U.S. patent number 11,440,954 [Application Number 16/863,971] was granted by the patent office on 2022-09-13 for optimized anti-tl1a antibodies.
This patent grant is currently assigned to Cedars-Sinai Medical Center, Prometheus Biosciences, Inc.. The grantee listed for this patent is Cedars-Sinai Medical Center, Prometheus Biosciences, Inc.. Invention is credited to Janine Bilsborough, Cindy T. Dickerson, Bradley Henkle, Patricia McNeeley, Stephan R. Targan, J. Monty Watkins, Jeffry D. Watkins.
United States Patent |
11,440,954 |
Watkins , et al. |
September 13, 2022 |
Optimized anti-TL1A antibodies
Abstract
Described herein are humanized anti-TL1A antibodies and
pharmaceutical compositions for the treatment of inflammatory bowel
disease (IBD), such as Crohn's Disease (CD) and ulcerative colitis
(UC).
Inventors: |
Watkins; Jeffry D. (Encinitas,
CA), Dickerson; Cindy T. (Encinitas, CA), Watkins; J.
Monty (Encinitas, CA), McNeeley; Patricia (Encinitas,
CA), Bilsborough; Janine (Adelaide, AU), Henkle;
Bradley (West Hollywood, CA), Targan; Stephan R. (Santa
Monica, CA) |
Applicant: |
Name |
City |
State |
Country |
Type |
Prometheus Biosciences, Inc.
Cedars-Sinai Medical Center |
San Diego
Los Angeles |
CA
CA |
US
US |
|
|
Assignee: |
Prometheus Biosciences, Inc.
(San Diego, CA)
Cedars-Sinai Medical Center (Los Angeles, CA)
|
Family
ID: |
1000006554760 |
Appl.
No.: |
16/863,971 |
Filed: |
April 30, 2020 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20200255510 A1 |
Aug 13, 2020 |
|
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
16694814 |
Nov 25, 2019 |
10689439 |
|
|
|
PCT/US2019/028987 |
Apr 24, 2019 |
|
|
|
|
62662605 |
Apr 25, 2018 |
|
|
|
|
62756494 |
Nov 6, 2018 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12N
15/09 (20130101); A61P 1/04 (20180101); C07K
16/28 (20130101); C07K 16/241 (20130101); C12N
5/10 (20130101); C07K 16/2875 (20130101); C12N
15/63 (20130101); A61K 9/0019 (20130101); A61P
1/00 (20180101); C07K 16/24 (20130101); C07K
2317/76 (20130101); C07K 2317/565 (20130101); A61K
39/3955 (20130101); C07K 2317/24 (20130101); C07K
2317/732 (20130101); C07K 2317/90 (20130101); C07K
2317/56 (20130101); C07K 2317/71 (20130101); C07K
2317/734 (20130101); A61K 2039/505 (20130101) |
Current International
Class: |
C07K
16/24 (20060101); C12N 15/63 (20060101); C12N
15/09 (20060101); C07K 16/28 (20060101); C12N
5/10 (20060101); A61K 9/00 (20060101); A61P
1/00 (20060101); A61P 1/04 (20060101); C07K
14/525 (20060101); A61K 39/00 (20060101); A61K
39/395 (20060101); C07K 14/715 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
|
|
|
|
|
2462165 |
|
May 2016 |
|
EP |
|
2638069 |
|
Jan 2018 |
|
EP |
|
WO-9614328 |
|
May 1996 |
|
WO |
|
WO-0066608 |
|
Nov 2000 |
|
WO |
|
WO-0204643 |
|
Jan 2002 |
|
WO |
|
WO-03068821 |
|
Aug 2003 |
|
WO |
|
WO-2004035537 |
|
Apr 2004 |
|
WO |
|
WO-2004050836 |
|
Jun 2004 |
|
WO |
|
WO-2006017173 |
|
Feb 2006 |
|
WO |
|
WO-2007005608 |
|
Jan 2007 |
|
WO |
|
WO-2008106579 |
|
Sep 2008 |
|
WO |
|
WO-2009064854 |
|
May 2009 |
|
WO |
|
WO-2009064854 |
|
Jul 2009 |
|
WO |
|
WO-2012161856 |
|
Nov 2012 |
|
WO |
|
WO-2013044298 |
|
Apr 2013 |
|
WO |
|
WO-2014051109 |
|
Apr 2014 |
|
WO |
|
WO-2014106602 |
|
Jul 2014 |
|
WO |
|
WO-2014160463 |
|
Oct 2014 |
|
WO |
|
WO-2014197849 |
|
Dec 2014 |
|
WO |
|
WO-2015073580 |
|
May 2015 |
|
WO |
|
WO-2017049024 |
|
Mar 2017 |
|
WO |
|
WO-2017076878 |
|
May 2017 |
|
WO |
|
WO-2017077715 |
|
May 2017 |
|
WO |
|
WO-2017106383 |
|
Jun 2017 |
|
WO |
|
WO-2017189983 |
|
Nov 2017 |
|
WO |
|
WO-2017196663 |
|
Nov 2017 |
|
WO |
|
WO-2018081074 |
|
May 2018 |
|
WO |
|
WO-2019121906 |
|
Jun 2019 |
|
WO |
|
WO-2019209995 |
|
Oct 2019 |
|
WO |
|
WO-2020011964 |
|
Jan 2020 |
|
WO |
|
WO-2020011966 |
|
Jan 2020 |
|
WO |
|
WO-2020011968 |
|
Jan 2020 |
|
WO |
|
WO-2020011970 |
|
Jan 2020 |
|
WO |
|
WO-2020011972 |
|
Jan 2020 |
|
WO |
|
WO-2020011974 |
|
Jan 2020 |
|
WO |
|
WO-2020011976 |
|
Jan 2020 |
|
WO |
|
WO-2020086758 |
|
Apr 2020 |
|
WO |
|
WO-2021081365 |
|
Apr 2021 |
|
WO |
|
WO-2021260577 |
|
Dec 2021 |
|
WO |
|
Other References
Jaye et al. Isolation of a human anti-haemophilic factor IX cDNA
clone using a 52-base synthetic oligonucleotide probe deduced from
the amino acid sequence of bovine factor IX. Nucleic Acids Res
11(8): 2325-2335, 1983. cited by examiner .
Lewin, B. Genes IV, Oxford: Oxford University Press, 1990; pp.
118-120. cited by examiner .
Sambrook et al. Molecular Cloning A Laboratory Manual, 2nd edition,
Cold Spring Harbor, N.Y., 1989, pp. 2.43-2.84. cited by examiner
.
PCT/US2019/028987 International Preliminary Report on Patentability
dated Oct. 27, 2020. cited by applicant .
PCT/US2020/057119 International Search Report and Written Opinion
dated Mar. 10, 2021. cited by applicant .
Prometheus Biosciences, Inc. Form S-1 Registration Statement as
filed with the Securities and Exchange Commission on Feb. 19, 2021
(246 pages). cited by applicant .
Scheepers et al.: Ability To Develop Broadly Neutralizing HIV-1
Antibodies Is Not Restricted by the Germline Ig Gene Repertoire. J
Immunol. 194(9):4371-4378 (2015). cited by applicant .
Adams et al., Two-stage genome-wide methylation profiling in
childhood-onset Crohn's Disease implicates epigenetic alterations
at the VMP1/MIR21 and HLA loci. Inflamm Bowel Dis. 20(10):1784-1793
(2014). cited by applicant .
Aiba et al., The role of TL1A and DR3 in autoimmune and
inflammatory diseases. Mediators Inflamm. 2013:#258164, 9 pages.
cited by applicant .
Al-Iazikani et al., Standard conformations for the canonical
structures of immunoglobulins. J. Molec. Biol. 273:927-948, 1997.
cited by applicant .
Australian Patent Application No. 2014241162 Office Action dated
Apr. 16, 2018. cited by applicant .
Bamias et al. Circulating levels of TNF-like cytokine 1A (TL1A) and
its decoy receptor 3 (DcR3) in rheumatoid arthritis. Clin Immunol
129:249-255, 2008. cited by applicant .
Bamias et al.: Expression, localization, and functional activity of
TL1A, a novel Th1-polarizing cytokine in inflammatory bowel
disease. Journal of Immunology 171(9):4868-4874 (2003). cited by
applicant .
Barrett et al., Constitutive TL1A Expression under Colitogenic
Condition Modulates the Severity and Location of Gut Mucosal
Inflammation and Induces Fibrostenosis, American Journal of
Pathology, 2012, vol. 180(2), pp. 636-649. cited by applicant .
Bauer et al., A genetic enrichment for mutations constructed by
oligodeoxynucleotide-directed mutagenesis. Gene, 37:73-81, 1985.
cited by applicant .
Benedict et al., Immunoglobulin Kappa light chain variable region,
Partial (Mus musculus). GenBank: AAD39789.1, Jul. 26, 2016, 1 page.
cited by applicant .
Bird et al.: Single-chain antigen-binding proteins; Science,
242:423-42, 1988. cited by applicant .
Brennan et al. Preparation of bispecific antibodies by chemical
recombination of monoclonal immunoglobulin G1 fragments. Science
229:81-83, 1985. cited by applicant .
Brummell et al. Probing the combining site of an anti-carbohydrate
antibody by saturation-mutagenesis: Role of the heavy-chain CDR3
residues. Biochem. 32: 1180-1187, 1993 . cited by applicant .
Burks et al., In vitro scanning saturation mutagenesis of an
antibody binding pocket. Proc. Natl. Acad. Sci. USA 94:412-417,
1997. cited by applicant .
Camoglio et al.: Altered expression of interfero-gamma and
interleukin-4 in inflammatory bowel disease; Inflamm Bowel Dis.,
4(4): 285-290; Abstract only (1998). cited by applicant .
Clarke et al. An anti-TL1A antibody for the treatment of asthma and
inflammatory bowel disease. MAbs 10(4):664-677 (2018). cited by
applicant .
Craik, Charles. Use of oligonucleotides for site-specific
mutagenesis. BioTechniques 1985 :12-19, 1985 . cited by applicant
.
Erpenbeck et al. Segmental allergen challenge in patients with
atopic asthma leads to increased IL-9 expression in bronchoalveolar
lavage fluid lymphocytes. J Allergy Clin Immunol 111(6):1319-1327,
2003. cited by applicant .
European Patent Application No. 14797214.5 Office Action dated Apr.
19, 2018. cited by applicant .
Fang et al., Essential role of TNF receptor superfamily 25
(TNFRS25) in the development of allergic lung inflammation. J.Exp.
Med., 205(5):1037-1048, 2008. cited by applicant .
Fitzpatrick, LR, Novel Pharmacological Approaches for Inflammatory
Bowl Disease: Targeting Key Intracellular Pathways and the
IL-23/IL-17 Axis, International Journal of Inflammation, vol. 2012,
pp. 1-8. cited by applicant .
Fransen et al., Inflammatory bowel disease: the genetic background
and beyond. University of Groningen PhD Dissertation
http://www.rug.nl/research/portal/files/12805965/Complete_dissertation.pd-
f (2014). cited by applicant .
Heusch et al., IL-9 exacerbates colitis induced by CD4+CD45RBhigh T
cells transfer, via directed activation of in vivo
antigen-experienced T cells. Cytokine 56:PS1-056, p. 31 (2011).
cited by applicant .
Hodgson, John. Making monoclonals in microbes. Bio/Technology
9:421-425, 1991. cited by applicant .
Holliger et al.: Engineered antibody fragments and the rise of
single domains. Nat. Biotechnol. 23(9):1126-36 (2005). cited by
applicant .
Hsu et al. The tale of TL1A in inflammation. Mucosal Immunol
4(4):368-370, 2011. cited by applicant .
Hundorean et al., Functional relevance of T helper 17 (Th17) cells
and the IL-17 cytokine family in inflammatory bowel disease.
Inflammatory Bowel Disease 18:180-186 (2012). cited by applicant
.
Huse et al. Generation of a large combinatorial library of the
immunoglobulin repertoire in phage lambda. Science 246:1275-1281,
1989. cited by applicant .
Huston et al. Protein engineering of antibody binding sites:
Recovery of specific activity in an anti-digoxin single-chain Fv
analogue produced in Escherichia coli. Proc. Natl. Acad. Sci. USA
85:5879-5883, 1988 . cited by applicant .
International Application No. PCT/US2017/058019 International
Preliminary Report on Patentability dated Apr. 30, 2019. cited by
applicant .
Jones et al. Replacing the complementarity-determining regions in a
human antibody with those from a mouse. Nature 321:522-525, 1986 .
cited by applicant .
Kakuta et al., Su1746 Rare Variants of TNFSF15 Are Significantly
Associated With Crohn's Disease in Non-Jewish Caucasian Independent
of the Known Common Susceptibility SNPs, Gastroenterology, 144(5):
S-466, 2013. cited by applicant .
Kim et al. Effects of IL-9 blockade on chronic airway inflammation
of murine asthma models. Allergy: Eur J Allergy Clin Immunol Suppl
96(67):448, Nov. 2012. cited by applicant .
Kim et al. Effects of interleukin-9 blockade on chronic airway
inflammation in murine asthma models. Allergy Asthma Immunol Res
5(4):197-206, 2013. cited by applicant .
Kobayashi et al. Tryptophan H33 plays an important role in
pyrimidine (6-4) pyrimidone photoproduct binding by a high-affinity
antibody. Protein Eng. 12(10):879-884, 1999. cited by applicant
.
Koga et al., Transanal Delivery of Angiotensin Converting Enzyme
Inhibitor Prevents Colonic Fibrosis in a Mouse Colitis Model:
Development of a Unique Mode of Treatment, Surgery, 144(2):259-268,
2008. cited by applicant .
Kohler et al.:Derivation of specific antibody-producing tissue
culture and tumor lines by cell fusion. Eur. J. Immunol. 6: 511-519
(1976). cited by applicant .
McGovern et al., Genetics of inflammatory bowel diseases.
Gastroenterology 149(5):1163-1173 (2015). cited by applicant .
Meylan et al., The TNF-family cytokine TL1A drives IL-13 dependent
small intestinal inflammation. Muscosal Immunol., 4(2):172-185,
2011. cited by applicant .
Migone et al., TL1A is a TNF-like Ligand for DR3 and TR6/DcR3 and
Functions as a T cell Costimulator, Immunity, 16:479-492, 2002.
cited by applicant .
Morimoto et al. Single-step purification of F(ab')2 fragments of
mouse monoclonal antibodies (immunoglobulins G1) by hydrophobic
interaction high performance liquid chromatography using TSKgel
Phenyl-5PW. Journal of Biochemical and Biophysical Methods
24:107-117, 1993. cited by applicant .
Nalleweg et al., Inflammatory bowel disease patients failing
anti-TNF therapy show activation of the Th9/TH17 pathway.
Gastroenterol 142(5)(Suppl1):S867-868; Abstract No. Tu1878 (2012).
cited by applicant .
Takedatsu et al.: TL1A (TNFSF15) Regulates the Development of
Chronic Colitis By Modulating both T helper (TH) 1 and TH17
Activation; Gastroenterology; HHS Public Access; 135(2): 552-567
(2008). cited by applicant .
Nowak et al., IL-9 as a mediator of Th17-driven inflammatory
disease. Journal of Experimental Medicine 206(8):1653-1660 (2009).
cited by applicant .
Oh et al., A randomized, controlled trial to evaluate the effect of
an anti-interleukin-9 monoclonal antibody in adults with
uncontrolled asthma. Respiratory Research 14:93 (2013). cited by
applicant .
Parente et al., Bowel Ultrasound in Assessment of Crohn's Disease
and Detection of Related Small Bowel Strictures: A Prospective
Comparative Study Versus X Ray and Intraoperative Findings, Gut,
50: 490-495, 2002. cited by applicant .
PCT/US2009/069541 International Search Report dated Mar. 4, 2010.
cited by applicant .
PCT/US2017/023082 International Search Report and Written Opinion
dated Aug. 15, 2017. cited by applicant .
PCT/US2017/058019 International Search Report and Written Opinion
dated Feb. 15, 2018. cited by applicant .
PCT/US2019/028987 International Search Report and Written Opinion
dated Oct. 29, 2019. cited by applicant .
Pinchuk et al., Human Colonic Myofibroblast Promote Expansion of
CD4+ CD25high Foxp3+ Regulatory T Cells, Gastroenterology,
140(7):2019-2030, pp. 1-19, and p. 8, 2011. cited by applicant
.
Queen et al. A humanized antibody that binds to the interleukin 2
receptor. Proc. Natl. Acad Sci USA 86:10029-10032, 1989. cited by
applicant .
R&D datasheet for human/mouse TL1A/TNFSF15 antibody, catalog
No. MAB7441; clone #293327 (Feb. 7, 2018). cited by applicant .
Reichwald et al. TL1A induces TCR independent IL-6 and TNF-alpha
production and growth of PLZF leukocytes. PLOS ONE 9(1):e85793,
2013. cited by applicant .
Richard et al. The TNF-family cytokine TL1A: from lymphocyte
costimulator to disease co-conspirator. J Leukocyte Biol 98:333-345
2015. cited by applicant .
Riechmann et al. Reshaping human antibodies for therapy. Nature,
332.6162:323-7 (1988). cited by applicant .
Rothe et al., The human combinatorial antibody library HuCAL GOLD
combines diversification of all six CDRs according to the natural
immune system with a novel display method for efficient selection
of high-affinity antibodies. J. Mol. Bio. 376:1182-1200, 2008 .
cited by applicant .
Shih et al. Reversal of murine colitis and fibrosis by neutralizing
TL1A antibody: potential novel therapy to alter natural history of
Crohn's disease. Gastroenterol 142(5):S84, Abstract #357, 2012.
cited by applicant .
Shih et at, Constitutive TL1A (TNFSF15) Expression on Lymphoid or
Myeloid Cells Leads to Mild Intestinal Inflammation and Fibrosis,
PLOS One, 6(1), pp. 1-16, 2011. cited by applicant .
Spinelli et al., Intestinal Fibrosis in Crohn's Disease: Medical
Treatment or Surgery?, Current Drug Targets, 11(2):242-248, 2010.
cited by applicant .
Strober et at, Proinflammatoly Cytokines in Pathogenesis of
Inflammatory Bowel Diseases, Gastroenterology, 140(6):1756-1767,
2011. cited by applicant .
Tomlinson et al.: Methods for generating multivalent and bispecific
antibody fragments. Methods Enzymol 326:461-479 (2000). cited by
applicant .
UniprotKB Database, Q8NI17 (IL31R_Human), Retrieved online Sep. 5,
2019. Retrieved from<url
https://www.uniprot.org/uniprot/Q8NI17>. Jul. 31,
2019</url>. cited by applicant .
U.S. Appl. No. 16/384,521 Office Action dated May 28, 2020. cited
by applicant .
U.S. Appl. No. 14/779,893 Final Office Action dated Apr. 26, 2019.
cited by applicant .
U.S. Appl. No. 14/779,893 Office Action dated Sep. 12, 2019. cited
by applicant .
U.S. Appl. No. 15/245,875 Office Action dated Jun. 12, 2018. cited
by applicant .
U.S. Appl. No. 15/792,266 Office Action dated Aug. 6, 2018. cited
by applicant .
Verhoeyen et al. Reshaping human antibodies: Grafting an
antilysozyme activity. Science 239:1534-1536 (1988). cited by
applicant .
Walder et al. Oligodeoxynucleotide-directed mutagenesis using the
yeast transformation system. Gene 42:133-139, 1986. cited by
applicant .
Ward et al. Binding activities of a repertoire of single
immunoglobulin variable domains secreted from Escherichia coli.
Nature 334:544-54, 1989 . cited by applicant .
European Patent Application No. 17865031.3 Extended European Search
Report dated Jun. 5, 2020. cited by applicant .
Shih et al.: Inhibition of a novel fibrogenic factor TI 1a reverses
established colonic fibrosis. Mucosal Immunol., 7(6):1492-1503,
2014. cited by applicant .
