated with the
expression level of the reporter gene. The reporter gene may be
transcribed together with the nucleic acid(s) encoding said ABM as
a single mRNA molecule; their respective coding sequences may be
linked either by an internal ribosome entry site (IRES) or by a
cap-independent translation enhancer (CITE). The reporter gene may
be translated together with at least one nucleic acid encoding an
ABM disclosed herein such that a single polypeptide chain is
formed. The nucleic acids encoding an ABM of the present invention
may be operatively linked to the reporter gene under the control of
a single promoter, such that the nucleic acid encoding the ABM and
the reporter gene are transcribed into an RNA molecule which is
alternatively spliced into two separate messenger RNA (mRNA)
molecules; one of the resulting mRNAs is translated into said
reporter protein, and the other is translated into the ABM.
[0167] Methods which are well known to those skilled in the art can
be used to construct expression vectors containing the coding
sequence of an anti-CEA ABM provided herein along with appropriate
transcriptional/translational control signals. These methods
include in vitro recombinant DNA techniques, synthetic techniques
and in vivo recombination/genetic recombination. See, for example,
the techniques described in Maniatis et al., MOLECULAR CLONING: A
LABORATORY MANUAL, Cold Spring Harbor Laboratory, N.Y. (1989) and
Ausubel et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, Greene
Publishing Associates and Wiley Interscience, N.Y (1989).
[0168] A variety of host-expression vector systems may be utilized
to express the coding sequence of the ABMs of the present
invention. Preferably, mammalian cells are used as host cell
systems transfected with recombinant plasmid DNA or cosmid DNA
expression vectors containing the coding sequence of the protein of
interest and the coding sequence of the fusion polypeptide. Most
preferably, CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO
myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells
or hybridoma cells, other mammalian cells, yeast cells, insect
cells, or plant cells are used as host cell system. Some examples
of expression systems and selection methods are described in the
following references and references cited therein: Borth et al.,
Biotechnol. Bioen. 71(4):266-73 (2000-2001), in Werner et al.,
Arzneimittelforschung/Drug Res. 48(8):870-80 (1998), in Andersen
and Krummen, Curr. Op. Biotechnol. 13:117-123 (2002), in Chadd and
Chamow, Curr. Op. Biotechnol. 12:188-194 (2001), and in Giddings,
Curr. Op. Biotechnol. 12: 450-454 (2001).
[0169] In alternate embodiments, other eukaryotic host cell systems
may be used, including yeast cells transformed with recombinant
yeast expression vectors containing the coding sequence of an ABM
of the present invention, such as the expression systems taught in
U.S. Pat. Appl. No. 60/344,169 and WO 03/056914 (methods for
producing human-like glycoprotein in a non-human eukaryotic host
cell) (the contents of each of which are incorporated by reference
in their entirety); insect cell systems infected with recombinant
virus expression vectors (e.g., baculovirus) containing the coding
sequence of an anti-CEA; plant cell systems infected with
recombinant virus expression vectors (e.g., cauliflower mosaic
virus, CaMV; tobacco mosaic virus, TMV) or transformed with
recombinant plasmid expression vectors (e.g., Ti plasmid)
containing the coding sequence of the ABM of the invention
including, but not limited to, the expression systems taught in
U.S. Pat. No. 6,815,184 (methods for expression and secretion of
biologically active polypeptides from genetically engineered
duckweed); WO 2004/057002 (production of glycosylated proteins in
bryophyte plant cells by introduction of a glycosyl transferase
gene) and WO 2004/024927 (methods of generating extracellular
heterologous non-plant protein in moss protoplast); and U.S. Pat.
Appl. Nos. 60/365,769, 60/368,047, and WO 2003/078614 (glycoprotein
processing in transgenic plants comprising a functional mammalian
GnTIII enzyme) (the contents of each of which is herein
incorporated by reference in its entirety); or animal cell systems
infected with recombinant virus expression vectors (e.g.,
adenovirus, vaccinia virus) including cell lines engineered to
contain multiple copies of the DNA encoding a chimeric anti-CEA ABM
either stably amplified (CHO/dhfr) or unstably amplified in
double-minute chromosomes (e.g., murine cell lines). In one
embodiment, the vector comprising the polynucleotide(s) encoding
the ABM of the invention is polycistronic. Also, in one embodiment
the ABM discussed above is an antibody or a fragment thereof. In
one embodiment, the ABM is an affinity matured antibody. In one
embodiment, the ABM is a stability matured antibody.
[0170] Stable expression is generally preferred to transient
expression because it typically achieves more reproducible results
and also is more amenable to large-scale production; however, it is
within the skill of one in the art to determine whether transient
expression is better for a particular situation. Rather than using
expression vectors which contain viral origins of replication, host
cells can be transformed with the respective coding nucleic acids
controlled by appropriate expression control elements (e.g.,
promoter, enhancer, sequences, transcription terminators,
polyadenylation sites, etc.), and a selectable marker. Following
the introduction of foreign DNA, engineered cells may be allowed to
grow for 1-2 days in an enriched media, and then are switched to a
selective media. The selectable marker in the recombinant plasmid
confers resistance to the selection and allows selection of cells
which have stably integrated the plasmid into their chromosomes and
grow to form foci which in turn can be cloned and expanded into
cell lines.
[0171] A number of selection systems may be used, including, but
not limited to, the herpes simplex virus thymidine kinase (Wigler
et al., Cell 11:223 (1977)), hypoxanthine-guanine
phosphoribosyltransferase (Szybalska & Szybalski, Proc. Natl.
Acad. Sci. USA 48:2026 (1962)), and adenine
phosphoribosyltransferase (Lowy et al., Cell 22:817 (1980)) genes,
which can be employed in tk.sup.-, hgprt.sup.- or aprt.sup.- cells,
respectively. Also, antimetabolite resistance can be used as the
basis of selection for dhfr, which confers resistance to
methotrexate (Wigler et al., Natl. Acad. Sci. USA 77:3567 (1989);
O'Hare et al., Proc. Natl. Acad. Sci. USA 78:1527 (1981)); gpt,
which confers resistance to mycophenolic acid (Mulligan & Berg,
Proc. Natl. Acad. Sci. USA 78:2072 (1981)); neo, which confers
resistance to the aminoglycoside G-418 (Colberre-Garapin et al., J.
Mol. Biol. 150:1 (1981)); and hygro, which confers resistance to
hygromycin (Santerre et al., Gene 30:147 (1984) genes. Recently,
additional selectable genes have been described, namely trpB, which
allows cells to utilize indole in place of tryptophan; hisD, which
allows cells to utilize histinol in place of histidine (Hartman
& Mulligan, Proc. Natl. Acad. Sci. USA 85:8047 (1988)); the
glutamine synthase system; and ODC (ornithine decarboxylase) which
confers resistance to the ornithine decarboxylase inhibitor,
2-(difluoromethyl)-DL-ornithine, DFMO (McConlogue, in: Current
Communications in Molecular Biology, Cold Spring Harbor Laboratory
ed. (1987)).
[0172] The present invention is further directed to a method for
modifying the glycosylation profile of the ABMs of the present
invention that are produced by a host cell, comprising expressing
in said host cell a nucleic acid encoding an ABM of the invention
and a nucleic acid encoding a polypeptide with a
glycosyltransferase activity, or a vector comprising such nucleic
acids. Genes with glycosyltransferase activity include
.beta.(1,4)-N-acetylglucosaminyltransferase III (GnTII),
a-mannosidase II (ManII), .beta.(1,4)-galactosyltransferase (GalT),
.beta.(1,2)-N-acetylglucosaminyltransferase I (GnTI), and
.beta.(1,2)-N-acetylglucosaminyltransferase II (GnTII). In one
embodiment, a combination of genes with glycosyltransferase
activity are expressed in the host cell (e.g., GnTIII and Man II).
Likewise, the method also encompasses expression of one or more
polynucleotide(s) encoding the ABM in a host cell in which a
glycosyltransferase gene has been disrupted or otherwise
deactivated (e.g., a host cell in which the activity of the gene
encoding .alpha.1-6 core fucosyltransferase has been knocked out).
In another embodiment, the ABMs of the present invention can be
produced in a host cell that further expresses a polynucleotide
encoding a polypeptide having GnTIII activity to modify the
glycosylation pattern. In a specific embodiment, the polypeptide
having GnTIII activity is a fusion polypeptide comprising the Golgi
localization domain of a Golgi resident polypeptide. In another
preferred embodiment, the expression of the ABMs of the present
invention in a host cell that expresses a polynucleotide encoding a
polypeptide having GnTIII activity results in ABMs with increased
Fc receptor binding affinity and increased effector function.
Accordingly, in one embodiment, the present invention is directed
to a host cell comprising (a) an isolated nucleic acid comprising a
sequence encoding a polypeptide having GnTIII activity; and (b) an
isolated polynucleotide encoding an ABM of the present invention,
such as a chimeric, primatized or humanized antibody that binds
human CEA. In a preferred embodiment, the polypeptide having GnTIII
activity is a fusion polypeptide comprising the catalytic domain of
GnTIII and the Golgi localization domain is the localization domain
of mannosidase II. Methods for generating such fusion polypeptides
and using them to produce antibodies with increased effector
functions are disclosed in U.S. Provisional Pat. Appl. No.
60/495,142 and U.S. Pat. Appl. Publ. No. 2004/0241817, the entire
contents of which are expressly incorporated herein by reference.
In a particular embodiment, the modified ABM produced by the host
cell has an IgG constant region or a fragment thereof comprising
the Fc region. In another particular embodiment the ABM is a
humanized antibody or a fragment thereof comprising an Fc
region.
[0173] The ABMs with altered glycosylation produced by the host
cells of the invention typically exhibit increased Fc receptor
binding affinity and/or increased effector function as a result of
the modification of the host cell (e.g., by expression of a
glycosyltransferase gene). Preferably, the increased Fc receptor
binding affinity is increased binding to a Fc.gamma. activating
receptor, such as the Fc.gamma.RIIIa receptor. The increased
effector function is preferably an increase in one or more of the
following: increased antibody-dependent cellular cytotoxicity,
increased antibody-dependent cellular phagocytosis (ADCP),
increased cytokine secretion, increased immune-complex-mediated
antigen uptake by antigen-presenting cells, increased Fc-mediated
cellular cytotoxicity, increased binding to NK cells, increased
binding to macrophages, increased binding to polymorphonuclear
cells (PMNs), increased binding to monocytes, increased
crosslinking of target-bound antibodies, increased direct signaling
inducing apoptosis, increased dendritic cell maturation, and
increased T cell priming.
Generation and Use of ABMs Having Increased Effector Function
Including Antibody-Dependent Cellular Cytotoxicity
[0174] In one aspect, the present invention provides glycoforms of
anti-CEA ABMs (e.g., variant ABMs) having increased effector
function, including antibody-dependent cellular cytotoxicity.
Glycosylation engineering of antibodies has been previously
described. See, e.g., U.S. Pat. No. 6,602,684, incorporated herein
by reference in its entirety. Methods of producing ABMs from host
cells that have altered activity of genes involved in glycosylation
are also described herein in detail (See, e.g, preceding section
entitled "Expression Vectors and Host Cells"). Increases in ADCC of
the ABMs of the present invention is also achieved by increasing
affinity of the antigen binding molecule for membrane-bound CEA,
for example by affinity maturation or other methods of improving
affinity (see Tang et al., J. Immunol. 2007, 179:2815-2823).
Combinations of these approaches are also encompassed by the
present invention.
[0175] Clinical trials of unconjugated monoclonal antibodies (mAbs)
for the treatment of some types of cancer have recently yielded
encouraging results. Dillman, Cancer Biother. & Radiopharm.
12:223-25 (1997); Deo et al., Immunology Today 18:127 (1997). A
chimeric, unconjugated IgG1 has been approved for low-grade or
follicular B-cell non-Hodgkin's lymphoma. Dillman, Cancer Biother.
& Radiopharm. 12:223-25 (1997), while another unconjugated mAb,
a humanized IgG1 targeting solid breast tumors, has also showed
promising results in phase III clinical trials. Deo et al.,
Immunology Today 18:127 (1997). The antigens of these two mAbs are
highly expressed in their respective tumor cells and the antibodies
mediate potent tumor destruction by effector cells in vitro and in
vivo. In contrast, many other unconjugated mAbs with fine tumor
specificities cannot trigger effector functions of sufficient
potency to be clinically useful. Frost et al., Cancer 80:317-33
(1997); Surfus et al., J. Immunother. 19:184-91 (1996). For some of
these weaker mAbs, adjunct cytokine therapy is currently being
tested. Addition of cytokines can stimulate antibody-dependent
cellular cytotoxicity (ADCC) by increasing the activity and number
of circulating lymphocytes. Frost et al., Cancer 80:317-33 (1997);
Surfus et al., J. Immunother. 19:184-91 (1996). ADCC, a lytic
attack on targeted cells, is triggered upon binding of leukocyte
receptors to the constant region (Fc) of antibodies. Deo et al.,
Immunology Today 18:127 (1997).
[0176] A different, but complementary, approach to increase ADCC
activity of unconjugated IgG1 s is to engineer the Fc region of the
antibody. Protein engineering studies have shown that Fc.gamma.Rs
interact with the lower hinge region of the IgG CH2 domain. Lund et
al., J. Immunol. 157:4963-69 (1996). However, Fc.gamma.R binding
also requires the presence of oligosaccharides covalently attached
at the conserved Asn 297 in the CH2 region. Lund et al., J.
Immunol. 157:4963-69 (1996); Wright and Morrison, Trends Biotech.
15:26-31 (1997), suggesting that either oligosaccharide and
polypeptide both directly contribute to the interaction site or
that the oligosaccharide is required to maintain an active CH2
polypeptide conformation. Modification of the oligosaccharide
structure can therefore be explored as a means to increase the
affinity of the interaction.
[0177] An IgG molecule carries two N-linked oligosaccharides in its
Fc region, one on each heavy chain. As any glycoprotein, an
antibody is produced as a population of glycoforms which share the
same polypeptide backbone but have different oligosaccharides
attached to the glycosylation sites. The oligosaccharides normally
found in the Fc region of serum IgG are of complex bi-antennary
type (Wormald et al., Biochemistry 36:130-38 (1997), with a low
level of terminal sialic acid and bisecting N-acetylglucosamine
(GlcNAc), and a variable degree of terminal galactosylation and
core fucosylation. Some studies suggest that the minimal
carbohydrate structure required for Fc.gamma.R binding lies within
the oligosaccharide core. Lund et al., J. Immunol. 157:4963-69
(1996)
[0178] The mouse- or hamster-derived cell lines used in industry
and academia for production of unconjugated therapeutic mAbs
normally attach the required oligosaccharide determinants to Fc
sites. IgGs expressed in these cell lines lack, however, the
bisecting GlcNAc found in low amounts in serum IgGs. Lifely et al.,
Glycobiology 318:813-22 (1995). In contrast, it was recently
observed that a rat myeloma-produced, humanized IgG1 (CAMPATH-1H)
carried a bisecting GlcNAc in some of its glycoforms. Lifely et
al., Glycobiology 318:813-22 (1995). The rat cell-derived antibody
reached a similar maximal in vitro ADCC activity as CAMPATH-1H
antibodies produced in standard cell lines, but at significantly
lower antibody concentrations.
[0179] The CAMPATH antigen is normally present at high levels on
lymphoma cells, and this chimeric mAb has high ADCC activity in the
absence of a bisecting GlcNAc. Lifely et al., Glycobiology
318:813-22 (1995). In the N-linked glycosylation pathway, a
bisecting GlcNAc is added by GnTIII. Schachter, Biochem. Cell Biol.
64:163-81 (1986).
[0180] Previous studies used a single, antibody-producing CHO cell
line that was previously engineered to express, in an
externally-regulated fashion, different levels of a cloned GnTIII
enzyme gene (Umana, P., et al., Nature Biotechnol. 17:176-180
(1999)). This approach established for the first time a rigorous
correlation between expression of a glycosyltransferase (e.g.,
GnTIII) and the ADCC activity of the modified antibody. Thus, the
invention contemplates a variant ABM (e.g., an affinity matured
ABM) that binds membrane-bound CEA, comprising an Fc region or
region equivalent to an Fc region having altered glycosylation
resulting from changing the expression level of a
glycosyltransferase gene in the ABM-producing host cell. In a
specific embodiment, the change in gene expression level is an
increase in GnTIII activity. Increased GnTIII activity results in
an increase in the percentage of bisected oligosaccharides, as well
as a decrease in the percentage of fucose residues, in the Fc
region of the ABM. This antibody, or fragment thereof, has
increased Fc receptor binding affinity and increased effector
function
[0181] The present invention is also directed to a method for
producing an anti-CEA ABM of the present invention having modified
oligosaccharides, comprising (a) culturing a host cell engineered
to express at least one nucleic acid encoding a polypeptide having
glycosyltransferase activity under conditions which permit the
production of an ABM according to the present invention, wherein
said polypeptide having glycosyltransferase activity is expressed
in an amount sufficient to modify the oligosaccharides in the Fc
region of said ABM produced by said host cell; and (b) isolating
said ABM. In one embodiment, the polypeptide having
glycosyltransferase activity is GnTIII. In another embodiment,
there are two polypeptides having glycosyltransferase activity. In
a particular embodiment, the two peptides having
glycosyltransferase activity are GnTIII and ManII In another
embodiment, the polypeptide having glycosltransferase activity is a
fusion polypeptide comprising the catalytic domain of GnTIII. In a
more specific embodiment, the fusion polypeptide further comprises
the Golgi localization domain of a Golgi resident polypeptide.
Preferably, the Golgi localization domain is the localization
domain of mannosidase II or GnTI. Alternatively, the Golgi
localization domain is selected from the group consisting of: the
localization domain of mannosidase I, the localization domain of
GnTII, and the localization domain of a 1-6 core
fucosyltransferase. The ABMs produced by the methods of the present
invention have increased Fc receptor binding affinity and/or
increased effector function. Generally, the increased effector
function is one or more of the following: increased Fc-mediated
cellular cytotoxicity (including increased antibody-dependent
cellular cytotoxicity), increased antibody-dependent cellular
phagocytosis (ADCP), increased cytokine secretion, increased
immune-complex-mediated antigen uptake by antigen-presenting cells,
increased binding to NK cells, increased binding to macrophages,
increased binding to monocytes, increased binding to
polymorphonuclear cells, increased direct signaling inducing
apoptosis, increased crosslinking of target-bound antibodies,
increased dendritic cell maturation, or increased T cell priming.
The increased Fc receptor binding affinity is preferably increased
binding to Fc activating receptors such as Fc.gamma.RIIIa. In a
particularly preferred embodiment the ABM is a humanized antibody
or a fragment thereof.
[0182] In one embodiment, the percentage of bisected N-linked
oligosaccharides in the Fc region of the ABM is at least about 10%
to about 100%, specifically at least about 50%, more specifically,
at least about 60%, at least about 70%, at least about 80%, or at
least about 90-95% of the total oligosaccharides. In yet another
embodiment, the antigen binding molecule or variant antigen binding
molecule produced by the methods of the invention has an increased
proportion of nonfucosylated oligosaccharides in the Fc region as a
result of the modification of its oligosaccharides by the methods
of the present invention. In one embodiment, the percentage of
nonfucosylated oligosaccharides is at least about 20% to about
100%, specifically at least about 50%, at least about 60% to about
70%, and more specifically, at least about 75%. The nonfucosylated
oligosaccharides may be of the hybrid or complex type. In yet
another embodiment, the antigen binding molecule or variant antigen
binding molecule produced by the methods of the invention has an
increased proportion of bisected oligosaccharides in the Fc region
as a result of the modification of its oligosaccharides by the
methods of the present invention. In one embodiment, the percentage
of bisected oligosaccharides is at least about 20% to about 100%,
specifically at least about 50%, at least about 60% to about 70%,
and more specifically, at least about 75%. In a particularly
preferred embodiment, the ABM produced by the host cells and
methods of the invention has an increased proportion of bisected,
nonfucosylated oligosaccharides in the Fc region. The bisected,
nonfucosylated oligosaccharides may be either hybrid or complex.
Specifically, the methods of the present invention may be used to
produce antigen binding molecules in which at least about 10% to
about 100%, specifically at least about 15%, more specifically at
least about 20% to about 50%, more specifically at least about 20%
to about 25%, and more specifically at least about 30% to about 35%
of the oligosaccharides in the Fc region of the antigen binding
molecule or variant antigen binding molecule are bisected,
nonfucosylated. The ABMs of the present invention may also comprise
an Fc region in which at least about 10% to about 100%,
specifically at least about 15%, more specifically at least about
20% to about 25%, and more specifically at least about 30% to about
35% of the oligosaccharides in the Fc region of the ABM are
bisected hybrid nonfucosylated.
[0183] In another embodiment, the present invention is directed to
an anti-CEA antigen binding molecule (e.g., a variant ABM))
engineered to have increased effector function and/or increased Fc
receptor binding affinity, produced by the methods of the
invention. The increased effector function can include, but is not
limited to one or more of the following: increased Fc-mediated
cellular cytotoxicity (including increased antibody-dependent
cellular cytotoxicity), increased antibody-dependent cellular
phagocytosis (ADCP), increased cytokine secretion, increased
immune-complex-mediated antigen uptake by antigen-presenting cells,
increased binding to NK cells, increased binding to macrophages,
increased binding to monocytes, increased binding to
polymorphonuclear cells, increased direct signaling inducing
apoptosis, increased crosslinking of target-bound antibodies,
increased dendritic cell maturation, or increased T cell priming.
In a preferred embodiment, the increased Fc receptor binding
affinity is increased binding to an Fc activating receptor, most
preferably Fc.gamma.RIIIa. In one embodiment, the antigen binding
molecule or variant antigen binding molecule is an antibody, an
antibody fragment containing the Fc region, or a fusion protein
that includes a region equivalent to the Fc region of an
immunoglobulin. In a particularly preferred embodiment, the antigen
binding molecule or variant antigen binding molecule is a humanized
affinity matured antibody.
[0184] The present invention further provides methods for the
generation and use of host cell systems for the production of
glycoforms of the ABMs of the present invention, having increased
Fc receptor binding affinity, preferably increased binding to Fc
activating receptors, and/or having increased effector functions,
including antibody-dependent cellular cytotoxicity. The
glycoengineering methodology that can be used with the ABMs of the
present invention has been described in greater detail in U.S. Pat.
No. 6,602,684, U.S. Pat. Appl. Publ. No. 2004/0241817 A1, U.S. Pat.
Appl. Publ. No. 2003/0175884 A1, Provisional U.S. Patent
Application No. 60/441,307 and WO 2004/065540, the entire contents
of each of which is incorporated herein by reference in its
entirety. The ABMs of the present invention can alternatively be
glycoengineered to have reduced fucose residues in the Fc region
according to the techniques disclosed in U.S. Pat. Appl. Pub. No.
2003/0157108 (Genentech), or in EP 1 176 195 A1, WO 03/084570, WO
03/085119 and U.S. Pat. Appl. Pub. Nos. 2003/0115614, 2004/093621,
2004/110282, 2004/110704, 2004/132140 (Kyowa). The contents of each
of these documents are herein incorporated by reference in their
entireties. Glycoengineered ABMs of the invention may also be
produced in expression systems that produce modified glycoproteins,
such as those taught in U.S. Pat. Appl. Pub. No. 60/344,169 and WO
03/056914 (GlycoFi, Inc.) or in WO 2004/057002 and WO 2004/024927
(Greenovation), the contents of each of which are hereby
incorporated by reference in their entirety.
Generation of Cell Lines for the Production of Proteins with
Altered Glycosylation Pattern
[0185] In one aspect, the present invention provides host cell
expression systems for the generation of the ABMs of the present
invention having modified glycosylation patterns. In particular,
the present invention provides host cell systems for the generation
of glycoforms of the ABMs of the present invention having an
improved therapeutic value. Therefore, the invention provides host
cell expression systems selected or engineered to express a
polypeptide having a glycosyltransferase activity. In a specific
embodiment, the glycosyltransferase activity is a GnTIII activity.
In one embodiment, the polypeptide having GnTIII activity is a
fusion polypeptide comprising the Golgi localization domain of a
heterologous Golgi resident polypeptide. Specifically, such host
cell expression systems may be engineered to comprise a recombinant
nucleic acid molecule encoding a polypeptide having GnTIII,
operatively linked to a constitutive or regulated promoter
system.
[0186] In one specific embodiment, the present invention provides a
host cell that has been engineered to express at least one nucleic
acid encoding a fusion polypeptide having GnTIII activity and
comprising the Golgi localization domain of a heterologous Golgi
resident polypeptide. In one aspect, the host cell is engineered
with a nucleic acid molecule comprising at least one gene encoding
a fusion polypeptide having GnTIII activity and comprising the
Golgi localization domain of a heterologous Golgi resident
polypeptide.
[0187] Generally, any type of cultured cell line, including the
cell lines discussed above, can be used as a background to engineer
the host cell lines of the present invention. In a preferred
embodiment, CHO cells, BHK cells, NS0 cells, SP2/0 cells, YO
myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells
or hybridoma cells, other mammalian cells, yeast cells, insect
cells, or plant cells are used as the background cell line to
generate the engineered host cells of the invention.
[0188] The invention is contemplated to encompass any engineered
host cells expressing a polypeptide having glycosyltransferase
activity, e.g., GnTIII activity, including a fusion polypeptide
that comprises the Golgi localization domain of a heterologous
Golgi resident polypeptide as defined herein.