Singapore Patent Application No. 11201903737P Search Report and
Written Opinion dated Jul. 17, 2020. cited by applicant .
Kabat numbering for IGHV1-46*02;
http://opig.stats.ox.ac.uk/webapps/newsabdab/sabpred/anarci;
accessed May 10, 2021. cited by applicant .
Kabat numbering for IGKV3-20*01;
http://opig.stats.ox.ac.uk/webapps/newsabdab/sabpred/anarci;
accessed May 10, 2021. cited by applicant .
Khanna et al.: Use of animal models in elucidating disease
pathogenesis in IBD. Semin Immunopathol. 36:541-551 (2014). cited
by applicant .
Rudikoff et al.: Single amino acid substitution altering
antigen-binding specificity. Proc Natl Acad Sci U S A. Mar.
1982;79(6):1979-1983. cited by applicant .
U.S. Appl. No. 17/110,004 First Action Interview Office Action
dated May 19, 2021. cited by applicant.
|
Primary Examiner: Bunner; Bridget E
Attorney, Agent or Firm: Wilson Sonsini Goodrich &
Rosati
Parent Case Text
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. application Ser. No.
16/694,814 filed Nov. 25, 2019, which is a continuation of
international PCT App. No. PCT/US19/28987 filed Apr. 24, 2019,
which claims the benefit of U.S. Provisional App. No. 62/662,605
filed on Apr. 25, 2018, and U.S. Provisional App. No. 62/756,494
filed on Nov. 6, 2018, which applications are incorporated by
reference herein in their entirety.
Claims
What is claimed is:
1. A nucleic acid encoding an antibody, or antigen binding portion
thereof, that binds to tumor necrosis factor-like ligand 1A (TL1A),
wherein the nucleic acid comprises: a nucleic acid sequence
encoding a heavy chain variable region comprising: (a) a nucleic
acid encoding a HCDR1 comprising the amino acid sequence set forth
by SEQ ID NO: 553; (b) a nucleic acid encoding a HCDR2 comprising
an amino acid sequence set forth by any one of SEQ ID NOs: 554 to
564, or 574 to 577; and (c) a nucleic acid encoding a HCDR3
comprising an amino acid sequence set forth by any one of SEQ ID
NOs: 565 to 568, or 579 to 581; and a nucleic acid sequence
encoding a light chain variable region comprising: (a) a nucleic
acid encoding a LCDR1 comprising an amino acid sequence set forth
by any one of SEQ ID NOs: 569 or 570; (b) a nucleic acid encoding a
LCDR2 comprising the amino acid sequence set forth by SEQ ID NO:
488; and (c) a nucleic acid encoding a LCDR3 comprising the amino
acid sequence set forth by SEQ ID NO: 572.
2. The nucleic acid encoding the antibody or antigen binding
portion of claim 1, wherein the HCDR2 comprises the amino acid
sequence as set forth by SEQ ID NO: 554.
3. The nucleic acid encoding the antibody or antigen binding
portion of claim 1, wherein the HCDR2 comprises the amino acid
sequence as set forth by SEQ ID NO: 555.
4. The nucleic acid encoding the antibody or antigen binding
portion of claim 1, wherein the HCDR3 comprises the amino acid
sequence as set forth by SEQ ID NO: 565.
5. The nucleic acid encoding the antibody or antigen binding
portion of claim 1, wherein the HCDR3 comprises the amino acid
sequence as set forth by SEQ ID NO: 566.
6. The nucleic acid encoding the antibody or antigen binding
portion of claim 1, wherein the LCDR1 comprises the amino acid
sequence as set forth by SEQ ID NO: 569.
7. The nucleic acid encoding the antibody or antigen binding
portion of claim 1, wherein the nucleic acid sequence encoding the
heavy chain variable region comprises a nucleic acid encoding for a
heavy chain constant region, and the nucleic acid sequence encoding
the light chain variable region comprises a nucleic acid encoding
for a light chain constant region.
8. A recombinant vector comprising the nucleic acid of claim 1.
9. The recombinant vector of claim 8, wherein the recombinant
vector is a plasmid.
10. The recombinant vector of claim 8, wherein the recombinant
vector is an expression vector.
11. A host cell comprising the recombinant vector of claim 8.
12. A nucleic acid encoding an antibody, or antigen binding portion
thereof, that binds to tumor necrosis factor-like ligand 1A (TL1A),
wherein the nucleic acid comprises: a nucleic acid sequence
encoding a heavy chain variable region comprising: (a) a nucleic
acid encoding a HCDR1 comprising the amino acid sequence set forth
by SEQ ID NO: 553; (b) a nucleic acid encoding a HCDR2 comprising
the amino acid sequence set forth by SEQ ID NO: 554; and (c) a
nucleic acid encoding a HCDR3 comprising the amino acid sequence
set forth by SEQ ID NO: 565; and a nucleic acid sequence encoding a
light chain variable region comprising: (a) a nucleic acid encoding
a LCDR1 comprising the amino acid sequence set forth by SEQ ID NO:
569; (b) a nucleic acid encoding a LCCR2 comprising the amino acid
sequence set forth by SEQ ID NO: 488; and (c) a nucleic acid
encoding a LCDR3 comprising the amino acid sequence set forth by
SEQ ID NO: 572.
13. The nucleic acid encoding the antibody or antigen binding
portion of claim 12, wherein the nucleic acid sequence encoding the
heavy chain variable region comprises a nucleic acid encoding for a
heavy chain constant region, and the nucleic acid sequence encoding
the light chain variable region comprises a nucleic acid encoding
for a light chain constant region.
14. A recombinant vector comprising the nucleic acid of claim
12.
15. The recombinant vector of claim 14, wherein the recombinant
vector is a plasmid.
16. The recombinant vector of claim 14, wherein the recombinant
vector is an expression vector.
17. A host cell comprising the recombinant vector of claim 14.
18. A method of producing an antibody, or an antigen binding
portion thereof, that binds to tumor necrosis factor-like ligand 1A
(TL1A), wherein the method comprises (a) providing a host cell
comprising an expression vector harboring a nucleic acid encoding
the antibody, or the antibody binding portion thereof, wherein the
nucleic acid comprises: a nucleic acid sequence encoding a heavy
chain variable region comprising: (i) a nucleic acid encoding a
HCDR1 comprising the amino acid sequence set forth by SEQ ID NO:
553; (ii) a nucleic acid encoding a HCDR2 comprising the amino acid
sequence set forth by SEQ ID NO: 554; and (iii) a nucleic acid
encoding a HCDR3 comprising the amino acid sequence set forth by
SEQ ID NO: 565; and a nucleic acid sequence encoding a light chain
variable region comprising: (i) a nucleic acid encoding a LCDR1
comprising the amino acid sequence set forth by SEQ ID NO: 569;
(ii) a nucleic acid encoding a LCCR2 comprising the amino acid
sequence set forth by SEQ ID NO: 488; and (iii) a nucleic acid
encoding a LCDR3 comprising the amino acid sequence set forth by
SEQ ID NO: 572; and (b) culturing the host cell under conditions to
induce expression of the antibody or antibody portion from the host
cell.
19. The method of claim 18, further comprising harvesting the
antibody or antigen binding portion.
20. The method of claim 19, further comprising purifying the
harvested antibody or antigen binding portion.
Description
SEQUENCE LISTING
The instant application contains a Sequence Listing which has been
submitted electronically in ASCII format and is hereby incorporated
by reference in its entirety. Said ASCII copy, created on May 13,
2019, is named 52388-735 601 SL.txt and is 162,559 bytes in
size.
BACKGROUND
Inflammatory bowel disease (IBD) refers to a collection of
intestinal disorders causing inflammatory conditions in the
gastrointestinal tract. The primary types of IBD are ulcerative
colitis (UC) and Crohn's Disease (CD). These diseases are
prevalent, with about 1.86 million people diagnosed globally with
UC, and about 1.3 million people diagnosed globally with CD.
Each of these forms has various subclinical phenotypes
characteristic of severe forms of IBD that are present in
sub-populations of CD and UC patients. One such condition is
obstructive Crohn's disease, which can result from long term
inflammation that may lead to the formation of scar tissue in the
intestinal wall (fibrostenosis) or swelling. Both outcomes can
cause narrowing, or obstruction, and are known as either fibrotic
or inflammatory strictures. Severe strictures can lead to blockage
of the intestine, leading to abdominal pain, bloating, nausea and
the inability to pass stool. As another example, penetrating
disease phenotypes characterized by bowel obstruction or internal
penetrating fistulas, or both, often resulting in complications
associated with IBD, including for e.g., intra-abdominal
sepsis.
Unfortunately, there are a limited number of therapies available
for IBD patients, and the development of new therapeutics has been
hampered by sub-optimal results in clinical trials. Existing
anti-inflammatory therapy such as steroids and tumor necrosis
factor (TNF) inhibitors are typically use as a first line treatment
for treating IBD. Unfortunately, a significant number of patients
experience a lack of response or a loss of response to existing
anti-inflammatory therapies, especially TNF-alpha inhibitors. While
the patient is treated with an anti-inflammatory therapy that is
ineffective, the disease worsens. Surgery, in the form of
structureplasty (reshaping of the intestine) or resection (removal
of the intestine), is the only treatment option for patients that
do not respond to first line therapies. Surgical treatments for IBD
are invasive, causing post-operative risks for an estimated third
of patients undergoing surgery, such as anastomotic leak,
infection, and bleeding.
The pathogenesis of IBD is thought to involve an uncontrolled
immune response that may be triggered by certain environmental
factors in a genetically susceptible host. The heterogeneity of
disease pathogenesis and clinical course, combined with the
variable response to treatment and its associated side effects,
suggests a targeted therapeutic approach to treating these diseases
is best treatment strategy. Yet there are very few targeted
therapies available to IBD patients, especially those patients who
may be non-responsive to existing IBD therapies (e.g., anti-TNFa
inhibitors). Accordingly, there is a need for novel therapeutics to
treat IBD that specifically target enzymes involved in IBD
pathogenesis.
SUMMARY
The present disclosure provides antibodies useful for the treatment
of IBD, including moderate to severe forms of IBD characterized by
subclinical phenotypes disclosed herein (e.g., refractory disease,
stricturing disease, penetrating disease). The antibodies described
herein possess superior therapeutic aspects compared to other Tumor
necrosis factor ligand 1A (TL1A) binding antibodies. Primarily, the
antibodies described herein possess high sequence homology to human
germline frameworks while still exhibiting high binding affinity,
express at high levels in bacterial and mammalian culture, and
possess fewer sequence liabilities, such as deamidation sites, that
lead to increased degradation and reduced therapeutic effect.
TL1A and nucleic acids encoding TL1A (Tumor Necrosis Factor Ligand
Superfamily Member 15 (TNFSF15) are provided Entrez Gene: 9966;
UniProtKB: O95150. TL1A is a proinflammatory molecule which
stimulates proliferation and effector functions of CD8 (+)
cytotoxic T cells as well as Th1, Th2, and Th17 cells in the
presence of TCR stimulation. TL1A is believed to be involved in the
pathogenesis of IBD by bridging the innate and adaptive immune
response, modulating adaptive immunity by augmenting Th1, Th2, and
Th17 effector cell function, and T-cell accumulation and
immunopathology of inflamed tissue.
Certain genotypes containing polymorphisms identified at the
TNFSF15 gene, are associated with, and therefore predictive of, a
risk of developing IBD (e.g., UC or CD), or a subclinical phenotype
of IBD. Expression of TL1A mRNA expression is enriched in patients
diagnosed with IBD who carry these risk genotypes. Therefore,
inhibiting TL1A expression and/or activity is a promising
therapeutic strategy in a variety of T cell-dependent autoimmune
diseases, including IBD (e.g., UC and CD).
In one aspect, provided herein, is an antibody or antigen-binding
fragment that specifically binds to TL1A, comprising: a heavy chain
variable region comprising: (a) an HCDR1 comprising an amino acid
sequence set forth by SEQ ID NO: 553; (b) an HCDR2 comprising an
amino acid sequence set forth by any one of SEQ ID NOs: 554 to 564
or 574 to 577; and (c) an HCDR3 comprising an amino acid sequence
set forth by any one of SEQ ID NOs: 565 to 568 or 578 to 581; and a
light chain variable region comprising: (d) an LCDR1 comprising an
amino acid sequence set forth by any one of SEQ ID NOs: 569 or 570;
(e) an LCCR2 comprising an amino acid sequence set forth by SEQ ID
NO: 488; and (f) an LCDR3 comprising an amino acid sequence set
forth by any one of SEQ ID NOs: 571 to 573 or 582 to 585. In
certain embodiments, the antibody or antigen binding fragment
comprises a human heavy chain framework region 1 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 545. In certain embodiments, the antibody or antigen binding
fragment comprises a human heavy chain framework region 2 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 546. In certain embodiments, the antibody or antigen
binding fragment comprises a human heavy chain framework region 3
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 547 or 586 to 588. In certain embodiments, the
antibody or antigen binding fragment comprises a human heavy chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, 99%
identical to that set forth is SEQ ID NO: 548. In certain
embodiments, the antibody or antigen binding fragment comprises a
human light chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 549.
In certain embodiments, the antibody or antigen binding fragment
comprises a human light chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 550. In certain embodiments, the antibody or antigen binding
fragment comprises a human light chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 551. In certain embodiments, the antibody or antigen
binding fragment comprises a human light chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 552. In certain embodiments, the antibody or
antigen binding fragment comprises: (a) a human heavy chain
framework region 1 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 545; (b) a human heavy chain
framework region 2 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 546; (c) a human heavy chain
framework region 3 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 547 or 586 to 588; (d) a human
heavy chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 548; (e) a human
light chain framework region 1 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 549; (f) a human
light chain framework region 2 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 550; (g) a human
light chain framework region 3 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 551; and (h) a human
light chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 552. In certain
embodiments, the antibody binds human TL1A with a stronger affinity
or a 2.times. stronger affinity compared to the L8 clone as
determined by ELISA, wherein the L8 clone comprises a heavy chain
variable region amino acid sequence as set forth by SEQ ID NO: 491,
and a light chain variable region amino acid sequence as set forth
by SEQ ID NO: 490. In certain embodiments, the antibody or
antigen-binding fragment is chimeric or humanized. In certain
embodiments, the antibody or antigen-binding fragment is an IgG
antibody. In certain embodiments, the antibody or antigen-binding
fragment comprises a Fab, F(ab).sub.2, a single-domain antibody, a
single chain variable fragment (scFv), or a nanobody. In certain
embodiments, the antibody or antigen-binding fragment comprises a
heavy chain constant region comprising an amino acid sequence as
set forth by SEQ ID NO: 542 or 543. In certain embodiments, the
antibody or antigen-binding fragment comprises a heavy chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 542. In certain embodiments, the antibody or
antigen-binding fragment comprises a light chain constant region
comprising an amino acid sequence as set forth by SEQ ID NO: 544.
In certain embodiments, the antibody or antigen-binding fragment
thereof inhibits TL1A induced secretion of interferon gamma from T
lymphocytes. In certain embodiments, the antibody or
antigen-binding fragment is a component of a pharmaceutical
composition comprising the antibody or antigen-binding fragment and
a pharmaceutically acceptable excipient, carrier, or diluent. In
certain embodiments, the pharmaceutical composition is formulated
for intravenous administration. In certain embodiments, antibody or
antigen-binding fragment or the pharmaceutical composition is for
use in treating inflammatory bowel disease, Crohn's disease, or
colitis. In certain embodiments, the antibody or antigen-binding
fragment is encoded by a nucleic acid. In certain embodiments, a
cell comprises the nucleic acid. In certain embodiments, the cell
is a eukaryotic cell. In certain embodiments, the cell is a Chinese
Hamster Ovary (CHO) cell. In certain embodiments, described herein,
is a method of treating an individual with inflammatory bowel
disease, Crohn's disease, or colitis comprising administering an
effective amount of the antibody or antigen-binding fragment or the
pharmaceutical composition to the individual, wherein the
individual is diagnosed with or suspected of being afflicted with
inflammatory bowel disease, Crohn's disease, or colitis. In certain
embodiments, the antibody or antigen-binding fragment or the
pharmaceutical composition is for use in preventing or reducing
interferon gamma secretion by T lymphocytes. In certain
embodiments, described herein is a method of preventing or reducing
interferon gamma secretion by T lymphocytes in an individual
comprising administering an effective amount of the antibody or
antigen-binding fragment or the pharmaceutical composition to the
individual. In certain embodiments, described herein, is a method
of preparing an inflammatory bowel disease, Crohn's disease, or
colitis treatment comprising incubating the cell comprising the
nucleic acid encoding the antibody or antigen-binding fragment into
a culture medium under conditions sufficient to secrete the
antibody or antigen-binding fragment into the culture medium. In
certain embodiments, the method further comprises subjecting the
culture medium to at least one purification step. In certain
embodiments, described herein, is a method of preparing an
inflammatory bowel disease, Crohn's disease, or colitis treatment
comprising admixing the antibody or antigen-binding fragment and a
pharmaceutically acceptable excipient, carrier, or diluent.
In another aspect, provided herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
559; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 567; and (d) a light chain variable region
comprising: (d) an LCDR1 comprising an amino acid sequence set
forth by SEQ ID NO: 569; (e) an LCCR2 comprising an amino acid
sequence set forth by SEQ ID NO: 488; and (f) an LCDR3 comprising
an amino acid sequence set forth by any one of SEQ ID NO: 573. In
certain embodiments, the antibody or antigen binding fragment
comprises a human heavy chain framework region 1 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 545. In certain embodiments, the antibody or antigen binding
fragment comprises a human heavy chain framework region 2 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 546. In certain embodiments, the antibody or antigen
binding fragment comprises a human heavy chain framework region 3
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 547 or 586 to 588. In certain embodiments, the
antibody or antigen binding fragment comprises a human heavy chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, 99%
identical to that set forth is SEQ ID NO: 548. In certain
embodiments, the antibody or antigen binding fragment comprises a
human light chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 549.
In certain embodiments, the antibody or antigen binding fragment
comprises a human light chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 550. In certain embodiments, the antibody or antigen binding
fragment comprises a human light chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 551. In certain embodiments, the antibody or antigen
binding fragment comprises a human light chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 552. In certain embodiments, the antibody or
antigen binding fragment comprises: (a) a human heavy chain
framework region 1 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 545; (b) a human heavy chain
framework region 2 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 546; (c) a human heavy chain
framework region 3 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 547 or 586 to 588; (d) a human
heavy chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 548; (e) a human
light chain framework region 1 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 549; (f) a human
light chain framework region 2 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 550; (g) a human
light chain framework region 3 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 551; and (h) a human
light chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 552. In certain
embodiments, the antibody or antigen-binding fragment that
specifically binds to TL1A, comprises: (a) a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 503; and (b) a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 502. In certain embodiments, the antibody binds human TL1A
with a stronger affinity or a 2.times. stronger affinity compared
to the L8 clone as determined by ELISA, wherein the L8 clone
comprises a heavy chain variable region amino acid sequence as set
forth by SEQ ID NO: 491, and a light chain variable region amino
acid sequence as set forth by SEQ ID NO: 490. In certain
embodiments, the antibody or antigen-binding fragment is chimeric
or humanized. In certain embodiments, the antibody or
antigen-binding fragment is an IgG antibody. In certain
embodiments, the antibody or antigen-binding fragment comprises a
Fab, F(ab).sub.2, a single-domain antibody, a single chain variable
fragment (scFv), or a nanobody. In certain embodiments, the
antibody or antigen-binding fragment comprises a heavy chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 542 or 543. In certain embodiments, the antibody or
antigen-binding fragment comprises a heavy chain constant region
comprising an amino acid sequence as set forth by SEQ ID NO: 542.
In certain embodiments, the antibody or antigen-binding fragment
comprises a light chain constant region comprising an amino acid
sequence as set forth by SEQ ID NO: 544. In certain embodiments,
the antibody or antigen-binding fragment thereof inhibits TL1A
induced secretion of interferon gamma from T lymphocytes. In
certain embodiments, the antibody or antigen-binding fragment is a
component of a pharmaceutical composition comprising the antibody
or antigen-binding fragment and a pharmaceutically acceptable
excipient, carrier, or diluent. In certain embodiments, the
pharmaceutical composition is formulated for intravenous
administration. In certain embodiments, antibody or antigen-binding
fragment or the pharmaceutical composition is for use in treating
inflammatory bowel disease, Crohn's disease, or colitis. In certain
embodiments, the antibody or antigen-binding fragment is encoded by
a nucleic acid. In certain embodiments, a cell comprises the
nucleic acid. In certain embodiments, the cell is a eukaryotic
cell. In certain embodiments, the cell is a Chinese Hamster Ovary
(CHO) cell. In certain embodiments, described herein, is a method
of treating an individual with inflammatory bowel disease, Crohn's
disease, or colitis comprising administering an effective amount of
the antibody or antigen-binding fragment or the pharmaceutical
composition to the individual, wherein the individual is diagnosed
with or suspected of being afflicted with inflammatory bowel
disease, Crohn's disease, or colitis. In certain embodiments, the
antibody or antigen-binding fragment or the pharmaceutical
composition is for use in preventing or reducing interferon gamma
secretion by T lymphocytes. In certain embodiments, described
herein is a method of preventing or reducing interferon gamma
secretion by T lymphocytes in an individual comprising
administering an effective amount of the antibody or
antigen-binding fragment or the pharmaceutical composition to the
individual. In certain embodiments, described herein, is a method
of preparing an inflammatory bowel disease, Crohn's disease, or
colitis treatment comprising incubating the cell comprising the
nucleic acid encoding the antibody or antigen-binding fragment into
a culture medium under conditions sufficient to secrete the
antibody or antigen-binding fragment into the culture medium. In
certain embodiments, the method further comprises subjecting the
culture medium to at least one purification step. In certain
embodiments, described herein, is a method of preparing an
inflammatory bowel disease, Crohn's disease, or colitis treatment
comprising admixing the antibody or antigen-binding fragment and a
pharmaceutically acceptable excipient, carrier, or diluent.
In another aspect, provided herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
563; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 568; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 572. In certain
embodiments, the antibody or antigen binding fragment comprises a
human heavy chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 545.
In certain embodiments, the antibody or antigen binding fragment
comprises a human heavy chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 546. In certain embodiments, the antibody or antigen binding
fragment comprises a human heavy chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 547 or 586 to 588. In certain embodiments, the
antibody or antigen binding fragment comprises a human heavy chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, 99%
identical to that set forth is SEQ ID NO: 548. In certain
embodiments, the antibody or antigen binding fragment comprises a
human light chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 549.
In certain embodiments, the antibody or antigen binding fragment
comprises a human light chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 550. In certain embodiments, the antibody or antigen binding
fragment comprises a human light chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 551. In certain embodiments, the antibody or antigen
binding fragment comprises a human light chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 552. In certain embodiments, the antibody or
antigen binding fragment comprises: (a) a human heavy chain
framework region 1 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 545; (b) a human heavy chain
framework region 2 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 546; (c) a human heavy chain
framework region 3 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 547 or 586 to 588; (d) a human
heavy chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 548; (e) a human
light chain framework region 1 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 549; (f) a human
light chain framework region 2 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 550; (g) a human
light chain framework region 3 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 551; and (h) a human
light chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 552. In certain
embodiments, the antibody or antigen-binding fragment that
specifically binds to TL1A, comprises: (a) a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 511; and (b) a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 510. In certain embodiments, the antibody binds human TL1A
with a stronger affinity or a 2.times. stronger affinity compared
to the L8 clone as determined by ELISA, wherein the L8 clone
comprises a heavy chain variable region amino acid sequence as set
forth by SEQ ID NO: 491, and a light chain variable region amino
acid sequence as set forth by SEQ ID NO: 490. In certain
embodiments, the antibody or antigen-binding fragment is chimeric
or humanized. In certain embodiments, the antibody or
antigen-binding fragment is an IgG antibody. In certain
embodiments, the antibody or antigen-binding fragment comprises a
Fab, F(ab).sub.2, a single-domain antibody, a single chain variable
fragment (scFv), or a nanobody. In certain embodiments, the
antibody or antigen-binding fragment comprises a heavy chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 542 or 543. In certain embodiments, the antibody or
antigen-binding fragment comprises a heavy chain constant region
comprising an amino acid sequence as set forth by SEQ ID NO: 542.
In certain embodiments, the antibody or antigen-binding fragment
comprises a light chain constant region comprising an amino acid
sequence as set forth by SEQ ID NO: 544. In certain embodiments,
the antibody or antigen-binding fragment thereof inhibits TL1A
induced secretion of interferon gamma from T lymphocytes. In
certain embodiments, the antibody or antigen-binding fragment is a
component of a pharmaceutical composition comprising the antibody
or antigen-binding fragment and a pharmaceutically acceptable
excipient, carrier, or diluent. In certain embodiments, the
pharmaceutical composition is formulated for intravenous
administration. In certain embodiments, antibody or antigen-binding
fragment or the pharmaceutical composition is for use in treating
inflammatory bowel disease, Crohn's disease, or colitis. In certain
embodiments, the antibody or antigen-binding fragment is encoded by
a nucleic acid. In certain embodiments, a cell comprises the
nucleic acid. In certain embodiments, the cell is a eukaryotic
cell. In certain embodiments, the cell is a Chinese Hamster Ovary
(CHO) cell. In certain embodiments, described herein, is a method
of treating an individual with inflammatory bowel disease, Crohn's
disease, or colitis comprising administering an effective amount of
the antibody or antigen-binding fragment or the pharmaceutical
composition to the individual, wherein the individual is diagnosed
with or suspected of being afflicted with inflammatory bowel
disease, Crohn's disease, or colitis. In certain embodiments, the
antibody or antigen-binding fragment or the pharmaceutical
composition is for use in preventing or reducing interferon gamma
secretion by T lymphocytes. In certain embodiments, described
herein is a method of preventing or reducing interferon gamma
secretion by T lymphocytes in an individual comprising
administering an effective amount of the antibody or
antigen-binding fragment or the pharmaceutical composition to the
individual. In certain embodiments, described herein, is a method
of preparing an inflammatory bowel disease, Crohn's disease, or
colitis treatment comprising incubating the cell comprising the
nucleic acid encoding the antibody or antigen-binding fragment into
a culture medium under conditions sufficient to secrete the
antibody or antigen-binding fragment into the culture medium. In
certain embodiments, the method further comprises subjecting the
culture medium to at least one purification step. In certain
embodiments, described herein, is a method of preparing an
inflammatory bowel disease, Crohn's disease, or colitis treatment
comprising admixing the antibody or antigen-binding fragment and a
pharmaceutically acceptable excipient, carrier, or diluent.
In another aspect, provided herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
555; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 566; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 572. In certain
embodiments, the antibody or antigen binding fragment comprises a
human heavy chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 545.
In certain embodiments, the antibody or antigen binding fragment
comprises a human heavy chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 546. In certain embodiments, the antibody or antigen binding
fragment comprises a human heavy chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 547 or 586 to 588. In certain embodiments, the
antibody or antigen binding fragment comprises a human heavy chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, 99%
identical to that set forth is SEQ ID NO: 548. In certain
embodiments, the antibody or antigen binding fragment comprises a
human light chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 549.
In certain embodiments, the antibody or antigen binding fragment
comprises a human light chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 550. In certain embodiments, the antibody or antigen binding
fragment comprises a human light chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 551. In certain embodiments, the antibody or antigen
binding fragment comprises a human light chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 552. In certain embodiments, the antibody or
antigen binding fragment comprises: (a) a human heavy chain
framework region 1 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 545; (b) a human heavy chain
framework region 2 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 546; (c) a human heavy chain
framework region 3 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 547 or 586 to 588; (d) a human
heavy chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 548; (e) a human
light chain framework region 1 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 549; (f) a human
light chain framework region 2 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 550; (g) a human
light chain framework region 3 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 551; and (h) a human
light chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 552. In certain
embodiments, the antibody or antigen-binding fragment that
specifically binds to TL1A, comprises: (a) a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 493; and (b) a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 492. In certain embodiments, the antibody binds human TL1A
with a stronger affinity or a 2.times. stronger affinity compared
to the L8 clone as determined by ELISA, wherein the L8 clone
comprises a heavy chain variable region amino acid sequence as set
forth by SEQ ID NO: 491, and a light chain variable region amino
acid sequence as set forth by SEQ ID NO: 490. In certain
embodiments, the antibody or antigen-binding fragment is chimeric
or humanized. In certain embodiments, the antibody or
antigen-binding fragment is an IgG antibody. In certain
embodiments, the antibody or antigen-binding fragment comprises a
Fab, F(ab).sub.2, a single-domain antibody, a single chain variable
fragment (scFv), or a nanobody. In certain embodiments, the
antibody or antigen-binding fragment comprises a heavy chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 542 or 543. In certain embodiments, the antibody or
antigen-binding fragment comprises a heavy chain constant region
comprising an amino acid sequence as set forth by SEQ ID NO: 542.
In certain embodiments, the antibody or antigen-binding fragment
comprises a light chain constant region comprising an amino acid
sequence as set forth by SEQ ID NO: 544. In certain embodiments,
the antibody or antigen-binding fragment thereof inhibits TL1A
induced secretion of interferon gamma from T lymphocytes. In
certain embodiments, the antibody or antigen-binding fragment is a
component of a pharmaceutical composition comprising the antibody
or antigen-binding fragment and a pharmaceutically acceptable
excipient, carrier, or diluent. In certain embodiments, the
pharmaceutical composition is formulated for intravenous
administration. In certain embodiments, antibody or antigen-binding
fragment or the pharmaceutical composition is for use in treating
inflammatory bowel disease, Crohn's disease, or colitis. In certain
embodiments, the antibody or antigen-binding fragment is encoded by
a nucleic acid. In certain embodiments, a cell comprises the
nucleic acid. In certain embodiments, the cell is a eukaryotic
cell. In certain embodiments, the cell is a Chinese Hamster Ovary
(CHO) cell. In certain embodiments, described herein, is a method
of treating an individual with inflammatory bowel disease, Crohn's
disease, or colitis comprising administering an effective amount of
the antibody or antigen-binding fragment or the pharmaceutical
composition to the individual, wherein the individual is diagnosed
with or suspected of being afflicted with inflammatory bowel
disease, Crohn's disease, or colitis. In certain embodiments, the
antibody or antigen-binding fragment or the pharmaceutical
composition is for use in preventing or reducing interferon gamma
secretion by T lymphocytes. In certain embodiments, described
herein is a method of preventing or reducing interferon gamma
secretion by T lymphocytes in an individual comprising
administering an effective amount of the antibody or
antigen-binding fragment or the pharmaceutical composition to the
individual. In certain embodiments, described herein, is a method
of preparing an inflammatory bowel disease, Crohn's disease, or
colitis treatment comprising incubating the cell comprising the
nucleic acid encoding the antibody or antigen-binding fragment into
a culture medium under conditions sufficient to secrete the
antibody or antigen-binding fragment into the culture medium. In
certain embodiments, the method further comprises subjecting the
culture medium to at least one purification step. In certain
embodiments, described herein, is a method of preparing an
inflammatory bowel disease, Crohn's disease, or colitis treatment
comprising admixing the antibody or antigen-binding fragment and a
pharmaceutically acceptable excipient, carrier, or diluent.
In another aspect, provided herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
558; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 566; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 572. In certain
embodiments, the antibody or antigen binding fragment comprises a
human heavy chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 545.
In certain embodiments, the antibody or antigen binding fragment
comprises a human heavy chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 546. In certain embodiments, the antibody or antigen binding
fragment comprises a human heavy chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 547 or 586 to 588. In certain embodiments, the
antibody or antigen binding fragment comprises a human heavy chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, 99%
identical to that set forth is SEQ ID NO: 548. In certain
embodiments, the antibody or antigen binding fragment comprises a
human light chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 549.
In certain embodiments, the antibody or antigen binding fragment
comprises a human light chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 550. In certain embodiments, the antibody or antigen binding
fragment comprises a human light chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 551. In certain embodiments, the antibody or antigen
binding fragment comprises a human light chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 552. In certain embodiments, the antibody or
antigen binding fragment comprises: (a) a human heavy chain
framework region 1 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 545; (b) a human heavy chain
framework region 2 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 546; (c) a human heavy chain
framework region 3 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 547 or 586 to 588; (d) a human
heavy chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 548; (e) a human
light chain framework region 1 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 549; (f) a human
light chain framework region 2 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 550; (g) a human
light chain framework region 3 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 551; and (h) a human
light chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 552. In certain
embodiments, the antibody or antigen-binding fragment that
specifically binds to TL1A, comprises: (a) a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 501; and (b) a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 500. In certain embodiments, the antibody binds human TL1A
with a stronger affinity or a 2.times. stronger affinity compared
to the L8 clone as determined by ELISA, wherein the L8 clone
comprises a heavy chain variable region amino acid sequence as set
forth by SEQ ID NO: 491, and a light chain variable region amino
acid sequence as set forth by SEQ ID NO: 490. In certain
embodiments, the antibody or antigen-binding fragment is chimeric
or humanized. In certain embodiments, the antibody or
antigen-binding fragment is an IgG antibody. In certain
embodiments, the antibody or antigen-binding fragment comprises a
Fab, F(ab).sub.2, a single-domain antibody, a single chain variable
fragment (scFv), or a nanobody. In certain embodiments, the
antibody or antigen-binding fragment comprises a heavy chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 542 or 543. In certain embodiments, the antibody or
antigen-binding fragment comprises a heavy chain constant region
comprising an amino acid sequence as set forth by SEQ ID NO: 542.
In certain embodiments, the antibody or antigen-binding fragment
comprises a light chain constant region comprising an amino acid
sequence as set forth by SEQ ID NO: 544. In certain embodiments,
the antibody or antigen-binding fragment thereof inhibits TL1A
induced secretion of interferon gamma from T lymphocytes. In
certain embodiments, the antibody or antigen-binding fragment is a
component of a pharmaceutical composition comprising the antibody
or antigen-binding fragment and a pharmaceutically acceptable
excipient, carrier, or diluent. In certain embodiments, the
pharmaceutical composition is formulated for intravenous
administration. In certain embodiments, antibody or antigen-binding
fragment or the pharmaceutical composition is for use in treating
inflammatory bowel disease. In certain embodiments, the antibody or
antigen-binding fragment is encoded by a nucleic acid. In certain
embodiments, a cell comprises the nucleic acid. In certain
embodiments, the cell is a eukaryotic cell. In certain embodiments,
the cell is a Chinese Hamster Ovary (CHO) cell. In certain
embodiments, described herein, is a method of treating an
individual with inflammatory bowel disease, Crohn's disease, or
colitis comprising administering an effective amount of the
antibody or antigen-binding fragment or the pharmaceutical
composition to the individual, wherein the individual is diagnosed
with or suspected of being afflicted with inflammatory bowel
disease, Crohn's disease, or colitis. In certain embodiments, the
antibody or antigen-binding fragment or the pharmaceutical
composition is for use in preventing or reducing interferon gamma
secretion by T lymphocytes. In certain embodiments, described
herein is a method of preventing or reducing interferon gamma
secretion by T lymphocytes in an individual comprising
administering an effective amount of the antibody or
antigen-binding fragment or the pharmaceutical composition to the
individual. In certain embodiments, described herein, is a method
of preparing an inflammatory bowel disease, Crohn's disease, or
colitis treatment comprising incubating the cell comprising the
nucleic acid encoding the antibody or antigen-binding fragment into
a culture medium under conditions sufficient to secrete the
antibody or antigen-binding fragment into the culture medium. In
certain embodiments, the method further comprises subjecting the
culture medium to at least one purification step. In certain
embodiments, described herein, is a method of preparing an
inflammatory bowel disease, Crohn's disease, or colitis treatment
comprising admixing the antibody or antigen-binding fragment and a
pharmaceutically acceptable excipient, carrier, or diluent.
In another aspect, provided herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
564; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 568; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 572. In certain
embodiments, the antibody or antigen binding fragment comprises a
human heavy chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 545.
In certain embodiments, the antibody or antigen binding fragment
comprises a human heavy chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 546. In certain embodiments, the antibody or antigen binding
fragment comprises a human heavy chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 547 or 586 to 588. In certain embodiments, the
antibody or antigen binding fragment comprises a human heavy chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, 99%
identical to that set forth is SEQ ID NO: 548. In certain
embodiments, the antibody or antigen binding fragment comprises a
human light chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, 99% identical to that set forth is SEQ ID NO: 549.
In certain embodiments, the antibody or antigen binding fragment
comprises a human light chain framework region 2 that is at least
90%, 95%, 96%, 97%, 98%, 99% identical to that set forth is SEQ ID
NO: 550. In certain embodiments, the antibody or antigen binding
fragment comprises a human light chain framework region 3 that is
at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set forth
is SEQ ID NO: 551. In certain embodiments, the antibody or antigen
binding fragment comprises a human light chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, 99% identical to that set
forth is SEQ ID NO: 552. In certain embodiments, the antibody or
antigen binding fragment comprises: (a) a human heavy chain
framework region 1 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 545; (b) a human heavy chain
framework region 2 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 546; (c) a human heavy chain
framework region 3 that is at least 90%, 95%, 97%, or 98% identical
to that set forth is SEQ ID NO: 547 or 586 to 588; (d) a human
heavy chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 548; (e) a human
light chain framework region 1 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 549; (f) a human
light chain framework region 2 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 550; (g) a human
light chain framework region 3 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 551; and (h) a human
light chain framework region 4 that is at least 90%, 95%, 97%, or
98% identical to that set forth is SEQ ID NO: 552. In certain
embodiments, the antibody or antigen-binding fragment that
specifically binds to TL1A, comprises: (a) a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 515; and (b) a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 514. In certain embodiments, the antibody binds human TL1A
with a stronger affinity or a 2.times. stronger affinity compared
to the L8 clone as determined by ELISA, wherein the L8 clone
comprises a heavy chain variable region amino acid sequence as set
forth by SEQ ID NO: 491, and a light chain variable region amino
acid sequence as set forth by SEQ ID NO: 490. In certain
embodiments, the antibody or antigen-binding fragment is chimeric
or humanized. In certain embodiments, the antibody or
antigen-binding fragment is an IgG antibody. In certain
embodiments, the antibody or antigen-binding fragment comprises a
Fab, F(ab).sub.2, a single-domain antibody, a single chain variable
fragment (scFv), or a nanobody. In certain embodiments, the
antibody or antigen-binding fragment comprises a heavy chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 542 or 543. In certain embodiments, the antibody or
antigen-binding fragment comprises a heavy chain constant region
comprising an amino acid sequence as set forth by SEQ ID NO: 542.