[0189] One or several nucleic acids encoding a polypeptide having
glycosyltransferase activity, e.g., GnTIII activity, may be
expressed under the control of a constitutive promoter or,
alternately, a regulated expression system. Such systems are well
known in the art, and include the systems discussed above. If
several different nucleic acids encoding fusion polypeptides having
glycosyltransferase activity, e.g., GnTIII activity, and comprising
the Golgi localization domain of a heterologous Golgi resident
polypeptide are comprised within the host cell system, some of them
may be expressed under the control of a constitutive promoter,
while others are expressed under the control of a regulated
promoter. Expression levels of the fusion polypeptides having
glycosyltransferase activity, e.g., GnTIII activity, are determined
by methods generally known in the art, including Western blot
analysis, Northern blot analysis, reporter gene expression analysis
or measurement of glycosyltransferase activity, e.g., GnTIII
activity. Alternatively, a lectin may be employed which binds to
biosynthetic products of the GnTIII, for example, E.sub.4-PHA
lectin. Alternatively, a functional assay which measures the
increased Fc receptor binding or increased effector function
mediated by antibodies produced by the cells engineered with the
nucleic acid encoding a polypeptide with glycosyltransferase
activity, e.g., GnTIII activity, may be used.
Identification of Transfectants or Transformants that Express the
Protein Having a Modified Glycosylation Pattern
[0190] The host cells which contain the coding sequence of a
variant anti-CEA ABM (e.g., a humanized, affinity matured and/or
stability matured ABM) and which express the biologically active
gene products may be identified by at least four general
approaches; (a) DNA-DNA or DNA-RNA hybridization; (b) the presence
or absence of "marker" gene functions; (c) assessing the level of
transcription as measured by the expression of the respective mRNA
transcripts in the host cell; and (d) detection of the gene product
as measured by immunoassay or by its biological activity.
[0191] In the first approach, the presence of the coding sequence
of a variant anti-CEA and/or the coding sequence of the polypeptide
having glycosyltransferase (e.g., GnTIII) activity can be detected
by DNA-DNA or DNA-RNA hybridization using probes comprising
nucleotide sequences that are homologous to the respective coding
sequences, respectively, or portions or derivatives thereof.
[0192] In the second approach, the recombinant expression
vector/host system can be identified and selected based upon the
presence or absence of certain "marker" gene functions (e.g.,
thymidine kinase activity, resistance to antibiotics, resistance to
methotrexate, transformation phenotype, occlusion body formation in
baculovirus, etc.). For example, if the coding sequence of the ABM
of the invention, or a fragment thereof, and/or the coding sequence
of the polypeptide having glycosyltransferase (e.g., GnTIII)
activity are inserted within a marker gene sequence of the vector,
recombinants containing the respective coding sequences can be
identified by the absence of the marker gene function.
Alternatively, a marker gene can be placed in tandem with the
coding sequences under the control of the same or different
promoter used to control the expression of the coding sequences.
Expression of the marker in response to induction or selection
indicates expression of the coding sequence of the ABM of the
invention and/or the coding sequence of the polypeptide having
glycosyltransferase (e.g., GnTIII) activity.
[0193] In the third approach, transcriptional activity for the
coding region of the ABM of the invention, or a fragment thereof,
and/or the coding sequence of the polypeptide having
glycosyltransferase (e.g., GnTIII) activity can be assessed by
hybridization assays. For example, RNA can be isolated and analyzed
by Northern blot using a probe homologous to the coding sequences
of the ABM of the invention, or a fragment thereof, and/or the
coding sequence of the polypeptide having glycosyltransferase
(e.g., GnTIII) activity or particular portions thereof.
Alternatively, total nucleic acids of the host cell may be
extracted and assayed for hybridization to such probes.
[0194] In the fourth approach, the expression of the protein
products can be assessed immunologically, for example by Western
blots, immunoassays such as radioimmuno-precipitation,
enzyme-linked immunoassays and the like. The ultimate test of the
success of the expression system, however, involves the detection
of the biologically active gene products.
Therapeutic Applications and Methods of Using Anti-CEA of Antigen
Binding Molecules
[0195] The invention is also directed to a method for targeting in
vivo or in vitro cells expressing CEA. Cells that express CEA may
be targeted for therapeutic purposes (e.g., to treat a disorder by
targeting CEA-expressing cells for destruction by the immune
system). In one embodiment, the present invention is directed to a
method for targeting cells expressing CEA in a subject comprising
administering to the subject a composition comprising an ABM of the
invention. Cells that express CEA may also be targeted for
diagnostic purposes (e.g., to determine if they are expressing CEA,
either normally or abnormally) Thus, the invention is also directed
to methods for detecting the presence of CEA or a cell expressing
CEA, either in vivo or in vitro. One method of detecting CEA
expression according to the present invention comprises contacting
a sample to be tested, optionally with a control sample, with an
ABM of the present invention, under conditions that allow for
formation of a complex between the ABM and CEA. The complex
formation is then detected (e.g., by ELISA or other methods known
in the art). When using a control sample with the test sample, any
statistically significant difference in the formation of ABM-CEA
complexes when comparing the test and control samples is indicative
of the presence of CEA in the test sample.
[0196] In one aspect, ABMs and/or variant ABMs of the present
invention can be used target cells in vivo or in vitro that express
CEA. The cells expressing CEA can be targeted for diagnostic or
therapeutic purposes. In one aspect, the ABMs of the present
invention can be used to detect the presence of CEA in a sample.
CEA is abnormally expressed (e.g., overexpressed) in many human
tumors compared to non-tumor tissue of the same cell type. Thus,
the ABMs and/or variant ABMs of the invention are particularly
useful in the prevention of tumor formation, eradication of tumors
and inhibition of tumor growth or metastasis. The ABMs and/or
variant ABMs of the invention also act to arrest the cell cycle,
cause apoptosis of the target cells (e.g., tumor cells), and
inhibit angiogenesis and/or differentiation of target cells. The
ABMs and/or variant ABMs of the invention can be used to treat any
tumor expressing CEA. Particular malignancies that can be treated
with the ABMs and/or variant ABMs of the invention include, but are
not limited to, colorectal cancer, non-small cell lung cancer,
gastric cancer, pancreatic cancer and breast cancer.
[0197] The anti-CEA ABMs and/or variant ABMs disclosed herein can
be used alone to inhibit tumor growth or kill tumor cells. For
example, the anti-CEA ABMs can bind to CEA that is on the membrane
or cell surface of cancerous cells and elicit, e.g., ADCC or other
effector mediated killing of the cancerous cells. The anti-CEA ABMs
and/or variant ABMs can be humanized, specifically, affinity and/or
stability matured, more specifically, glycoengineered, stability,
and affinity matured.
[0198] The ABMs and/or variant ABMs can alternatively be used alone
in order to block the activity of the CEA antigen, particularly by
physically interfering with its binding of another compound. For
example, the antigen binding molecules and variant antigen binding
molecules can be used to block CEA mediated cell-adhesion.
[0199] The anti-CEA ABMs and/or variant ABMs of the invention are
administered to a mammal, preferably a human, in a pharmaceutically
acceptable dosage form such as those discussed below, including
those that may be administered to a human intravenously as a bolus
or by continuous infusion over a period of time, by intramuscular,
intraperitoneal, intra-cerebrospinal, subcutaneous,
intra-articular, intrasynovial, intrathecal, oral, topical, or
inhalation routes. The ABMs also are suitably administered by intra
tumoral, peritumoral, intralesional, or perilesional routes, to
exert local as well as systemic therapeutic effects. The
intraperitoneal route is expected to be particularly useful, for
example, in the treatment of colorectal tumors.
[0200] For the treatment of disease, the appropriate dosage of ABM
and/or variant ABM will depend on the type of disease to be
treated, the severity and course of the disease, previous therapy,
the patient's clinical history and response to the antibody, and
the discretion of the attending physician. The ABM is suitably
administered to the patient at one time or over a series of
treatments.
[0201] The present invention provides a method for selectively
killing tumor cells expressing CEA. This method comprises reacting
the antigen binding molecules or the conjugates (e.g., the
immunotoxin) of the invention with said tumor cells. These tumor
cells may be from a human carcinoma including colorectal carcinoma,
non-small cell lung carcinoma (NSCLC), gastric carcinoma,
pancreatic carcinoma and breast carcinoma.
[0202] In one embodiment, the present invention provides a method
inhibiting CEA-mediated cell adhesion of a tumor cell. This method
comprises contacting said tumor cell with the antigen binding
molecules or variant antigen binding molecules of the invention or
the conjugates thereof. These tumor cells may be from human cells,
including colorectal cancer cells, non-small cell lung cancer cells
(NSCLC), gastric cancer cells, pancreatic cancer cells and breast
cancer cells.
[0203] Additionally, this invention provides a method of treating
carcinomas (for example, human carcinomas) in vivo. This method
comprises administering to a subject a pharmaceutically effective
amount of a composition containing at least one of the antigen
binding molecules or the immunoconjugates (e.g., the immunotoxin)
of the invention.
[0204] In a further aspect, the invention is directed to a method
for treating cancers characterized by CEA over-expression,
including but not limited to colorectal cancer cells, NSCLC
(non-small cell lung cancer), gastric cancer cells, pancreatic
cancer cells and breast cancer cells, by administering a
therapeutically effective amount of the anti-CEA antigen binding
molecules or variant antigen binding molecules disclosed
herein.
[0205] In a further embodiment, the invention is directed to a
method for inducing tumor tissue regression in a subject using
anti-CEA antigen binding molecules or variant antigen binding
molecules disclosed herein. Non-limiting examples of the tumor
tissue includes colorectal tumor, non-small cell lung tumor,
gastric tumor, pancreatic tumor and breast tumor. In a particular
embodiment, the tumor tissue is a colorectal tumor.
[0206] In accordance with the practice of this invention, the
subject may be a human, equine, porcine, bovine, murine, canine,
feline, and avian subjects. Other warm blooded animals are also
included in this invention.
[0207] The invention further provides methods for inhibiting the
growth of tumor cells, treating a tumor in a subject, and treating
a proliferative type disease in a subject. These methods comprise
administering to the subject an effective amount of the composition
of the invention.
[0208] In another aspect, the invention is directed to the use of
the anti-CEA antigen binding molecules or variant antigen binding
molecules disclosed herein for the manufacture of a medicament for
treating a disease related to abnormal CEA expression. In a
particular embodiment, the disease is a cancer that overexpresses
CEA, including but not limited to colorectal tumor, non-small cell
lung tumor, gastric tumor, pancreatic tumor and breast tumor. In a
particular embodiment, the tumor is a colorectal tumor.
Anti-CEA Antigen Binding Molecule Conjugates
[0209] The invention also provides immunoconjugates comprising an
anti-CEA ABM or variant ABM herein conjugated to one or more
cytotoxic agents, such as chemotherapeutic agents or drugs, growth
inhibitory agents, toxins (e.g., protein toxins, enzymatically
active toxins of bacterial, fungal, plant, or animal origin, or
fragments thereof), or radioactive isotopes.
[0210] In one embodiment, an immunoconjugate is an antibody-drug
conjugate (ADC) in which an antibody is conjugated to one or more
drugs, including but not limited to a maytansinoid (see U.S. Pat.
Nos. 5,208,020, 5,416,064 and European Patent EP 0 425 235 B1); an
auristatin such as monomethylauristatin drug moieties DE and DF
(MMAE and MMAF) (see U.S. Pat. Nos. 5,635,483 and 5,780,588, and
7,498,298); a dolastatin; a calicheamicin or derivative thereof
(see U.S. Pat. Nos. 5,712,374, 5,714,586, 5,739,116, 5,767,285,
5,770,701, 5,770,710, 5,773,001, and 5,877,296; Hinman et al.,
Cancer Res. 53:3336-3342 (1993); and Lode et al., Cancer Res.
58:2925-2928 (1998)); an anthracycline such as daunomycin or
doxorubicin (see Kratz et al., Current Med. Chem. 13:477-523
(2006); Jeffrey et al., Bioorganic & Med. Chem. Letters
16:358-362 (2006); Torgov et al., Bioconj. Chem. 16:717-721 (2005);
Nagy et al., Proc. Natl. Acad. Sci. USA 97:829-834 (2000);
Dubowchik et al., Bioorg. & Med. Chem. Letters 12:1529-1532
(2002); King et al., J. Med. Chem. 45:4336-4343 (2002); and U.S.
Pat. No. 6,630,579); methotrexate; vindesine; a taxane such as
docetaxel, paclitaxel, larotaxel, tesetaxel, and ortataxel; a
trichothecene; and CC1065.
[0211] In another embodiment, an immunoconjugate comprises an
anti-CEA ABM or variant ABM as described herein conjugated to an
enzymatically active toxin or fragment thereof, including but not
limited to diphtheria A chain, nonbinding active fragments of
diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa),
ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin,
Aleurites fordii proteins, dianthin proteins, Phytolaca americana
proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor,
curcin, crotin, Sapaonaria officinalis inhibitor, gelonin,
mitogellin, restrictocin, phenomycin, enomycin, and the
tricothecenes.
[0212] In another embodiment, an immunoconjugate comprises an
anti-CEA ABM or variant ABM as described herein conjugated to a
radioactive atom to form a radioconjugate. A variety of radioactive
isotopes are available for the production of radioconjugates.
Examples include At.sup.211, I.sup.131, I.sup.125, Y.sup.90,
Re.sup.186, Re.sup.188, Sm.sup.153, Bi.sup.212, P.sup.32, P.sup.212
and radioactive isotopes of Lu. When the radioconjugate is used for
detection, it may comprise a radioactive atom for scintigraphic
studies, for example tc99m or 1123, or a spin label for nuclear
magnetic resonance (NMR) imaging (also known as magnetic resonance
imaging, mri), such as iodine-123 again, iodine-131, indium-111,
fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium,
manganese or iron.
[0213] Conjugates of an antibody and cytotoxic agent may be made
using a variety of bifunctional protein coupling agents such as
N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP),
succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate
(SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters
(such as dimethyl adipimidate HCl), active esters (such as
disuccinimidyl suberate), aldehydes (such as glutaraldehyde),
bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine),
bis-diazonium derivatives (such as
bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as
toluene 2,6-diisocyanate), and bis-active fluorine compounds (such
as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin
immunotoxin can be prepared as described in Vitetta et al., Science
238:1098 (1987). Carbon-14-labeled
1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid
(MX-DTPA) is an exemplary chelating agent for conjugation of
radionucleotide to the antibody. See WO94/11026. The linker may be
a "cleavable linker" facilitating release of a cytotoxic drug in
the cell. For example, an acid-labile linker, peptidase-sensitive
linker, photolabile linker, dimethyl linker or disulfide-containing
linker (Chari et al., Cancer Res. 52:127-131 (1992); U.S. Pat. No.
5,208,020) may be used.
[0214] The immunuoconjugates or ADCs herein expressly contemplate,
but are not limited to such conjugates prepared with cross-linker
reagents including, but not limited to, BMPS, EMCS, GMBS, HBVS,
LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS,
sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, and
sulfo-SMPB, and SVSB (succinimidyl-(4-vinylsulfone)benzoate) which
are commercially available (e.g., from Pierce Biotechnology, Inc.,
Rockford, Ill., U.S.A).
Compositions, Formulations, Dosages, and Routes of
Administration
[0215] In one aspect, the present invention is directed to
pharmaceutical compositions comprising the anti-CEA ABMs or variant
ABMs of the present invention and a pharmaceutically acceptable
carrier. The present invention is further directed to the use of
such pharmaceutical compositions in the method of treatment of
disease, such as cancer, or in the manufacture of a medicament for
the treatment of disease, such as cancer. Specifically, the present
invention is directed to a method for the treatment of disease, and
more particularly, for the treatment of cancer, the method
comprising administering a therapeutically effective amount of the
pharmaceutical composition of the invention.
[0216] In one aspect, the present invention encompasses
pharmaceutical compositions, combinations and methods for treating
human carcinomas, for example colorectal carcinoma. For example,
the invention includes pharmaceutical compositions for use in the
treatment of human carcinomas comprising a pharmaceutically
effective amount of an antibody of the present invention and a
pharmaceutically acceptable carrier.
[0217] The ABM compositions of the invention can be administered
using conventional modes of administration including, but not
limited to, intravenous, intraperitoneal, oral, intralymphatic or
administration directly into the tumor. Intravenous administration
is preferred.
[0218] In one aspect of the invention, therapeutic formulations
containing the ABMs of the invention are prepared for storage by
mixing an antibody having the desired degree of purity with
optional pharmaceutically acceptable carriers, excipients or
stabilizers (Remington's Pharmaceutical Sciences 16th edition,
Osol, A. Ed. (1980)), in the form of lyophilized formulations or
aqueous solutions. Acceptable carriers, excipients, or stabilizers
are nontoxic to recipients at the dosages and concentrations
employed.
[0219] The formulations to be used for in vivo administration must
be sterile. This is readily accomplished by filtration through
sterile filtration membranes.
[0220] The most effective mode of administration and dosage regimen
for the pharmaceutical compositions of this invention depends upon
the severity and course of the disease, the patient's health and
response to treatment and the judgment of the treating physician.
Accordingly, the dosages of the compositions should be titrated to
the individual patient. Nevertheless, an effective dose of the
compositions of this invention will generally be in the range of
from about 0.01 to about 2000 mg/kg.
[0221] The molecules described herein may be in a variety of dosage
forms which include, but are not limited to, liquid solutions or
suspensions, tablets, pills, powders, suppositories, polymeric
microcapsules or microvesicles, liposomes, and injectable or
infusible solutions. The preferred form depends upon the mode of
administration and the therapeutic application.
[0222] The composition comprising an ABM of the present invention
will be formulated, dosed, and administered in a fashion consistent
with good medical practice. Factors for consideration in this
context include the particular disease or disorder being treated,
the particular mammal being treated, the clinic condition of the
individual patient, the cause of the disease or disorder, the site
of delivery of the agent, the method of administration, the
scheduling of administration, and other factors known to medical
practitioners. The therapeutically effective amount of the
antagonist to be administered will be governed by such
considerations.
Articles of Manufacture
[0223] In another aspect of the invention, an article of
manufacture containing materials useful for the treatment,
prevention and/or diagnosis of the disorders described above is
provided. The article of manufacture comprises a container and a
label or package insert on or associated with the container.
Suitable containers include, for example, bottles, vials, syringes,
IV solution bags, etc. The containers may be formed from a variety
of materials such as glass or plastic. The container holds a
composition which is by itself or combined with another composition
effective for treating, preventing and/or diagnosing the condition
and may have a sterile access port (for example the container may
be an intravenous solution bag or a vial having a stopper
pierceable by a hypodermic injection needle). At least one active
agent in the composition is an antibody of the invention. The label
or package insert indicates that the composition is used for
treating the condition of choice. Moreover, the article of
manufacture may comprise (a) a first container with a composition
contained therein, wherein the composition comprises an antibody of
the invention; and (b) a second container with a composition
contained therein, wherein the composition comprises a further
cytotoxic or otherwise therapeutic agent. The article of
manufacture in this embodiment of the invention may further
comprise a package insert indicating that the compositions can be
used to treat a particular condition. Alternatively, or
additionally, the article of manufacture may further comprise a
second (or third) container comprising a
pharmaceutically-acceptable buffer, such as bacteriostatic water
for injection (BWFI), phosphate-buffered saline, Ringer's solution
and dextrose solution. It may further include other materials
desirable from a commercial and user standpoint, including other
buffers, diluents, filters, needles, and syringes.
[0224] It is understood that any of the above articles of
manufacture may include an immunoconjugate of the invention in
place of or in addition to an anti-CEA ABM.
[0225] The examples below explain the invention in more detail. The
following preparations and examples are given to enable those
skilled in the art to more clearly understand and to practice the
present invention. The present invention, however, is not limited
in scope by the exemplified embodiments, which are intended as
illustrations of single aspects of the invention only, and methods
which are functionally equivalent are within the scope of the
invention. Indeed, various modifications of the invention in
addition to those described herein will become apparent to those
skilled in the art from the foregoing description and accompanying
drawings. Such modifications are intended to fall within the scope
of the appended claims.
EXAMPLES
[0226] Unless otherwise specified, references to the numbering of
specific amino acid residue positions in the following Examples are
according to the Kabat numbering system.
Example 1
Generation of Affinity Maturation Libraries
[0227] H1/H2 Library
[0228] For generation of an affinity maturation library randomized
in the HCDR1 and HCDR2 region, triplets encoding positions F32 G33
in CDR1 and positions W50 N52 T52a K52b T54 E56 T58 in CDR2 were
randomized. In a first step, a DNA fragment (fragment 1) was
amplified using pMS22 as a template and primers MS-43 (SEQ ID NO:
123) and EAB-679 (SEQ ID NO: 127) which contains the randomized
CDR1 positions (FIG. 11). Using the same template, primers MS-56
(SEQ ID NO: 126) and MS-52 (SEQ ID NO: 124) amplified a second
fragment (fragment 2) which has an overlapping region with the
3'end of fragment 1. Amplification conditions included an initial
5-min 94.degree. C. incubation step followed by 25 cycles, each
consisting of a 1-min 94.degree. C. denaturation, a 1-min
55.degree. C. annealing, and a 20-sec and 50-sec 72.degree. C.
elongation step, for fragment 1 and fragment2, respectively. A
final 10-min 72.degree. C. incubation step was performed at the
end. Both fragments were purified on an agarose gel. An overlapping
extension PCR with fragment 1 and 2 using primers MS-43 (SEQ ID NO:
123) and EAB-680 (SEQ ID NO: 128), which harboured randomized
positions of CDR2, generated a fragment with both CDRs randomized
(fragment 3). For the assembly of fragments 1 and 2, equimolar
amounts of fragment 1 and fragment 2 were used. Amplification
conditions included an initial 5-min 94.degree. C. incubation step
followed by 5 cycles without primers, each cycle consisting of a
1-min 94.degree. C. denaturation, a 1-min 55.degree. C. annealing,
and a 40-sec 72.degree. C. elongation step. After the addition of
the outer primers, 20 additional cycles were performed using the
same parameters. A fourth fragment (fragment 4) which overlaps with
the 3' region of fragment 3 was PCR-amplified using again pMS22 as
a template and primers MS-55 (SEQ ID NO: 125) and MS-52 (SEQ ID NO:
124). After gel purification, a final overlap extension PCR using
fragment 3 and 4 as templates and primers MS-43 and MS-52 generated
a fragment containing CL and parts of VH. For this, equimolar
amounts of fragment 3 and fragment 4 were used. Amplification
conditions included an initial 5-min 94.degree. C. incubation step
followed by 5 cycles without primers, each cycle consisting of a
1-min 94.degree. C. denaturation, a 1-min 55.degree. C. annealing,
and a 80-sec 72.degree. C. elongation step. After the addition of
the outer primers, 20 additional cycles were performed using the
same parameters. The resulting fragment was then gel-purified and
ligated with pMS22 after NcoI/NheI digestion.
L1/L2 Library
[0229] For the generation of an affinity maturation library
randomized in the LCDR1 and LCDR2 region, triplets encoding
positions Q27, N28, V29, G30 T31 N32 in CDR1 and positions Y49 S50
Y53 R54 Y55 S56 in CDR2 were randomized. In a first step, a DNA
fragment (fragment 1) was amplified using pMS22 as a template and
primers EAB-685 (SEQ ID NO: 129) and EAB-681 (SEQ ID NO: 133) which
contains the randomized CDR1 positions (FIG. 12). Using the same
template, primers EAB-686 (SEQ ID NO: 130) and EAB-687 (SEQ ID NO:
131) amplified a second fragment (fragment 2) which has an
overlapping region with the 3'end of fragment 1. Amplification
conditions included an initial 5-min 94.degree. C. incubation step
followed by 25 cycles, each consisting of a 1-min 94.degree. C.
denaturation, a 1-min 55.degree. C. annealing, and a 60-sec
72.degree. C. elongation step, for fragment 1 and fragment2,
respectively. A final 10-min 72.degree. C. incubation step was
performed at the end. Both fragments were purified on a agarose
gel. An overlapping extension PCR with fragment 1 and 2 using
primers EAB-685 (SEQ ID NO: 129) and EAB-682 (SEQ ID NO: 134),
which harboured randomized positions of CDR2, generated a fragment
with both CDRs randomized (fragment 3). For the assembly of
fragments 1 and 2, equimolar amounts of fragment 1 and fragment 2
were used. Amplification conditions included an initial 5-min
94.degree. C. incubation step followed by 5 cycles without primers,
each cycle consisting of a 1-min 94.degree. C. denaturation, a
1-min 55.degree. C. annealing, and a 60-sec 72.degree. C.
elongation step. After the addition of the outer primers, 20
additional cycles were performed using the same parameters. A
fourth fragment (fragment 4) which overlaps with the 3' region of
fragment 3 was PCR-amplified using again pMS22 as a template and
primers EAB-688 (SEQ ID NO: 132) and EAB-687 (SEQ ID NO: 131).
After gel purification, a final overlap extension PCR using
fragment 3 and 4 as templates and primers EAB-685 (SEQ ID NO: 129)
and EAB-687 (SEQ ID NO: 131) generated a fragment containing VL and
parts of CL. For this, equimolar amounts of fragment 3 and fragment
4 were used. Amplification conditions included an initial 5-min
94.degree. C. incubation step followed by 5 cycles without primers,
each cycle consisting of a 1-min 94.degree. C. denaturation, a
1-min 55.degree. C. annealing, and a 80-sec 72.degree. C.
elongation step. After the addition of the outer primers, 20
additional cycles were performed using the same parameters. This
fragment was then ligated with pMS22 after HindIII/SacI
digestion.
H3 Libraries
[0230] For the generation of affinity maturation libraries
randomized in the HCDR3 region, triplets encoding positions W95,
D96, F97, Y98, D99, Y100, V100a, E100b, A100c, and M100d were
randomized in two different approaches: (1) randomization of the
entire segment (H3 full library) or (2) individual randomization of
each position resulting in ten sublibraries. Sublibraries
containing clones with individually randomized positions were
pooled after transformation into bacteria (H3 pooled library). For
the randomization of the HCDR3 region, fragments were PCR-amplified
using a primer that annealed in the 3 `end of CL and primers that
harbour the randomized sequences of HCDR3 (FIG. 13). An overlap
extension PCR was then performed with a second fragment that
overlaps with the 3` end of fragment 1, and comprises the end of VH
and the 5' region of CH1. The assembled fragments were then ligated
into pMS22 after SacI/NheI digestion. For the generation of the H3
pooled library, ten DNA fragments were separately PCR-amplified
using each of primers AC7-AC16 (SEQ ID NO: 135; SEQ ID NO: 144) in
combination with primer EAB-749 (SEQ ID NO: 146). For the
generation of the L3 full library, primers AC17 (SEQ ID NO: 145)
and EAB-749 (SEQ ID NO: 146) were used. Plasmid pMS22 was used as a
template. Amplification conditions included an initial 5-min
94.degree. C. incubation step followed by 25 cycles, each
consisting of a 1-min 94.degree. C. denaturation, a 1-min
55.degree. C. annealing, and a 36-sec 72.degree. C. elongation
step, followed by a final 10-mM 72.degree. C. incubation step. This
resulted in about 580 bp long fragments which were purified on an
agarose gel. For the overlap extension PCR, a second fragment was
amplified using either primer EAB-750 (SEQ ID NO: 147) or EAB-751
(SEQ ID NO: 148) in combination with EAB-752 (SEQ ID NO: 149).
While primer EAB-750 SEQ ID NO: 147) had an overlapping sequence
with randomization primers AC7-11 (SEQ ID NO: 139), EAB-751 (SEQ ID
NO: 148) shared sequence homologies with randomization primers
AC12-17 (SEQ ID NOs: 140-145). Amplification conditions included an
initial 5-min 94.degree. C. incubation step followed by 25 cycles,
each consisting of a 1-min 94.degree. C. denaturation, a 1-min
55.degree. C. annealing, and a 12-sec 72.degree. C. elongation
step, followed by a final 10-mM 72.degree. C. incubation step. The
resulting fragments were about 180 bp long. For the assembly of
both fragments, equimolar amounts of fragment 1 and the
corresponding fragment 2 were used. Amplification conditions
included an initial 5-min 94.degree. C. incubation step followed by
5 cycles without primers, each cycle consisting of a 1-min
94.degree. C. denaturation, a 1-min 55.degree. C. annealing, and a
60-sec 72.degree. C. elongation step. After the addition of the
outer primers EAB-749 (SEQ ID NO: 146) and EAB-752 (SEQ ID NO:
149), 20 additional cycles were performed using the same
parameters. At the end, a final 10-min 72.degree. C. incubation
step was performed. The gel-purified fragments were then ligated
into pMS22 after SacI/NheI-digestion and purified ligations were
transformed into TG1 bacteria by electropration.
L3 Libraries
[0231] For the generation of affinity maturation libraries
randomized in the CDR3 region of the light chain, triplets encoding
positions Y91, Y92, T93, Y94, and L95a were either randomized
throughout the segment (L3 full library) or individually resulting
in five sublibraries. Sublibraries containing clones with
individually randomized positions were pooled after transformation
into bacteria (L3 pooled library). For the generation of the five
sublibraries, five DNA fragments were PCR-amplified using each of
primers AC1-AC5 (SEQ ID NOs: 150-154) in combination with primer
MS43 (SEQ ID NO: 123). For the generation of the L3 full library,
primer combination AC6 (SEQ ID NO: 155) and MS43 (SEQ ID NO: 123)
were used (FIG. 14). Plasmid pMS22 was used as a template.
Amplification conditions included an initial 5-min 94.degree. C.
incubation step followed by 25 cycles, each consisting of a 1-min
94.degree. C. denaturation, a 1-min 55.degree. C. annealing, and a
25-sec 72.degree. C. elongation step, followed by a final 10-min
72.degree. C. incubation step. The resulting fragments which
encompass positions 1-104 of the VL domain were purified on an
agarose gel and used as a template for an additional PCR
amplification. All reactions were performed with primer EAB-746
(SEQ ID NO: 156) which has an overlapping sequence with the
randomization primers and MS43 (SEQ ID NO: 123) using the same
conditions described above. The purified fragments as well as pMS22
were digested with NcoI/XhoI. For all five sublibraries, 0.5 .mu.g
insert were ligated with 0.5 .mu.g pAC16. For the L3 full library,
ligation was performed with 9.8 .mu.g insert and 9.8 .mu.g pMS22.
Purified ligations were transformed into TG1 bacteria by
electroporation.
Generation of the Antigens
[0232] Because both murine and humanized PR1A3 antibodies recognize
only membrane bound but not shed soluble human CEA, a recombinant
chimeric protein which contains the epitope of PR1A3 was generated
for in vitro affinity maturation of humanized PR1A3 (SEQ ID NO:7
and 8). Generation of this hybrid protein was performed as
described in Steward et al., 1999. In brief, DNA sequence of the B
domain of human biliary glycoprotein (BGP) was replaced with the
sequence of the human CEA-B3 domain, which contains the epitope of
PR1A3. As a result, the sequence encodes a hybrid protein which
comprises the N and A1 domains of BGP, the B3 domain of CEA and the
A2 domain of BGP (N-A1-B3-A2, huNABA). This fusion product was then
either linked to the Fc portion of human IgG1 (huNABA-Fc) (Steward
et al., Cancer Immunol Immunother, 47:299-306, 1999) or fused to a
sequence encoding the precision protease cleavage site, an avi tag
and a (His)6 tag (huNABA-avi-his) (SEQ ID NO:158). huNABA-Fc was
purified from the supernatant of a stably transfected CHO cell line
using a protein A column huNABA-avi-his (SEQ ID NO: 158) was
transiently transfected into HEK 293 cells, stably expressing the
EBV-derived protein EBNA. A simultaneously co-transfected plasmid
encoding a biotin ligase allowed avi tag-specific biotinlylation in
vivo. The protein was then purified by immobilized metal affinity
chromatography (IMAC) followed by gel filtration.
Affinity Maturation of Humanized PR1A3
[0233] Generation of affinity-matured humanized PR1A3 Fabs was
carried out by phage display using standard protocols (Silacci et
al, Proteomics, 5(9):2340-2350, 2005). Selections with all affinity
maturation libraries were carried out in solution according to the
following procedure: 1. binding of .about.1012 phagemid particles
of each affinity maturation libraries to 100 nM biotinylated
huNABA-avi-his for 0.5 h in a total volume of 1 ml, 2. capture of
biotinylated huNABA-avi-his and specifically bound phage particles
by addition of 5.4.times.10.sup.7 streptavidin-coated magnetic
beads for 10 mM, 3. washing of beads using 5-10.times.1 ml
PBS/Tween20 and 5-10.times.1 ml PBS, 4. elution of phage particles
by addition of 1 ml 100 mM TEA (triethylamine) for 10 mM and
neutralization by adding 500 ul 1M Tris/HCl pH 7.4 and 5.
re-infection of exponentially growing E. coli TG1 bacteria,
infection with helper phage VCSM13 and subsequent PEG/NaCl
precipitation of phagemid particles to be used in subsequent
selection rounds. Selections were carried out over 3-5 rounds using
either constant or decreasing (from 10.sup.-7M to
2.times.10.sup.-9M) antigen concentrations. In round 2, capture of
antigen:phage complexes was performed using neutravidin plates
instead of streptavidin beads. Specific binders were identified by
ELISA as follows: 100 ul of 10 nM biotinylated huNABA-avi-his per
well were coated on neutravidin plates. Fab-containing bacterial
supernatants were added and binding Fabs were detected via their
Flag-tags by using an anti-Flag/HRP secondary antibody.
ELISA-positive clones were bacterially expressed as soluble Fab
fragments in 96-well format and supernatants were subjected to a
kinetic screening experiment by SPR-analysis using BIACORE T100.
Clones expressing Fabs with the highest affinity constants were
identified and the corresponding phagemids were sequenced.
Purification of Fabs and Measurement of the Kinetic Parameters
[0234] For the exact analysis of the kinetic parameters, Fabs were
purified from bacterial cultures. A 500 ml culture was inoculated
and induced with 1 mM IPTG at an OD600 0.9. Bacteria were incubated
at 25.degree. C. overnight and harvested by centrifugation. After
the incubation of the resuspended pellet for 20 mM in 25 ml PPB
buffer (30 mM Tris-HCl pH8, 1 mM EDTA, 20% sucrose), bacteria were
centrifuged again and the supernatant was harvested. This
incubation step was repeated once with 25 ml of a 5 mM MgSO.sub.4
solution. The supernatants of both incubation steps were pooled,
filtered and loaded on a IMAC column (His gravitrap, GE
Healthcare). Subsequently, the column was washed with 40 volumes.
After the elution (500 mM NaCl, 500 mM Imidazole, 20 mM
NaH.sub.2PO.sub.4 pH 7.4) the eluate was re-buffered using PD10
columns (GE Healthcare). The kinetic parameters of the purified
Fabs were then studied by SPR-analysis in a dilution row that
ranged from 200 nM to 6.25 nM.
Example 2
[0235] The PR1A3 antibody was chimerized to have a human IgG1/kappa
constant region, and expressed using the GylcoMab technology in
order to have a high degree of afucosylated sugars in the Fc. The
glycoengineered and non-glycoengineered antibodies were compared at
a effector to target ratio of 25:1. The maximal amount of antibody
dependent target cell killing was doubled by glycoengineering of
the Fc region (FIG. 2). A further increase in cell killing was
achieved by increasing the effector to target ratio (FIG. 2).
[0236] PR1A3 was humanized using frameworks identical to human
germline sequences. The IMGT sequence IGHV7-4-1*02 (Accession No.
X62110) was the acceptor for VH humanized and IMGT_hVK_1_39
(Accession No. X59315) was the acceptor for VL humanization. A
humanized PR1A3 antibody comprising a heavy chain variable region
construct CH7A and a light chain variable region construct CL1A
showed satisfactory binding to human colon carcinoma cells as
measured by flow cytometry (FIG. 3).
[0237] Affinity maturation of PR1A3 by phage display was performed
using standard protocols as described in detail in Example 1,
herein. The parent humanized PR1A3 antibody that was used for
affinity maturation comprises a heavy chain variable region
construct CH7A and a light chain variable region construct CL1A.
Tables 3-6 below show the libraries used for affinity maturation.
For the L1/L2 library, positions Valine 29, Alanine50, or Serine51
within the CDRs were kept constant. For the H1/H2 library,
positions Isoleucine51, Glycine55, or Alanine57 within the CDRs
were kept constant (FIGS. 4 and 5).
[0238] An affinity matured heavy chain variable region construct,
CH7A rF9, and an affinity matured light chain variable region
construct, CL1A rH11, were paired with the parent light chain
variable region construct and the heavy chain variable region
construct, respectively, and with each other. All antibodies were
converted into human IgG1/kappa and binding to the CEA-positive
cell-line MKN45 was measured by flow cytometry. Antibodies
comprising either one affinity matured heavy or light chain
variable regions or both affinity matured heavy or light chain
variable regions showed improved binding characteristic as compared
to the humanized parent antibody (FIG. 6). FIGS. 6, 10 and 15 show
several examples where the matured light and heavy chains
independently contribute to increased affinity. The parental
antibody CH7A CL1A has the lowest signal intensity, as well as the
highest EC50 value in FIGS. 6 and 15. The matured light chain
shifts the EC50 values to lower numbers, whereas the matured heavy
chains (rF9 in FIG. 6, and rB9 in FIG. 15) shift the total
fluorescence signal intensity in a flow-cytometry measurement. FIG.
10 shows the individual contributions of heavy and light chain
measured by Biacore methodology. The combination of these two
chains increases the affinity even further. Additionally, as shown
in FIG. 16, improvement of affinity leads to improvement of ADCC
characteristics.
[0239] The binding affinities of the affinity matured heavy and
light chain CDRs were determined by Biacore and listed FIGS. 34A
and B.
[0240] FIG. 35 summarizes the affinity constants of the various
affinity matured antibody sequences. The parental antibody PR1A3 is
listed as well as several light chain and heavy chain combinations
of matured and non matured sequences. All values were obtained by
Biacore technology by measuring the association (k.sub.on) and
dissociation (k.sub.off) rate constants of the various soluble
antibody constructs in Fab format on a Biacore chip with
immobilized NABA-avi-his reagent (SEQ ID NO 158) as the antigen.
The affinity constant is labeled with KD.
Example 3
[0241] The acceptor framework used in generating the affinity
matured anti-CEA antibodies described in Example 2 was of the human
VH7 class. In order to increase stability, a more stable acceptor
framework sequence was used as the basis for stability engineering
of the antibody. Based on sequence homology of the murine antibody
PR1A3, and the assumption that VH1 derived sequences should have a
higher intrinsic stability than VH7, or the even numbers of the
human VH clans (Ewert, S., Huber, T., Honegger, A. and Pluckthun,
A. (2003) J. Mol. Biol., 325, 531-553), the sequence IGHV-1-18; Acc
No.:M99641 was used as the new acceptor framework. Conventional CDR
loop-grafting of the PR1A3 antibody lead to construct CH1A (SEQ ID
NO: 279). Unfortunately, this molecule did not show significant
binding activity towards the CEA antigen. The binding activity of
this construct was compared to the binding activity of the chimeric
antibody PR1A3 harbouring mouse-derived variable domains at various
concentrations. BxPC3 cells were used for specific binding of the
antibodies to CEA and binding intensity was measured by FACS
analysis. FIG. 17.
[0242] In order to recover binding affinity, several back-mutations
were introduced into the CH1A sequence to generated new heavy
chains CH1A1 (SEQ ID NO: 257), CH1A2 (SEQ ID NO:258), CH1A3 (SEQ ID
NO: 259), and CH1A4 (SEQ ID NO: 260). CH1A1 includes the M69F/T71L
double point mutation. The latter three variants have the entire
frameworks 1, 2, or 3, respectively, replaced by the murine
counterpart. FIG. 18 shows the binding of those constructs when
paired with the 2F1 light chain (SEQ ID NO: 209). In this assay,
cell binding of the CH1A-based antibody variants to CEA-expressing
MKN-45 cells was analyzed at various concentrations. The
affinity-matured light chain 2F1 was identical for all antibodies
tested except for the parental antibody where the original light
chain CL1A was used. Mean fluorescence was determined by FACS
analysis. FIG. 19 shows the stabilities of those constructs when
paired with the 2F1 light chain, as measured by dynamic light
scattering (DLS) of the samples. The DLS assay was performed using
1 mg/ml of the antibodies in a buffer of 20 mM Histidine and 140 mM
NaCl at pH 6.0. The assay was conducted starting at 25.degree. C.
with an incremental temperature increase of 0.05.degree. C./min up
to 70.degree. C. All antibodies tested in this assay had 2F1 as the
light chain.
[0243] Since CH1A1 still keeps the original stability, and also
shows some significant (but somewhat less than CH1A4, that was of
highest affinity, but lowest stability) binding, this construct was
chosen for further optimization of binding. New heavy chains CH1A1A
(SEQ ID NO: 261), CH1A1B (SEQ ID NO: 262), CH1A1C (SEQ ID NO: 263),
CH1A1D (SEQ ID NO: 264), CH1A1E (SEQ ID NO: 265), CH1A1F (SEQ ID
NO: 266), and CH1A1G (SEQ ID NO: 267) were generated. These are
essentially variants of CH1A1 with only a few backmutations in the
FR1 and the FR3 region. FIGS. 20 and 21 shows that their affinities
are all comparable, albeit still slightly inferior to the VH7 based
humanized construct CH7A. FIG. 20 shows Proteon (Biacore)
sensorgrams obtained for the binding of the CH1A1-based framework
variants to the CEA antigen-harbouring chimeric protein NABA.
Biotinylated NABA was immobilized on a Neutravidin-coated chip and
antibodies were used as analytes at concentrations of 100, 50, 25,
12.5, 6.25, and 0 nM. The precursor clone CH1A1 and the parental
antibody CH7A were included for direct comparison. The light chain
2F1 was identical for all antibodies tested. FIG. 21 shows the
binding intensity of the seven CH1A1-based variants carrying
additional framework mutations. Antibodies were incubated with the
CEA-expressing MKN45 cells in a concentration series and binding
intensity was measured by FACS analysis. The precursor clone CH was
included for direct comparison. All antibodies tested in this assay
had 2F1 as the light chain Variants CH1A1A and CH1A1B were chosen
as final variants of the VH1 based humanization based on their
comparatively better purification yields and monomeric
behavior.
Example 4
[0244] The residues of CDR-H3 that were selected in the affinity
maturation process were individually introduced into the PR1A3
sequence to test for increased antibody stability. FIG. 36.
[0245] FIG. 22 shows the Surface Plasmon Resonance (SPR)
measurements of the affinity (as measured in the bivalent form) of
each antibody towards the CEA antigen (NABA reagent as described by
Stewart et al. Cancer Immunol Immunother (1999) 47:299-306). Shown
in FIG. 22 are Proteon (Biacore) sensorgrams obtained for the
binding of the CDR-H3 antibody variants to the CEA
antigen-harbouring chimeric protein NABA. Biotinylated NABA was
immobilized on a Neutravidin-coated chip and antibodies were used
as analytes at concentrations of 100, 50, 25, 12.5, 6.25, and 0 nM.
The affinity matured precursor clone 5HFF12 and the parental
antibody CH7A were included for direct comparison. All antibodies
tested in this assay had 2F1 as the light chain. Variant CH7A
(W95Y) shows no measurable activity in this assay, all the other
variants exhibit an affinity to target within a factor of ten of
each other. The relative affinity of each, measured in the bivalent
form, is as follows: 5HFF12>CH7A (Y98A/D99Y)>CH7A
(Y98A)>CH7A>CH7A (E102Q)>CH7A (D99Y)>CH7A
(D99H)>CH7A (A103T)>CH7A (V101F)>CH7A (W95Y).
[0246] DLS analysis was performed on the antibodies as compared
with their precursor 5HFF12 and the parental antibody harbouring
heavy chain CH7A. The light chain 2F1 was identical for all
antibodies tested in this experiment. FIGS. 23 and 24. The results
of this analysis provided the following ranking in stabilities:
CH7A (D99Y)>CH7A (Y98A/D99Y)>CH7A (V101F)>CH7A
(D99H)>CH7A (A103T)>CH7A (W95Y)>CH7Ax2F1
(=PR1A3)>5HFF12. The DLS assay was performed using 1 mg/ml of
the antibodies in a buffer of 20 mM Histidine and 140 mM NaCl at pH
6.0. The assay was conducted starting at 25.degree. C. with an
incremental temperature increase of 0.05.degree. C./min up to
70.degree. C.
[0247] The double mutant (Y98A/D99Y) (SEQ ID NO: 223) was chosen
for further stability engineering as it exhibited high stability
while retaining high affinity for the CEA target.
Example 5
[0248] The double mutant (Y98A/D99Y) of the humanized PR1A3
derivative CH7A was introduced into the final variants of the VH1
based humanization-constructs CH1A1A, and CH1A1B.
[0249] Proteon (Biacore) sensorgrams were obtained for the binding
of the combined framework and CDR-H3 variants to the CEA
antigen-harbouring chimeric protein NABA. Biotinylated NABA was
immobilized on a Neutravidin-coated chip and antibodies were used
as analytes at concentrations of 100, 50, 25, 12.5, 6.25, and 0 nM.
The precursor clone 5L1A10 and the parental antibody CH7A were
included for direct comparison. All antibodies tested in this assay
had 2F1 as the light chain FIG. 25.
Example 6
[0250] The CH1A1A and CH1A1B constructs exhibited ADCC activity.
ADCC mediated by CH1A1A and CH1A1B variants, their precursor
variant CH1A1, and the parental CH7A variant was measured after 4h
by lactate dehydrogenase release using MKN45 cells as target cells
(T) and human PBMC as effector cells (E) at E:T ratios of 25:1.
Lactate dehydrogenase release is proportionate to target cell lysis
and shown as percent cytotoxicity. FIG. 26. ADCC activity for these
variants was confirmed as measured by calceine release using MKN45
cells as target cells (T) and human PBMC as effector cells (E) at
E:T ratios of 25:1. Calceine release is proportionate to target
cell lysis and shown as percent cytotoxicity with mean.+-.standard
deviation values. FIG. 27.
[0251] ADCC activity was also observed for CH1A1A and CH1A1B
constructs containing the affinity matured CDRH3 with the double
mutation Y98A/D99Y. ADCC mediated by the combined framework and
CDR-H3 variants and the parental antibody harbouring CH7A was
measured after 24h by lactate dehydrogenase release using MKN45
cells (FIG. 28) or LS174T (FIG. 29) as target cells and human PBMC
as effector cells at E:T ratios of 5:1. While MKN45 cells express
CEA at high levels, the expression of CEA is medium in LS174T
cells. Lactate dehydrogenase release is proportionate to target
cell lysis and shown as percent cytotoxicity. All antibodies tested
in these ADCC assays had 2F1 as the light chain
[0252] FIG. 38 shows the amino acid sequence alignments of VH
regions of various stability matured anti-CEA antibodies.
Example 7
[0253] The glycoengineered version of anti-CEA antibody comprising
the CH1A1A (98/99) heavy chain and 2F1 light chain was tested for
efficacy in a colorectal carcinoma xenograft model in SCID mice
transgenic for human CD16. The model assay conditions are set forth
below. The results of the assay indicate that this anti-CEA
antibody provides a survival benefit as compared to a vehicle
control. FIG. 30.
[0254] Animals: CD16 Scid transgenic female mice; age 7-9 weeks at
start of experiment (Charles River France) were maintained under
specific-pathogen-free condition with daily cycles of 12 h light/12
h darkness according to committed guidelines (GV-Solas; Felasa;
TierschG). Experimental study protocol was reviewed and approved by
local government authorities. After arrival, animals were
maintained for one week for acclimation and for observation.
Continuous health monitoring was carried out on a regular
basis.
[0255] Cell culture and application: LS174T cells (human colon
carcinoma cells; European Collection of Cell Culture) were cultured
in DMEM medium containing 10% FCS (PAA Laboratories, Austria). The
cells were cultured at 37.degree. C. in a water-saturated
atmosphere at 5% CO2. In vitro passage 24 was used for
intra-splenic injection, at a viability of 97%.
[0256] Tumor cell injection: At day of injection, LS174T tumor
cells were harvested using trypsin-EDTA (Gibco, Switzerland) from
culture flasks (Greiner Bio-One) and transferred into 50 ml culture
medium, washed once and resuspended in AIM V (Gibco, Switzerland).
A small incision was made at the left abdomal site of anesthetized
SCID/beige mouse. The skin and the muscle were opened and thirty
microliters (3.times.10.sup.6 LS174T cells in AimV medium) of cell
suspension was injected in the apex of the spleen. The muscle first
and then the abdominal skin were sutured with a absorbable sutures
(Monosyn.RTM. 3-0, Braun).
[0257] Treatment: All anti-CEA antibodies and the corresponding
vehicle were administered i.v. once weekly. Three dosings in total.
625 ug of antibody was administered per injection per mouse. The
antibody dilutions were prepared freshly from stock before use and
formulated in 20 mM Histidine, 140 mM NaCl, pH6.0 at 4.38 mg/ml
antibody concentration.
Example 8
[0258] The glycoengineered version of anti-CEA antibody comprising
CH1A1A (Y98A/D99Y) heavy chain and 2F1 light chain was tested for
efficacy in an A549 lung carcinoma xenograft model in scid mice
transgenic for human CD16. The model assay conditions are set forth
below. The results of the assay indicate that this anti-CEA
antibody provides a dose-dependant survival benefit as compared to
a vehicle control. FIG. 31.
[0259] Animals: Thirty five CD16 Scid transgenic female mice; age
7-9 weeks at start of experiment (Charles River France) were
maintained under specific-pathogen-free condition with daily cycles
of 12 h light/12 h darkness according to committed guidelines
(GV-Solas; Felasa; TierschG). Experimental study protocol was
reviewed and approved by local government authorities. After
arrival, animals were maintained for one week for acclimation and
for observation. Continuous health monitoring was carried out on a
regular basis.
[0260] Cell culture and application: A549 cells (human NSCLC cells;
American Tissue Culture collection) were cultured in DMEM medium
containing 10% FCS (PAA Laboratories, Austria). The cells were
cultured at 37.degree. C. in a water-saturated atmosphere at 5%
CO2.
[0261] Treatment: Mice were injected i.v. on study day 0 with
1.times.10.sup.6 A549 cells. The antibody started on study day 7
and continued with 2 more weekly injections.