In certain embodiments, the antibody or antigen-binding fragment
comprises a light chain constant region comprising an amino acid
sequence as set forth by SEQ ID NO: 544. In certain embodiments,
the antibody or antigen-binding fragment thereof inhibits TL1A
induced secretion of interferon gamma from T lymphocytes. In
certain embodiments, the antibody or antigen-binding fragment is a
component of a pharmaceutical composition comprising the antibody
or antigen-binding fragment and a pharmaceutically acceptable
excipient, carrier, or diluent. In certain embodiments, the
pharmaceutical composition is formulated for intravenous
administration. In certain embodiments, antibody or antigen-binding
fragment or the pharmaceutical composition is for use in treating
inflammatory bowel disease, Crohn's disease, or colitis. In certain
embodiments, the antibody or antigen-binding fragment is encoded by
a nucleic acid. In certain embodiments, a cell comprises the
nucleic acid. In certain embodiments, the cell is a eukaryotic
cell. In certain embodiments, the cell is a Chinese Hamster Ovary
(CHO) cell. In certain embodiments, described herein, is a method
of treating an individual with inflammatory bowel disease, Crohn's
disease, or colitis comprising administering an effective amount of
the antibody or antigen-binding fragment or the pharmaceutical
composition to the individual, wherein the individual is diagnosed
with or suspected of being afflicted with inflammatory bowel
disease, Crohn's disease, or colitis. In certain embodiments, the
antibody or antigen-binding fragment or the pharmaceutical
composition is for use in preventing or reducing interferon gamma
secretion by T lymphocytes. In certain embodiments, described
herein is a method of preventing or reducing interferon gamma
secretion by T lymphocytes in an individual comprising
administering an effective amount of the antibody or
antigen-binding fragment or the pharmaceutical composition to the
individual. In certain embodiments, described herein, is a method
of preparing an inflammatory bowel disease, Crohn's disease, or
colitis treatment comprising incubating the cell comprising the
nucleic acid encoding the antibody or antigen-binding fragment into
a culture medium under conditions sufficient to secrete the
antibody or antigen-binding fragment into the culture medium. In
certain embodiments, the method further comprises subjecting the
culture medium to at least one purification step. In certain
embodiments, described herein, is a method of preparing an
inflammatory bowel disease, Crohn's disease, or colitis treatment
comprising admixing the antibody or antigen-binding fragment and a
pharmaceutically acceptable excipient, carrier, or diluent.
In another aspect, provided herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: (a) a heavy chain variable region comprising an HCDR1,
an HCDR2, and an HCDR3 from any one of SEQ ID NOs: 491, 493, 495,
497, 499,
501,503,505,507,509,511,513,515,517,519,521,523,525,527,529,531,533,535,
537, 539, or 541; and (b) a light chain variable region comprising
n LCDR1, an LCDR2, and an LCDR3 from any one of SEQ ID NOs: 490,
492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516,
518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, or 540;
wherein the CDRs are defined by the Kabat, Chothia, or IMGT method
or a combination thereof. In certain embodiments, the antibody or
antigen-binding fragment that specifically binds to TL1A,
comprises: (a) a heavy chain variable region comprising an amino
acid sequence at least about 85%, 90%, 95%, 97%, 98%, 99%, or 100%
identical to any one of SEQ ID NOs: 491, 493, 495, 497, 499, 501,
503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527,
529, 531, 533, 535, 537, 539, or 541; and (b) a light chain
variable region comprising an amino acid sequence at least about
85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to any one of SEQ
ID NOs: 490, 492, 494, 496, 498, 500, 502, 504, 506, 508, 510, 512,
514, 516, 518, 520, 522, 524, 526, 528, 530, 532, 534, 536, 538, or
540. In certain embodiments, the antibody binds human TL with a
stronger affinity or a 2.times. stronger affinity compared to the
L8 clone as determined by ELISA, wherein the L8 clone comprises a
heavy chain variable region amino acid sequence as set forth by SEQ
ID NO: 491, and a light chain variable region amino acid sequence
as set forth by SEQ ID NO: 490. In certain embodiments, the
antibody or antigen-binding fragment is chimeric or humanized. In
certain embodiments, the antibody or antigen-binding fragment is an
IgG antibody. In certain embodiments, the antibody or
antigen-binding fragment comprises a Fab, F(ab).sub.2, a
single-domain antibody, a single chain variable fragment (scFv), or
a nanobody. In certain embodiments, the antibody or antigen-binding
fragment comprises a heavy chain constant region comprising an
amino acid sequence as set forth by SEQ ID NO: 542 or 543. In
certain embodiments, the antibody or antigen-binding fragment
comprises a heavy chain constant region comprising an amino acid
sequence as set forth by SEQ ID NO: 542. In certain embodiments,
the antibody or antigen-binding fragment comprises a light chain
constant region comprising an amino acid sequence as set forth by
SEQ ID NO: 544. In certain embodiments, the antibody or
antigen-binding fragment is a component of a pharmaceutical
composition comprising the antibody or antigen-binding fragment and
a pharmaceutically acceptable excipient, carrier, or diluent. In
certain embodiments, the pharmaceutical composition is formulated
for intravenous administration.
In another aspect, described herein, is a method of treating a
disease or a condition in an individual possessing a risk variant
associated with the disease or the condition, the method comprising
administering an effective amount of an antibody or antigen-binding
fragment of this disclosure to the individual possessing a risk
variant, wherein the disease or the condition comprises at least
one of an inflammatory bowel disease (IBD), Crohn's disease (CD),
or colitis. In certain embodiments, the individual possesses a
plurality of risk variants. In certain embodiments, the plurality
of risk variants is at least 3, 4, 5, or 10 risk variants. In
certain embodiments, the risk variant of the plurality of risk
variants is associated with a subclinical phenotype of the disease
or the condition. In certain embodiments, the disease or the
condition is a severe form of the at least one of the IBD, the CD,
or the colitis.
BRIEF DESCRIPTION OF THE FIGURES
Exemplary embodiments are illustrated in referenced figures. It is
intended that the embodiments and figures disclosed herein are to
be considered illustrative rather than restrictive.
FIG. 1 depicts the results of a filter lift assay performed as a
qualitative assessment of chimeric 5C3D11 Fab expression and
antigen binding. Section A of the filter shows expression of heavy
chain 5C3D11, section B of the filter shows expression of light
chain 5C3D11, and section C of the filter show binding of 5C3D11
Fab binding to human TL antigen.
FIG. 2 depicts binding of chimeric 5C3D11 and humanized clone 12835
antibodies to human TL1A by enzyme-linked immunosorbent assay
(ELISA).
FIG. 3 depicts the results of a capture filter lift assay
demonstrating high sensitivity and high binding strength of
chimeric 5C3D11 for human TL1A.
FIG. 4A depicts the results of an ELISA showing binding of
CDR-grafted antibody clones 18-7, 21-3 and humanized clone 12835 to
human TL1A.
FIG. 4B depicts the results of an ELISA showing binding of
CDR-grafted antibody L8 to human TL1A as compared to binding of
humanized clone 12835 to human TL1A.
FIG. 5 depicts the results of an ELISA demonstrating the strong
binding of immobilized Fabs (chimeric 5C3D11, humanized clone
12835, clone 18-7, clone 21-3, and CDR graft clone L8) to soluble
human TL1A antigen.
FIG. 6A depicts the results of an ELISA demonstrating increased
affinity of anti-TL1A antibodies having heavy chain CDR3 mutations
H3-7 (V102M)--SEQ ID NOS: 44, 38, H3-7 (V102K)--SEQ ID NOS: 43, 38,
and H3-7 (V102Q)--SEQ ID NOS: 45, 38 and humanized clone 12835 to
human TL1A as compared to CDR graft (clone L8).
FIG. 6B depicts the results of an ELISA demonstrating increased
affinity of an anti-TL1A antibody having a heavy chain CDR3
mutation H3-7 (V102W)--SEQ ID NOS: 46, 38, and humanized clone
12835 to human TL1A as compared to CDR graft clone L8.
FIG. 7A depicts the results of an ELISA demonstrating increased
affinity of anti-TL1A antibodies having light chain CDR3 mutations
L3-4 (S92D)--SEQ ID NOS: 47, 40, L3-4 (S92E)--SEQ ID NOS: 48, 40,
L3-4 (S92H)--SEQ ID NOS: 49, 40, L3-4 (S92N)--SEQ ID NOS: 50, 40,
and humanized clone 12835, to human TL1A as compared to CDR graft
clone L8.
FIG. 7B depicts the results of an ELISA demonstrating increased
affinity of an anti-TL1A antibody having a light chain CDR3
mutation L3-4 (S92Q)--SEQ ID NOS: 51, 40, and humanized clone 12835
to human TL1A as compared to CDR graft clone L8.
FIGS. 8A, 8B and 8C depict ELISAs demonstrating binding of Fabs
comprising 5C3D11 CDR variants grafted on human heavy chain
germline IGH1-46*02 and human light chain germline IGKV3-20*01 to
immobilized human TL1A.
FIGS. 9A and 9B depict ELISAs demonstrating binding of Fabs
comprising 5C3D11 CDR variants grafted on human heavy chain
germline IGH1-3*01 and human light chain germline IGKV3-20*01 to
immobilized human TL1A.
FIGS. 10A and 10B depict ELISAs demonstrating binding of
immobilized Fabs comprising 5C3D11 CDR variants grafted on human
heavy chain germline IGH1-46*02 and human light chain germline
IGKV3-20*01 to soluble, biotinylated human TL1A.
FIGS. 11A and 11B depict ELISAs demonstrating binding of
immobilized Fabs comprising 5C3D11 CDR variants grafted on human
heavy chain germline IGH1-46*02 and human light chain germline
IGKV3-20*01 to soluble, biotinylated human TL1A.
FIGS. 12A and 12B depict ELISAs demonstrating binding of
immobilized Fabs comprising 5C3D11 CDR variants grafted on human
heavy chain germline IGH1-46*02 and human light chain germline
IGKV3-20*01 to soluble, biotinylated human TL1A.
FIGS. 13A and 13B depict ELISAs demonstrating binding of
immobilized Fabs comprising 5C3D11 CDR variants grafted on human
heavy chain germline IGH1-3*01 and human light chain germline
IGKV3-20*01 to soluble, biotinylated human TL1A.
FIGS. 14A and 14B demonstrate binding of Fabs comprising 5C3D11 CDR
variants to membrane-associated human TL1A.
FIGS. 15A and 15B depict lack of binding of Fabs comprising 5C3D11
CDR variants grafted on human heavy chain germline IGH1-46*02 and
human light chain germline IGKV3-20*01 to TRAIL.
FIGS. 16A and 16B depict lack of binding of Fabs comprising 5C3D11
CDR variants grafted on human heavy chain germline IGH1-46*02 and
human light chain germline IGKV3-20*01 to LIGHT.
FIGS. 17A and 17B depict lack of binding of Fabs comprising 5C3D11
CDR variants grafted on human heavy chain germline IGH1-46*02 and
human light chain germline IGKV3-20*01 to Fas.
FIG. 18 depicts lack of binding of Fabs comprising 5C3D11 CDR
variants grafted on human heavy chain germline IGH1-3*01 and human
light chain germline IGKV3-20*01 to TRAIL.
FIG. 19 depicts lack of binding of Fabs comprising 5C3D11 CDR
variants grafted on human heavy chain germline IGH1-3*01 and human
light chain germline IGKV3-20*01 to LIGHT.
FIG. 20 depicts lack of binding of Fabs comprising 5C3D11 CDR
variants grafted on human heavy chain germline IGH1-3*01 and human
light chain germline IGKV3-20*01 to Fas.
FIGS. 21A and 21B depict ELISAs demonstrating binding of heavy and
light chain variable regions comprising 5C3D11 CDR variants with an
IgG1 heavy chain (modified) and kappa light chain constant region
(21A), or with an IgG2 heavy chain and kappa light chain constant
region (21B) to immobilized human TL1A.
FIGS. 22A and 22B depict ELISAs demonstrating binding of soluble,
biotinylated human TL1A to immobilized heavy and light chain
variable regions comprising 5C3D11 CDR variants with an IgG1 heavy
chain (modified) and kappa light chain constant region (22A), or
with an IgG2 heavy chain and kappa light chain constant region
(22B).
FIG. 23 demonstrates maintenance of binding of heavy and light
chain variable regions comprising 5C3D11 CDR variants with an IgG1
heavy chain (modified) and kappa light chain constant region, or
with an IgG2 heavy chain and kappa light chain constant region to a
membrane-associated form of human TL1A.
FIGS. 24A, 24B, and 24C depict ELISAs demonstrating lack of binding
of heavy and light chain variable regions comprising 5C3D11 CDR
variants with an IgG1 heavy chain (modified) and kappa light chain
constant region, or with an IgG2 heavy chain and kappa light chain
constant region to TNFSF family members Fas (24A), TRAIL (24B), or
LIGHT (24C).
FIG. 25 demonstrates the inhibition of cynomolgus TL1A induced
IFN-.gamma. production in whole blood by humanized Ig constructs
comprising 5C3D11 CDR variants grafted on human heavy chain
germline IGH1-46*02 and human light chain germline IGKV3-20*01 with
an IgG1 heavy chain (modified) and kappa light chain constant
region.
FIGS. 26A, 26B, and 26C illustrate inhibition of TL1A induced
IFN-.gamma. production, by antibodies described herein, from human
whole blood. Shown are results from 3 different donors (26A),
(26B), and (26C), antibody concentration (nanomolar) is shown on
the x-axis.
FIGS. 27A, 27B, and 27C illustrate inhibition of TL1A induced
IFN-.gamma. production, by antibodies described herein, from
cynomolgus monkey whole blood. Shown are results from 3 different
donors (27A), (27B), and (27C), antibody concentration (nanomolar)
is shown on the x-axis.
DESCRIPTION OF THE INVENTION
Tumor necrosis factor-like protein 1A (TL1A) has been associated
with the development and severity of severe inflammatory bowel
disease (IBD), including severe forms of colitis and Crohn's
Disease (CD). In addition, preclinical and human genetic
association data suggests that TL1A is a potential therapeutic
target in Crohn's disease. The present disclosure describes
optimized antibodies against TL1A, and offers a novel therapeutic
for the treatment of IBD.
Described herein, in one aspect, is an antibody or antigen-binding
fragment that specifically binds TL1A, comprising: a heavy chain
variable region comprising: (a) an HCDR1 comprising an amino acid
sequence set forth by SEQ ID NO: 553; (b) an HCDR2 comprising an
amino acid sequence set forth by any one of SEQ ID NOs: 554 to 564
or 574 to 577; and (c) an HCDR3 comprising an amino acid sequence
set forth by any one of SEQ ID NOs: 565 to 568 or 578 to 581; and a
light chain variable region comprising: (d) an LCDR1 comprising an
amino acid sequence set forth by any one of SEQ ID NOs: 569 or 570;
(e) an LCCR2 comprising an amino acid sequence set forth by SEQ ID
NO: 488; and (f) an LCDR3 comprising an amino acid sequence set
forth by any one of SEQ ID NOs: 571 to 573 or 582 to 585.
Described herein, in another aspect, is an antibody or
antigen-binding fragment that specifically binds TL1A, comprising:
(a) a heavy chain variable region comprising an amino acid sequence
at least about 90% identical to any one of SEQ ID NOs: 491, 493,
495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519,
521, 523, 525, 527, 529, 531, 533, 535, 537, 539, or 541; and (b) a
light chain variable region comprising an amino acid sequence at
least about 90% identical to any one of SEQ ID NOs: 490, 492, 494,
496, 498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520,
522, 524, 526, 528, 530, 532, 534, 536, 538, or 540.
In some embodiments, an antibody refers to an immunoglobulin
molecule that recognizes and specifically binds to a target, such
as a protein, polypeptide, peptide, carbohydrate, polynucleotide,
lipid, or combinations of the foregoing through at least one
antigen recognition site within the variable region of the
immunoglobulin molecule. In some embodiments, an antibody includes
intact polyclonal antibodies, intact monoclonal antibodies,
antibody fragments (such as Fab, Fab', F(ab).sub.2, and Fv
fragments), single chain Fv (scFv) mutants, a CDR-grafted antibody,
multispecific antibodies, chimeric antibodies, humanized
antibodies, human antibodies, fusion proteins comprising an antigen
determination portion of an antibody, and any other modified
immunoglobulin molecule comprising an antigen recognition site so
long as the antibodies exhibit the desired biological activity. An
antibody can be of any the five major classes of immunoglobulins:
IgA, IgD, IgE, IgG, and IgM, or subclasses (isotypes) thereof (e.g.
IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2), based on the identity of
their heavy-chain constant domains referred to as alpha, delta,
epsilon, gamma, and mu, respectively. The different classes of
immunoglobulins have different and well-known subunit structures
and three-dimensional configurations. Antibodies can be naked or
conjugated to other molecules such as toxins, radioisotopes,
etc.
In some embodiments, one or more amino acid modifications may be
introduced into the Fc region of an antibody provided herein,
thereby generating an Fc region variant. An Fc region herein is a
C-terminal region of an immunoglobulin heavy chain that contains at
least a portion of the constant region. An Fc region includes
native sequence Fc regions and variant Fc regions. The Fc region
variant may comprise a human Fc region sequence (e.g., a human
IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid
modification (e.g., a substitution) at one or more amino acid
positions.
In some embodiments, the antibodies of this disclosure have reduced
antibody-dependent cell-mediated cytotoxicity (ADCC) or the reduced
ability to fix complement. This is desirable in situations where
inhibition of target function is desired, but activation of
downstream immune responses may create unwanted side effects. Some
Fc regions have a natural lack of effector function (e.g., IgG2,
SEQ ID NO: 543), and some Fc regions can comprise mutations that
reduce effector functions (e.g., a modified IgG1, SEQ ID NO: 542).
In certain embodiments, the antibodies of this disclosure have
reduced effector function. In certain embodiments, the antibodies
of this disclosure comprise an IgG2 constant region as set forth in
SEQ ID NO: 543. In certain embodiments, the antibodies of this
disclosure comprise a modified IgG1 constant region as set forth in
SEQ ID NO: 542.
In some embodiments, the antibodies of this disclosure are variants
that possess some but not all effector functions, which make it a
desirable candidate for applications in which the half-life of the
antibody in vivo is important yet certain effector functions (such
as complement and ADCC) are unnecessary or deleterious. In vitro
and/or in vivo cytotoxicity assays can be conducted to confirm the
reduction/depletion of CDC and/or ADCC activities. For example, Fc
receptor (FcR) binding assays can be conducted to ensure that the
antibody lacks FcyR binding (hence likely lacking ADCC activity),
but retains FcRn binding ability. Non-limiting examples of in vitro
assays to assess ADCC activity of a molecule of interest is
described in U.S. Pat. Nos. 5,500,362 and 5,821,337. Alternatively,
non-radioactive assays methods may be employed (e.g., ACTI.TM. and
CytoTox 96.RTM. non-radioactive cytotoxicity assays). Useful
effector cells for such assays include peripheral blood mononuclear
cells (PBMC), monocytes, macrophages, and Natural Killer (NK)
cells.
Antibodies can have increased half-lives and/or improved binding to
the neonatal Fc receptor (FcRn) (See e.g., US 2005/0014934). Such
antibodies can comprise an Fc region with one or more substitutions
therein which improve binding of the Fc region to FcRn, and include
those with substitutions at one or more of Fc region residues: 238,
256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360,
362, 376, 378, 380, 382, 413, 424 or 434 according to the EU
numbering system (See e.g., U.S. Pat. No. 7,371,826). Other
examples of Fc region variants are also contemplated (See e.g.,
Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. Nos.
5,648,260 and 5,624,821; and WO94/29351). In certain embodiments,
the antibodies of this disclosure have increased serum half-life as
a result of alternations to the Fc region. In certain embodiments,
the alterations comprise M252Y/S254T/T256E mutations to IgG1, or
M428L/N434S mutations to IgG1.
In some embodiments, an antibody comprises an antigen-binding
fragment that refers to a portion of an antibody having antigenic
determining variable regions of an antibody. Examples of antibody
fragments include, but are not limited to Fab, Fab', F(ab').sub.2,
and Fv fragments, linear antibodies, single chain antibodies, and
multispecific antibodies formed from antibody fragments.
In some embodiments, a humanized antibody refers to forms of
non-human (e.g., murine) antibodies having specific immunoglobulin
chains, chimeric immunoglobulins, or fragments thereof that contain
minimal non-human (e.g., murine) sequences. In a non-limiting
example, a humanized antibody comprises less than about 40%
non-human sequence in the variable region. In some cases, a
humanized antibody comprises less than about 20% non-human sequence
in a full length antibody sequence. In some cases, humanized
antibodies are human immunoglobulins in which residues from the
complementarity determining region (CDR) are replaced by residues
from the CDR of a non-human species (e.g., mouse, rat, rabbit,
hamster) that have the desired specificity, affinity, and
capability.
In some embodiments, chimeric antibodies refer to antibodies
wherein the sequence of the immunoglobulin molecule is derived from
two or more species. As a non-limiting example, the variable region
of both light and heavy chains corresponds to the variable region
of antibodies derived from one species of mammals (e.g., mouse,
rat, rabbit, etc) with the desired specificity, affinity, and
capability while the constant regions are homologous to the
sequences in antibodies derived from another (usually human) to
avoid eliciting an immune response in that species.
As used herein the term "about" means within 10% of the stated
amount.
As used herein "risk variant" means any genetic sequence, typically
a DNA sequence, of an individual that increases that individuals
risk for developing a phenotype (e.g. inflammatory bowel disease,
Crohn's disease, colitis, or subclinical phenotype thereof). Risk
variants include without limitation single nucleotide polymorphisms
(SNPs), indels of any length, short tandem repeats (STRs), and
chromosol translocations, duplications, or deletions. Said risk
variants include those variants that are associated with severe
forms of inflammatory bowel disease, Crohn's disease, or colitis.
Said risk variants include those variants which may indicate that
an individual may be refractory to treatment with any current
therapy for inflammatory bowel disease, Crohn's disease, or
colitis. As contemplated herein risk variants can be used to inform
a treatment decision with any of the antibodies described
herein.
The terms "complementarity determining region," and "CDR," which
are synonymous with "hypervariable region" or "HVR," are known in
the art to refer to non-contiguous sequences of amino acids within
antibody variable regions, which confer antigen specificity and/or
binding affinity. In general, there are three CDRs in each heavy
chain variable region (CDR-H1, CDR-H2, CDR-H3) and three CDRs in
each light chain variable region (CDR-L1, CDR-L2, CDR-L3).
"Framework regions" and "FR" are known in the art to refer to the
non-CDR portions of the variable regions of the heavy and light
chains. In general, there are four FRs in each full-length heavy
chain variable region (FR-H1, FR-H2, FR-H3, and FR-H4), and four
FRs in each full-length light chain variable region (FR-L1, FR-L2,
FR-L3, and FR-L4). The precise amino acid sequence boundaries of a
given CDR or FR can be readily determined using any of a number of
well-known schemes, including those described by Kabat et al.
(1991), "Sequences of Proteins of Immunological Interest," 5th Ed.
Public Health Service, National Institutes of Health, Bethesda, MD
("Kabat" numbering scheme), Al-Lazikani et al., (1997) JMB
273,927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol.
Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact
analysis and binding site topography," J. Mol. Biol. 262, 732-745."
("Contact" numbering scheme); Lefranc M P et al., "IMGT unique
numbering for immunoglobulin and T cell receptor variable domains
and Ig superfamily V-like domains," Dev Comp Immunol, 2003 January;
27(1):55-77 ("IMGT" numbering scheme); Honegger A and Pluckthun A,
"Yet another numbering scheme for immunoglobulin variable domains:
an automatic modeling and analysis tool," J Mol Biol, 2001 Jun. 8;
309(3):657-70, ("Aho" numbering scheme); and Whitelegg N R and Rees
A R, "WAM: an improved algorithm for modelling antibodies on the
WEB," Protein Eng. 2000 December; 13(12):819-24 ("AbM" numbering
scheme. In certain embodiments, the CDRs of the antibodies
described herein can be defined by a method selected from Kabat,
Chothia, IMGT, Aho, AbM, or combinations thereof.
In some embodiments, an antibody that specifically binds to a
protein indicates that the antibody reacts or associates more
frequently, more rapidly, with greater duration, with greater
affinity, or with some combination of the above to the protein than
with alternative substances, including unrelated proteins.
In some embodiments, the terms "polypeptide," "peptide," and
"protein" are used interchangeably herein to refer to polymers of
amino acids of any length. The polymer may be linear or branched,
it may comprise modified amino acids, and it may be interrupted by
non-amino acids. The terms also encompass an amino acid polymer
that has been modified naturally or by intervention; for example,
disulfide bond formation, glycosylation, lipidation, acetylation,
phosphorylation, or any other manipulation or modification, such as
fusion with another polypeptide and/or conjugation, e.g., with a
labeling component. Also included within the definition are, for
example, polypeptides containing one or more analogs of an amino
acid (for example, unnatural amino acids, etc.), as well as other
modifications known in the art.
In some embodiments, "polynucleotide," or "nucleic acid," as used
interchangeably herein, refer to polymers of nucleotides of any
length, and include DNA and RNA. The nucleotides can be
deoxyribonucleotides, ribonucleotides, modified nucleotides or
bases, and/or their analogs, or any substrate that can be
incorporated into a polymer by DNA or RNA polymerase. A
polynucleotide may comprise modified nucleotides, such as, but not
limited to methylated nucleotides and their analogs or
non-nucleotide components. Modifications to the nucleotide
structure may be imparted before or after assembly of the polymer.
A polynucleotide may be further modified after polymerization, such
as by conjugation with a labeling component.
Percent (%) sequence identity with respect to a reference
polypeptide sequence is the percentage of amino acid residues in a
candidate sequence that are identical with the amino acid residues
in the reference polypeptide sequence, after aligning the sequences
and introducing gaps, if necessary, to achieve the maximum percent
sequence identity, and not considering any conservative
substitutions as part of the sequence identity. Alignment for
purposes of determining percent amino acid sequence identity can be
achieved in various ways that are known for instance, using
publicly available computer software such as BLAST, BLAST-2, ALIGN
or Megalign (DNASTAR) software. Appropriate parameters for aligning
sequences are able to be determined, including algorithms needed to
achieve maximal alignment over the full length of the sequences
being compared. For purposes herein, however, % amino acid sequence
identity values are generated using the sequence comparison
computer program ALIGN-2. The ALIGN-2 sequence comparison computer
program was authored by Genentech, Inc., and the source code has
been filed with user documentation in the U.S. Copyright Office,
Washington D.C., 20559, where it is registered under U.S. Copyright
Registration No. TXU510087. The ALIGN-2 program is publicly
available from Genentech, Inc., South San Francisco, Calif, or may
be compiled from the source code. The ALIGN-2 program should be
compiled for use on a UNIX operating system, including digital UNIX
V4.0D. All sequence comparison parameters are set by the ALIGN-2
program and do not vary.
In situations where ALIGN-2 is employed for amino acid sequence
comparisons, the % amino acid sequence identity of a given amino
acid sequence A to, with, or against a given amino acid sequence B
(which can alternatively be phrased as a given amino acid sequence
A that has or comprises a certain % amino acid sequence identity
to, with, or against a given amino acid sequence B) is calculated
as follows: 100 times the fraction X/Y, where X is the number of
amino acid residues scored as identical matches by the sequence
alignment program ALIGN-2 in that program's alignment of A and B,
and where Y is the total number of amino acid residues in B. It
will be appreciated that where the length of amino acid sequence A
is not equal to the length of amino acid sequence B, the % amino
acid sequence identity of A to B will not equal the % amino acid
sequence identity of B to A. Unless specifically stated otherwise,
all % amino acid sequence identity values used herein are obtained
as described in the immediately preceding paragraph using the
ALIGN-2 computer program.
In some embodiments, the terms "individual" or "subject" are used
interchangeably and refer to any animal, including, but not limited
to, humans, non-human primates, rodents, and domestic and game
animals, which is to be the recipient of a particular treatment.
Primates include chimpanzees, cynomolgus monkeys, spider monkeys,
and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks,
ferrets, rabbits and hamsters. Domestic and game animals include
cows, horses, pigs, deer, bison, buffalo, feline species, e.g.,
domestic cat, canine species, e.g., dog, fox, wolf, avian species,
e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and
salmon. In various embodiments, a subject can be one who has been
previously diagnosed with or identified as suffering from or having
a condition in need of treatment. In certain embodiments, the
subject is a human. In various other embodiments, the subject
previously diagnosed with or identified as suffering from or having
a condition may or may not have undergone treatment for a
condition. In yet other embodiments, a subject can also be one who
has not been previously diagnosed as having a condition (i.e., a
subject who exhibits one or more risk factors for a condition). A
"subject in need" of treatment for a particular condition can be a
subject having that condition, diagnosed as having that condition,
or at risk of developing that condition. In some embodiments, the
subject is a "patient," that has been diagnosed with a disease or
condition described herein.
In some embodiments, the term "therapeutically effective amount"
refers to an amount of an antibody, polypeptide, polynucleotide,
small organic molecule, or other drug effective to "treat" a
disease or disorder in a subject or mammal. In some cases,
therapeutically effective amount of the drug reduces the severity
of symptoms of the disease or disorder. In some instances, the
disease or disorder comprises inflammatory bowel disease (IBD),
Crohn's disease (CD), or ulcerative colitis (UC). In some
instances, the IBD, CD, and/or UC are severe or medically
refractory forms of the IBD, CD, and/or UC. Non-limiting examples
of symptoms of IBD, CD, and/or UC include, but are not limited to,
diarrhea, fever, fatigue, abdominal pain, abdominal cramping,
inflammation, ulceration, nausea, vomiting, bleeding, blood in
stool, reduced appetite, and weight loss.
In some embodiments, the terms, "treat" or "treating" as used
herein refer to both therapeutic treatment and prophylactic or
preventative measures, wherein the object is to prevent or slow
down (lessen) the targeted pathologic condition, prevent the
pathologic condition, pursue or obtain good overall survival, or
lower the chances of the individual developing the condition even
if the treatment is ultimately unsuccessful. In some aspects
provided herein, subjects in need of treatment include those
already with a disease or condition, as well as those susceptible
to develop the disease or condition or those in whom the disease or
condition is to be prevented. The disease or condition may comprise
an inflammatory disease or condition, fibrostenotic or fibrotic
disease, thiopurine toxicity or disease related to thiopurine
toxicity, non-response to anti-TNF therapy, steroids or
immunomodulators.
Anti-TL1A Antibodies
Various embodiments provide antibodies that bind to TL1A. In some
embodiments, the antibodies specifically bind to soluble TL1A. In
some embodiments, the antibodies specifically bind to membrane
bound TL1A. In some embodiments, an anti-TL1A antibody is provided
having a heavy chain comprising four heavy chain framework regions
(HCFR) and three heavy chain complementarity-determining regions
(HCDR): HCFR1, HCDR1, HCFR2, HCDR2, HCFR3, HCDR3, and HCFR4; and a
light chain comprising four light chain framework regions (LCFR)
and three light chain complementarity-determining regions (LCDR):
LCFR1, LCDR1, LCFR2, LCDR2, LCFR3, LCDR3, and LCFR4. An anti-TL1A
antibody may comprise any region provided herein, for example, as
provided in Tables 1, 2, 3, the examples, and SEQ ID NOs: 1 to 54,
490 to 588. In some embodiments, an anti-TL1A antibody comprises a
variable domain, e.g., as provided herein, with one or more CDR
mutations as shown in Table 2, or 19 to 22. In some embodiments, an
anti-TL1A antibody comprises one or more CDRs comprising a sequence
shown in Tables 19 to 22.
In certain embodiments, the anti-TL1A antibody comprises CDRs
corresponding to those set forth in Tables 19 to 22. In certain
embodiments, the anti-TL1A antibody or antigen binding fragment
comprises a heavy chain variable region comprising: (a) an HCDR1
comprising the amino acid sequence set forth by SEQ ID NO: 484
(DTYMH); (b) an HCDR2 comprising the amino acid sequence set forth
by SEQ ID NO: 485 (PASGH); and (c) an HCDR3 comprising the amino
acid sequence set forth by SEQ ID NO: 486 (SGGLPD); and a light
chain variable region comprising (d) an LCDR1 comprising the amino
acid sequence set forth by SEQ ID NO: 487 (ASSSVSYMY); (e) an LCCR2
comprising the amino acid sequence set forth by SEQ ID NO: 488
(ATSNLAS); and (f) an LCDR3 comprising the amino acid sequence set
forth by SEQ ID NO: 489 (GNPRT).
In certain embodiments, described herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by any one of
SEQ ID NOs: 554 to 564 or 574 to 577; and (c) an HCDR3 comprising
an amino acid sequence set forth by any one of SEQ ID NOs: 565 to
568 or 578 to 581; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by any one
of SEQ ID NOs: 569 or 570; (e) an LCCR2 comprising an amino acid
sequence set forth by SEQ ID NO: 488; and (f) an LCDR3 comprising
an amino acid sequence set forth by any one of SEQ ID NOs: 571 to
573 or 582 to 585.
In certain embodiments, described herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
559; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 567; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 573. In certain
embodiments, described herein, is an antibody or antigen-binding
fragment that specifically binds to TL1A, comprising: a heavy chain
variable region comprising an amino acid sequence at least about
85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 503;
and a light chain variable region comprising an amino acid sequence
at least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to
SEQ ID NO: 502. In certain embodiments, the antibody or antigen
binding fragment comprises a kappa light chain constant region and
IgG1 heavy chain constant region. In certain embodiments, the
antibody or antigen binding fragment comprises a kappa light chain
constant region and IgG2 heavy chain constant region.
In certain embodiments, described herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
563; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 568; and a light chain variable region comprising (d)
an LCDR1 comprising an amino acid sequence set forth by SEQ ID NO:
569; (e) an LCCR2 comprising an amino acid sequence set forth by
SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid sequence
set forth by any one of SEQ ID NO: 572. In certain embodiments,
described herein, is an antibody or antigen-binding fragment that
specifically binds to TL1A, comprising: a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 511; and a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 510. In certain embodiments, the antibody or antigen binding
fragment comprises a kappa light chain constant region and IgG1
heavy chain constant region. In certain embodiments, the antibody
or antigen binding fragment comprises a kappa light chain constant
region and IgG2 heavy chain constant region.
In certain embodiments, described herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
555; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 566; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 572. In certain
embodiments, described herein, is an antibody or antigen-binding
fragment that specifically binds to TL1A, comprising: a heavy chain
variable region comprising an amino acid sequence at least about
85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 493;
and a light chain variable region comprising an amino acid sequence
at least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to
SEQ ID NO: 492. In certain embodiments, the antibody or antigen
binding fragment comprises a kappa light chain constant region and
IgG1 heavy chain constant region. In certain embodiments, the
antibody or antigen binding fragment comprises a kappa light chain
constant region and IgG2 heavy chain constant region.
In certain embodiments, described herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
558; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 566; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by SEQ ID NO: 572. In certain embodiments,
described herein, is an antibody or antigen-binding fragment that
specifically binds to TL1A, comprising: a heavy chain variable
region comprising an amino acid sequence at least about 85%, 90%,
95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 501; and a
light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ
ID NO: 500. In certain embodiments, the antibody or antigen binding
fragment comprises a kappa light chain constant region and IgG1
heavy chain constant region. In certain embodiments, the antibody
or antigen binding fragment comprises a kappa light chain constant
region and IgG2 heavy chain constant region.
In certain embodiments, described herein, is an antibody or
antigen-binding fragment that specifically binds to TL1A,
comprising: a heavy chain variable region comprising: (a) an HCDR1
comprising an amino acid sequence set forth by SEQ ID NO: 553; (b)
an HCDR2 comprising an amino acid sequence set forth by SEQ ID NO:
564; and (c) an HCDR3 comprising an amino acid sequence set forth
by SEQ ID NO: 568; and a light chain variable region comprising:
(d) an LCDR1 comprising an amino acid sequence set forth by SEQ ID
NO: 569; (e) an LCCR2 comprising an amino acid sequence set forth
by SEQ ID NO: 488; and (f) an LCDR3 comprising an amino acid
sequence set forth by any one of SEQ ID NO: 572. In certain
embodiments, described herein, is an antibody or antigen-binding
fragment that specifically binds to TL1A, comprising: a heavy chain
variable region comprising an amino acid sequence at least about
85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 515;
and a light chain variable region comprising an amino acid sequence
at least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to
SEQ ID NO: 514. In certain embodiments, the antibody or antigen
binding fragment comprises a kappa light chain constant region and
IgG1 heavy chain constant region. In certain embodiments, the
antibody or antigen binding fragment comprises a kappa light chain
constant region and IgG2 heavy chain constant region.
In certain embodiments, the anti-TL1A antibody or antigen binding
fragment comprises a heavy chain variable region comprising n
HCDR1, an HCDR2, and an HCDR3 from any one of SEQ ID NOs: 491, 493,
495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517, 519,
521, 523, 525, 527, 529, 531, 533, 535, 537, 539, or 541; and a
light chain variable region comprising n LCDR1, an LCDR2, and an
LCDR3 from any one of SEQ ID NOs: 490, 492, 494, 496, 498, 500,
502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526,
528, 530, 532, 534, 536, 538, or 540, wherein the CDRs are defined
by Kabat method, the IMGT method, the Chothia method or a
combination thereof. In certain embodiments, the anti-TL1A antibody
or antigen binding fragment comprises a heavy chain variable region
comprising an amino acid sequence at least about 85%, 90%, 95%,
97%, 98%, 99%, or 100% identical to any one of SEQ ID NOs: 491,
493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517,
519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, or 541; and
a light chain variable region comprising an amino acid sequence at
least about 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to any
one of SEQ ID NOs: 490, 492, 494, 496, 498, 500, 502, 504, 506,
508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, 530, 532,
534, 536, 538, or 540.
In certain embodiments, the anti-TL1A antibody or antigen binding
fragment comprises a heavy chain variable region comprising: (a) a
human heavy chain framework region 1 that is at least 90%, 95%,
96%, 97%, 98%, or 99% identical to that set forth is SEQ ID NO:
545; (b) an HCDR1 comprising the amino acid sequence set forth by
SEQ ID NO: 484 (DTYMH); (c) a human heavy chain framework region 2
that is at least 90%, 95%, 96%, 97%, 98%, or 99% identical to that
set forth is SEQ ID NO: 546; (d) an HCDR2 comprising the amino acid
sequence set forth by SEQ ID NO: 485 (PASGH); (e) a human heavy
chain framework region 3 that is at least 90%, 95%, 96%, 97%, 98%,
or 99% identical to that set forth is SEQ ID NO: 547 or 586 to 588;
(f) an HCDR3 comprising the amino acid sequence set forth by SEQ ID
NO: 486 (SGGLPD); and (g) a human heavy chain framework region 4
that is at least 90%, 95%, 96%, 97%, 98%, or 99% identical to that
set forth is SEQ ID NO: 548; and a light chain variable region
comprising (h) a human light chain framework region 1 that is at
least 90%, 95%, 96%, 97%, 98%, or 99% identical to that set forth
is SEQ ID NO: 549; (i) an LCDR1 comprising the amino acid sequence
set forth by SEQ ID NO: 487 (ASSSVSYMY); (j) a human light chain
framework region 2 that is at least 90%, 95%, 96%, 97%, 98%, or 99%
identical to that set forth is SEQ ID NO: 550; (k) an LCCR2
comprising the amino acid sequence set forth by SEQ ID NO: 488
(ATSNLAS); (l) a human light chain framework region 3 that is at
least 90%, 95%, 96%, 97%, 98%, or 99% identical to that set forth
is SEQ ID NO: 551; (m) an LCDR3 comprising the amino acid sequence
set forth by SEQ ID NO: 489 (GNPRT); and (n) a human light chain
framework region 4 that is at least 90%, 95%, 96%, 97%, 98%, or 99%
identical to that set forth is SEQ ID NO: 552.