Treatment Groups:
[0262] Mice 35 SCID-CD16Tg mice, N=7 per group. Cells--A549 cells 5
Mio/mouse
Compound and Therapy Schedule
Vehicle 3q7d
[0263] CH1A1A(Y98A/D99Y).times.2F1 (500 ug) 3q7d (25 mg/kg)
CH1A1A(Y98A/D99Y).times.2F1 (200 ug) 3q7d (10 mg/kg)
CH1A1A(Y98A/D99Y).times.2F1 (100 ug) 3q7d (5 mg/kg)
CH1A1A(Y98A/D99Y).times.2F1 (50 ug) 3q7d (1 mg/kg)
[0264] Although the foregoing invention has been described in some
detail by way of illustration and example for purposes of clarity
of understanding, the descriptions and examples should not be
construed as limiting the scope of the invention. The disclosures
of all patent and scientific literature cited herein are expressly
incorporated in their entirety by reference.
Sequence CWU 1 SEQUENCE LISTING <160> NUMBER OF SEQ ID
NOS: 281 <210> SEQ ID NO 1 <211> LENGTH: 5 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Heavy Chain CDR1 Kabat
<400> SEQUENCE: 1 Glu Phe Gly Met Asn 1 5 <210> SEQ ID
NO 2 <211> LENGTH: 5 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Heavy Chain CDR1 Kabat <400> SEQUENCE: 2
Glu Tyr Gly Met Asn 1 5 <210> SEQ ID NO 3 <211> LENGTH:
5 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR1 Kabat <400> SEQUENCE: 3 Glu Tyr Ser Met Asn 1 5
<210> SEQ ID NO 4 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain Variable Region
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (32)..(32) <223> OTHER INFORMATION: Xaa = Tyr or
Phe <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (33)..(33) <223> OTHER INFORMATION: Xaa
= Ser or Gly <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (35)..(35) <223> OTHER
INFORMATION: Xaa = Asn or ser <220> FEATURE: <221>
NAME/KEY: MISC_FEATURE <222> LOCATION: (50)..(50) <223>
OTHER INFORMATION: Xaa = Trp or Tyr <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (55)..(55)
<223> OTHER INFORMATION: Xaa = Thr, Ser, or Asn <220>
FEATURE: <221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(59)..(59) <223> OTHER INFORMATION: Xaa = Thr or Asn
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (61)..(61) <223> OTHER INFORMATION: Xaa = Val or
Met <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (101)..(101) <223> OTHER INFORMATION:
Xaa = Phe or Ala <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (102)..(102) <223> OTHER
INFORMATION: Xaa = Val, Phe, Ser, Tyr, or Ala <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(103)..(103) <223> OTHER INFORMATION: Xaa = Asp, His, Trp,
Glu or Tyr <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (105)..(105) <223> OTHER INFORMATION:
Xaa = Val, Phe, or Leu <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (106)..(106) <223> OTHER
INFORMATION: Xaa = Glu, Lys or Gln <220> FEATURE: <221>
NAME/KEY: MISC_FEATURE <222> LOCATION: (107)..(107)
<223> OTHER INFORMATION: Xaa = Ala or Thr <220>
FEATURE: <221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(108)..(108) <223> OTHER INFORMATION: Xaa = Met or Leu
<400> SEQUENCE: 4 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu
Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser
Gly Tyr Thr Phe Thr Glu Xaa 20 25 30 Xaa Met Xaa Trp Val Arg Gln
Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Xaa Ile Asn Thr
Lys Xaa Gly Glu Ala Xaa Tyr Xaa Glu Glu Phe 50 55 60 Lys Gly Arg
Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu
Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Trp Asp Xaa Xaa Xaa Tyr Xaa Xaa Xaa Xaa Asp Tyr Trp Gly
100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210>
SEQ ID NO 5 <211> LENGTH: 5 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Heavy Chain CDR1 Kabat <400> SEQUENCE: 5
Glu Phe Gly Met Ser 1 5 <210> SEQ ID NO 6 <211> LENGTH:
7 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR1 Chothia <400> SEQUENCE: 6 Gly Tyr Thr Phe Thr Glu Phe 1
5 <210> SEQ ID NO 7 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 Chothia <400>
SEQUENCE: 7 Gly Tyr Thr Phe Thr Glu Tyr 1 5 <210> SEQ ID NO 8
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 AbM <400> SEQUENCE: 8 Gly Tyr
Thr Phe Thr Glu Phe Gly Met Asn 1 5 10 <210> SEQ ID NO 9
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 AbM <400> SEQUENCE: 9 Gly Tyr
Thr Phe Thr Glu Tyr Gly Met Asn 1 5 10 <210> SEQ ID NO 10
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 AbM <400> SEQUENCE: 10 Gly Tyr
Thr Phe Thr Glu Tyr Ser Met Asn 1 5 10 <210> SEQ ID NO 11
<211> LENGTH: 108 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain Variable Region <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (27)..(27)
<223> OTHER INFORMATION: Xaa = Gln, Ala, Lys, or His
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (28)..(28) <223> OTHER INFORMATION: Xaa = Asn, Ala,
Tyr, Ile, Lys, or Thr <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (29)..(29) <223> OTHER
INFORMATION: Xaa = Val, Ala, Gly, or Met <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (30)..(30)
<223> OTHER INFORMATION: Xaa = Gly, Ser, Thr, or Leu
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (31)..(31) <223> OTHER INFORMATION: Xaa = Thr, Asn,
Pro, or Ala <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (32)..(32) <223> OTHER INFORMATION: Xaa
= Asn or Tyr <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (46)..(46) <223> OTHER
INFORMATION: Xaa = Pro or Leu <220> FEATURE: <221>
NAME/KEY: MISC_FEATURE <222> LOCATION: (50)..(50) <223>
OTHER INFORMATION: Xaa = Ser, Leu, or Trp <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (53)..(53)
<223> OTHER INFORMATION: Xaa = Tyr, Asn, or His <220>
FEATURE: <221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(54)..(54) <223> OTHER INFORMATION: Xaa = Arg, Leu, Pro, or
His <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (55)..(55) <223> OTHER INFORMATION: Xaa
= Tyr, Ser, Gln, Lys, Phe, Pro, or Glu <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (56)..(56)
<223> OTHER INFORMATION: Xaa = Ser, Gly, Ile, or Arg
<400> SEQUENCE: 11 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Xaa Ala Ser
Xaa Xaa Xaa Xaa Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 12 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 AbM <400>
SEQUENCE: 12 Gly Tyr Thr Phe Thr Glu Phe Gly Met Ser 1 5 10
<210> SEQ ID NO 13 <211> LENGTH: 17 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 Kabat <400>
SEQUENCE: 13 Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe Lys 1 5 10 15 Gly <210> SEQ ID NO 14 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 Kabat <400> SEQUENCE: 14 Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Ile Glu Glu Phe Lys 1 5 10 15 Gly <210>
SEQ ID NO 15 <211> LENGTH: 17 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 Kabat <400>
SEQUENCE: 15 Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu
Glu Phe Lys 1 5 10 15 Gly <210> SEQ ID NO 16 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 Kabat <400> SEQUENCE: 16 Tyr Ile Asn Thr Lys Asn
Gly Glu Ala Asn Tyr Val Glu Glu Phe Lys 1 5 10 15 Gly <210>
SEQ ID NO 17 <211> LENGTH: 17 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 Kabat <400>
SEQUENCE: 17 Trp Ile Asn Thr Lys Asn Gly Glu Ala Thr Tyr Ile Glu
Glu Phe Lys 1 5 10 15 Gly <210> SEQ ID NO 18 <211>
LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 Chothia <400> SEQUENCE: 18 Asn Thr Lys Thr Gly Glu
Ala Thr 1 5 <210> SEQ ID NO 19 <211> LENGTH: 8
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR2 Chothia <400> SEQUENCE: 19 Asn Thr Lys Ser Gly Glu Ala
Thr 1 5 <210> SEQ ID NO 20 <211> LENGTH: 8 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Heavy Chain CDR2 Chothia
<400> SEQUENCE: 20 Asn Thr Lys Asn Gly Glu Ala Asn 1 5
<210> SEQ ID NO 21 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 21 Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr 1 5 10
<210> SEQ ID NO 22 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 22 Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr 1 5 10
<210> SEQ ID NO 23 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 23 Tyr Ile Asn Thr Lys Asn Gly Glu Ala Asn 1 5 10
<210> SEQ ID NO 24 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 24 Trp Ile Asn Thr Lys Asn Gly Glu Ala Thr 1 5 10
<210> SEQ ID NO 25 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 25 Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 26 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 26 Trp Asp Phe Tyr His Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 27 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 27 Trp Asp Phe Val Asp Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 28 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 28 Trp Asp Phe Tyr Trp Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 29 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 29 Trp Asp Ala Phe Glu Tyr Val Lys Ala Leu Asp Tyr 1 5 10
<210> SEQ ID NO 30 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 30 Trp Asp Phe Phe Glu Tyr Phe Lys Thr Met Asp Tyr 1 5 10
<210> SEQ ID NO 31 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 31 Trp Asp Phe Phe Tyr Tyr Val Gln Thr Met Asp Tyr 1 5 10
<210> SEQ ID NO 32 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 32 Trp Asp Phe Ser Tyr Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 33 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 33 Trp Asp Phe Ala His Tyr Phe Gln Thr Met Asp Tyr 1 5 10
<210> SEQ ID NO 34 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 34 Trp Asp Phe Ala Tyr Tyr Phe Gln Thr Met Asp Tyr 1 5 10
<210> SEQ ID NO 35 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 35 Trp Asp Phe Ala Tyr Tyr Leu Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 36 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 36 Lys Ala Ser Gln Asn Val Gly Thr Asn Val Ala 1 5 10
<210> SEQ ID NO 37 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 37 Lys Ala Ser Ala Asn Val Gly Asn Asn Val Ala 1 5 10
<210> SEQ ID NO 38 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 38 Lys Ala Ser Lys Asn Val Gly Thr Asn Val Ala 1 5 10
<210> SEQ ID NO 39 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 39 Lys Ala Ser Ala Ala Val Gly Thr Tyr Val Ala 1 5 10
<210> SEQ ID NO 40 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 40 Lys Ala Ser Gln Tyr Ala Ser Thr Asn Val Ala 1 5 10
<210> SEQ ID NO 41 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 41 Lys Ala Ser His Asn Val Gly Thr Asn Val Ala 1 5 10
<210> SEQ ID NO 42 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 42 Lys Ala Ser Gln Ile Met Gly Pro Asn Val Ala 1 5 10
<210> SEQ ID NO 43 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 43 Lys Ala Ser Gln Ile Val Gly Thr Asn Val Ala 1 5 10
<210> SEQ ID NO 44 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 44 Lys Ala Ser Gln Lys Val Leu Thr Asn Val Ala 1 5 10
<210> SEQ ID NO 45 <211> LENGTH: 11 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 45 Lys Ala Ser Gln Thr Val Ser Ala Asn Val Ala 1 5 10
<210> SEQ ID NO 46 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 46 Ser Ala Ser Tyr Arg Tyr Ser 1 5 <210> SEQ ID NO
47 <211> LENGTH: 8 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Light Chain CDR2 <400> SEQUENCE: 47 Tyr
Leu Ala Ser Asn Leu Ser Gly 1 5 <210> SEQ ID NO 48
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 48 Tyr Leu Ala
Ser Tyr Pro Gln Ile 1 5 <210> SEQ ID NO 49 <211>
LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Light
Chain CDR2 <400> SEQUENCE: 49 Tyr Ser Ala Ser Tyr Arg Lys Arg
1 5 <210> SEQ ID NO 50 <211> LENGTH: 8 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Light Chain CDR2
<400> SEQUENCE: 50 Tyr Trp Ala Ser Tyr Arg Tyr Ser 1 5
<210> SEQ ID NO 51 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 51 Tyr Ser Ala Ser His Arg Tyr Ser 1 5 <210> SEQ ID
NO 52 <211> LENGTH: 8 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Light Chain CDR2 <400> SEQUENCE: 52 Tyr
Leu Ala Ser Tyr His Glu Ser 1 5 <210> SEQ ID NO 53
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 53 Tyr Ser Ala
Ser His Arg Pro Ser 1 5 <210> SEQ ID NO 54 <211>
LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Light
Chain CDR2 <400> SEQUENCE: 54 Tyr Leu Ala Ser Tyr Arg Tyr Ser
1 5 <210> SEQ ID NO 55 <211> LENGTH: 8 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Light Chain CDR2
<400> SEQUENCE: 55 Tyr Leu Ala Ser Tyr Arg Tyr Arg 1 5
<210> SEQ ID NO 56 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR3 <400>
SEQUENCE: 56 His Gln Tyr Tyr Thr Tyr Pro Leu Phe Thr 1 5 10
<210> SEQ ID NO 57 <211> LENGTH: 30 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 <400>
SEQUENCE: 57 ggatacacct tcactgagtt tggaatgaac 30 <210> SEQ ID
NO 58 <211> LENGTH: 30 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Heavy Chain CDR1 <400> SEQUENCE: 58
ggatacacct tcactgagta tggtatgaac 30 <210> SEQ ID NO 59
<211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 <400> SEQUENCE: 59 ggatacacct
tcactgagta ttctatgaac 30 <210> SEQ ID NO 60 <400>
SEQUENCE: 60 000 <210> SEQ ID NO 61 <211> LENGTH: 30
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR1 <400> SEQUENCE: 61 ggatacacct tcactgagtt tggaatgagc 30
<210> SEQ ID NO 62 <211> LENGTH: 51 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 <400>
SEQUENCE: 62 tggataaaca ccaaaactgg agaggcaaca tatgttgaag agtttaaggg
a 51 <210> SEQ ID NO 63 <211> LENGTH: 51 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Heavy Chain CDR2
<400> SEQUENCE: 63 tggataaaca ccaaaactgg agaggcaaca
tatattgaag agtttaaggg a 51 <210> SEQ ID NO 64 <211>
LENGTH: 51 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 <400> SEQUENCE: 64 tggataaaca ccaaaagtgg
agaggcaaca tatgttgaag agtttaaggg a 51 <210> SEQ ID NO 65
<211> LENGTH: 51 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR2 <400> SEQUENCE: 65 tatataaaca
ccaaaaatgg agaggcaaac tatgttgaag agtttaaggg a 51 <210> SEQ ID
NO 66 <211> LENGTH: 51 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Heavy Chain CDR2 <400> SEQUENCE: 66
tggataaaca ccaaaaatgg agaggcaaca tatattgaag agtttaaggg a 51
<210> SEQ ID NO 67 <211> LENGTH: 36 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 67 tgggacttct atgattacgt ggaggctatg gactac 36 <210>
SEQ ID NO 68 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 68 tgggacttct atcattacgt ggaggctatg gactac 36 <210>
SEQ ID NO 69 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 69 tgggacttcg tggattacgt ggaggctatg gactac 36 <210>
SEQ ID NO 70 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 70 tgggacttct attggtacgt ggaggctatg gactac 36 <210>
SEQ ID NO 71 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 71 tgggacgcct ttgagtacgt gaaggcgctg gactac 36 <210>
SEQ ID NO 72 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 72 tgggatttct ttgagtattt taagactatg gactac 36 <210>
SEQ ID NO 73 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 73 tgggactttt tttattacgt gcagactatg gactac 36 <210>
SEQ ID NO 74 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 74 tgggattttt cttattacgt tgaggcgatg gactac 36 <210>
SEQ ID NO 75 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 75 tgggactttg ctcattactt tcagactatg gactac 36 <210>
SEQ ID NO 76 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 76 tgggacttcg cttattactt tcagactatg gactac 36 <210>
SEQ ID NO 77 <211> LENGTH: 36 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 77 tgggatttcg cgtattacct tgaggctatg gactac 36 <210>
SEQ ID NO 78 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 78 aaggccagtc agaatgtggg tactaatgtt gcc 33 <210>
SEQ ID NO 79 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 79 aaggccagtg ccaatgtggg taataatgtt gcc 33 <210>
SEQ ID NO 80 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 80 aaggccagta agaatgtggg gactaatgtt gcg 33 <210>
SEQ ID NO 81 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 81 aaggccagtg cggctgtggg tacgtatgtt gcg 33 <210>
SEQ ID NO 82 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 82 aaggccagtc agatagcgag tactaatgtt gcc 33 <210>
SEQ ID NO 83 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 83 aaggccagtc acaatgtggg taccaacgtt gcg 33 <210>
SEQ ID NO 84 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 84 aaggccagtc agattatggg tcctaatgtt gcg 33 <210>
SEQ ID NO 85 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 85 aaggccagtc aaattgtggg tactaatgtt gcg 33 <210>
SEQ ID NO 86 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 86 aaggccagtc agaaggtgct tactaatgtt gcg 33 <210>
SEQ ID NO 87 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 87 aaggccagtc agactgtgag tgctaatgtt gcg 33 <210>
SEQ ID NO 88 <211> LENGTH: 24 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 88 tattcggcat cctaccgcta cagt 24 <210> SEQ ID NO 89
<211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 89 tatttggcct
ccaacctctc cggt 24 <210> SEQ ID NO 90 <211> LENGTH: 24
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 90 tacctggcat cctaccccca gatt 24
<210> SEQ ID NO 91 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 91 tattcggcat cctaccgcaa aagg 24 <210> SEQ ID NO 92
<211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 92 tattgggcat
cctaccgcta tagt 24 <210> SEQ ID NO 93 <211> LENGTH: 24
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 93 tattcggcat cccaccggta cagt 24
<210> SEQ ID NO 94 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 94 tatttggcat cctaccacga aagt 24 <210> SEQ ID NO 95
<211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 95 tattcggcat
cccaccgtcc cagt 24 <210> SEQ ID NO 96 <211> LENGTH: 24
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 96 tatttggcat cctaccgcta cagt 24
<210> SEQ ID NO 97 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 97 tatttggcat cctaccgcta caga 24 <210> SEQ ID NO 98
<211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR3 <400> SEQUENCE: 98 caccaatatt
acacctatcc tctattcacg 30 <210> SEQ ID NO 99 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: PR1A3
VH <400> SEQUENCE: 99 Gln Val Lys Leu Gln Gln Ser Gly Pro Glu
Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Lys
Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Tyr 65 70 75 80
Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Lys Tyr Phe Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 100 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pEM1496 huPR1A3 VH <400>
SEQUENCE: 100 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 101 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7A <400> SEQUENCE: 101 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 102 <211> LENGTH: 98
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: IGHV7-4-1*02
<400> SEQUENCE: 102 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Asn Thr Gly Asn Pro Thr Tyr Ala Gln Gly Phe 50 55 60 Thr Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg <210> SEQ ID NO 103 <211> LENGTH: 110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 VL
<400> SEQUENCE: 103 Asp Ile Val Met Thr Gln Ser Gln Arg Phe
Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Val Thr Cys Lys
Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser
Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80
Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Met Lys Arg Thr 100
105 110 <210> SEQ ID NO 104 <211> LENGTH: 109
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pEM1495 huPR1A3
VL <400> SEQUENCE: 104 Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys
Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala
Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser
Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70
75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro
Leu 85 90 95 Phe Ser Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105 <210> SEQ ID NO 105 <211> LENGTH: 108
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CL1A
<400> SEQUENCE: 105 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser
Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 106 <211> LENGTH: 95 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: IMGT_hVK_1_39 <400> SEQUENCE:
106 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser
Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr
Tyr Cys Gln Gln Ser Tyr Ser Thr Pro 85 90 95 <210> SEQ ID NO
107 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7 rF9 <400> SEQUENCE: 107 Gln Val Gln
Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 108 <211> LENGTH:
110 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CLA1 rH11
<400> SEQUENCE: 108 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Thr Val Ser Ala Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser
Tyr Arg Tyr Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 109 <400> SEQUENCE: 109 000
<210> SEQ ID NO 110 <400> SEQUENCE: 110 000 <210>
SEQ ID NO 111 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 VH <400> SEQUENCE: 111
caggtgaagc tgcagcagtc aggacctgag ttgaagaagc ctggagagac agtcaagatc
60 tcctgcaagg cttctggata taccttcaca gaattcggaa tgaactgggt
gaagcaggct 120 cctggaaagg gtttaaagtg gatgggctgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgccttc
tctttggaga cctctgccac cactgcctat 240 ttgcagatca acaacctcaa
aaatgaggac acggctaaat atttctgtgc tcgatgggat 300 ttctatgact
atgttgaagc tatggactac tggggccaag ggaccaccgt gaccgtctcc 360 tca 363
<210> SEQ ID NO 112 <211> LENGTH: 362 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pEM1496 <400> SEQUENCE: 112
caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagttggaat gaactgggtg
cgacaggccc 120 ctggacaagg gcttgagtgg atgggatgga taaacaccaa
aactggagag gcaacatatg 180 ttgaagagtt taagggacgg tttgtcttct
ccttggacac ctctgtcagc acggcatatc 240 tgcagatcag cagcctaaag
gctgacgaca ctgccgtgta ttactgtgcg agatgggact 300 tctatgatta
cgtggaggct atggactact ggggccaagg gaccacggtc accgtctcct 360 ca 362
<210> SEQ ID NO 113 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A <400> SEQUENCE: 113
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttctatgatt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 114 <211> LENGTH: 294 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: IGHV7-4-1*02 <400> SEQUENCE:
114 caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact agctatgcta
tgaattgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg
atcaacacca acactgggaa cccaacgtat 180 gcccagggct tcacaggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaggac actgccgtgt attactgtgc gaga 294 <210>
SEQ ID NO 115 <211> LENGTH: 330 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 VL <400> SEQUENCE: 115
gatatcgtga tgacccagtc tcaaagattc atgtccacat cagtaggaga cagggtcagc
60 gtcacctgca aggccagtca gaatgtgggt actaatgttg cctggtatca
acagaaacca 120 ggacaatccc ctaaagcact gatttactcg gcatcctacc
ggtacagtgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat
ttcactctca ccatcagcaa tgtacagtct 240 gaagacttgg cggagtattt
ctgtcaccaa tattacacct atcctctatt cacgttcggc 300 tcggggacaa
agttggaaat gaaacgtacg 330 <210> SEQ ID NO 116 <211>
LENGTH: 327 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
pEM1495 <400> SEQUENCE: 116 gacatccaga tgactcagag cccaagcagc
ctgagcgcca gcgtgggtga cagagtgacc 60 atcacctgta aggccagtca
gaatgtgggt actaatgttg cctggtacca gcagaagcca 120 ggtaaggctc
caaagctgct gatctactcg gcatcctacc ggtacagtgg tgtgccaagc 180
agattcagcg gtagcggtag cggtaccgac ttcaccttca ccatcagcag cctccagcca
240 gaggacatcg ccacctacta ctgccaccaa tattacacct atcctctatt
cagcttcggc 300 caagggacca aggtggaaat caaacgt 327 <210> SEQ ID
NO 117 <211> LENGTH: 324 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CL1A <400> SEQUENCE: 117 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtca gaatgtgggt actaatgttg cctggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctattcg gcatcctacc gctacagtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat caag 324
<210> SEQ ID NO 118 <211> LENGTH: 285 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: IMGT_hVK_1_39 <400> SEQUENCE:
118 gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga
cagagtcacc 60 atcacttgcc gggcaagtca gagcattagc agctatttaa
attggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct
gcatccagtt tgcaaagtgg ggtcccatca 180 aggttcagtg gcagtggatc
tgggacagat ttcactctca ccatcagcag tctgcaacct 240 gaagattttg
caacttacta ctgtcaacag agttacagta cccct 285 <210> SEQ ID NO
119 <211> LENGTH: 369 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7 rF9 <400> SEQUENCE: 119 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtatggta tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacga aatctggaga
ggcaacctat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatgatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tcagctagc 369
<210> SEQ ID NO 120 <211> LENGTH: 330 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CLA1 rH11 <400> SEQUENCE: 120
gatatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc
60 atcacttgca aggccagtca gactgtgagt gctaatgttg cgtggtatca
gcagaaacca 120 gggaaagcac ctaagctcct gatctacttg gcatcctacc
gctacagagg agtcccatca 180 aggttcagtg gcagtggatc tgggacagat
ttcactctca ccatcagcag tctgcaacct 240 gaagatttcg caacttacta
ctgtcaccaa tattacacct atcctctatt cacgtttggc 300 cagggcacca
agctcgagat caagcgtacg 330 <210> SEQ ID NO 121 <211>
LENGTH: 987 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: IgG1
<400> SEQUENCE: 121 accaagggcc catcggtctt ccccctggca
ccctcctcca agagcacctc tgggggcaca 60 gcggccctgg gctgcctggt
caaggactac ttccccgaac cggtgacggt gtcgtggaac 120 tcaggcgccc
tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 180
tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc
240 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagcaga
gcccaaatct 300 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg
aactcctggg gggaccgtca 360 gtcttcctct tccccccaaa acccaaggac
accctcatga tctcccggac ccctgaggtc 420 acatgcgtgg tggtggacgt
gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 480 gacggcgtgg
aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 540
taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac
600 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
ctccaaagcc 660 aaagggcagc cccgagaacc acaggtgtac accctgcccc
catcccggga tgagctgacc 720 aagaaccagg tcagcctgac ctgcctggtc
aaaggcttct atcccagcga catcgccgtg 780 gagtgggaga gcaatgggca
gccggagaac aactacaaga ccacgcctcc cgtgctggac 840 tccgacggct
ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 900
gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag
960 agcctctccc tgtctccggg taaatga 987 <210> SEQ ID NO 122
<211> LENGTH: 328 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: IgG1 <400> SEQUENCE: 122 Thr Lys Gly Pro Ser Val
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr 1 5 10 15 Ser Gly Gly Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro 20 25 30 Glu Pro
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val 35 40 45
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser 50
55 60 Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
Ile 65 70 75 80 Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp
Lys Lys Ala 85 90 95 Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
Pro Pro Cys Pro Ala 100 105 110 Pro Glu Leu Leu Gly Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys Pro 115 120 125 Lys Asp Thr Leu Met Ile Ser
Arg Thr Pro Glu Val Thr Cys Val Val 130 135 140 Val Asp Val Ser His
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 145 150 155 160 Asp Gly
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 165 170 175
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 180
185 190 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala 195 200 205 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro 210 215 220 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
Arg Asp Glu Leu Thr 225 230 235 240 Lys Asn Gln Val Ser Leu Thr Cys
Leu Val Lys Gly Phe Tyr Pro Ser 245 250 255 Asp Ile Ala Val Glu Trp
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 260 265 270 Lys Thr Thr Pro
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 275 280 285 Ser Lys
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 290 295 300
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 305
310 315 320 Ser Leu Ser Leu Ser Pro Gly Lys 325 <210> SEQ ID
NO 123 <211> LENGTH: 42 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer MS-43 <400> SEQUENCE: 123
ccagccggcc atggccgata tccagatgac ccagtctcca tc 42 <210> SEQ
ID NO 124 <211> LENGTH: 26 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer MS-52 <400> SEQUENCE: 124
gaagaccgat gggcctttgg tgctag 26 <210> SEQ ID NO 125
<211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer MS-55 <400> SEQUENCE: 125 gcaacatatg
ttgaagagtt taagggacgg 30 <210> SEQ ID NO 126 <211>
LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: MS-56
<400> SEQUENCE: 126 atgaactggg tgcgacaggc ccctg 25
<210> SEQ ID NO 127 <211> LENGTH: 56 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-679 <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (26)..(26)
<223> OTHER INFORMATION: m is a or c <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (27)..(28)
<223> OTHER INFORMATION: n is a, c, g, or t <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(30)..(30) <223> OTHER INFORMATION: w is a or t <400>
SEQUENCE: 127 caggggcctg tcgcacccag ttcatmnnaw actcagtgaa
ggtgtatcca gaagcc 56 <210> SEQ ID NO 128 <211> LENGTH:
81 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer EAB-680
<400> SEQUENCE: 128 ccgtccctta aactcttcaa cataggttgc
ctctccagtt ttggtgttta tccatcccat 60 ccactcaagc ccttgtccag g 81
<210> SEQ ID NO 129 <211> LENGTH: 50 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-685 <400> SEQUENCE:
129 cagctatgac catgattacg ccaagcttgc atgcaaattc tatttcaagg 50
<210> SEQ ID NO 130 <211> LENGTH: 27 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-686 <400> SEQUENCE:
130 gttgcgtggt atcagcagaa accaggg 27 <210> SEQ ID NO 131
<211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer EAB-687 <400> SEQUENCE: 131 gctctttgtg
acgggcgagc tcaggccctg atgg 34 <210> SEQ ID NO 132 <211>
LENGTH: 35 <212> TYPE: DNA <213> ORGANISM: Artificial
sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
EAB-688 <400> SEQUENCE: 132 ggagtcccat caaggttcag tggcagtgga
tctgg 35 <210> SEQ ID NO 133 <211> LENGTH: 70
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer EAB-681
<400> SEQUENCE: 133 cctggtttct gctgatacca cgcaacatta
gtacccacat tctgactggc cttgcaagtg 60 atggtgactc 70 <210> SEQ
ID NO 134 <211> LENGTH: 77 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer EAB-682 <400> SEQUENCE: 134
ctgccactga accttgatgg gactccactg tagcggtagg atgccgaata gatcaggagc
60 ttaggtgctt tccctgg 77 <210> SEQ ID NO 135 <211>
LENGTH: 64 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
AC7 <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (37)..(37) <223> OTHER INFORMATION: m
is a or c <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (38)..(39) <223> OTHER INFORMATION: n
is a, c, g, or t <400> SEQUENCE: 135 ccagtagtcc atagcctcca
cgtaatcata gaagtcmnnt ctcgcacagt aatacacggc 60 agtg 64 <210>
SEQ ID NO 136 <211> LENGTH: 64 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer AC8 <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (34)..(34)
<223> OTHER INFORMATION: m is a or c <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (35)..(36)
<223> OTHER INFORMATION: n is a, c, g, or t <400>
SEQUENCE: 136 ccagtagtcc atagcctcca cgtaatcata gaamnnccat
ctcgcacagt aatacacggc 60 agtg 64 <210> SEQ ID NO 137