TABLE-US-00001 TABLE 1 Exemplary anti-TL1A antibodies Murine HC -
HC - LC - LC - FR Back Clone DNA protein DNA protein Mutations
Murine 5C3D11 SEQ ID SEQ ID SEQ ID SEQ ID NA NO: 1 NO: 3 NO: 4 NO:
6 Murine 5C3D11 SEQ ID SEQ ID SEQ ID SEQ ID NA (codon optimized)
NO: 2 NO: 3 NO: 5 NO: 6 Chimeric 5C3D11 SEQ ID SEQ ID SEQ ID SEQ ID
NA NO: 2 NO: 3 NO: 5 NO: 6 12835 SEQ ID SEQ ID SEQ ID SEQ ID 9
(humanized NO: 25 NO: 26 NO: 27 NO: 28 5C3D11) 18-7 (CDR- SEQ ID
SEQ ID SEQ ID SEQ ID 2 grafted LC) NO: 35 NO: 36 NO: 37 NO: 38 21-3
(CDR- SEQ ID SEQ ID SEQ ID SEQ ID 2 grafted HC) NO: 39 NO: 40 NO:
41 NO: 42 L8 (CDR graft) SEQ ID SEQ ID SEQ ID SEQ ID 0 NO: 39 NO:
40 NO: 37 NO: 38
TABLE-US-00002 TABLE 2 Exemplary anti-TL1A CDR sequences SEQ ID CDR
NO Sequence Definitions H1 150
GFX.sub.1X.sub.2X.sub.3DX.sub.4X.sub.5X.sub.6H X.sub.1 = D or E
X.sub.2 = I, L, P, or V X.sub.3 = G, Q, S, or V X.sub.4 = A, S, T
X.sub.5 = F or Y X.sub.6 = I, L, or M H2 12
RX.sub.1X.sub.2PX.sub.3X.sub.4X.sub.5HX.sub.6K X.sub.1 = I or L
X.sub.7X.sub.8PKFX.sub.9X.sub.10 X.sub.2 = D or E X.sub.3 = A or E
X.sub.4 = G or S X.sub.5 = A or G X.sub.6 = I, L, T, or V X.sub.7 =
I, L, M, S, T, V, or Y X.sub.8 = D, I, N, R, or S X.sub.9 = Q or R
X.sub.10 = A, D, E, G, H, K, L, M, N, P, R, S, T, or V H3 152
X.sub.1X.sub.2GX.sub.3PX.sub.4X.sub.5 X.sub.1 = L or S X.sub.2 = A
or G X.sub.3 = A, L, or M X.sub.4 = D or E X.sub.5 = K, M, Q, R, S,
T, V, or W L1 18 X.sub.1ASSSVX.sub.2X.sub.3X.sub.4X.sub.5 X.sub.1 =
G, R, or W X.sub.2 = I or S X.sub.3 = F or Y X.sub.4 = L or M
X.sub.5 = R or Y L2 21 AX.sub.1X.sub.2X.sub.3LX.sub.4S X.sub.1 = K
or T X.sub.2 = E, P, or S X.sub.3 = L, N, or P X.sub.4 = A or T L3
155 X.sub.1QX.sub.2X.sub.3X.sub.4X.sub.5PRX.sub.6 X.sub.1 = H, N,
Q, or S X.sub.2 = F, H, I, P, R, S, W, or Y X.sub.3 = D, E, H, N,
Q, S, or V X.sub.4 = A, D, G, Q, or S X.sub.5 = D, F, H, K, L, M,
N, Q, R, S, or T
In various embodiments, an anti-TL1A antibody binds specifically to
the same region of TL1A or binds specifically to a region of TL1A
that overlaps with the region of TL1A to which an antibody
comprising a heavy chain comprising a sequence at least about 90%,
92%, 95%, 98%, or 100% identical to SEQ ID NO: 3 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 6, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 26 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 28, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 36 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 38, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 40 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 42, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 40 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 38, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 503 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 502, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 511 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 510, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 493 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 492, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 501 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 500, binds specifically. In various
embodiments, an anti-TL1A antibody binds specifically to the same
region of TL1A or binds specifically to a region of TL1A that
overlaps with the region of TL1A to which an antibody comprising a
heavy chain comprising a sequence at least about 90%, 92%, 95%,
98%, or 100% identical to SEQ ID NO: 515 and a light chain
comprising a sequence at least about 90%, 92%, 95%, 98%, or 100%
identical to SEQ ID NO: 514, binds specifically.
In various embodiments, the anti-TL1A antibody or fragment has a
binding affinity to TL1A of at least about 1E.sup.-7, 1E.sup.-8,
1E.sup.-9, 1E.sup.-10, or 1E.sup.-11. In some cases, the binding
affinity is from about 1E.sup.-9 to about 1E.sup.-11.
Various embodiments provide for an anti-TL1A antibody that binds to
the same region of a TL1A protein or portion thereof as a reference
antibody, e.g., any anti-TL1A antibody described herein. In some
embodiments, the reference antibody comprises the heavy chain CDRs
of SEQ ID NOS: 150, 12, and 152 and the light chain CDRs of SEQ ID
NOS: 18, 21, and 155.
Non-limiting methods for determining whether an anti-TL1A antibody
(i.e. test antibody) binds to the same region of a TL1A protein or
portion thereof as an antibody described herein are provided. An
exemplary embodiment comprises a competition assay. For instance,
the method comprises determining whether the test antibody can
compete with binding between the reference antibody and the TL1A
protein or portion thereof, or determining whether the reference
antibody can compete with binding between the test antibody and the
TL1A protein or portion thereof. Exemplary methods include use of
surface plasmon resonance to evaluate whether an anti-TL1A antibody
can compete with the binding between TL1A and another anti-TL1A
antibody. In some cases, surface plasmon resonance is utilized in
the competition assay. Non-limiting methods are described in the
examples.
The TL1A antibodies described herein bind to specific regions or
epitopes of human TL1A. These regions are demonstrated herein as
useful to inhibit interferon gamma secretion from T lymphocytes. In
certain embodiments, disclosed herein are antibodies that compete
for binding TL1A with the antibodies described herein. In certain
embodiments, disclosed herein are antibodies that bind the same
epitope of TL1A bound by the antibodies described herein. In
certain embodiments, disclosed herein are antibodies that bind a
discrete epitope that overlaps with an epitope of TL1A bound by an
antibody described herein. In certain embodiments, disclosed herein
are antibodies that bind the same epitope of TL1A, overlap with the
an epitope of TL1A by one or more amino acid residues, or that
compete for binding to an epitope of TL1A with an antibody or
fragment thereof that comprises a heavy chain variable region
comprising the amino acid sequence of SEQ ID NO: 503; and a light
chain variable region comprising the amino acid of SEQ ID NO: 502.
In certain embodiments, disclosed herein are antibodies that bind
the same epitope of TL1A, overlap with the an epitope of TL1A by
one or more amino acid residues, or that compete for binding to an
epitope of TL1A with an antibody or fragment thereof that comprises
a heavy chain variable region comprising the amino acid sequence of
SEQ ID NO: 511; and a light chain variable region comprising the
amino acid of SEQ ID NO: 510. In certain embodiments, disclosed
herein are antibodies that bind the same epitope of TL1A, overlap
with the an epitope of TL1A by one or more amino acid residues, or
that compete for binding to an epitope of TL1A with an antibody or
fragment thereof that comprises a heavy chain variable region
comprising the amino acid sequence of SEQ ID NO: 493; and a light
chain variable region comprising the amino acid of SEQ ID NO: 492.
In certain embodiments, disclosed herein are antibodies that bind
the same epitope of TL1A, overlap with the an epitope of TL1A by
one or more amino acid residues, or that compete for binding to an
epitope of TL1A with an antibody or fragment thereof that comprises
a heavy chain variable region comprising the amino acid sequence of
SEQ ID NO: 501; and a light chain variable region comprising the
amino acid of SEQ ID NO: 500. In certain embodiments, disclosed
herein are antibodies that bind the same epitope of TL1A, overlap
with the an epitope of TL1A by one or more amino acid residues, or
that compete for binding to an epitope of TL1A with an antibody or
fragment thereof that comprises the amino acid sequence of SEQ ID
NO: 515; and a light chain variable region comprising the amino
acid of SEQ ID NO: 514.
Methods of Generating Antibodies
Various embodiments provide for an antibody that is generated using
a polypeptide or a nucleotide sequence. In some embodiments, the
antibody is a monoclonal antibody. In some embodiments, the
antibody is a human antibody or a humanized antibody. In some
embodiments, the antibody is an antibody fragment. For example, the
antibody is a Fab, an scFv, or a (Fab).sub.2. In some embodiments,
the antibody is a chimeric antibody.
The antibodies described herein can be assayed for specific binding
by any method known in the art. The immunoassays which can be used
include, but are not limited to, competitive and non-competitive
assay systems using techniques such as BIAcore analysis, FACS
analysis, immunofluorescence, immunocytochemistry, Western blots,
radioimmunoassays, ELISA, "sandwich" immunoassays,
immunoprecipitation assays, precipitation reactions, gel diffusion
precipitin reactions, immunodiffusion assays, agglutination assays,
complement-fixation assays, immunoradiometric assays, fluorescent
immunoassays, and protein A immunoassays. Such assays are provided
in for e.g., Ausubel et al., eds, 1994, Current Protocols in
Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New
York.
In various embodiments, the anti-TL1A antibody is an antagonist of
a TL1A receptor, such as, but not limited to, DR3 and TR6/DcR3. In
certain embodiments, the antibody inhibits at least about 10%, at
least about 20%, at least about 30%, at least about 50%, at least
about 75%, at least about 90%, or about 100% of one or more
activity of the bound TL1A receptor. In certain embodiments, the
antibodies inhibit TL1A activation as measured by interferon gamma
release in human blood. In certain embodiments, the antibody
inhibits interferon gamma release in human blood at an IC.sub.50 of
between about 1 nanomolar and about 100 picomolar. In certain
embodiments, the antibody inhibits interferon gamma release in
human blood at an IC.sub.50 of between about 500 picomolar and
about 100 picomolar. In certain embodiments, the antibody inhibits
interferon gamma release in human blood at an IC.sub.50 of about
500 picomolar. In certain embodiments, the antibody inhibits
interferon gamma release in human blood at an IC.sub.50 of about
250 picomolar.
In various embodiments, monoclonal antibodies are prepared using
methods known in the art, such as, but not limited to the hybridoma
method, where a host animal is immunized, as described above to
elicit the production by lymphocytes of antibodies that will
specifically bind to an immunizing antigen (Kohler and Milstein
(1975) Nature 256:495). Hybridomas produce monoclonal antibodies
directed specifically against a chosen antigen. The monoclonal
antibodies are purified from the culture medium or ascites fluid by
techniques known in the art, when propagated either in vitro or in
vivo.
In some embodiments, monoclonal antibodies are made using
recombinant DNA methods as described in U.S. Pat. No. 4,816,567.
The polynucleotides encoding a monoclonal antibody are isolated
from mature B-cells or hybridoma cells. The isolated
polynucleotides encoding the heavy and light chains are then cloned
into suitable expression vectors, which when transfected into host
cells (e.g., E. coli cells, simian COS cells, Chinese hamster ovary
(CHO) cells, or myeloma cells) generate monoclonal antibodies. The
polynucleotide(s) encoding a monoclonal antibody can further be
modified in a number of different manners using recombinant DNA
technology to generate alternative antibodies.
In various embodiments, a chimeric antibody, a molecule in which
different portions are derived from different animal species, such
as those having a variable region derived from a murine monoclonal
antibody and a human immunoglobulin constant region (e.g.,
humanized antibodies) can be generated. Chimeric antibodies can be
produced using various techniques such as those set forth in
Morrison et al., Proc. Natl. Acad. Sci. 81:851-855 (1984);
Neuberger et al., Nature 312:604-608 (1984); Takeda et al., Nature
314:452-454 (1985).
In some embodiments, the anti-TL1A monoclonal antibody is a
humanized antibody, to reduce antigenicity and HAMA (human
anti-mouse antibody) responses when administered to a human
subject. Humanized antibodies can be produced using various
techniques known in the art. For example, an antibody is humanized
by (1) determining the nucleotide and predicted amino acid sequence
of the starting antibody light and heavy variable domains; (2)
designing the humanized antibody, e.g., deciding which antibody
framework region to use during the humanizing process; (3) the
actual humanizing methodologies/techniques; and (4) the
transfection and expression of the humanized antibody (see, e.g.,
U.S. Pat. Nos. 5,585,089; 6,835,823; 6,824,989). In various
embodiments, a humanized antibody can be further optimized to
decrease potential immunogenicity, while maintaining functional
activity, for therapy in humans.
Humanized antibodies can also be made in transgenic mice containing
human immunoglobulin loci that are capable, upon immunization, of
producing the full repertoire of human antibodies in the absence of
endogenous immunoglobulin production. This approach is described in
U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126;
5,633,425; and 5,661,016. A humanized antibody may also be obtained
by a novel genetic engineering approach that enables production of
affinity-matured human-like polyclonal antibodies in large animals
such as, for example, rabbits and mice. (See, e.g. U.S. Pat. No.
6,632,976).
A fully humanized antibody may be created by first designing a
variable region amino acid sequence that contains non-human, e.g.,
rodent-derived CDRs, embedded in human-derived framework sequences.
The non-human CDRs provide the desired specificity. Accordingly, in
some cases these residues are included in the design of the
reshaped variable region essentially unchanged. In some cases,
modifications should therefore be restricted to a minimum and
closely watched for changes in the specificity and affinity of the
antibody. On the other hand, framework residues in theory can be
derived from any human variable region. A human framework sequences
should be chosen, which is equally suitable for creating a reshaped
variable region and for retaining antibody affinity, in order to
create a reshaped antibody which shows an acceptable or an even
improved affinity. The human framework may be of germline origin,
or may be derived from non-germline (e.g. mutated or affinity
matured) sequences. Genetic engineering techniques well known to
those in the art, for example, but not limited to, phage display of
libraries of human antibodies, transgenic mice, human-human
hybridoma, hybrid hybridoma, B cell immortalization and cloning,
single-cell RT-PCR or HuRAb Technology, may be used to generate a
humanized antibody with a hybrid DNA sequence containing a human
framework and a non-human CDR. Methods to obtain "humanized
antibodies" are well known to those skilled in the art. (e.g., U.S.
Pat. Nos. 5,861,155, 6,479,284, 6,407,213, 5,624,821, US2003166871,
US20020078757, Queen et al., Proc. Natl. Acad Sci USA,
86:10029-10032 (1989) and Hodgson et al., Bio/Technology, 9:421
(1991)).
In certain embodiments, the anti-TL1A antibody is a human antibody.
Human antibodies can be directly prepared using various techniques
known in the art. Immortalized human B lymphocytes immunized in
vitro or isolated from an immunized individual that produce an
antibody directed against a target antigen can be generated (See,
e.g., Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan
R. Liss, p. 77 (1985); Boerner et al., 1991, J. Immunol., 147
(1):86-95; and U.S. Pat. No. 5,750,373). A human antibody can be
selected from a phage library. Techniques for the generation and
use of antibody phage libraries are described in U.S. Pat. Nos.
5,969,108, 6,172,197, 5,885,793, 6,521,404; 6,544,731; 6,555,313;
6,582,915; 6,593,081; 6,300,064; 6,653,068; 6,706,484; and
7,264,963; and Rothe et al., 2007, J. Mol. Bio.,
doi:10.1016/j.jmb.2007.12.018.
Chimeric, humanized and human antibodies may be produced by
recombinant expression. Recombinant polynucleotide constructs
typically include an expression control sequence operably linked to
the coding sequences of antibody chains, including
naturally-associated or heterologous promoter regions. In certain
embodiments, it may be desirable to generate amino acid sequence
variants of these humanized antibodies, particularly where these
improve the binding affinity or other biological properties of the
antibody.
In certain embodiments, an antibody fragment is used to treat
and/or ameliorate IBD. Various techniques are known for the
production of antibody fragments. Generally, these fragments are
derived via proteolytic digestion of intact antibodies (for example
Morimoto et al., 1993, Journal of Biochemical and Biophysical
Methods 24:107-117; Brennan et al., 1985, Science, 229:81). Fab,
Fv, and scFv antibody fragments can all be expressed in and
secreted from E. coli or other host cells, thus allowing the
production of large amounts of these fragments. Other techniques
for the production of antibody fragments will be apparent to the
skilled practitioner.
According to the present disclosure, techniques can be adapted for
the production of single-chain antibodies specific to TL1A (see
e.g., U.S. Pat. No. 4,946,778). In addition, methods can be adapted
for the construction of Fab expression libraries (see e.g., Huse,
et al., Science 246:1275-1281 (1989)) to allow rapid and effective
identification of monoclonal Fab fragments with the desired
specificity for TL1A, or derivatives, fragments, analogs or
homologs thereof. Antibody fragments may be produced by techniques
in the art including, but not limited to: (a) a F(ab').sub.2
fragment produced by pepsin digestion of an antibody molecule; (b)
a Fab fragment generated by reducing the disulfide bridges of an
F(ab').sub.2 fragment, (c) a Fab fragment generated by the
treatment of the antibody molecule with papain and a reducing
agent, and (d) Fv fragments.
Also provided herein are modified antibodies comprising any type of
variable region that provides for the association of the antibody
with TL1A. Those skilled in the art will appreciate that the
modified antibodies may comprise antibodies (e.g., full-length
antibodies or immunoreactive fragments thereof) in which at least a
fraction of one or more of the constant region domains has been
deleted or otherwise altered so as to provide desired biochemical
characteristics such as decreasing TL1A. In certain embodiments,
the variable regions in both the heavy and light chains are altered
by at least partial replacement of one or more CDRs and, if
necessary, by partial framework region replacement and sequence
changing. In some embodiments, the replaced CDRs may be derived
from an antibody of the same class, subclass, from an antibody of a
different class, for instance, from an antibody from a different
species and/or a combination thereof In some embodiments, the
constant region of the modified antibodies will comprise a human
constant region. Modifications to the constant region compatible
with this disclosure comprise additions, deletions or substitutions
of one or more amino acids in one or more domains.
In various embodiments, the expression of an antibody or
antigen-binding fragment thereof as described herein can occur in
either prokaryotic or eukaryotic cells. Suitable hosts include
bacterial or eukaryotic hosts, including yeast, insects, fungi,
bird and mammalian cells either in vivo, or in situ, or host cells
of mammalian, insect, bird or yeast origin. The mammalian cell or
tissue can be of human, primate, hamster, rabbit, rodent, cow, pig,
sheep, horse, goat, dog or cat origin, but any other mammalian cell
may be used. In other embodiments, the antibody or antigen-fragment
thereof as described herein may be transfected into the host.
In some embodiments, the expression vectors are transfected into
the recipient cell line for the production of the chimeric,
humanized, or composite human antibodies described herein. In
various embodiments, mammalian cells can be useful as hosts for the
production of antibody proteins, which can include, but are not
limited to cells of fibroblast origin, such as Vero (ATCC CRL 81)
or CHO-K1 (ATCC CRL 61) cells, HeLa cells and L cells. Exemplary
eukaryotic cells that can be used to express polypeptides include,
but are not limited to, COS cells, including COS 7 cells; 293
cells, including 293-6E cells; CHO cells, including CHO--S and DG44
cells; PER.C6.TM. cells (Crucell); and NSO cells. In some
embodiments, a particular eukaryotic host cell is selected based on
its ability to make desired post-translational modifications to the
heavy chains and/or light chains.
A number of suitable host cell lines capable of secreting intact
heterologous proteins have been developed in the art, and include,
but are not limited to CHO cell lines, various COS cell lines, HeLa
cells, L cells and multiple myeloma cell lines.
An expression vector carrying a chimeric, humanized, or composite
human antibody construct, antibody or antigen-binding fragment
thereof as described herein can be introduced into an appropriate
host cell by any of a variety of suitable means, depending on the
type of cellular host including, but not limited to transformation,
transfection, lipofection, conjugation, electroporation, direct
microinjection, and microprojectile bombardment, as known to one of
ordinary skill in the art. Expression vectors for these cells can
include expression control sequences, such as an origin of
replication sites, a promoter, an enhancer and necessary processing
information sites, such as ribosome binding sites, RNA splice
sites, polyadenylation sites, and transcriptional terminator
sequences.
In various embodiments, yeast can also be utilized as hosts for the
production of the antibody molecules or peptides described herein.
In various other embodiments, bacterial strains can also be
utilized as hosts for the production of the antibody molecules or
peptides described herein. Examples of bacterial strains include,
but are not limited to E. coli, Bacillus species, enterobacteria,
and various Pseudomonas species.
In some embodiments, one or more antibodies or antigen-binding
fragments thereof as described herein can be produced in vivo in an
animal that has been engineered (transgenic) or transfected with
one or more nucleic acid molecules encoding the polypeptides,
according to any suitable method. For production of transgenic
animals, transgenes can be microinjected into fertilized oocytes,
or can be incorporated into the genome of embryonic stem cells, and
the nuclei of such cells transferred into enucleated oocytes. Once
expressed, antibodies can be purified according to standard
procedures of the art, including HPLC purification, column
chromatography, gel electrophoresis and the like (see generally,
Scopes, Protein Purification (Springer-Verlag, NY, 1982)).
Once expressed in the host, the whole antibodies,
antibody-fragments (e.g., individual light and heavy chains), or
other immunoglobulin forms of the present disclosure can be
recovered and purified by known techniques, e.g., immunoabsorption
or immunoaffinity chromatography, chromatographic methods such as
HPLC (high performance liquid chromatography), ammonium sulfate
precipitation, gel electrophoresis, or any combination of these.
See generally, Scopes, PROTEIN PURIF. (Springer-Verlag, NY, 1982).
Substantially pure immunoglobulins of at least about 90% to 95%
homogeneity are advantageous, as are those with 98% to 99% or more
homogeneity, particularly for pharmaceutical uses. Once purified,
partially or to homogeneity as desired, a humanized or composite
human antibody can then be used therapeutically or in developing
and performing assay procedures, immunofluorescent stainings, etc.
See generally, Vols. I & II Immunol. Meth. (Lefkovits &
Pernis, eds., Acad. Press, NY, 1979 and 1981).
Various embodiments provide for a genetic construct comprising a
nucleic acid encoding an anti-TL1A antibody or fragment provided
herein. Genetic constructs of the antibody can be in the form of
expression cassettes, which can be suitable for expression of the
encoded anti-TL1A antibody or fragment. The genetic construct may
be introduced into a host cell with or without being incorporated
in a vector. For example, the genetic construct can be incorporated
within a liposome or a virus particle. Alternatively, a purified
nucleic acid molecule can be inserted directly into a host cell by
methods known in the art. The genetic construct can be introduced
directly into cells of a host subject by transfection, infection,
electroporation, cell fusion, protoplast fusion, microinjection or
ballistic bombardment.
Various embodiments provide a recombinant vector comprising the
genetic construct of an antibody provided herein. The recombinant
vector can be a plasmid, cosmid or phage. The recombinant vectors
can include other functional elements; for example, a suitable
promoter to initiate gene expression.
Various embodiments provide a host cell comprising a genetic
construct and/or recombinant vector described herein.
Various host systems are also advantageously employed to express
recombinant protein. Examples of suitable mammalian host cell lines
include the COS-7 lines of monkey kidney cells, and other cell
lines capable of expressing an appropriate vector including, for
example, L cells, C127, 3T3, Chinese hamster ovary (CHO), HeLa and
BHK cell lines. Mammalian expression vectors can comprise
nontranscribed elements such as an origin of replication, a
suitable promoter and enhancer linked to the gene to be expressed,
and other 5' or 3' flanking nontranscribed sequences, and 5' or 3'
nontranslated sequences, such as necessary ribosome binding sites,
a polyadenylation site, splice donor and acceptor sites, and
transcriptional termination sequences.
The proteins produced by a transformed host can be purified
according to any suitable method. Such standard methods include
chromatography (e.g., ion exchange, affinity and sizing column
chromatography), centrifugation, differential solubility, or by any
other standard technique for protein purification. Affinity tags
such as hexahistidine (SEQ ID NO: 589), maltose binding domain,
influenza coat sequence and glutathione-S-transferase can be
attached to the protein to allow easy purification by passage over
an appropriate affinity column. Isolated proteins can also be
physically characterized using such techniques as proteolysis,
nuclear magnetic resonance and x-ray crystallography. Recombinant
protein produced in bacterial culture can be isolated. Methods
known in the art for purifying antibodies and other proteins also
include, for example, those described in U.S. Patent Publication
No. 2008/0177048, and 2009/0187005.
One of skill will recognize that individual substitutions,
deletions or additions to a nucleic acid, peptide, polypeptide, or
protein sequence which alters a single amino acid or a small
percentage of amino acids in the encoded sequence is a
"conservatively modified variant" where the alteration results in
the substitution of an amino acid with a chemically similar amino
acid and retain the ability to specifically bind the target
antigen. Such conservatively modified variants are in addition to
and do not exclude polymorphic variants, interspecies homologs, and
alleles consistent with the disclosure.
A given amino acid can be replaced by a residue having similar
physiochemical characteristics, e.g., substituting one aliphatic
residue for another (such as He, Val, Leu, or Ala for one another),
or substitution of one polar residue for another (such as between
Lys and Arg; Glu and Asp; or Gln and Asn). Other such conservative
substitutions, e.g., substitutions of entire regions having similar
hydrophobicity characteristics, are well known. Polypeptides
comprising conservative amino acid substitutions can be tested in
any one of the assays described herein to confirm that a desired
activity, e.g. antigen-binding activity and specificity of a native
or reference polypeptide is retained.
Particular conservative substitutions include, for example; Ala
into Gly or into Ser; Arg into Lys; Asn into Gin or into H is; Asp
into Glu; Cys into Ser; Gin into Asn; Glu into Asp; Gly into Ala or
into Pro; His into Asn or into Gin; lie into Leu or into Val; Leu
into lie or into Val; Lys into Arg, into Gin or into Glu; Met into
Leu, into Tyr or into lie; Phe into Met, into Leu or into Tyr; Ser
into Thr; Thr into Ser; Trp into Tyr; Tyr into Trp; and/or Phe into
Val, into lie or into Leu.
In some embodiments, the antibody and/or antigen-binding fragment
thereof described herein can be a variant of a sequence described
herein, e.g., a conservative substitution variant of an antibody
polypeptide. In some embodiments, the variant is a conservatively
modified variant. A variant may refer to a polypeptide
substantially homologous to a native or reference polypeptide, but
which has an amino acid sequence different from that of the native
or reference polypeptide because of one or a plurality of
deletions, insertions or substitutions. Variant
polypeptide-encoding DNA sequences encompass sequences that
comprise one or more additions, deletions, or substitutions of
nucleotides when compared to a native or reference DNA sequence,
but that encode a variant protein or fragment thereof that retains
activity, e.g., antigen-specific binding activity for the relevant
target polypeptide.
Alterations of the native amino acid sequence can be accomplished
by any of a number of techniques known to one of skill in the art.
Mutations can be introduced at particular loci or by
oligonucleotide-directed site-specific mutagenesis procedures.
Techniques for making such alterations are very well established
and include, for example, those disclosed by Walder et al. (Gene
42: 133, 1986); Bauer et al. (Gene 37:73, 1985); Craik
(BioTechniques, January 1985, 12-19); Smith et al. (Genetic
Engineering: Principles and Methods, Plenum Press, 1981); and U.S.
Pat. Nos. 4,518,584 and 4,737,462.
Nucleic acid molecules encoding amino acid sequence variants of
antibodies are prepared by a variety of methods known in the art.
These methods include, but are not limited to, preparation by
oligonucleotide-mediated (or site-directed) mutagenesis, PCR
mutagenesis, and cassette mutagenesis of an earlier prepared
variant or a non-variant version of the antibody. A nucleic acid
sequence encoding at least one antibody, portion or polypeptide as
described herein can be recombined with vector DNA in accordance
with conventional techniques, including but not limited to,
blunt-ended or staggered-ended termini for ligation and restriction
enzyme digestion. Techniques for such manipulations are disclosed,
e.g., by Maniatis et al., Molecular Cloning, Lab. Manual (Cold
Spring Harbor Lab. Press, NY, 1982 and 1989), and can be used to
construct nucleic acid sequences which encode a monoclonal antibody
molecule or antigen-binding region.
In some embodiments, a nucleic acid encoding an antibody or
antigen-binding fragment thereof as described herein is comprised
by a vector. In some of the aspects described herein, a nucleic
acid sequence encoding an antibody or antigen-binding fragment
thereof as described herein, or any module thereof, is operably
linked to a vector. The term "vector," as used herein, refers to a
nucleic acid construct designed for delivery to a host cell or for
transfer between different host cells. As used herein, a vector can
be viral or non-viral. The term "vector" encompasses any genetic
element that is capable of replication when associated with the
proper control elements and that can transfer gene sequences to
cells. A vector can include, but is not limited to, a cloning
vector, an expression vector, a plasmid, phage, transposon, cosmid,
chromosome, virus, virion, etc.
As used herein, the term "expression vector" refers to a vector
that directs expression of an RNA or polypeptide from sequences
linked to transcriptional regulatory sequences on the vector. The
term "expression" refers to the cellular processes involved in
producing RNA and proteins and as appropriate, secreting proteins,
including where applicable, but not limited to, for example,
transcription, transcript processing, translation and protein
folding, modification and processing. "Expression products" include
RNA transcribed from a gene, and polypeptides obtained by
translation of mRNA transcribed from a gene. The term "gene" means
the nucleic acid sequence which is transcribed (DNA) to RNA in
vitro or in vivo when operably linked to appropriate regulatory
sequences. The gene may or may not include regions preceding and
following the coding region, e.g., 5' untranslated (5'UTR) or
"leader" sequences and 3' UTR or "trailer" sequences, as well as
intervening sequences (introns) between individual coding segments
(exons).
As used herein, the term "viral vector" refers to a nucleic acid
vector construct that includes at least one element of viral origin
and has the capacity to be packaged into a viral vector particle.
The viral vector can contain the nucleic acid encoding an antibody
or antigen-binding portion thereof as described herein in place of
non-essential viral genes. The vector and/or particle may be
utilized for the purpose of transferring any nucleic acids into
cells either in vitro or in vivo. Numerous forms of viral vectors
are known in the art.
By "recombinant vector," it is meant that the vector includes a
heterologous nucleic acid sequence, or "transgene" that is capable
of expression in vivo.
Pharmaceutical Compositions, Administration and Dosage
The anti-TL1A antibodies provided are useful in a variety of
applications including, but not limited to, therapeutic treatment
methods, such as the treatment of IBD. The methods of use may be in
vitro, ex vivo, or in vivo methods. In certain embodiments, the
anti-TL1A antibody is an antagonist for TL1A receptors.
In certain embodiments, the disease treated with anti-TL1A antibody
or TL1A receptor antagonist is IBD, CD, UC and/or MR-UC.
In various embodiments, the pharmaceutical compositions are
formulated for delivery via any route of administration. "Route of
administration" may refer to any administration pathway known in
the art, including but not limited to aerosol, nasal, oral,
transmucosal, transdermal or parenteral.
"Transdermal" administration may be accomplished using a topical
cream or ointment or by means of a transdermal patch.
"Parenteral" refers to a route of administration that is generally
associated with injection, including intraorbital, infusion,
intraarterial, intracapsular, intracardiac, intradermal,
intramuscular, intraperitoneal, intrapulmonary, intraspinal,
intrastemal, intrathecal, intrauterine, intravenous, subarachnoid,
subcapsular, subcutaneous, transmucosal, or transtracheal. Via the
parenteral route, the compositions may be in the form of solutions
or suspensions for infusion or for injection, or as lyophilized
powders.
Via the enteral route, the pharmaceutical compositions can be in
the form of tablets, gel capsules, sugar-coated tablets, syrups,
suspensions, solutions, powders, granules, emulsions, microspheres
or nanospheres or lipid vesicles or polymer vesicles allowing
controlled release.
Via the topical route, the pharmaceutical compositions are
formulated for treating the skin and mucous membranes and are in
the form of ointments, creams, milks, salves, powders, impregnated
pads, solutions, gels, sprays, lotions or suspensions. They can
also be in the form of microspheres or nanospheres or lipid
vesicles or polymer vesicles or polymer patches and hydrogels
allowing controlled release. These topical-route compositions can
be either in anhydrous form or in aqueous form depending on the
clinical indication.
Via the ocular route, they may be in the form of eye drops.
In various embodiments, an agent can be administered intravenously
by injection or by gradual infusion over time. Given an appropriate
formulation for a given route, for example, agents useful in the
methods and compositions described herein can be administered
intravenously, intranasally, by inhalation, intraperitoneally,
intramuscularly, subcutaneously, intracavity, and can be delivered
by peristaltic means, if desired, or by other means known by those
skilled in the art. In particular embodiments, compounds used
herein are administered orally, intravenously or intramuscularly to
a patient having IBD, CD, UC and/or MR-UC.
The pharmaceutical compositions can also contain any
pharmaceutically acceptable carrier. "Pharmaceutically acceptable
carrier" refers to a pharmaceutically acceptable material,
composition, or vehicle that is involved in carrying or
transporting a compound of interest from one tissue, organ, or
portion of the body to another tissue, organ, or portion of the
body. For example, the carrier may be a liquid or solid filler,
diluent, excipient, solvent, or encapsulating material, or a
combination thereof. Each component of the carrier must be
"pharmaceutically acceptable" in that it must be compatible with
the other ingredients of the formulation. It must also be suitable
for use in contact with any tissues or organs with which it may
come in contact, meaning that it must not carry a risk of toxicity,
irritation, allergic response, immunogenicity, or any other
complication that excessively outweighs its therapeutic
benefits.
In various embodiments, provided are pharmaceutical compositions
including a pharmaceutically acceptable excipient along with a
therapeutically effective amount of an anti-TL1A antibody.
"Pharmaceutically acceptable excipient" means an excipient that is
useful in preparing a pharmaceutical composition that is generally
safe, non-toxic, and desirable, and includes excipients that are
acceptable for veterinary use as well as for human pharmaceutical
use. The active ingredient can be mixed with excipients which are
pharmaceutically acceptable and compatible with the active
ingredient and in amounts suitable for use in therapeutic methods
described herein. Such excipients may be solid, liquid, semisolid,
or, in the case of an aerosol composition, gaseous. Suitable
excipients are, for example, starch, glucose, lactose, sucrose,
gelatin, malt, rice, flour, chalk, silica gel, sodium stearate,
glycerol monostearate, talc, sodium chloride, dried skim milk,
water, saline, dextrose, propylene glycol, glycerol, ethanol,
mannitol, polysorbate or the like and combinations thereof. In
addition, if desired, the composition can contain minor amounts of
auxiliary substances such as wetting or emulsifying agents, pH
buffering agents and the like which enhance or maintain the
effectiveness of the active ingredient. Therapeutic compositions as
described herein can include pharmaceutically acceptable salts.
Pharmaceutically acceptable salts include the acid addition salts
formed with inorganic acids such as, for example, hydrochloric or
phosphoric acids, organic acids, for example, acetic, tartaric or
mandelic, salts formed from inorganic bases such as, for example,
sodium, potassium, ammonium, calcium or ferric hydroxides, and
salts formed from organic bases such as isopropylamine,
trimethylamine, 2-ethylamino ethanol, histidine, procaine and the
like. Liquid compositions can contain liquid phases in addition to
and in the exclusion of water, for example, glycerin, vegetable
oils such as cottonseed oil, and water-oil emulsions.
Physiologically tolerable carriers are well known in the art. The
amount of an active agent (i.e. antibody or fragment thereof) used
that will be effective in the treatment of a particular disorder or
condition will depend on the nature of the disorder or condition,
and can be determined by one of skill in the art with standard
clinical techniques.
The pharmaceutical compositions can also be encapsulated, tableted
or prepared in an emulsion or syrup for oral administration.
Pharmaceutically acceptable solid or liquid carriers may be added
to enhance or stabilize the composition, or to facilitate
preparation of the composition. Liquid carriers include syrup,
peanut oil, olive oil, glycerin, saline, alcohols and water. Solid
carriers include starch, lactose, calcium sulfate, dihydrate, terra
alba, magnesium stearate or stearic acid, talc, pectin, acacia,
agar or gelatin. The carrier may also include a sustained release
material such as glyceryl monostearate or glyceryl distearate,
alone or with a wax.
The pharmaceutical preparations are made following the conventional
techniques of pharmacy involving milling, mixing, granulation, and
compressing, when necessary, for tablet forms; or milling, mixing
and filling for hard gelatin capsule forms. When a liquid carrier
is used, the preparation will be in the form of a syrup, elixir,
emulsion or an aqueous or non-aqueous suspension. Such a liquid
formulation may be administered directly p.o. or filled into a soft
gelatin capsule.
The pharmaceutical compositions may be delivered in a
therapeutically effective amount. The precise therapeutically
effective amount is that amount of the composition that will yield
the most effective results in terms of efficacy of treatment in a
given subject. This amount will vary depending upon a variety of
factors, including but not limited to the characteristics of
therapeutic compound (including activity, pharmacokinetics,
pharmacodynamics, and bioavailability), the physiological condition
of the subject (including age, sex, disease type and stage, general
physical condition, responsiveness to a given dosage, and type of
medication), the nature of the pharmaceutically acceptable carrier
or carriers in the formulation, and the route of administration.
One skilled in the clinical and pharmacological arts will be able
to determine a therapeutically effective amount through routine
experimentation, for instance, by monitoring a subject's response
to administration of a compound and adjusting the dosage
accordingly. For additional guidance, see Remington: The Science
and Practice of Pharmacy (Gennaro ed. 20th edition, Williams &
Wilkins PA, USA) (2000).
Typical dosages of an effective anti-TL1A antibody can be as
indicated to the skilled artisan by the in vitro responses or
responses in animal models. Such dosages typically can be reduced
by up to about one order of magnitude in concentration or amount
without losing the relevant biological activity. Thus, the actual
dosage will depend upon the judgment of the physician, the
condition of the patient, and the effectiveness of therapeutic
method based, for example, on the in vitro responsiveness of the
relevant primary cultured cells or histocultured tissue sample,
such as biological samples obtained, or the responses observed in
the appropriate animal models.
For the treatment of the disease, the appropriate dosage of an
antibody depends on the type of disease to be treated, the severity
and course of the disease, the responsiveness of the disease,
whether the antibody is administered for therapeutic or
preventative purposes, previous therapy, and patient's clinical
history. The dosage can also be adjusted by the individual
physician in the event of any complication and at the discretion of
the treating physician. The administering physician can determine
optimum dosages, dosing methodologies and repetition rates. The
TL1A antibody can be administered one time or over a series of
treatments lasting from several days to several months, or until a
cure is effected or a diminution of the disease state is achieved
(e.g., treatment or amelioration of IBD). The duration of treatment
depends upon the subject's clinical progress and responsiveness to
therapy. In certain embodiments, dosage is from 0.01 .mu.g to 100
mg per kg of body weight, and can be given once or more daily,
weekly, monthly or yearly. For systemic administration, subjects
can be administered a therapeutic amount, such as, e.g. about 0.1
mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10
mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg,
or more. In certain embodiments, a therapeutic amount is selected
from about 1, 3, 10, 30, 100, 300, 600 and 800 milligrams
administered as a flat dosage. In certain embodiments, a
therapeutic amount is about 1, 2, 3, 4, 5, 6, 7, 8, or 9 milligrams
administered as a flat dosage. In certain embodiments, a
therapeutic amount is about 10, 20, 30, 40, 50, 60, 70, 80, or 90,
milligrams administered as a flat dosage. In certain embodiments, a
therapeutic amount is about 100, 150, 200, 250, 300, 350, 400, 450,
500, 550, 600, 650, 700, 750, 800, 850 or 900, milligrams
administered as a flat dosage. In certain embodiments, a
therapeutic amount is about 5 to about 30 milligrams per kilogram.
In certain embodiments, a therapeutic amount is about 5 to about 30
milligrams per kilogram dosed every week or every other week. In
certain embodiments, a therapeutic amount is about 5, 10, 15, 20,
25, or 30 milligrams per kilogram. In certain embodiments, a
therapeutic amount is about 5, 10, 15, 20, 25, or 30 milligrams per
kilogram dosed every week or every other week.