<211> LENGTH: 64 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer AC9 <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (31)..(31) <223> OTHER
INFORMATION: m is a or c <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (32)..(33) <223> OTHER
INFORMATION: n is a, c, g, or t <400> SEQUENCE: 137
ccagtagtcc atagcctcca cgtaatcata mnngtcccat ctcgcacagt aatacacggc
60 agtg 64 <210> SEQ ID NO 138 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC10
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (28)..(28) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (29)..(30) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 138 ccagtagtcc atagcctcca cgtaatcmnn
gaagtcccat ctcgcacagt aatacacggc 60 agtg 64 <210> SEQ ID NO
139 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC11 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (25)..(25) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (26)..(27) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 139
ccagtagtcc atagcctcca cgtamnnata gaagtcccat ctcgcacagt aatacacggc
60 agtg 64 <210> SEQ ID NO 140 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC12
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (37)..(37) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (38)..(39) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 140 cgtggtccct tggccccagt agtccatagc
ctccacmnna tcatagaagt cccatctcgc 60 acag 64 <210> SEQ ID NO
141 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC13 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (34)..(34) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (35)..(36) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 141
cgtggtccct tggccccagt agtccatagc ctcmnngtaa tcatagaagt cccatctcgc
60 acag 64 <210> SEQ ID NO 142 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC14
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (31)..(31) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (32)..(33) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 142 cgtggtccct tggccccagt agtccatagc
mnncacgtaa tcatagaagt cccatctcgc 60 acag 64 <210> SEQ ID NO
143 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC15 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (28)..(28) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (29)..(30) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 143
cgtggtccct tggccccagt agtccatmnn ctccacgtaa tcatagaagt cccatctcgc
60 acag 64 <210> SEQ ID NO 144 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC16
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (25)..(25) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (26)..(27) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 144 cgtggtccct tggccccagt agtcmnnagc
ctccacgtaa tcatagaagt cccatctcgc 60 acag 64 <210> SEQ ID NO
145 <211> LENGTH: 77 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC17 <400> SEQUENCE: 145 cgtggtccct
tggccccagt agtccatagc ctccacgtaa tcatagaagt cccatctcgc 60
acagtaatac acggcag 77 <210> SEQ ID NO 146 <211> LENGTH:
25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer EAB-749
<400> SEQUENCE: 146 ccatcagggc ctgagctcgc ccgtc 25
<210> SEQ ID NO 147 <211> LENGTH: 30 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-750 <400> SEQUENCE:
147 cgtggaggct atggactact ggggccaagg 30 <210> SEQ ID NO 148
<211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer EAB-751 <400> SEQUENCE: 148 gactactggg
gccaagggac cacggtcac 29 <210> SEQ ID NO 149 <211>
LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
EAB-752 <400> SEQUENCE: 149 ggtcagggcg cctgagttcc acg 23
<210> SEQ ID NO 150 <211> LENGTH: 60 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer AC1 <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (37)..(37)
<223> OTHER INFORMATION: m is a or c <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (38)..(39)
<223> OTHER INFORMATION: n is a, c, g, or t <400>
SEQUENCE: 150 ggtgccctgg ccaaacgtga atagaggata ggtgtamnnt
tggtgacagt agtaagttgc 60 <210> SEQ ID NO 151 <211>
LENGTH: 60 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
AC2 <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (34)..(34) <223> OTHER INFORMATION: m
is a or c <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (35)..(36) <223> OTHER INFORMATION: n
is a, c, g, or t <400> SEQUENCE: 151 ggtgccctgg ccaaacgtga
atagaggata ggtmnnatat tggtgacagt agtaagttgc 60 <210> SEQ ID
NO 152 <211> LENGTH: 60 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC3 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (31)..(31) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (32)..(33) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 152
ggtgccctgg ccaaacgtga atagaggata mnngtaatat tggtgacagt agtaagttgc
60 <210> SEQ ID NO 153 <211> LENGTH: 60 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Primer AC4 <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(28)..(28) <223> OTHER INFORMATION: m is a or c <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(29)..(30) <223> OTHER INFORMATION: n is a, c, g, or t
<400> SEQUENCE: 153 ggtgccctgg ccaaacgtga atagaggmnn
ggtgtaatat tggtgacagt agtaagttgc 60 <210> SEQ ID NO 154
<211> LENGTH: 60 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer AC5 <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (22)..(22) <223> OTHER
INFORMATION: m is a or c <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (23)..(24) <223> OTHER
INFORMATION: n is a, c, g, or t <400> SEQUENCE: 154
ggtgccctgg ccaaacgtga amnnaggata ggtgtaatat tggtgacagt agtaagttgc
60 <210> SEQ ID NO 155 <211> LENGTH: 60 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequences <220>
FEATURE: <223> OTHER INFORMATION: Primer AC6 <400>
SEQUENCE: 155 ggtgccctgg ccaaacgtga atagaggata ggtgtaatat
tggtgacagt agtaagttgc 60 <210> SEQ ID NO 156 <211>
LENGTH: 36 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
EAB-746 <400> SEQUENCE: 156 cgcttgatct cgagcttggt gccctggcca
aacgtg 36 <210> SEQ ID NO 157 <400> SEQUENCE: 157 000
<210> SEQ ID NO 158 <211> LENGTH: 428 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pETR6592 <400> SEQUENCE: 158
Gln Leu Thr Thr Glu Ser Met Pro Phe Asn Val Ala Glu Gly Lys Glu 1 5
10 15 Val Leu Leu Leu Val His Asn Leu Pro Gln Gln Leu Phe Gly Tyr
Ser 20 25 30 Trp Tyr Lys Gly Glu Arg Val Asp Gly Asn Arg Gln Ile
Val Gly Tyr 35 40 45 Ala Ile Gly Thr Gln Gln Ala Thr Pro Gly Pro
Ala Asn Ser Gly Arg 50 55 60 Glu Thr Ile Tyr Pro Asn Ala Ser Leu
Leu Ile Gln Asn Val Thr Gln 65 70 75 80 Asn Asp Thr Gly Phe Tyr Thr
Leu Gln Val Ile Lys Ser Asp Leu Val 85 90 95 Asn Glu Glu Ala Thr
Gly Gln Phe His Val Tyr Pro Glu Leu Pro Lys 100 105 110 Pro Ser Ile
Ser Ser Asn Asn Ser Asn Pro Val Glu Asp Lys Asp Ala 115 120 125 Met
Ala Phe Thr Cys Glu Pro Glu Thr Gln Asp Thr Thr Tyr Leu Trp 130 135
140 Trp Ile Asn Asn Gln Ser Leu Pro Val Ser Pro Arg Leu Gln Leu Ser
145 150 155 160 Asn Gly Asn Arg Thr Leu Thr Leu Leu Ser Val Thr Arg
Asn Asp Thr 165 170 175 Gly Pro Tyr Glu Cys Glu Ile Gln Asn Pro Val
Ser Ala Asn Arg Ser 180 185 190 Asp Pro Val Thr Leu Asn Val Thr Tyr
Gly Pro Asp Thr Pro Thr Ile 195 200 205 Ser Pro Pro Asp Ser Ser Tyr
Leu Ser Gly Ala Asn Leu Asn Leu Ser 210 215 220 Cys His Ser Ala Ser
Asn Pro Ser Pro Gln Tyr Ser Trp Arg Ile Asn 225 230 235 240 Gly Ile
Pro Gln Gln His Thr Gln Val Leu Phe Ile Ala Lys Ile Thr 245 250 255
Pro Asn Asn Asn Gly Thr Tyr Ala Cys Phe Val Ser Asn Leu Ala Thr 260
265 270 Gly Arg Asn Asn Ser Ile Val Lys Ser Ile Thr Val Ser Ala Leu
Ser 275 280 285 Pro Val Val Ala Lys Pro Gln Ile Lys Ala Ser Lys Thr
Thr Val Thr 290 295 300 Gly Asp Lys Asp Ser Val Asn Leu Thr Cys Ser
Thr Asn Asp Thr Gly 305 310 315 320 Ile Ser Ile Arg Trp Phe Phe Lys
Asn Gln Ser Leu Pro Ser Ser Glu 325 330 335 Arg Met Lys Leu Ser Gln
Gly Asn Ile Thr Leu Ser Ile Asn Pro Val 340 345 350 Lys Arg Glu Asp
Ala Gly Thr Tyr Trp Cys Glu Val Phe Asn Pro Ile 355 360 365 Ser Lys
Asn Gln Ser Asp Pro Ile Met Leu Asn Val Asn Tyr Asn Ala 370 375 380
Leu Pro Gln Glu Asn Leu Ile Asn Val Asp Leu Glu Val Leu Phe Gln 385
390 395 400 Gly Pro Gly Ser Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys
Ile Glu 405 410 415 Trp His Glu Ala Arg Ala His His His His His His
420 425 <210> SEQ ID NO 159 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PMS22
<400> SEQUENCE: 159 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 ttctatgatt acgtggaggc tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 160
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer 1C8 <400> SEQUENCE: 160 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatcatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 161 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: 3E1 <400> SEQUENCE: 161
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttcgtggatt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 162 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: 2D7 <400> SEQUENCE: 162
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttctattggt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 163 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Affinity Matured Heavy Chain
<400> SEQUENCE: 163 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 tttgctcatt actttcagac tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 164
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Affinity Matured Heavy Chain <400> SEQUENCE: 164
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttcgcttatt
actttcagac tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 165 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Affinity Matured Heavy Chain
<400> SEQUENCE: 165 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggat 300 ttcgcgtatt accttgaggc tatggactac tggggccaag
ggaccacgat caccgtctcc 360 tca 363 <210> SEQ ID NO 166
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H3 Full (5) 19 <400> SEQUENCE: 166 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgagctgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgctgtgt attactgtgc gagatgggac 300 gcctttgagt acgtgaaggc
gctggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 167 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H3 Full (5) 8 <400> SEQUENCE:
167 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact gagtttggaa
tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg
ataaacacca aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggat 300
ttctttgagt attttaagac tatggactac tggggccaag ggaccacggt caccgtctcc
360 tca 363 <210> SEQ ID NO 168 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H3 Full (5) 28
<400> SEQUENCE: 168 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 tttttttatt acgtgcagac tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 169
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H3 Full (5) 27 <400> SEQUENCE: 169 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggat 300 ttttcttatt acgttgaggc
gatggactac tggggccaag ggaccacagt caccgtctcc 360 tca 363 <210>
SEQ ID NO 170 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H4E9 Heavy Chain <400>
SEQUENCE: 170 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc
ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata caccttcact
gagtttggta tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg
gatgggatgg ataaatacca aaactggaga ggcaacttat 180 attgaagagt
ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240
ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac
300 ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt
caccgtctcc 360 tca 363 <210> SEQ ID NO 171 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: pAC14
(B9) <400> SEQUENCE: 171 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggta tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaagtggaga ggcaacctat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 ttctatgatt acgtggaggc tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 172
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC15 (F9) <400> SEQUENCE: 172 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtatggta tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacga aatctggaga
ggcaacctat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatgatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 173 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 2 <400> SEQUENCE:
173 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact gagtattcta
tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatac
ataaacacca aaaatggaga ggcaaactat 180 gttgaagagt ttaagggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac 300
ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc
360 tca 363 <210> SEQ ID NO 174 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H1/H2 (5) 11
<400> SEQUENCE: 174 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtatggta tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaaatggaga ggcaacctat 180
attgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 ttctatgatt acgtggaggc tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 175
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H1/H2 (5) 13 <400> SEQUENCE: 175 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggta tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatat ataaacacca aaaatggaga
ggcaaactat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggacg
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatgatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 176 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 14 <400> SEQUENCE:
176 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact gagtatggta
tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatat
ataaacacca aaaatggaga ggcaaactat 180 gttgaagagt ttaagggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac 300
ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc
360 tca 363 <210> SEQ ID NO 177 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H3 Full (5) 19
<400> SEQUENCE: 177 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgagctgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgctgtgt attactgtgc
gagatgggac 300 gcctttgagt acgtgaaggc gctggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 178
<211> LENGTH: 330 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC21 (3A1) <400> SEQUENCE: 178 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtgc caatgtgggt aataatgttg cctggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctatttg gcctccaacc gctccggtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 179 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC19 (2C6)
<400> SEQUENCE: 179 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtaa
gaatgtgggg actaatgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagcccct gatctacctg gcatcctacc cccagattgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcccctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 180 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC18 (2F1) <400> SEQUENCE: 180 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtgc ggctgtgggt acgtatgttg cgtggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctattcg gcatcctacc gcaaaagggg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 181 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC23 (2F11)
<400> SEQUENCE: 181 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
gatagcgagt actaatgttg cctggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctattgg gcatcctacc gctatagtgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 182 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: H4E9 light chain <400> SEQUENCE: 182
gatatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc
60 atcacttgca aggccagtca gaatgtgggt actaatgttg cctggtatca
gcagaaacca 120 gggaaagcac ctaagcccct gatctattcg gcatcctacc
gctacagtgg agtcccatca 180 aggttcagtg gcagtggatc tgggacagat
ttcactctca ccatcagcag tctgcaacct 240 gaagatttcg caacttacta
ctgtcaccaa tattacacct atcctctatt cacgtttggc 300 cagggcacca
agctcgagat caagcgtacg 330 <210> SEQ ID NO 183 <211>
LENGTH: 330 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: L2D2
<400> SEQUENCE: 183 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
caatgtgggt accaacgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctattcg gcatcccacc ggtacagtgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 184 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC6 (C1) <400> SEQUENCE: 184 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtca gattatgggt cctaatgttg cgtggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctatttg gcatcctacc acgaaagtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 185 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC7 (E10)
<400> SEQUENCE: 185 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
aattgtgggt actaatgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctattcg gcatcccacc gtcccagtgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 186 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC12 (H7) <400> SEQUENCE: 186 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtca gaaggtgctt actaatgttg cgtggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctatttg gcatcctacc gctacagtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 187 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC13 (H11)
<400> SEQUENCE: 187 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
gactgtgagt gctaatgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctacttg gcatcctacc gctacagagg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 188 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: PMS22 <400> SEQUENCE: 188 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 189 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: 1C8 <400>
SEQUENCE: 189 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr His Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 190 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: 3E1 <400> SEQUENCE: 190 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Val Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 191 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: 2D7 <400>
SEQUENCE: 191 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr Trp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 192 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Affinity Matured Heavy Chain <400>
SEQUENCE: 192 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Ala His Tyr Phe Gln Thr Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 193 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Affinity Matured Heavy Chain <400>
SEQUENCE: 193 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Ala Tyr Tyr Phe Gln Thr Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 194 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Affinity Matured Heavy Chain <400>
SEQUENCE: 194 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Ala Tyr Tyr Leu Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Ile Thr Val Ser Ser 115 120 <210> SEQ ID
NO 195 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: H3 Full (5) 19 <400> SEQUENCE: 195 Gln Val
Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser
Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Ala Phe Glu
Tyr Val Lys Ala Leu Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 196 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: H3
Full (5) 8 <400> SEQUENCE: 196 Gln Val Gln Leu Val Gln Ser
Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55
60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr
65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Phe Glu Tyr Phe Lys Thr Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 197 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: H3 Full (5) 28 <400>
SEQUENCE: 197 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Phe Tyr Tyr Val Gln Thr Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 198 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: H3 Full (5) 27 <400> SEQUENCE: 198 Gln Val
Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser
Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ser Tyr
Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 199 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: H4E9
Heavy Chain <400> SEQUENCE: 199 Gln Val Gln Leu Val Gln Ser
Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Ile Glu Glu Phe 50 55
60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr
65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 200 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: pAC14 (B9) <400>
SEQUENCE: 200 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Ser
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 201 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC15 (F9) <400> SEQUENCE: 201 Gln Val Gln
Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 202 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H1/H2 (5) 2
<400> SEQUENCE: 202 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 Ser Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn
Thr Lys Asn Gly Glu Ala Asn Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 203 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 11 <400> SEQUENCE:
203 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Asn Gly Glu Ala
Thr Tyr Ile Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu
Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu
Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 204
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H1/H2 (5) 13 <400> SEQUENCE: 204 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Tyr Ile Asn Thr Lys Asn Gly Glu Ala Asn Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Ala Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 205 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H1/H2 (5) 14
<400> SEQUENCE: 205 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn
Thr Lys Asn Gly Glu Ala Asn Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 206 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H3 Full (5) 19 <400> SEQUENCE:
206 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu
Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu
Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Ala Phe Glu Tyr Val Lys Ala Leu Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 207
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC21 (3A1) <400> SEQUENCE: 207 Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Ala Asn Val Gly Asn Asn 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35
40 45 Tyr Leu Ala Ser Asn Arg Ser Gly Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr
Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 208
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC19 (2C6) <400> SEQUENCE: 208 Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Lys Asn Val Gly Thr Asn 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu Ile 35
40 45 Tyr Leu Ala Ser Tyr Pro Gln Ile Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr
Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 209
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC18 (2F1) <400> SEQUENCE: 209 Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Ala Ala Val Gly Thr Tyr 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35
40 45 Tyr Ser Ala Ser Tyr Arg Lys Arg Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr
Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 210
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC23 (2F11) <400> SEQUENCE: 210 Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Gln Ile Ala Ser Thr Asn 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35
40 45 Tyr Trp Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr
Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 211
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H4E9 light chain <400> SEQUENCE: 211 Asp Ile Gln
Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp
Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25
30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu Ile
35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln
Tyr Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys
Leu Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 212
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: L2D2 <400> SEQUENCE: 212 Asp Ile Gln Met Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr
Ile Thr Cys Lys Ala Ser His Asn Val Gly Thr Asn 20 25 30 Val Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45
Tyr Ser Ala Ser His Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50
55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr
Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr 100 105 110 <210> SEQ ID NO 213 <211>
LENGTH: 110 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: pAC6
(C1) <400> SEQUENCE: 213 Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys
Lys Ala Ser Gln Ile Met Gly Pro Asn 20 25 30 Val Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala
Ser Tyr His Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70
75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro
Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
Thr 100 105 110 <210> SEQ ID NO 214 <211> LENGTH: 110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC7 (E10)
<400> SEQUENCE: 214 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Ile Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser
His Arg Pro Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 215 <211> LENGTH: 110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC12 (H7)
<400> SEQUENCE: 215 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Lys Val Leu Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser
Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 216 <211> LENGTH: 110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC13 (H11)
<400> SEQUENCE: 216 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Thr Val Ser Ala Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser
Tyr Arg Tyr Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 217 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 (Y98A)
Heavy Chain CDR3 <400> SEQUENCE: 217 Trp Asp Phe Ala Asp Tyr
Val Glu Ala Met Asp Tyr 1 5 10 <210> SEQ ID NO 218
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 (D99Y) Heavy Chain CDR3 <400> SEQUENCE:
218 Trp Asp Phe Tyr Tyr Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 219 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 (D99H) Heavy Chain CDR3
<400> SEQUENCE: 219 Trp Asp Phe Tyr His Tyr Val Glu Ala Met
Asp Tyr 1 5 10 <210> SEQ ID NO 220 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 (V101F)
Heavy Chain CDR3 <400> SEQUENCE: 220 Trp Asp Phe Tyr Asp Tyr
Phe Glu Ala Met Asp Tyr 1 5 10 <210> SEQ ID NO 221
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 (E101aQ) Heavy Chain CDR3 <400> SEQUENCE:
221 Trp Asp Phe Tyr Asp Tyr Val Gln Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 222 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 (A103T) Heavy Chain CDR3
<400> SEQUENCE: 222 Trp Asp Phe Tyr Asp Tyr Val Glu Thr Met
Asp Tyr 1 5 10 <210> SEQ ID NO 223 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 (Y98A /
D99Y) Heavy Chain CDR3 <400> SEQUENCE: 223 Trp Asp Phe Ala
Tyr Tyr Val Glu Ala Met Asp Tyr 1 5 10 <210> SEQ ID NO 224
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 (W95Y) Heavy Chain CDR3 <400> SEQUENCE:
224 Tyr Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 225 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (Y98A) Heavy Chain Construct
<400> SEQUENCE: 225 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Ala Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 226 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (D99Y) Heavy Chain Construct
<400> SEQUENCE: 226 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Tyr Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 227 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (D99H) Heavy Chain Construct
<400> SEQUENCE: 227 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr His Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 228 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (V101F) Heavy Chain Construct
<400> SEQUENCE: 228 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Phe Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 229 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (E102Q) Heavy Chain Construct
<400> SEQUENCE: 229 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Gln Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 230 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (A103T) Heavy Chain Construct
<400> SEQUENCE: 230 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Thr Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 231 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (Y98A / D99Y) Heavy Chain
Construct <400> SEQUENCE: 231 Gln Val Gln Leu Val Gln Ser Gly
Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65
70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ala Tyr Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 232 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (W95Y) Heavy Chain
Construct <400> SEQUENCE: 232 Gln Val Gln Leu Val Gln Ser Gly
Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65
70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Tyr Asp Phe Tyr Asp Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 233 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH1A1A (Y98A) <400>
SEQUENCE: 233 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr
Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu
Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Ala Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 234 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH1A1A (D99Y) <400> SEQUENCE: 234 Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser
Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Tyr
Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 235 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1A
(D99H) <400> SEQUENCE: 235 Gln Val Gln Leu Val Gln Ser Gly
Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr His Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 236 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH1A1A (V101F) <400>
SEQUENCE: 236 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr
Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu
Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr Asp Tyr Phe Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 237 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH1A1A (E102Q) <400> SEQUENCE: 237 Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser
Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp
Tyr Val Gln Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 238 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1A
(A103T) <400> SEQUENCE: 238 Gln Val Gln Leu Val Gln Ser Gly
Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Thr Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 239 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH1A1A (Y98A / D99Y)
<400> SEQUENCE: 239 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Ala Tyr Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 240 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1A (W95Y) <400> SEQUENCE:
240 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr
Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Asp
Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 241
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1B (Y98A) <400> SEQUENCE: 241 Gln Val Lys
Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr
Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ala Asp Tyr
Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 242 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1B (D99Y)
<400> SEQUENCE: 242 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Tyr Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 243 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B (D99H) <400> SEQUENCE:
243 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr
Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr His Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 244
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1B (V101F) <400> SEQUENCE: 244 Gln Val Lys
Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr
Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Phe Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 245 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1B (E102Q)
<400> SEQUENCE: 245 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Gln Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 246 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B (A103T) <400> SEQUENCE:
246 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr
Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Asp Tyr Val Glu Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 247
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1B (Y98A / D99Y) <400> SEQUENCE: 247 Gln Val
Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser
Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ala Tyr
Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 248 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1B
(W95Y) <400> SEQUENCE: 248 Gln Val Lys Leu Gln Gln Ser Gly
Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Tyr Asp Phe Tyr Asp Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 249 <211> LENGTH: 363 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (Y98A) Heavy Chain
Construct <400> SEQUENCE: 249 caggtgcagc tggtgcaatc
tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg
cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat
180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag
cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt
attactgtgc gagatgggac 300 ttcgctgatt acgtggaggc tatggactac
tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO
250 <211> LENGTH: 363 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7A (D99Y) Heavy Chain Construct <400>
SEQUENCE: 250 caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc
ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata caccttcact
gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg
gatgggatgg ataaacacca aaactggaga ggcaacatat 180 gttgaagagt
ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240
ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac
300 ttctattatt acgtggaggc tatggactac tggggccaag ggaccacggt
caccgtctcc 360 tca 363 <210> SEQ ID NO 251 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH7A
(D99H) Heavy Chain Construct <400> SEQUENCE: 251 caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatcatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 252 <211> LENGTH: 364 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (V101F) Heavy Chain Construct
<400> SEQUENCE: 252 ccaggtgcag ctggtgcaat ctgggtctga
gttgaagaag cctggggcct cagtgaaggt 60 ttcctgcaag gcttctggat
acaccttcac tgagtttgga atgaactggg tgcgacaggc 120 ccctggacaa
gggcttgagt ggatgggatg gataaacacc aaaactggag aggcaacata 180
tgttgaagag tttaagggac ggtttgtctt ctccttggac acctctgtca gcacggcata
240 tctgcagatc agcagcctaa aggctgaaga cactgccgtg tattactgtg
cgagatggga 300 cttctatgat tacttcgagg ctatggacta ctggggccaa
gggaccacgg tcaccgtctc 360 ctca 364 <210> SEQ ID NO 253
<211> LENGTH: 364 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH7A (E102Q) Heavy Chain Construct <400>
SEQUENCE: 253 ccaggtgcag ctggtgcaat ctgggtctga gttgaagaag
cctggggcct cagtgaaggt 60 ttcctgcaag gcttctggat acaccttcac
tgagtttgga atgaactggg tgcgacaggc 120 ccctggacaa gggcttgagt
ggatgggatg gataaacacc aaaactggag aggcaacata 180 tgttgaagag
tttaagggac ggtttgtctt ctccttggac acctctgtca gcacggcata 240
tctgcagatc agcagcctaa aggctgaaga cactgccgtg tattactgtg cgagatggga
300 cttctatgat tacgtgcagg ctatggacta ctggggccaa gggaccacgg
tcaccgtctc 360 ctca 364 <210> SEQ ID NO 254 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH7A
(A103T) Heavy Chain Construct <400> SEQUENCE: 254 caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatgatt acgtggagac
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 255 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (Y98A / D99Y) Heavy Chain
Construct <400> SEQUENCE: 255 caggtgcagc tggtgcaatc
tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg
cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat
180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag
cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt
attactgtgc gagatgggac 300 ttcgcttatt acgtggaggc tatggactac
tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO
256 <211> LENGTH: 363 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7A (W95Y) Heavy Chain Construct <400>
SEQUENCE: 256 caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc
ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata caccttcact
gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg
gatgggatgg ataaacacca aaactggaga ggcaacatat 180 gttgaagagt
ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240
ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatacgac
300 ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt
caccgtctcc 360 tca 363 <210> SEQ ID NO 257 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1
<400> SEQUENCE: 257 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Leu Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 258 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A2 <400> SEQUENCE: 258 Gln
Val Lys Leu Gln Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10
15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 259 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A3
<400> SEQUENCE: 259 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Lys
Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 260 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A4 <400> SEQUENCE: 260 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr
Ser Ala Thr Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn
Glu Asp Thr Ala Lys Tyr Phe Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 261 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1A
<400> SEQUENCE: 261 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 262 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B <400> SEQUENCE: 262 Gln
Val Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 263 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1C
<400> SEQUENCE: 263 Gln Val Gln Leu Val Gln Ser Gly Pro Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 264 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1D <400> SEQUENCE: 264 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10
15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 265 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1E
<400> SEQUENCE: 265 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 266 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1F <400> SEQUENCE: 266 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Leu Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 267 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1G
<400> SEQUENCE: 267 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Ile Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 268 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1 <400> SEQUENCE: 268
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccttc
accctggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 269 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A2 <400> SEQUENCE: 269
caggtcaaac tgcagcagag cggccctgag ctgaagaaac ccggcgagac agtgaagatc
60 agctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggca 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accaccgaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 270 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A3 <400> SEQUENCE: 270
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggcgccag cgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
caagcaggcc 120 cctggcaagg gcctgaagtg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accaccgaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 271 <211> LENGTH: 362 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A4 <400> SEQUENCE: 271
aggtgcagct ggtgcagtct ggcgccgaag tgaagaaacc tggcgccagc gtgaaggtgt
60 cctgcaaggc cagcggctac accttcaccg agttcggcat gaactgggtc
cgacaggcac 120 caggccaggg cctcgaatgg atgggctgga tcaacaccaa
gaccggcgag gccacctacg 180 tggaagagtt caagggcaga ttcgccttca
gcctggaaac cagcgccacc accgcctacc 240 tgcagatcaa caacctgaag
aacgaggata ccgccaagta cttctgcgcc agatgggact 300 tctacgatta
cgtggaagcc atggactact ggggccaggg caccaccgtg accgtgtcta 360 gc 362
<210> SEQ ID NO 272 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1A <400> SEQUENCE: 272
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccttc
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 273 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B <400> SEQUENCE: 273
caggtgaagc tgcagcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 274 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1C <400> SEQUENCE: 274
caggtgcagc tggtgcagtc tggccccgaa ctgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 275 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1D <400> SEQUENCE: 275
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagagac tgtgaagatc
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 276 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1E <400> SEQUENCE: 276
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag attcaccatg
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 277 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1F <400> SEQUENCE: 277
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 ctggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 278 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1G <400> SEQUENCE: 278
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaaatcc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 279 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A <400> SEQUENCE: 279 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 280 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A
<400> SEQUENCE: 280 caggtgcagc tggtgcagtc tggcgctgag
gtgaagaagc ctggcgcctc ggtgaaggtc 60 tcctgcaagg cctctggtta
cacatttacg gaattcggga tgaattgggt cagacaagca 120 cctgggcaag
ggctcgagtg gatgggatgg ataaacacga agacaggcga ggccacctac 180
gtagaagagt tcaaaggaag ggtcacgatg accacagata cttctacctc tactgcgtat
240 atggaactac ggagcttgcg tagcgatgac acagcggtgt actattgtgc
tcgatgggat 300 ttctatgact atgttgaagc tatggactac tggggccaag
ggaccaccgt gaccgtctcc 360 tca 363 <210> SEQ ID NO 281
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 <400> SEQUENCE: 281 Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys
Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly
Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe
50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr
Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu
Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser
Ser 115 120
1 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 281
<210> SEQ ID NO 1 <211> LENGTH: 5 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 Kabat <400>
SEQUENCE: 1 Glu Phe Gly Met Asn 1 5 <210> SEQ ID NO 2
<211> LENGTH: 5 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 Kabat <400> SEQUENCE: 2 Glu Tyr
Gly Met Asn 1 5 <210> SEQ ID NO 3 <211> LENGTH: 5
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR1 Kabat <400> SEQUENCE: 3 Glu Tyr Ser Met Asn 1 5
<210> SEQ ID NO 4 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain Variable Region
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (32)..(32) <223> OTHER INFORMATION: Xaa = Tyr or
Phe <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (33)..(33) <223> OTHER INFORMATION: Xaa
= Ser or Gly <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (35)..(35) <223> OTHER
INFORMATION: Xaa = Asn or ser <220> FEATURE: <221>
NAME/KEY: MISC_FEATURE <222> LOCATION: (50)..(50) <223>
OTHER INFORMATION: Xaa = Trp or Tyr <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (55)..(55)
<223> OTHER INFORMATION: Xaa = Thr, Ser, or Asn <220>
FEATURE: <221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(59)..(59) <223> OTHER INFORMATION: Xaa = Thr or Asn
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (61)..(61) <223> OTHER INFORMATION: Xaa = Val or
Met <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (101)..(101) <223> OTHER INFORMATION:
Xaa = Phe or Ala <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (102)..(102) <223> OTHER
INFORMATION: Xaa = Val, Phe, Ser, Tyr, or Ala <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(103)..(103) <223> OTHER INFORMATION: Xaa = Asp, His, Trp,
Glu or Tyr <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (105)..(105) <223> OTHER INFORMATION:
Xaa = Val, Phe, or Leu <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (106)..(106) <223> OTHER
INFORMATION: Xaa = Glu, Lys or Gln <220> FEATURE: <221>
NAME/KEY: MISC_FEATURE <222> LOCATION: (107)..(107)
<223> OTHER INFORMATION: Xaa = Ala or Thr <220>
FEATURE: <221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(108)..(108) <223> OTHER INFORMATION: Xaa = Met or Leu
<400> SEQUENCE: 4 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu
Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser
Gly Tyr Thr Phe Thr Glu Xaa 20 25 30 Xaa Met Xaa Trp Val Arg Gln
Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Xaa Ile Asn Thr
Lys Xaa Gly Glu Ala Xaa Tyr Xaa Glu Glu Phe 50 55 60 Lys Gly Arg
Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu
Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Trp Asp Xaa Xaa Xaa Tyr Xaa Xaa Xaa Xaa Asp Tyr Trp Gly
100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210>
SEQ ID NO 5 <211> LENGTH: 5 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Heavy Chain CDR1 Kabat <400> SEQUENCE: 5
Glu Phe Gly Met Ser 1 5 <210> SEQ ID NO 6 <211> LENGTH:
7 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR1 Chothia <400> SEQUENCE: 6 Gly Tyr Thr Phe Thr Glu Phe 1
5 <210> SEQ ID NO 7 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 Chothia <400>
SEQUENCE: 7 Gly Tyr Thr Phe Thr Glu Tyr 1 5 <210> SEQ ID NO 8
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 AbM <400> SEQUENCE: 8 Gly Tyr
Thr Phe Thr Glu Phe Gly Met Asn 1 5 10 <210> SEQ ID NO 9
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 AbM <400> SEQUENCE: 9 Gly Tyr
Thr Phe Thr Glu Tyr Gly Met Asn 1 5 10 <210> SEQ ID NO 10
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 AbM <400> SEQUENCE: 10 Gly Tyr
Thr Phe Thr Glu Tyr Ser Met Asn 1 5 10 <210> SEQ ID NO 11
<211> LENGTH: 108 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain Variable Region <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (27)..(27)
<223> OTHER INFORMATION: Xaa = Gln, Ala, Lys, or His
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (28)..(28) <223> OTHER INFORMATION: Xaa = Asn, Ala,
Tyr, Ile, Lys, or Thr <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (29)..(29) <223> OTHER
INFORMATION: Xaa = Val, Ala, Gly, or Met <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (30)..(30)
<223> OTHER INFORMATION: Xaa = Gly, Ser, Thr, or Leu
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (31)..(31) <223> OTHER INFORMATION: Xaa = Thr, Asn,
Pro, or Ala <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (32)..(32) <223> OTHER INFORMATION: Xaa
= Asn or Tyr <220> FEATURE: <221> NAME/KEY:
MISC_FEATURE <222> LOCATION: (46)..(46) <223> OTHER
INFORMATION: Xaa = Pro or Leu <220> FEATURE: <221>
NAME/KEY: MISC_FEATURE <222> LOCATION: (50)..(50)
<223> OTHER INFORMATION: Xaa = Ser, Leu, or Trp <220>
FEATURE: <221> NAME/KEY: MISC_FEATURE <222> LOCATION:
(53)..(53) <223> OTHER INFORMATION: Xaa = Tyr, Asn, or His
<220> FEATURE: <221> NAME/KEY: MISC_FEATURE <222>
LOCATION: (54)..(54) <223> OTHER INFORMATION: Xaa = Arg, Leu,
Pro, or His <220> FEATURE: <221> NAME/KEY: MISC_FEATURE
<222> LOCATION: (55)..(55) <223> OTHER INFORMATION: Xaa
= Tyr, Ser, Gln, Lys, Phe, Pro, or Glu <220> FEATURE:
<221> NAME/KEY: MISC_FEATURE <222> LOCATION: (56)..(56)
<223> OTHER INFORMATION: Xaa = Ser, Gly, Ile, or Arg
<400> SEQUENCE: 11 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Xaa Xaa Xaa Xaa Xaa Xaa 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Xaa Ala Ser
Xaa Xaa Xaa Xaa Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 12 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 AbM <400>
SEQUENCE: 12 Gly Tyr Thr Phe Thr Glu Phe Gly Met Ser 1 5 10
<210> SEQ ID NO 13 <211> LENGTH: 17 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 Kabat <400>
SEQUENCE: 13 Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe Lys 1 5 10 15 Gly <210> SEQ ID NO 14 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 Kabat <400> SEQUENCE: 14 Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Ile Glu Glu Phe Lys 1 5 10 15 Gly <210>
SEQ ID NO 15 <211> LENGTH: 17 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 Kabat <400>
SEQUENCE: 15 Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu
Glu Phe Lys 1 5 10 15 Gly <210> SEQ ID NO 16 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 Kabat <400> SEQUENCE: 16 Tyr Ile Asn Thr Lys Asn
Gly Glu Ala Asn Tyr Val Glu Glu Phe Lys 1 5 10 15 Gly <210>
SEQ ID NO 17 <211> LENGTH: 17 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 Kabat <400>
SEQUENCE: 17 Trp Ile Asn Thr Lys Asn Gly Glu Ala Thr Tyr Ile Glu
Glu Phe Lys 1 5 10 15 Gly <210> SEQ ID NO 18 <211>
LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 Chothia <400> SEQUENCE: 18 Asn Thr Lys Thr Gly Glu
Ala Thr 1 5 <210> SEQ ID NO 19 <211> LENGTH: 8
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR2 Chothia <400> SEQUENCE: 19 Asn Thr Lys Ser Gly Glu Ala
Thr 1 5 <210> SEQ ID NO 20 <211> LENGTH: 8 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Heavy Chain CDR2 Chothia
<400> SEQUENCE: 20 Asn Thr Lys Asn Gly Glu Ala Asn 1 5
<210> SEQ ID NO 21 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 21 Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr 1 5 10
<210> SEQ ID NO 22 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 22 Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr 1 5 10
<210> SEQ ID NO 23 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 23 Tyr Ile Asn Thr Lys Asn Gly Glu Ala Asn 1 5 10
<210> SEQ ID NO 24 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 AbM <400>
SEQUENCE: 24 Trp Ile Asn Thr Lys Asn Gly Glu Ala Thr 1 5 10
<210> SEQ ID NO 25 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 25 Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 26 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR3 <400>
SEQUENCE: 26 Trp Asp Phe Tyr His Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 27 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 27 Trp Asp Phe Val Asp Tyr Val Glu Ala
Met Asp Tyr 1 5 10 <210> SEQ ID NO 28 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 28 Trp Asp Phe Tyr Trp Tyr Val Glu Ala
Met Asp Tyr 1 5 10 <210> SEQ ID NO 29 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 29 Trp Asp Ala Phe Glu Tyr Val Lys Ala
Leu Asp Tyr 1 5 10 <210> SEQ ID NO 30 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 30 Trp Asp Phe Phe Glu Tyr Phe Lys Thr
Met Asp Tyr 1 5 10 <210> SEQ ID NO 31 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 31 Trp Asp Phe Phe Tyr Tyr Val Gln Thr
Met Asp Tyr 1 5 10 <210> SEQ ID NO 32 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 32 Trp Asp Phe Ser Tyr Tyr Val Glu Ala
Met Asp Tyr 1 5 10 <210> SEQ ID NO 33 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 33 Trp Asp Phe Ala His Tyr Phe Gln Thr
Met Asp Tyr 1 5 10 <210> SEQ ID NO 34 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 34 Trp Asp Phe Ala Tyr Tyr Phe Gln Thr
Met Asp Tyr 1 5 10 <210> SEQ ID NO 35 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR3 <400> SEQUENCE: 35 Trp Asp Phe Ala Tyr Tyr Leu Glu Ala
Met Asp Tyr 1 5 10 <210> SEQ ID NO 36 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 36 Lys Ala Ser Gln Asn Val Gly Thr Asn
Val Ala 1 5 10 <210> SEQ ID NO 37 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 37 Lys Ala Ser Ala Asn Val Gly Asn Asn
Val Ala 1 5 10 <210> SEQ ID NO 38 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 38 Lys Ala Ser Lys Asn Val Gly Thr Asn
Val Ala 1 5 10 <210> SEQ ID NO 39 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 39 Lys Ala Ser Ala Ala Val Gly Thr Tyr
Val Ala 1 5 10 <210> SEQ ID NO 40 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 40 Lys Ala Ser Gln Tyr Ala Ser Thr Asn
Val Ala 1 5 10 <210> SEQ ID NO 41 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 41 Lys Ala Ser His Asn Val Gly Thr Asn
Val Ala 1 5 10 <210> SEQ ID NO 42 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 42 Lys Ala Ser Gln Ile Met Gly Pro Asn
Val Ala 1 5 10 <210> SEQ ID NO 43 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 43 Lys Ala Ser Gln Ile Val Gly Thr Asn
Val Ala 1 5 10 <210> SEQ ID NO 44 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 44 Lys Ala Ser Gln Lys Val Leu Thr Asn
Val Ala 1 5 10 <210> SEQ ID NO 45 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 45 Lys Ala Ser Gln Thr Val Ser Ala Asn
Val Ala 1 5 10 <210> SEQ ID NO 46 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 46
Ser Ala Ser Tyr Arg Tyr Ser 1 5 <210> SEQ ID NO 47
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 47 Tyr Leu Ala
Ser Asn Leu Ser Gly 1 5 <210> SEQ ID NO 48 <211>
LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Light
Chain CDR2 <400> SEQUENCE: 48 Tyr Leu Ala Ser Tyr Pro Gln Ile
1 5 <210> SEQ ID NO 49 <211> LENGTH: 8 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Light Chain CDR2
<400> SEQUENCE: 49 Tyr Ser Ala Ser Tyr Arg Lys Arg 1 5
<210> SEQ ID NO 50 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 50 Tyr Trp Ala Ser Tyr Arg Tyr Ser 1 5 <210> SEQ ID
NO 51 <211> LENGTH: 8 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Light Chain CDR2 <400> SEQUENCE: 51 Tyr
Ser Ala Ser His Arg Tyr Ser 1 5 <210> SEQ ID NO 52
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 52 Tyr Leu Ala
Ser Tyr His Glu Ser 1 5 <210> SEQ ID NO 53 <211>
LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Light
Chain CDR2 <400> SEQUENCE: 53 Tyr Ser Ala Ser His Arg Pro Ser
1 5 <210> SEQ ID NO 54 <211> LENGTH: 8 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Light Chain CDR2
<400> SEQUENCE: 54 Tyr Leu Ala Ser Tyr Arg Tyr Ser 1 5
<210> SEQ ID NO 55 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 55 Tyr Leu Ala Ser Tyr Arg Tyr Arg 1 5 <210> SEQ ID
NO 56 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Light Chain CDR3 <400> SEQUENCE: 56 His
Gln Tyr Tyr Thr Tyr Pro Leu Phe Thr 1 5 10 <210> SEQ ID NO 57
<211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR1 <400> SEQUENCE: 57 ggatacacct
tcactgagtt tggaatgaac 30 <210> SEQ ID NO 58 <211>
LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR1 <400> SEQUENCE: 58 ggatacacct tcactgagta
tggtatgaac 30 <210> SEQ ID NO 59 <211> LENGTH: 30
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Heavy Chain
CDR1 <400> SEQUENCE: 59 ggatacacct tcactgagta ttctatgaac 30
<210> SEQ ID NO 60 <400> SEQUENCE: 60 000 <210>
SEQ ID NO 61 <211> LENGTH: 30 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR1 <400>
SEQUENCE: 61 ggatacacct tcactgagtt tggaatgagc 30 <210> SEQ ID
NO 62 <211> LENGTH: 51 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Heavy Chain CDR2 <400> SEQUENCE: 62
tggataaaca ccaaaactgg agaggcaaca tatgttgaag agtttaaggg a 51
<210> SEQ ID NO 63 <211> LENGTH: 51 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Heavy Chain CDR2 <400>
SEQUENCE: 63 tggataaaca ccaaaactgg agaggcaaca tatattgaag agtttaaggg
a 51 <210> SEQ ID NO 64 <211> LENGTH: 51 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Heavy Chain CDR2
<400> SEQUENCE: 64 tggataaaca ccaaaagtgg agaggcaaca
tatgttgaag agtttaaggg a 51 <210> SEQ ID NO 65 <211>
LENGTH: 51 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Heavy
Chain CDR2 <400> SEQUENCE: 65 tatataaaca ccaaaaatgg
agaggcaaac tatgttgaag agtttaaggg a 51 <210> SEQ ID NO 66
<211> LENGTH: 51 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR2 <400> SEQUENCE: 66 tggataaaca
ccaaaaatgg agaggcaaca tatattgaag agtttaaggg a 51 <210> SEQ ID
NO 67
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 67 tgggacttct
atgattacgt ggaggctatg gactac 36 <210> SEQ ID NO 68
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 68 tgggacttct
atcattacgt ggaggctatg gactac 36 <210> SEQ ID NO 69
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 69 tgggacttcg
tggattacgt ggaggctatg gactac 36 <210> SEQ ID NO 70
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 70 tgggacttct
attggtacgt ggaggctatg gactac 36 <210> SEQ ID NO 71
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 71 tgggacgcct
ttgagtacgt gaaggcgctg gactac 36 <210> SEQ ID NO 72
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 72 tgggatttct
ttgagtattt taagactatg gactac 36 <210> SEQ ID NO 73
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 73 tgggactttt
tttattacgt gcagactatg gactac 36 <210> SEQ ID NO 74
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 74 tgggattttt
cttattacgt tgaggcgatg gactac 36 <210> SEQ ID NO 75
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 75 tgggactttg
ctcattactt tcagactatg gactac 36 <210> SEQ ID NO 76
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 76 tgggacttcg
cttattactt tcagactatg gactac 36 <210> SEQ ID NO 77
<211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Heavy Chain CDR3 <400> SEQUENCE: 77 tgggatttcg
cgtattacct tgaggctatg gactac 36 <210> SEQ ID NO 78
<211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR1 <400> SEQUENCE: 78 aaggccagtc
agaatgtggg tactaatgtt gcc 33 <210> SEQ ID NO 79 <211>
LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Light
Chain CDR1 <400> SEQUENCE: 79 aaggccagtg ccaatgtggg
taataatgtt gcc 33 <210> SEQ ID NO 80 <211> LENGTH: 33
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR1 <400> SEQUENCE: 80 aaggccagta agaatgtggg gactaatgtt gcg
33 <210> SEQ ID NO 81 <211> LENGTH: 33 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Light Chain CDR1
<400> SEQUENCE: 81 aaggccagtg cggctgtggg tacgtatgtt gcg 33
<210> SEQ ID NO 82 <211> LENGTH: 33 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 82 aaggccagtc agatagcgag tactaatgtt gcc 33 <210>
SEQ ID NO 83 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 83 aaggccagtc acaatgtggg taccaacgtt gcg 33 <210>
SEQ ID NO 84 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 84 aaggccagtc agattatggg tcctaatgtt gcg 33 <210>
SEQ ID NO 85 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 85 aaggccagtc aaattgtggg tactaatgtt gcg 33 <210>
SEQ ID NO 86 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 86 aaggccagtc agaaggtgct tactaatgtt gcg 33 <210>
SEQ ID NO 87 <211> LENGTH: 33 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR1 <400>
SEQUENCE: 87 aaggccagtc agactgtgag tgctaatgtt gcg 33
<210> SEQ ID NO 88 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 88 tattcggcat cctaccgcta cagt 24 <210> SEQ ID NO 89
<211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 89 tatttggcct
ccaacctctc cggt 24 <210> SEQ ID NO 90 <211> LENGTH: 24
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 90 tacctggcat cctaccccca gatt 24
<210> SEQ ID NO 91 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 91 tattcggcat cctaccgcaa aagg 24 <210> SEQ ID NO 92
<211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 92 tattgggcat
cctaccgcta tagt 24 <210> SEQ ID NO 93 <211> LENGTH: 24
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 93 tattcggcat cccaccggta cagt 24
<210> SEQ ID NO 94 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 94 tatttggcat cctaccacga aagt 24 <210> SEQ ID NO 95
<211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR2 <400> SEQUENCE: 95 tattcggcat
cccaccgtcc cagt 24 <210> SEQ ID NO 96 <211> LENGTH: 24
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Light Chain
CDR2 <400> SEQUENCE: 96 tatttggcat cctaccgcta cagt 24
<210> SEQ ID NO 97 <211> LENGTH: 24 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Light Chain CDR2 <400>
SEQUENCE: 97 tatttggcat cctaccgcta caga 24 <210> SEQ ID NO 98
<211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Light Chain CDR3 <400> SEQUENCE: 98 caccaatatt
acacctatcc tctattcacg 30 <210> SEQ ID NO 99 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: PR1A3
VH <400> SEQUENCE: 99 Gln Val Lys Leu Gln Gln Ser Gly Pro Glu
Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Lys
Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Tyr 65 70 75 80
Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Lys Tyr Phe Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 100 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pEM1496 huPR1A3 VH <400>
SEQUENCE: 100 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 101 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7A <400> SEQUENCE: 101 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 102 <211> LENGTH: 98
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: IGHV7-4-1*02
<400> SEQUENCE: 102 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45
Gly Trp Ile Asn Thr Asn Thr Gly Asn Pro Thr Tyr Ala Gln Gly Phe 50
55 60 Thr Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala
Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg <210> SEQ ID NO 103 <211>
LENGTH: 110 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: PR1A3
VL <400> SEQUENCE: 103 Asp Ile Val Met Thr Gln Ser Gln Arg
Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Val Thr Cys
Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln
Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala
Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70
75 80 Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Tyr Tyr Thr Tyr Pro
Leu 85 90 95 Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Met Lys Arg
Thr 100 105 110 <210> SEQ ID NO 104 <211> LENGTH: 109
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pEM1495 huPR1A3
VL <400> SEQUENCE: 104 Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys
Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala
Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser
Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70
75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro
Leu 85 90 95 Phe Ser Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105 <210> SEQ ID NO 105 <211> LENGTH: 108
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CL1A
<400> SEQUENCE: 105 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser
Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 106 <211> LENGTH: 95 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: IMGT_hVK_1_39 <400> SEQUENCE:
106 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser
Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr
Tyr Cys Gln Gln Ser Tyr Ser Thr Pro 85 90 95 <210> SEQ ID NO
107 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7 rF9 <400> SEQUENCE: 107 Gln Val Gln
Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 108 <211> LENGTH:
110 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CLA1 rH11
<400> SEQUENCE: 108 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Thr Val Ser Ala Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser
Tyr Arg Tyr Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 109 <400> SEQUENCE: 109 000
<210> SEQ ID NO 110 <400> SEQUENCE: 110 000 <210>
SEQ ID NO 111 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 VH <400> SEQUENCE: 111
caggtgaagc tgcagcagtc aggacctgag ttgaagaagc ctggagagac agtcaagatc
60 tcctgcaagg cttctggata taccttcaca gaattcggaa tgaactgggt
gaagcaggct 120 cctggaaagg gtttaaagtg gatgggctgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgccttc
tctttggaga cctctgccac cactgcctat 240 ttgcagatca acaacctcaa
aaatgaggac acggctaaat atttctgtgc tcgatgggat 300 ttctatgact
atgttgaagc tatggactac tggggccaag ggaccaccgt gaccgtctcc 360 tca 363
<210> SEQ ID NO 112 <211> LENGTH: 362 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pEM1496 <400> SEQUENCE: 112
caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagttggaat gaactgggtg
cgacaggccc 120 ctggacaagg gcttgagtgg atgggatgga taaacaccaa
aactggagag gcaacatatg 180
ttgaagagtt taagggacgg tttgtcttct ccttggacac ctctgtcagc acggcatatc
240 tgcagatcag cagcctaaag gctgacgaca ctgccgtgta ttactgtgcg
agatgggact 300 tctatgatta cgtggaggct atggactact ggggccaagg
gaccacggtc accgtctcct 360 ca 362 <210> SEQ ID NO 113
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH7A <400> SEQUENCE: 113 caggtgcaat tggtgcaatc
tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg
cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat
180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag
cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt
attactgtgc gagatgggac 300 ttctatgatt acgtggaggc tatggactac
tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO
114 <211> LENGTH: 294 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: IGHV7-4-1*02 <400> SEQUENCE: 114
caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact agctatgcta tgaattgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg atcaacacca
acactgggaa cccaacgtat 180 gcccagggct tcacaggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaggac actgccgtgt attactgtgc gaga 294 <210> SEQ ID NO 115
<211> LENGTH: 330 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 VL <400> SEQUENCE: 115 gatatcgtga
tgacccagtc tcaaagattc atgtccacat cagtaggaga cagggtcagc 60
gtcacctgca aggccagtca gaatgtgggt actaatgttg cctggtatca acagaaacca
120 ggacaatccc ctaaagcact gatttactcg gcatcctacc ggtacagtgg
agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactctca
ccatcagcaa tgtacagtct 240 gaagacttgg cggagtattt ctgtcaccaa
tattacacct atcctctatt cacgttcggc 300 tcggggacaa agttggaaat
gaaacgtacg 330 <210> SEQ ID NO 116 <211> LENGTH: 327
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pEM1495
<400> SEQUENCE: 116 gacatccaga tgactcagag cccaagcagc
ctgagcgcca gcgtgggtga cagagtgacc 60 atcacctgta aggccagtca
gaatgtgggt actaatgttg cctggtacca gcagaagcca 120 ggtaaggctc
caaagctgct gatctactcg gcatcctacc ggtacagtgg tgtgccaagc 180
agattcagcg gtagcggtag cggtaccgac ttcaccttca ccatcagcag cctccagcca
240 gaggacatcg ccacctacta ctgccaccaa tattacacct atcctctatt
cagcttcggc 300 caagggacca aggtggaaat caaacgt 327 <210> SEQ ID
NO 117 <211> LENGTH: 324 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CL1A <400> SEQUENCE: 117 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtca gaatgtgggt actaatgttg cctggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctattcg gcatcctacc gctacagtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat caag 324
<210> SEQ ID NO 118 <211> LENGTH: 285 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: IMGT_hVK_1_39 <400> SEQUENCE:
118 gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga
cagagtcacc 60 atcacttgcc gggcaagtca gagcattagc agctatttaa
attggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgct
gcatccagtt tgcaaagtgg ggtcccatca 180 aggttcagtg gcagtggatc
tgggacagat ttcactctca ccatcagcag tctgcaacct 240 gaagattttg
caacttacta ctgtcaacag agttacagta cccct 285 <210> SEQ ID NO
119 <211> LENGTH: 369 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7 rF9 <400> SEQUENCE: 119 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtatggta tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacga aatctggaga
ggcaacctat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatgatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tcagctagc 369
<210> SEQ ID NO 120 <211> LENGTH: 330 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CLA1 rH11 <400> SEQUENCE: 120
gatatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc
60 atcacttgca aggccagtca gactgtgagt gctaatgttg cgtggtatca
gcagaaacca 120 gggaaagcac ctaagctcct gatctacttg gcatcctacc
gctacagagg agtcccatca 180 aggttcagtg gcagtggatc tgggacagat
ttcactctca ccatcagcag tctgcaacct 240 gaagatttcg caacttacta
ctgtcaccaa tattacacct atcctctatt cacgtttggc 300 cagggcacca
agctcgagat caagcgtacg 330 <210> SEQ ID NO 121 <211>
LENGTH: 987 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: IgG1
<400> SEQUENCE: 121 accaagggcc catcggtctt ccccctggca
ccctcctcca agagcacctc tgggggcaca 60 gcggccctgg gctgcctggt
caaggactac ttccccgaac cggtgacggt gtcgtggaac 120 tcaggcgccc
tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 180
tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc
240 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagcaga
gcccaaatct 300 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg
aactcctggg gggaccgtca 360 gtcttcctct tccccccaaa acccaaggac
accctcatga tctcccggac ccctgaggtc 420 acatgcgtgg tggtggacgt
gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 480 gacggcgtgg
aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 540
taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac
600 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
ctccaaagcc 660 aaagggcagc cccgagaacc acaggtgtac accctgcccc
catcccggga tgagctgacc 720 aagaaccagg tcagcctgac ctgcctggtc
aaaggcttct atcccagcga catcgccgtg 780 gagtgggaga gcaatgggca
gccggagaac aactacaaga ccacgcctcc cgtgctggac 840 tccgacggct
ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 900
gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag
960 agcctctccc tgtctccggg taaatga 987 <210> SEQ ID NO 122
<211> LENGTH: 328 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: IgG1 <400> SEQUENCE: 122 Thr Lys Gly Pro Ser Val
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr 1 5 10 15 Ser Gly Gly Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro 20 25 30 Glu Pro
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val 35 40 45
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser 50
55 60
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile 65
70 75 80 Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
Lys Ala 85 90 95 Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro
Pro Cys Pro Ala 100 105 110 Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
Leu Phe Pro Pro Lys Pro 115 120 125 Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro Glu Val Thr Cys Val Val 130 135 140 Val Asp Val Ser His Glu
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 145 150 155 160 Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 165 170 175 Tyr
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 180 185
190 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
195 200 205 Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln Pro 210 215 220 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu Thr 225 230 235 240 Lys Asn Gln Val Ser Leu Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser 245 250 255 Asp Ile Ala Val Glu Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr 260 265 270 Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 275 280 285 Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 290 295 300 Ser
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 305 310
315 320 Ser Leu Ser Leu Ser Pro Gly Lys 325 <210> SEQ ID NO
123 <211> LENGTH: 42 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer MS-43 <400> SEQUENCE: 123
ccagccggcc atggccgata tccagatgac ccagtctcca tc 42 <210> SEQ
ID NO 124 <211> LENGTH: 26 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer MS-52 <400> SEQUENCE: 124
gaagaccgat gggcctttgg tgctag 26 <210> SEQ ID NO 125
<211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer MS-55 <400> SEQUENCE: 125 gcaacatatg
ttgaagagtt taagggacgg 30 <210> SEQ ID NO 126 <211>
LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: MS-56
<400> SEQUENCE: 126 atgaactggg tgcgacaggc ccctg 25
<210> SEQ ID NO 127 <211> LENGTH: 56 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-679 <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (26)..(26)
<223> OTHER INFORMATION: m is a or c <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (27)..(28)
<223> OTHER INFORMATION: n is a, c, g, or t <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(30)..(30) <223> OTHER INFORMATION: w is a or t <400>
SEQUENCE: 127 caggggcctg tcgcacccag ttcatmnnaw actcagtgaa
ggtgtatcca gaagcc 56 <210> SEQ ID NO 128 <211> LENGTH:
81 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer EAB-680
<400> SEQUENCE: 128 ccgtccctta aactcttcaa cataggttgc
ctctccagtt ttggtgttta tccatcccat 60 ccactcaagc ccttgtccag g 81
<210> SEQ ID NO 129 <211> LENGTH: 50 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-685 <400> SEQUENCE:
129 cagctatgac catgattacg ccaagcttgc atgcaaattc tatttcaagg 50
<210> SEQ ID NO 130 <211> LENGTH: 27 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-686 <400> SEQUENCE:
130 gttgcgtggt atcagcagaa accaggg 27 <210> SEQ ID NO 131
<211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer EAB-687 <400> SEQUENCE: 131 gctctttgtg
acgggcgagc tcaggccctg atgg 34 <210> SEQ ID NO 132 <211>
LENGTH: 35 <212> TYPE: DNA <213> ORGANISM: Artificial
sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
EAB-688 <400> SEQUENCE: 132 ggagtcccat caaggttcag tggcagtgga
tctgg 35 <210> SEQ ID NO 133 <211> LENGTH: 70
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer EAB-681
<400> SEQUENCE: 133 cctggtttct gctgatacca cgcaacatta
gtacccacat tctgactggc cttgcaagtg 60 atggtgactc 70 <210> SEQ
ID NO 134 <211> LENGTH: 77 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer EAB-682 <400> SEQUENCE: 134
ctgccactga accttgatgg gactccactg tagcggtagg atgccgaata gatcaggagc
60 ttaggtgctt tccctgg 77 <210> SEQ ID NO 135 <211>
LENGTH: 64 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
AC7 <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (37)..(37) <223> OTHER INFORMATION: m
is a or c <220> FEATURE: <221> NAME/KEY: misc_feature
<222> LOCATION: (38)..(39) <223> OTHER INFORMATION: n
is a, c, g, or t <400> SEQUENCE: 135 ccagtagtcc atagcctcca
cgtaatcata gaagtcmnnt ctcgcacagt aatacacggc 60 agtg 64 <210>
SEQ ID NO 136 <211> LENGTH: 64 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer AC8 <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (34)..(34)
<223> OTHER INFORMATION: m is a or c <220> FEATURE:
<221> NAME/KEY: misc_feature <222> LOCATION: (35)..(36)
<223> OTHER INFORMATION: n is a, c, g, or t
<400> SEQUENCE: 136 ccagtagtcc atagcctcca cgtaatcata
gaamnnccat ctcgcacagt aatacacggc 60 agtg 64 <210> SEQ ID NO
137 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC9 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (31)..(31) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (32)..(33) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 137
ccagtagtcc atagcctcca cgtaatcata mnngtcccat ctcgcacagt aatacacggc
60 agtg 64 <210> SEQ ID NO 138 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC10
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (28)..(28) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (29)..(30) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 138 ccagtagtcc atagcctcca cgtaatcmnn
gaagtcccat ctcgcacagt aatacacggc 60 agtg 64 <210> SEQ ID NO
139 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC11 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (25)..(25) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (26)..(27) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 139
ccagtagtcc atagcctcca cgtamnnata gaagtcccat ctcgcacagt aatacacggc
60 agtg 64 <210> SEQ ID NO 140 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC12
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (37)..(37) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (38)..(39) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 140 cgtggtccct tggccccagt agtccatagc
ctccacmnna tcatagaagt cccatctcgc 60 acag 64 <210> SEQ ID NO
141 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC13 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (34)..(34) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (35)..(36) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 141
cgtggtccct tggccccagt agtccatagc ctcmnngtaa tcatagaagt cccatctcgc
60 acag 64 <210> SEQ ID NO 142 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC14
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (31)..(31) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (32)..(33) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 142 cgtggtccct tggccccagt agtccatagc
mnncacgtaa tcatagaagt cccatctcgc 60 acag 64 <210> SEQ ID NO
143 <211> LENGTH: 64 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC15 <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (28)..(28) <223>
OTHER INFORMATION: m is a or c <220> FEATURE: <221>
NAME/KEY: misc_feature <222> LOCATION: (29)..(30) <223>
OTHER INFORMATION: n is a, c, g, or t <400> SEQUENCE: 143
cgtggtccct tggccccagt agtccatmnn ctccacgtaa tcatagaagt cccatctcgc
60 acag 64 <210> SEQ ID NO 144 <211> LENGTH: 64
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer AC16
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (25)..(25) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (26)..(27) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 144 cgtggtccct tggccccagt agtcmnnagc
ctccacgtaa tcatagaagt cccatctcgc 60 acag 64 <210> SEQ ID NO
145 <211> LENGTH: 77 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Primer AC17 <400> SEQUENCE: 145 cgtggtccct
tggccccagt agtccatagc ctccacgtaa tcatagaagt cccatctcgc 60
acagtaatac acggcag 77 <210> SEQ ID NO 146 <211> LENGTH:
25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Primer EAB-749
<400> SEQUENCE: 146 ccatcagggc ctgagctcgc ccgtc 25
<210> SEQ ID NO 147 <211> LENGTH: 30 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-750 <400> SEQUENCE:
147 cgtggaggct atggactact ggggccaagg 30 <210> SEQ ID NO 148
<211> LENGTH: 29 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer EAB-751 <400> SEQUENCE: 148 gactactggg
gccaagggac cacggtcac 29 <210> SEQ ID NO 149 <211>
LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: Primer
EAB-752 <400> SEQUENCE: 149 ggtcagggcg cctgagttcc acg 23
<210> SEQ ID NO 150 <211> LENGTH: 60 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer AC1
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (37)..(37) <223> OTHER INFORMATION: m is a or c
<220> FEATURE: <221> NAME/KEY: misc_feature <222>
LOCATION: (38)..(39) <223> OTHER INFORMATION: n is a, c, g,
or t <400> SEQUENCE: 150 ggtgccctgg ccaaacgtga atagaggata
ggtgtamnnt tggtgacagt agtaagttgc 60 <210> SEQ ID NO 151
<211> LENGTH: 60 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer AC2 <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (34)..(34) <223> OTHER
INFORMATION: m is a or c <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (35)..(36) <223> OTHER
INFORMATION: n is a, c, g, or t <400> SEQUENCE: 151
ggtgccctgg ccaaacgtga atagaggata ggtmnnatat tggtgacagt agtaagttgc
60 <210> SEQ ID NO 152 <211> LENGTH: 60 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Primer AC3 <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(31)..(31) <223> OTHER INFORMATION: m is a or c <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(32)..(33) <223> OTHER INFORMATION: n is a, c, g, or t
<400> SEQUENCE: 152 ggtgccctgg ccaaacgtga atagaggata
mnngtaatat tggtgacagt agtaagttgc 60 <210> SEQ ID NO 153
<211> LENGTH: 60 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Primer AC4 <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (28)..(28) <223> OTHER
INFORMATION: m is a or c <220> FEATURE: <221> NAME/KEY:
misc_feature <222> LOCATION: (29)..(30) <223> OTHER
INFORMATION: n is a, c, g, or t <400> SEQUENCE: 153
ggtgccctgg ccaaacgtga atagaggmnn ggtgtaatat tggtgacagt agtaagttgc
60 <210> SEQ ID NO 154 <211> LENGTH: 60 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Primer AC5 <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(22)..(22) <223> OTHER INFORMATION: m is a or c <220>
FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:
(23)..(24) <223> OTHER INFORMATION: n is a, c, g, or t
<400> SEQUENCE: 154 ggtgccctgg ccaaacgtga amnnaggata
ggtgtaatat tggtgacagt agtaagttgc 60 <210> SEQ ID NO 155
<211> LENGTH: 60 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequences <220> FEATURE: <223> OTHER
INFORMATION: Primer AC6 <400> SEQUENCE: 155 ggtgccctgg
ccaaacgtga atagaggata ggtgtaatat tggtgacagt agtaagttgc 60
<210> SEQ ID NO 156 <211> LENGTH: 36 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer EAB-746 <400> SEQUENCE:
156 cgcttgatct cgagcttggt gccctggcca aacgtg 36 <210> SEQ ID
NO 157 <400> SEQUENCE: 157 000 <210> SEQ ID NO 158
<211> LENGTH: 428 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pETR6592 <400> SEQUENCE: 158 Gln Leu Thr Thr Glu
Ser Met Pro Phe Asn Val Ala Glu Gly Lys Glu 1 5 10 15 Val Leu Leu
Leu Val His Asn Leu Pro Gln Gln Leu Phe Gly Tyr Ser 20 25 30 Trp
Tyr Lys Gly Glu Arg Val Asp Gly Asn Arg Gln Ile Val Gly Tyr 35 40
45 Ala Ile Gly Thr Gln Gln Ala Thr Pro Gly Pro Ala Asn Ser Gly Arg
50 55 60 Glu Thr Ile Tyr Pro Asn Ala Ser Leu Leu Ile Gln Asn Val
Thr Gln 65 70 75 80 Asn Asp Thr Gly Phe Tyr Thr Leu Gln Val Ile Lys
Ser Asp Leu Val 85 90 95 Asn Glu Glu Ala Thr Gly Gln Phe His Val
Tyr Pro Glu Leu Pro Lys 100 105 110 Pro Ser Ile Ser Ser Asn Asn Ser
Asn Pro Val Glu Asp Lys Asp Ala 115 120 125 Met Ala Phe Thr Cys Glu
Pro Glu Thr Gln Asp Thr Thr Tyr Leu Trp 130 135 140 Trp Ile Asn Asn
Gln Ser Leu Pro Val Ser Pro Arg Leu Gln Leu Ser 145 150 155 160 Asn
Gly Asn Arg Thr Leu Thr Leu Leu Ser Val Thr Arg Asn Asp Thr 165 170
175 Gly Pro Tyr Glu Cys Glu Ile Gln Asn Pro Val Ser Ala Asn Arg Ser
180 185 190 Asp Pro Val Thr Leu Asn Val Thr Tyr Gly Pro Asp Thr Pro
Thr Ile 195 200 205 Ser Pro Pro Asp Ser Ser Tyr Leu Ser Gly Ala Asn
Leu Asn Leu Ser 210 215 220 Cys His Ser Ala Ser Asn Pro Ser Pro Gln
Tyr Ser Trp Arg Ile Asn 225 230 235 240 Gly Ile Pro Gln Gln His Thr
Gln Val Leu Phe Ile Ala Lys Ile Thr 245 250 255 Pro Asn Asn Asn Gly
Thr Tyr Ala Cys Phe Val Ser Asn Leu Ala Thr 260 265 270 Gly Arg Asn
Asn Ser Ile Val Lys Ser Ile Thr Val Ser Ala Leu Ser 275 280 285 Pro
Val Val Ala Lys Pro Gln Ile Lys Ala Ser Lys Thr Thr Val Thr 290 295
300 Gly Asp Lys Asp Ser Val Asn Leu Thr Cys Ser Thr Asn Asp Thr Gly
305 310 315 320 Ile Ser Ile Arg Trp Phe Phe Lys Asn Gln Ser Leu Pro
Ser Ser Glu 325 330 335 Arg Met Lys Leu Ser Gln Gly Asn Ile Thr Leu
Ser Ile Asn Pro Val 340 345 350 Lys Arg Glu Asp Ala Gly Thr Tyr Trp
Cys Glu Val Phe Asn Pro Ile 355 360 365 Ser Lys Asn Gln Ser Asp Pro
Ile Met Leu Asn Val Asn Tyr Asn Ala 370 375 380 Leu Pro Gln Glu Asn
Leu Ile Asn Val Asp Leu Glu Val Leu Phe Gln 385 390 395 400 Gly Pro
Gly Ser Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu 405 410 415
Trp His Glu Ala Arg Ala His His His His His His 420 425 <210>
SEQ ID NO 159 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PMS22 <400> SEQUENCE: 159
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttctatgatt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 160 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Primer 1C8 <400> SEQUENCE: 160
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatcatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 161 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: 3E1 <400> SEQUENCE: 161
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttcgtggatt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 162 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: 2D7 <400> SEQUENCE: 162
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttctattggt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 163 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Affinity Matured Heavy Chain
<400> SEQUENCE: 163 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 tttgctcatt actttcagac tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 164
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Affinity Matured Heavy Chain <400> SEQUENCE: 164
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttcgcttatt
actttcagac tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 165 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Affinity Matured Heavy Chain
<400> SEQUENCE: 165 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggat 300 ttcgcgtatt accttgaggc tatggactac tggggccaag
ggaccacgat caccgtctcc 360 tca 363 <210> SEQ ID NO 166
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H3 Full (5) 19 <400> SEQUENCE: 166 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgagctgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgctgtgt attactgtgc gagatgggac 300 gcctttgagt acgtgaaggc
gctggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 167 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H3 Full (5) 8 <400> SEQUENCE:
167 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact gagtttggaa
tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg
ataaacacca aaactggaga ggcaacatat 180 gttgaagagt ttaagggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggat 300
ttctttgagt attttaagac tatggactac tggggccaag ggaccacggt caccgtctcc
360 tca 363 <210> SEQ ID NO 168 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H3 Full (5) 28
<400> SEQUENCE: 168 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 tttttttatt acgtgcagac tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 169
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H3 Full (5) 27 <400> SEQUENCE: 169 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggat 300 ttttcttatt acgttgaggc
gatggactac tggggccaag ggaccacagt caccgtctcc 360 tca 363 <210>
SEQ ID NO 170 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H4E9 Heavy Chain <400>
SEQUENCE: 170 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc
ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata caccttcact
gagtttggta tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg
gatgggatgg ataaatacca aaactggaga ggcaacttat 180 attgaagagt
ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240
ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac
300 ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt
caccgtctcc 360 tca 363
<210> SEQ ID NO 171 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pAC14 (B9) <400> SEQUENCE: 171
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtttggta tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacca
aaagtggaga ggcaacctat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttctatgatt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 172 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pAC15 (F9) <400> SEQUENCE: 172
caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt
60 tcctgcaagg cttctggata caccttcact gagtatggta tgaactgggt
gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatgg ataaacacga
aatctggaga ggcaacctat 180 gttgaagagt ttaagggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa
ggctgaagac actgccgtgt attactgtgc gagatgggac 300 ttctatgatt
acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363
<210> SEQ ID NO 173 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 2 <400> SEQUENCE:
173 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact gagtattcta
tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatac
ataaacacca aaaatggaga ggcaaactat 180 gttgaagagt ttaagggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac 300
ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc
360 tca 363 <210> SEQ ID NO 174 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H1/H2 (5) 11
<400> SEQUENCE: 174 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtatggta tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaaatggaga ggcaacctat 180
attgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 ttctatgatt acgtggaggc tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 175
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H1/H2 (5) 13 <400> SEQUENCE: 175 caggtgcaat
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggta tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatat ataaacacca aaaatggaga
ggcaaactat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggacg
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctatgatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 176 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 14 <400> SEQUENCE:
176 caggtgcaat tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60 tcctgcaagg cttctggata caccttcact gagtatggta
tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg gatgggatat
ataaacacca aaaatggaga ggcaaactat 180 gttgaagagt ttaagggacg
gtttgtcttc tccttggaca cctctgtcag cacggcatat 240 ctgcagatca
gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac 300
ttctatgatt acgtggaggc tatggactac tggggccaag ggaccacggt caccgtctcc
360 tca 363 <210> SEQ ID NO 177 <211> LENGTH: 363
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H3 Full (5) 19
<400> SEQUENCE: 177 caggtgcaat tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgagctgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgctgtgt attactgtgc
gagatgggac 300 gcctttgagt acgtgaaggc gctggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 178
<211> LENGTH: 330 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC21 (3A1) <400> SEQUENCE: 178 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtgc caatgtgggt aataatgttg cctggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctatttg gcctccaacc gctccggtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 179 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC19 (2C6)
<400> SEQUENCE: 179 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtaa
gaatgtgggg actaatgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagcccct gatctacctg gcatcctacc cccagattgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcccctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 180 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC18 (2F1) <400> SEQUENCE: 180 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtgc ggctgtgggt acgtatgttg cgtggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctattcg gcatcctacc gcaaaagggg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 181 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC23 (2F11)
<400> SEQUENCE: 181 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
gatagcgagt actaatgttg cctggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctattgg gcatcctacc gctatagtgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 182 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: H4E9 light chain <400> SEQUENCE: 182
gatatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc
60 atcacttgca aggccagtca gaatgtgggt actaatgttg cctggtatca
gcagaaacca 120 gggaaagcac ctaagcccct gatctattcg gcatcctacc
gctacagtgg agtcccatca 180 aggttcagtg gcagtggatc tgggacagat
ttcactctca ccatcagcag tctgcaacct 240 gaagatttcg caacttacta
ctgtcaccaa tattacacct atcctctatt cacgtttggc 300 cagggcacca
agctcgagat caagcgtacg 330 <210> SEQ ID NO 183 <211>
LENGTH: 330 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: L2D2
<400> SEQUENCE: 183 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
caatgtgggt accaacgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctattcg gcatcccacc ggtacagtgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 184 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC6 (C1) <400> SEQUENCE: 184 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtca gattatgggt cctaatgttg cgtggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctatttg gcatcctacc acgaaagtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 185 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC7 (E10)
<400> SEQUENCE: 185 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
aattgtgggt actaatgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctattcg gcatcccacc gtcccagtgg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 186 <211> LENGTH: 330 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC12 (H7) <400> SEQUENCE: 186 gatatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtgggaga cagagtcacc 60
atcacttgca aggccagtca gaaggtgctt actaatgttg cgtggtatca gcagaaacca
120 gggaaagcac ctaagctcct gatctatttg gcatcctacc gctacagtgg
agtcccatca 180 aggttcagtg gcagtggatc tgggacagat ttcactctca
ccatcagcag tctgcaacct 240 gaagatttcg caacttacta ctgtcaccaa
tattacacct atcctctatt cacgtttggc 300 cagggcacca agctcgagat
caagcgtacg 330 <210> SEQ ID NO 187 <211> LENGTH: 330
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC13 (H11)
<400> SEQUENCE: 187 gatatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtgggaga cagagtcacc 60 atcacttgca aggccagtca
gactgtgagt gctaatgttg cgtggtatca gcagaaacca 120 gggaaagcac
ctaagctcct gatctacttg gcatcctacc gctacagagg agtcccatca 180
aggttcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct
240 gaagatttcg caacttacta ctgtcaccaa tattacacct atcctctatt
cacgtttggc 300 cagggcacca agctcgagat caagcgtacg 330 <210> SEQ
ID NO 188 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: PMS22 <400> SEQUENCE: 188 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 189 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: 1C8 <400>
SEQUENCE: 189 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr His Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 190 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: 3E1 <400> SEQUENCE: 190 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Val Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 191 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: 2D7 <400>
SEQUENCE: 191 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu
Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu
Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Trp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 192
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Affinity Matured Heavy Chain <400> SEQUENCE: 192
Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5
10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu
Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr
Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp
Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe
Ala His Tyr Phe Gln Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr
Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 193
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Affinity Matured Heavy Chain <400> SEQUENCE: 193
Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5
10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu
Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr
Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp
Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe
Ala Tyr Tyr Phe Gln Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr
Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 194
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Affinity Matured Heavy Chain <400> SEQUENCE: 194
Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5
10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu
Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr
Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp
Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe
Ala Tyr Tyr Leu Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr
Thr Ile Thr Val Ser Ser 115 120 <210> SEQ ID NO 195
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H3 Full (5) 19 <400> SEQUENCE: 195 Gln Val Gln
Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Ala Phe Glu Tyr
Val Lys Ala Leu Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 196 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H3 Full (5) 8
<400> SEQUENCE: 196 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Phe Glu Tyr Phe Lys Thr Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 197 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H3 Full (5) 28 <400> SEQUENCE:
197 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu
Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu
Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Phe Tyr Tyr Val Gln Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 198
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H3 Full (5) 27 <400> SEQUENCE: 198 Gln Val Gln
Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Ser Tyr Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 199 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H4E9 Heavy Chain <400>
SEQUENCE: 199 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Ile Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val
Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile
Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 200 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC14 (B9) <400> SEQUENCE: 200 Gln Val Gln
Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 201 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC15 (F9)
<400> SEQUENCE: 201 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Ser Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 202 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 2 <400> SEQUENCE:
202 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Tyr 20 25 30 Ser Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Thr Lys Asn Gly Glu Ala
Asn Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu
Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu
Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 203
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H1/H2 (5) 11 <400> SEQUENCE: 203 Gln Val Gln Leu
Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val
Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Asn Gly Glu Ala Thr Tyr Ile Glu Glu
Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser
Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 204 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H1/H2 (5) 13
<400> SEQUENCE: 204 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn
Thr Lys Asn Gly Glu Ala Asn Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Ala Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 205 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: H1/H2 (5) 14 <400> SEQUENCE:
205 Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Tyr 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Thr Lys Asn Gly Glu Ala
Asn Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu
Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu
Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 206
<211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: H3 Full (5) 19
<400> SEQUENCE: 206 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Ser Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Ala Phe Glu Tyr Val Lys Ala Leu Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 207 <211> LENGTH: 110 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: pAC21 (3A1) <400> SEQUENCE:
207 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Ala Asn Val Gly
Asn Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser Asn Arg Ser Gly Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr
Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly
Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100 105 110 <210> SEQ
ID NO 208 <211> LENGTH: 110 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: pAC19 (2C6) <400> SEQUENCE: 208 Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Lys Asn Val Gly Thr Asn 20
25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu
Ile 35 40 45 Tyr Leu Ala Ser Tyr Pro Gln Ile Gly Val Pro Ser Arg
Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His
Gln Tyr Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 209
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC18 (2F1) <400> SEQUENCE: 209 Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Ala Ala Val Gly Thr Tyr 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35
40 45 Tyr Ser Ala Ser Tyr Arg Lys Arg Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr
Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 210
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: pAC23 (2F11) <400> SEQUENCE: 210 Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg
Val Thr Ile Thr Cys Lys Ala Ser Gln Ile Ala Ser Thr Asn 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35
40 45 Tyr Trp Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr
Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 211
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: H4E9 light chain <400> SEQUENCE: 211 Asp Ile Gln
Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp
Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25
30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Pro Leu Ile
35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln
Tyr Tyr Thr Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys
Leu Glu Ile Lys Arg Thr 100 105 110 <210> SEQ ID NO 212
<211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: L2D2 <400> SEQUENCE: 212 Asp Ile Gln Met Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr
Ile Thr Cys Lys Ala Ser His Asn Val Gly Thr Asn 20 25 30 Val Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45
Tyr Ser Ala Ser His Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50
55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr
Tyr Pro Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr 100 105 110 <210> SEQ ID NO 213 <211>
LENGTH: 110 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: pAC6
(C1) <400> SEQUENCE: 213 Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys
Lys Ala Ser Gln Ile Met Gly Pro Asn 20 25 30 Val Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala
Ser Tyr His Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70
75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro
Leu 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
Thr 100 105 110 <210> SEQ ID NO 214 <211> LENGTH:
110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC7 (E10)
<400> SEQUENCE: 214 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Ile Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser
His Arg Pro Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 215 <211> LENGTH: 110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC12 (H7)
<400> SEQUENCE: 215 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Lys Val Leu Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser
Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 216 <211> LENGTH: 110
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: pAC13 (H11)
<400> SEQUENCE: 216 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys
Ala Ser Gln Thr Val Ser Ala Asn 20 25 30 Val Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Leu Ala Ser
Tyr Arg Tyr Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu 85
90 95 Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr 100
105 110 <210> SEQ ID NO 217 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 (Y98A)
Heavy Chain CDR3 <400> SEQUENCE: 217 Trp Asp Phe Ala Asp Tyr
Val Glu Ala Met Asp Tyr 1 5 10 <210> SEQ ID NO 218
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 (D99Y) Heavy Chain CDR3 <400> SEQUENCE:
218 Trp Asp Phe Tyr Tyr Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 219 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 (D99H) Heavy Chain CDR3
<400> SEQUENCE: 219 Trp Asp Phe Tyr His Tyr Val Glu Ala Met
Asp Tyr 1 5 10 <210> SEQ ID NO 220 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 (V101F)
Heavy Chain CDR3 <400> SEQUENCE: 220 Trp Asp Phe Tyr Asp Tyr
Phe Glu Ala Met Asp Tyr 1 5 10 <210> SEQ ID NO 221
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 (E101aQ) Heavy Chain CDR3 <400> SEQUENCE:
221 Trp Asp Phe Tyr Asp Tyr Val Gln Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 222 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: PR1A3 (A103T) Heavy Chain CDR3
<400> SEQUENCE: 222 Trp Asp Phe Tyr Asp Tyr Val Glu Thr Met
Asp Tyr 1 5 10 <210> SEQ ID NO 223 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: PR1A3 (Y98A /
D99Y) Heavy Chain CDR3 <400> SEQUENCE: 223 Trp Asp Phe Ala
Tyr Tyr Val Glu Ala Met Asp Tyr 1 5 10 <210> SEQ ID NO 224
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 (W95Y) Heavy Chain CDR3 <400> SEQUENCE:
224 Tyr Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr 1 5 10
<210> SEQ ID NO 225 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (Y98A) Heavy Chain Construct
<400> SEQUENCE: 225 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Ala Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 226 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (D99Y) Heavy Chain Construct
<400> SEQUENCE: 226 Gln Val Gln Leu Val Gln Ser Gly Ser Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr
65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Tyr Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 227 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (D99H) Heavy Chain
Construct <400> SEQUENCE: 227 Gln Val Gln Leu Val Gln Ser Gly
Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65
70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr His Tyr Val Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 228 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (V101F) Heavy Chain
Construct <400> SEQUENCE: 228 Gln Val Gln Leu Val Gln Ser Gly
Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65
70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Phe Glu Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 229 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (E102Q) Heavy Chain
Construct <400> SEQUENCE: 229 Gln Val Gln Leu Val Gln Ser Gly
Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65
70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Gln Ala Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 230 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (A103T) Heavy Chain
Construct <400> SEQUENCE: 230 Gln Val Gln Leu Val Gln Ser Gly
Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65
70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Thr Met
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 231 <211> LENGTH: 121 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: CH7A (Y98A / D99Y) Heavy
Chain Construct <400> SEQUENCE: 231 Gln Val Gln Leu Val Gln
Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val
Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met
Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45
Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50
55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala
Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ala Tyr Tyr Val Glu Ala
Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser
115 120 <210> SEQ ID NO 232 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH7A (W95Y)
Heavy Chain Construct <400> SEQUENCE: 232 Gln Val Gln Leu Val
Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys
Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly
Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe
50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr
Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Asp Phe Tyr Asp Tyr Val Glu
Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser
Ser 115 120 <210> SEQ ID NO 233 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1A (Y98A)
<400> SEQUENCE: 233 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Ala Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 234
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1A (D99Y) <400> SEQUENCE: 234 Gln Val Gln
Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser Thr
Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Tyr Tyr
Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 235 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1A (D99H)
<400> SEQUENCE: 235 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr His Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 236 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1A (V101F) <400> SEQUENCE:
236 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr
Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Asp Tyr Phe Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 237
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1A (E102Q) <400> SEQUENCE: 237 Gln Val Gln
Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser Thr
Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Val Gln Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 238 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1A (A103T)
<400> SEQUENCE: 238 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Thr Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 239 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1A (Y98A / D99Y) <400>
SEQUENCE: 239 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr
Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro
Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr
Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr
Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu
Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Trp Asp Phe Ala Tyr Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105
110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 240 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH1A1A (W95Y) <400> SEQUENCE: 240 Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Thr Asp Thr Ser
Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Asp Phe Tyr Asp
Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 241 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1B
(Y98A) <400> SEQUENCE: 241 Gln Val Lys Leu Gln Gln Ser Gly
Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu
Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser
Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ala Asp Tyr Val
Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120 <210> SEQ ID NO 242 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1B (D99Y)
<400> SEQUENCE: 242 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Tyr Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 243 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B (D99H) <400> SEQUENCE:
243 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr
Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr His Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 244
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1B (V101F) <400> SEQUENCE: 244 Gln Val Lys
Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25
30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu
Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr
Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr
Phe Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 <210> SEQ ID NO 245 <211> LENGTH:
121 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1B (E102Q)
<400> SEQUENCE: 245 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Gln Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 246 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B (A103T) <400> SEQUENCE:
246 Gln Val Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
Glu Phe 20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala
Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr
Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp
Phe Tyr Asp Tyr Val Glu Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 247
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A1B (Y98A / D99Y) <400> SEQUENCE: 247 Gln Val
Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20
25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val
Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser
Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Ala Tyr
Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val
Thr Val Ser Ser 115 120 <210> SEQ ID NO 248 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1B
(W95Y) <400> SEQUENCE: 248 Gln Val Lys Leu Gln Gln Ser Gly
Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp
Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp
Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60
Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Arg Tyr Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100
105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ
ID NO 249 <211> LENGTH: 363 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH7A (Y98A) Heavy Chain Construct <400>
SEQUENCE: 249 caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc
ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata caccttcact
gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg
gatgggatgg ataaacacca aaactggaga ggcaacatat 180 gttgaagagt
ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240
ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac
300 ttcgctgatt acgtggaggc tatggactac tggggccaag ggaccacggt
caccgtctcc 360 tca 363 <210> SEQ ID NO 250 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH7A
(D99Y) Heavy Chain Construct <400> SEQUENCE: 250 caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatgggac 300 ttctattatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 251 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (D99H) Heavy Chain Construct
<400> SEQUENCE: 251 caggtgcagc tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 ttctatcatt acgtggaggc tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 252
<211> LENGTH: 364 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH7A (V101F) Heavy Chain Construct <400>
SEQUENCE: 252 ccaggtgcag ctggtgcaat ctgggtctga gttgaagaag
cctggggcct cagtgaaggt 60 ttcctgcaag gcttctggat acaccttcac
tgagtttgga atgaactggg tgcgacaggc 120 ccctggacaa gggcttgagt
ggatgggatg gataaacacc aaaactggag aggcaacata 180 tgttgaagag
tttaagggac ggtttgtctt ctccttggac acctctgtca gcacggcata 240
tctgcagatc agcagcctaa aggctgaaga cactgccgtg tattactgtg cgagatggga
300 cttctatgat tacttcgagg ctatggacta ctggggccaa gggaccacgg
tcaccgtctc 360 ctca 364 <210> SEQ ID NO 253 <211>
LENGTH: 364 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH7A
(E102Q) Heavy Chain Construct <400> SEQUENCE: 253 ccaggtgcag
ctggtgcaat ctgggtctga gttgaagaag cctggggcct cagtgaaggt 60
ttcctgcaag gcttctggat acaccttcac tgagtttgga atgaactggg tgcgacaggc
120 ccctggacaa gggcttgagt ggatgggatg gataaacacc aaaactggag
aggcaacata 180 tgttgaagag tttaagggac ggtttgtctt ctccttggac
acctctgtca gcacggcata 240 tctgcagatc agcagcctaa aggctgaaga
cactgccgtg tattactgtg cgagatggga 300 cttctatgat tacgtgcagg
ctatggacta ctggggccaa gggaccacgg tcaccgtctc 360 ctca 364
<210> SEQ ID NO 254 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH7A (A103T) Heavy Chain Construct
<400> SEQUENCE: 254 caggtgcagc tggtgcaatc tgggtctgag
ttgaagaagc ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata
caccttcact gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag
ggcttgagtg gatgggatgg ataaacacca aaactggaga ggcaacatat 180
gttgaagagt ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat
240 ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc
gagatgggac 300 ttctatgatt acgtggagac tatggactac tggggccaag
ggaccacggt caccgtctcc 360 tca 363 <210> SEQ ID NO 255
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH7A (Y98A / D99Y) Heavy Chain Construct <400>
SEQUENCE: 255 caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc
ctggggcctc agtgaaggtt 60 tcctgcaagg cttctggata caccttcact
gagtttggaa tgaactgggt gcgacaggcc 120 cctggacaag ggcttgagtg
gatgggatgg ataaacacca aaactggaga ggcaacatat 180 gttgaagagt
ttaagggacg gtttgtcttc tccttggaca cctctgtcag cacggcatat 240
ctgcagatca gcagcctaaa ggctgaagac actgccgtgt attactgtgc gagatgggac
300 ttcgcttatt acgtggaggc tatggactac tggggccaag ggaccacggt
caccgtctcc 360 tca 363 <210> SEQ ID NO 256 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH7A
(W95Y) Heavy Chain Construct <400> SEQUENCE: 256 caggtgcagc
tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc agtgaaggtt 60
tcctgcaagg cttctggata caccttcact gagtttggaa tgaactgggt gcgacaggcc
120 cctggacaag ggcttgagtg gatgggatgg ataaacacca aaactggaga
ggcaacatat 180 gttgaagagt ttaagggacg gtttgtcttc tccttggaca
cctctgtcag cacggcatat 240 ctgcagatca gcagcctaaa ggctgaagac
actgccgtgt attactgtgc gagatacgac 300 ttctatgatt acgtggaggc
tatggactac tggggccaag ggaccacggt caccgtctcc 360 tca 363 <210>
SEQ ID NO 257 <211> LENGTH: 121 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1 <400> SEQUENCE: 257 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Phe Thr Leu Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 258 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A2
<400> SEQUENCE: 258 Gln Val Lys Leu Gln Gln Ser Gly Pro Glu
Leu Lys Lys Pro Gly Glu 1 5 10 15 Thr Val Lys Ile Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45
Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50
55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala
Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala
Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser
115 120 <210> SEQ ID NO 259 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A3
<400> SEQUENCE: 259 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Lys
Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 260 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A4 <400> SEQUENCE: 260 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr
Ser Ala Thr Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn
Glu Asp Thr Ala Lys Tyr Phe Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 261 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1A
<400> SEQUENCE: 261 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Phe Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 262 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B <400> SEQUENCE: 262 Gln
Val Lys Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 263 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1C
<400> SEQUENCE: 263 Gln Val Gln Leu Val Gln Ser Gly Pro Glu
Leu Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 264 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1D <400> SEQUENCE: 264 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10
15 Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 265 <211>
LENGTH: 121 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1E
<400> SEQUENCE: 265 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Phe Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser
115 120 <210> SEQ ID NO 266 <211> LENGTH: 121
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: CH1A1F
<400> SEQUENCE: 266 Gln Val Gln Leu Val Gln Ser Gly Ala Glu
Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly Met Asn Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn
Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60 Lys Gly
Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80
Leu Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp
Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120
<210> SEQ ID NO 267 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1G <400> SEQUENCE: 267 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Ile Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 268 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A1
<400> SEQUENCE: 268 caggtgcagc tggtgcagtc tggcgccgaa
gtgaagaaac ctggagctag tgtgaaggtg 60 tcctgcaagg ccagcggcta
caccttcacc gagttcggca tgaactgggt ccgacaggct 120 ccaggccagg
gcctcgaatg gatgggctgg atcaacacca agaccggcga ggccacctac 180
gtggaagagt tcaagggcag agtgaccttc accctggaca ccagcaccag caccgcctac
240 atggaactgc ggagcctgag aagcgacgac accgccgtgt actactgcgc
cagatgggac 300 ttctacgatt acgtggaagc catggactac tggggccagg
gcaccaccgt gaccgtgtct 360 agc 363 <210> SEQ ID NO 269
<211> LENGTH: 363 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: CH1A2 <400> SEQUENCE: 269 caggtcaaac tgcagcagag
cggccctgag ctgaagaaac ccggcgagac agtgaagatc 60 agctgcaagg
ccagcggcta caccttcacc gagttcggca tgaactgggt ccgacaggca 120
ccaggccagg gcctcgaatg gatgggctgg atcaacacca agaccggcga ggccacctac
180 gtggaagagt tcaagggcag agtgaccatg accaccgaca ccagcaccag
caccgcctac 240 atggaactgc ggagcctgag aagcgacgac accgccgtgt
actactgcgc cagatgggac 300 ttctacgatt acgtggaagc catggactac
tggggccagg gcaccaccgt gaccgtgtct 360 agc 363 <210> SEQ ID NO
270 <211> LENGTH: 363 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH1A3 <400> SEQUENCE: 270 caggtgcagc
tggtgcagtc tggcgccgaa gtgaagaaac ctggcgccag cgtgaaggtg 60
tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt caagcaggcc
120 cctggcaagg gcctgaagtg gatgggctgg atcaacacca agaccggcga
ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg accaccgaca
ccagcaccag caccgcctac 240 atggaactgc ggagcctgag aagcgacgac
accgccgtgt actactgcgc cagatgggac 300 ttctacgatt acgtggaagc
catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363 <210>
SEQ ID NO 271 <211> LENGTH: 362 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A4 <400> SEQUENCE: 271
aggtgcagct ggtgcagtct ggcgccgaag tgaagaaacc tggcgccagc gtgaaggtgt
60 cctgcaaggc cagcggctac accttcaccg agttcggcat gaactgggtc
cgacaggcac 120 caggccaggg cctcgaatgg atgggctgga tcaacaccaa
gaccggcgag gccacctacg 180 tggaagagtt caagggcaga ttcgccttca
gcctggaaac cagcgccacc accgcctacc 240 tgcagatcaa caacctgaag
aacgaggata ccgccaagta cttctgcgcc agatgggact 300 tctacgatta
cgtggaagcc atggactact ggggccaggg caccaccgtg accgtgtcta 360 gc 362
<210> SEQ ID NO 272 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1A <400> SEQUENCE: 272
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccttc
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 273 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1B <400> SEQUENCE: 273
caggtgaagc tgcagcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 274 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1C <400> SEQUENCE: 274
caggtgcagc tggtgcagtc tggccccgaa ctgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 275 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1D <400> SEQUENCE: 275
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagagac tgtgaagatc
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120
ccaggccagg gcctcgaatg gatgggctgg atcaacacca agaccggcga ggccacctac
180 gtggaagagt tcaagggcag agtgaccatg accacggaca ccagcaccag
caccgcctac 240 atggaactgc ggagcctgag aagcgacgac accgccgtgt
actactgcgc cagatgggac 300 ttctacgatt acgtggaagc catggactac
tggggccagg gcaccaccgt gaccgtgtct 360 agc 363 <210> SEQ ID NO
276 <211> LENGTH: 363 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: CH1A1E <400> SEQUENCE: 276 caggtgcagc
tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg 60
tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt ccgacaggct
120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca agaccggcga
ggccacctac 180 gtggaagagt tcaagggcag attcaccatg accacggaca
ccagcaccag caccgcctac 240 atggaactgc ggagcctgag aagcgacgac
accgccgtgt actactgcgc cagatgggac 300 ttctacgatt acgtggaagc
catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363 <210>
SEQ ID NO 277 <211> LENGTH: 363 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1F <400> SEQUENCE: 277
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 ctggaactgc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 278 <211> LENGTH: 363 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A1G <400> SEQUENCE: 278
caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ctggagctag tgtgaaggtg
60 tcctgcaagg ccagcggcta caccttcacc gagttcggca tgaactgggt
ccgacaggct 120 ccaggccagg gcctcgaatg gatgggctgg atcaacacca
agaccggcga ggccacctac 180 gtggaagagt tcaagggcag agtgaccatg
accacggaca ccagcaccag caccgcctac 240 atggaaatcc ggagcctgag
aagcgacgac accgccgtgt actactgcgc cagatgggac 300 ttctacgatt
acgtggaagc catggactac tggggccagg gcaccaccgt gaccgtgtct 360 agc 363
<210> SEQ ID NO 279 <211> LENGTH: 121 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: CH1A <400> SEQUENCE: 279 Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10
15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe
20 25 30 Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Met 35 40 45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr
Val Glu Glu Phe 50 55 60 Lys Gly Arg Val Thr Met Thr Thr Asp Thr
Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr
Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr
Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 280 <211>
LENGTH: 363 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION: CH1A
<400> SEQUENCE: 280 caggtgcagc tggtgcagtc tggcgctgag
gtgaagaagc ctggcgcctc ggtgaaggtc 60 tcctgcaagg cctctggtta
cacatttacg gaattcggga tgaattgggt cagacaagca 120 cctgggcaag
ggctcgagtg gatgggatgg ataaacacga agacaggcga ggccacctac 180
gtagaagagt tcaaaggaag ggtcacgatg accacagata cttctacctc tactgcgtat
240 atggaactac ggagcttgcg tagcgatgac acagcggtgt actattgtgc
tcgatgggat 300 ttctatgact atgttgaagc tatggactac tggggccaag
ggaccaccgt gaccgtctcc 360 tca 363 <210> SEQ ID NO 281
<211> LENGTH: 121 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: PR1A3 <400> SEQUENCE: 281 Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys
Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30 Gly
Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe
50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr
Ala Tyr 65 70 75 80 Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu
Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser
Ser 115 120
* * * * *