Methods of Treatment
Various embodiments provide for methods of treating inflammatory
bowel disease (IBD), comprising administering an anti-TL1A antibody
described herein to a subject in need thereof In some embodiments,
the subject comprises one or more risk genotypes. In some
embodiments, the IBD is a severe form of IBD. Severe forms of IBD
may be characterized by subclinical phenotypes described
herein.
In various embodiments, provided herein is a method of treating
inflammatory bowel disease (IBD) in a subject in need thereof,
comprising: administering to the subject a therapeutically
effective amount of an antibody or an antigen-binding fragment that
specifically binds TL1A. In some embodiments, the anti-TL1A
antibody comprises a HCFR1 comprising SEQ ID NO: 545, or a sequence
that differs from SEQ ID NO: 545 by up to about 5, 4, 3, or 2 amino
acids. In some embodiments, the anti-TL1A antibody comprises a
HCDR1 selected from SEQ ID NOs: 9, 150, 484, and 553, or a sequence
that differs from a sequence selected from SEQ ID NOs: 9, 150, 484,
and 553 by up to about 5, 4, 3, or 2 amino acids. In some
embodiments, the anti-TL1A antibody comprises a HCFR2 comprising
SEQ ID NO: 546, or a sequence that differs from SEQ ID NO: 546 by
up to about 5, 4, 3, or 2 amino acids. In some embodiments, the
anti-TL1A antibody comprises a HCDR2 selected from SEQ ID NOs: 12,
554 to 564 and 574 to 577, or a sequence that differs from SEQ ID
NOs: 12, 554 to 564 and 574 to 577 by up to about 5, 4, 3, or 2
amino acids. In some embodiments, the anti-TL1A antibody comprises
a HCFR3 comprising a sequence selected from SEQ ID NOs: 547 and 586
to 588, or a sequence that differs from a sequence selected from
SEQ ID NOs: 547 and 586 to 588 by up to about 5, 4, 3, or 2 amino
acids. In some embodiments, the anti-TL1A antibody comprises a
HCDR3 selected from SEQ ID NOs: 15, 152, 565 to 568, and578 to 581,
or a sequence that differs from a sequence selected from SEQ ID
NOs: 15, 152, 565 to 568, and578 to 581 by up to about 5, 4, 3, or
2 amino acids. In some embodiments, the anti-TL1A antibody
comprises a HCFR4 comprising SEQ ID NO: 548, or a sequence that
differs from SEQ ID NO: 548 by up to about 5, 4, 3, or 2 amino
acids. In some embodiments, the anti-TL1A antibody comprises a
LCFR1 comprising SEQ ID NO: 549, or a sequence that differs from
SEQ ID NO: 549 by up to about 5, 4, 3, or 2 amino acids. In some
embodiments, the anti-TL1A antibody comprises a LCDR1 selected from
SEQ ID NOs: 487, 569 and 570, or a sequence that differs from SEQ
ID NOs: 487, 569 and 570 by up to about 5, 4, 3, or 2 amino acids.
In some embodiments, the anti-TL1A antibody comprises a LCFR2
comprising SEQ ID NO: 550, or a sequence that differs from SEQ ID
NO: 550 by up to about 5, 4, 3, or 2 amino acids. In some
embodiments, the anti-TL1A antibody comprises a LCDR2 comprising
SEQ ID NO: 488, or a sequence that differs from SEQ ID NO: 488 by
up to about 5, 4, 3, or 2 amino acids. In some embodiments, the
anti-TL1A antibody comprises a LCFR3 comprising SEQ ID NO: 551, or
a sequence that differs from SEQ ID NO: 551 by up to about 5, 4, 3,
or 2 amino acids. In some embodiments, the anti-TL1A antibody
comprises a LCDR3 selected from SEQ ID NOs: 571 to 573 and 582 to
585, or a sequence that differs from a sequence selected from SEQ
ID NOs: 571 to 573 and 582 to 585 by up to about 5, 4, 3, or 2
amino acids. In some embodiments, the anti-TL1A antibody comprises
a LCFR4 comprising SEQ ID NO: 552, or a sequence that differs from
SEQ ID NO: 552 by up to about 5, 4, 3, or 2 amino acids.
The subject disclosed herein can be a mammal, such as for example a
mouse, rat, guinea pig, rabbit, non-human primate, or farm animal.
In some instances, the subject is human. In some instances, the
subject is a patient who is diagnosed with IBD. In some instances,
the subject is not diagnosed with the IBD. In some instances, the
subject is suffering from a symptom related to a disease or
condition disclosed herein (e.g., abdominal pain, cramping,
diarrhea, rectal bleeding, fever, weight loss, fatigue, loss of
appetite, dehydration, and malnutrition, anemia, or ulcers).
In various embodiments, the subject is not responsive to induction
of an anti-TNF therapy (e.g., adalimumab, certolizumab, etanercept,
golimumab, infliximab) (anti-TNF non-response), or loses response
to said anti-TNF therapy after a period of time during treatment
(anti-TNF loss of response). In various embodiments, the subject is
at risk for developing anti-TNF non-response or anti-TNF loss of
response. In some embodiments, the subject is treated by
administering the anti-TL1A antibody disclosed herein to the
subject, provided the subject is at risk for developing, or suffers
from, anti-TNF non-response or anti-TNF loss of response.
In various other embodiments, the subject is determined to have an
increased TL1A expression. In some embodiments, the administration
of a therapeutically effective amount of an anti-TL1A antibody
causes a decrease in TL1A in the subject treated.
Methods disclosed herein provide methods of treating an
inflammatory bowel disease (IBD) in a subject by administering an
anti-TL1A antibody described herein to the subject. In various
embodiments, IBD is Crohn's Disease (CD) or ulcerative colitis
(UC). In some embodiments, the IBD is a severe form of IBD. In some
embodiments, the severe form of IBD is characterized by a
subclinical phenotype. In some embodiments, the IBD is a moderate
to severe form of IBD. In some embodiments, the IBD is a moderate
form of IBD.
Subclinical phenotypes of IBD may include, but are not limited to,
non-stricturing, stricturing, stricturing and penetrating, and
isolated internal penetrating, disease, and perianal CD (pCD).
Stricturing is the progressive narrowing of the intestine. Internal
penetrating disease creates abnormal passageways (fistulae) between
the bowel and other structures. pCD is a form of Crohn's disease
that causes inflammation around the anus.
The IBD may be refractory. The term "medically refractory," or
"refractory," as used herein, refers to the failure of a standard
treatment to induce remission of a disease. In some embodiments,
the disease comprises an inflammatory disease disclosed herein. A
non-limiting example of refractory inflammatory disease includes
refractory Crohn's disease, and refractory ulcerative colitis
(e.g., mrUC). Non-limiting examples of standard treatment include
glucocorticosteriods, anti-TNF therapy, anti-a4-b7 therapy
(vedolizumab), anti-IL12p40 therapy (ustekinumab), Thalidomide, and
Cytoxin. In some embodiments, the UC is medically refractory UC
(mrUC). In some embodiments, the CD is refractory.
Disclosed herein are methods of administering the anti-TL1A
antibody to a subject in need thereof. In various embodiments, the
antibody is a monoclonal antibody. In some embodiments, the
antibody is a human antibody. In various embodiments, the antibody
is a humanized antibody. In various embodiments, the antibody is a
neutralizing antibody.
In various aspects, the anti-TL1A antibody is administered to the
subject for treatment of an IBD described herein. In various other
embodiments, the anti-TL1A antibody is administered in a series of
treatments. In some embodiments, the anti-TL1A antibody and a
second IBD treatment may be administered in any order or
concurrently. In selected embodiments, the anti-TL1A antibody will
be administered to patients that have previously undergone
treatment with the second IBD treatment. In certain other
embodiments, the anti-TL1A antibody and the second IBD treatment
will be administered substantially simultaneously or concurrently.
For example, a subject may be given the anti-TL1A antibody while
undergoing a course of treatment with the second IBD treatment. In
certain embodiments, the anti-TL1A antibody will be administered
within 1 year of the treatment with the second IBD treatment. In
certain alternative embodiments, the anti-TL1A antibody will be
administered within 10, 8, 6, 4, or 2 months of any treatment with
the second IBD treatment. In certain other embodiments, the
anti-TL1A antibody will be administered within 4, 3, 2, or 1 week
of any treatment with the second IBD treatment. In some
embodiments, the anti-TL1A antibody will be administered within 5,
4, 3, 2, or 1 days of any treatment with the second IBD treatment.
It will further be appreciated that the two treatments may be
administered to the subject within a matter of hours or minutes
(i.e., simultaneously).
Other IBD treatments include, but are not limited to 1)
anti-inflammatory drugs (e.g., Aminosalicylates such as, but not
limited to sulfasalazine Azulfidine, 5-aminosalicylates,
Mesalamine, Asacol, Lialda, Rowasa, Canasa, balsalazide Colazal and
olsalazine, Dipentum); 2) corticosteroids (e.g., prednisone and
hydrocortisone); 3) immune system suppressors (e.g., Azathioprine,
Azasan, Imuran, mercaptopurine, Purinethol, Purixan, Cyclosporine,
Gengraf, Neoral and Sandimmune, Infliximab, Remicade, adalimumab,
Humira, golimumab, and Simponi, tumor necrosis factor (TNF)-alpha
inhibitors (e.g., Infliximab), Methotrexate, Rheumatrex,
Natalizumab, Tysabri, vedolizumab, Entyvio, Ustekinumab and
Stelara; 4) Antibiotics (e.g., Metronidazole, Flagyl,
Ciprofloxacin, Cipro); 5) Anti-diarrheal medications (e.g., fiber
supplements--Metamucil or Citrucel) or loperamide; 6) Pain
relievers (e.g.,Tylenol, ibuprofen, naproxen sodium and diclofenac
sodium); and 7) Surgery (e.g., removal of the colon, partial
digestive tract removal, colectomy, proctocolectomy and/or
strictureplasty). In some embodiments, these IBD treatments may be
administered in combination with the anti-TL1A antibody. Treatment
with an antibody can occur prior to, concurrently with, or
subsequent to administration of an IBD treatment. Combined
administration can include co-administration, either in a single
pharmaceutical formulation or using separate formulations, or
consecutive administration in either order but generally within a
time period such that all active agents can exert their biological
activities simultaneously. Any dosing schedules for such IBD
treatments can also be used as determined by the skilled
practitioner.
In some embodiments, the second IBD treatment comprises an
antibody. Thus, treatment can involve the combined administration
of antibodies provided herein with other antibodies against
additional IBD-associated antigens, such as, but not limited to
tumor necrosis factor (TNF)-alpha. Combined administration can
include co-administration, either in a single pharmaceutical
formulation or using separate formulations, or consecutive
administration in either order but generally within a time period
such that all active agents can exert their biological activities
simultaneously.
Kits
Further provided is a kit to treat IBD (e.g., CD, UC and/or mrUC).
The kit comprises of the antibodies described herein, which can be
used to perform the methods described herein. The kit is useful for
practicing the inventive method of providing treatment to an IBD,
CD, UC and/or mrUC patient by administering an anti-TL1A antibody.
The kit is an assemblage of materials or components, including at
least one of the inventive compositions. Thus, in some embodiments,
the kit contains a composition including anti-TL1A antibodies, for
the treatment of IBD, CD, UC and/or MR-UC, as described above. In
other embodiments, the kits contain all of the components necessary
and/or sufficient to perform a detection assay for TL1A, including
all controls, directions for performing assays, and any necessary
software for analysis and presentation of results.
The exact nature of the components configured in the inventive kit
depends on its intended purpose. For example, some embodiments are
configured for the purpose of treating IBD, CD, UC and/or MR-UC. In
one embodiment, the kit is configured particularly for the purpose
of treating mammalian subjects. In another embodiment, the kit is
configured particularly for the purpose of treating human subjects.
In further embodiments, the kit is configured for veterinary
applications, treating subjects such as, but not limited to, farm
animals, domestic animals, and laboratory animals.
Instructions for use may be included in the kit. "Instructions for
use" typically include a tangible expression describing the
technique to be employed in using the components of the kit to
effect a desired outcome, such as to treat or alleviate IBD, CD, UC
and/or MR-UC. Optionally, the kit also contains other useful
components, such as, diluents, buffers, pharmaceutically acceptable
carriers, syringes, catheters, applicators, pipetting or measuring
tools, bandaging materials or other useful paraphernalia as will be
readily recognized by those of skill in the art.
The materials or components assembled in the kit can be provided to
the practitioner stored in any convenient and suitable ways that
preserve their operability and utility. For example, the components
can be in dissolved, dehydrated, or lyophilized form; they can be
provided at room, refrigerated or frozen temperatures. The
components are typically contained in suitable packaging
material(s). As employed herein, the phrase "packaging material"
refers to one or more physical structures used to house the
contents of the kit, such as inventive compositions and the like.
The packaging material is constructed by well-known methods,
preferably to provide a sterile, contaminant-free environment. The
packaging materials employed in the kit are those customarily
utilized in gene expression assays and in the administration of
treatments. As used herein, the term "package" refers to a suitable
solid matrix or material such as glass, plastic, paper, foil, and
the like, capable of holding the individual kit components. Thus,
for example, a package can be a glass vial or prefilled syringes
used to contain suitable quantities of an inventive composition
containing anti-TL1A antibodies and/or primers and probes for TL1A.
The packaging material generally has an external label which
indicates the contents and/or purpose of the kit and/or its
components.
EXAMPLES
The following examples are illustrative of the embodiments
described herein and are not to be interpreted as limiting the
scope of this disclosure. To the extent that specific materials are
mentioned, it is merely for purposes of illustration and is not
intended to be limiting. One skilled in the art may develop
equivalent means or reactants without the exercise of inventive
capacity and without departing from the scope of this
disclosure.
Example 1: Generation and Characterization of Humanized anti-TL1A
Antibodies
A murine anti-TL1A antibody was humanized to reduce potential
immunogenicity. A first variant, 12835, was generated, which
consists of murine 5C3D11 CDRs (SEQ ID NOs: 9, 554, 15, 569, 488,
24) grafted into human variable region frameworks to generate a
heavy chain variable region comprising SEQ ID NO: 26 and a light
chain variable region comprising SEQ ID NO: 28. Unfortunately,
clone 12835 contains nine (9) framework back mutations (murine
framework residues), resulting in an incompletely humanized
variant. Complete humanization is important to reduce the chances
of a subject raising an immune response to an administered
antibody. Consequently, the goal was to generate humanized
antibodies comprising fewer murine framework residues while
retaining the functional activity of the parent 12835 antibody.
Unfortunately, using visual inspection of the sequence it is not
straight-forward to distinguish murine framework residues critical
for the antibody's function from those that are non-critical, and
thus which amino acid residues can be replaced by the corresponding
human framework residue. Therefore, as a first step, 12835 was
rehumanized by CDR grafting into the closest fully human germline
frameworks (IGV1-46*02 and IGKV3-20*01 as determined by the NCBI's
igblast tool). This clone is L8 and comprises the 5C3D11 and 12835
CDRs as defined by the combination of the Kabat, Chothia, and IMGT
methods (HCDR1, GFDIQDTYMH (SEQ ID NO: 9); HCDR2, RIDPASGHTKYDPKFQV
(SEQ ID NO: 554); HCDR3, SRSGGLPDV (SEQ ID NO: 590); LCDR1,
RASSSVSYMY (SEQ ID NO: 569); LCDR2, ATSNLAS (SEQ ID NO: 488);
LCDR3, QQWSGNPRT (SEQ ID NO: 24)).
In the present study, many variants of 12835 were made and tested
in order to identify a more human-like antibody that retains the
functional activity of the parent 12835 antibody. In the first
stage, variants containing significantly fewer murine framework
residues were identified. Subsequently, CDR libraries of 12835 were
combined with a fully human germline frameworks in order to
identify multiple variants that do not contain any murine framework
residues, yet retain the functional activity and/or affinity of the
parent 12835 antibody.
Cloning of Murine 5C3D11 and Humanized Construct 12835 into Phage
Expression System
DNA encoding the heavy and light chain variable regions of both
murine 5C3D11 and humanized 12835 was cloned into a phage
expression vector that contained human kappa light chain constant
domain and human G1 heavy chain constant domain 1. In addition, the
vector contains a his-tag and hemagglutinin A tag at the
carboxy-terminal end of the heavy chain to facilitate purification
and detection. Cloning of the murine variable regions into the
phage expression vector containing human constant domains results
in the expression of chimeric 5C3D11.
The murine 5C3D11 heavy chain variable region DNA (SEQ ID NO: 1)
and light chain variable region DNA (SEQ ID NO: 4) were codon
optimized for bacterial expression to generate SEQ ID NOS: 2 and 5,
respectively. The humanized 12835 heavy chain variable region DNA
was codon optimized to generate SEQ ID NO: 25 and the light chain
variable region DNA was codon optimized to generate SEQ ID NO:
27.
Expression and Quantitation of Fab in the Periplasmic Space of E.
coli.
Cloning was verified by expressing and quantitating Fab in the
periplasmic space of E. coli. Briefly, XL-0 bacteria were grown in
2X YT medium at 37.degree. C. until the culture reached a density
of 0.9-1.1 at OD600. Isopropyl .beta.-D-thiogalactoside was then
added to the cells to a final concentration of 1 mM and 3.0 mL of
culture was transferred to a 14 mL snap-top tube. Each tube was
transfected with 25 uL of high titer phage stock and the cultures
were placed in a shaker (225 rpm) at 37.degree. C. One hour later,
the temperature was shifted to 25.degree. C. and the cultures were
grown for an additional 14-16 h. The cells were collected by
centrifugation at 3900 rpm for 30 min in an Eppendorf 5810R
centrifuge (.about.3,200.times.g), the supernatant was decanted and
the cells were resuspended in 0.3 mL of lysis buffer (30 mM Tris,
pH 8.0, 2 mM EDTA, 20% sucrose, 2 mg/ml lysozyme, 5 U/mL DNase I)
and placed on ice for 15 min. The cell suspension was transferred
to a 1.5 mL tube and cell debris was pelleted by centrifugation at
15,000 rpm for 15 min in an Eppendorf 5424 microfuge
(.about.21,000.times.g). The supernatant was removed carefully
without disturbing the pellet and was stored at 4.degree. C. until
use.
In order to quantitate Fab expression, a 96-well Costar-3366 plate
was coated with 50 .mu.l/well of 2 .mu.g/ml sheep anti-human Fd
(Southern Biotech, Prod. #2046-01, Lot # A7212-VJO6) in PBS
overnight at 4.degree. C. The plate was washed three times with PBS
containing 0.05% Tween 20 (PBS-T) and 50 .mu.l/well of sample
dilutions was added. Sample dilutions were performed with PBS-T. A
standard curve was generated using human Fab (Rockland, Prod.
#009-01015, Lot # 38543) diluted serially 3-fold, beginning at 500
ng/ml. The plates were incubated 1 h at 25.degree. C., washed three
times with PBS-T, and incubated with 50 .mu.l/well of anti-kappa
HRP conjugate (Southern Biotech, Prod. #2060-05, Lot #
K3114-S506B), diluted 10,000-fold in PBS-T for 1 h at 25.degree. C.
The plate was washed three times with PBS-T, developed with 50
.mu.l/well 1-Step Ultra TMB-ELISA (Thermo Scientific, Prod. #34028,
Lot # SF2405221). The reaction was terminated by the addition of 2
N H.sub.2SO.sub.4 and the A650 and A450 were determined before and
after addition of H.sub.2SO.sub.4, respectively, using a Spectramax
plate reader.
Characterization of Chimeric 5C3D11 and 12835--Filter Lift
Assay
A filter lift assay was developed to facilitate characterization of
heavy and light chain expression and to verify functional activity
of the Fab constructs through binding to biotinylated antigen. With
filter lift assays bacterial lawns are infected with phage under
conditions where each phage produces a distinct plaque (zone of
slower growing bacteria). Nitrocellulose filters are placed on the
lawns, capturing expressed Fab. Subsequently, the filters can be
probed with biotinylated antigen and/or reagents directed against
immunoglobulins or peptide tags.
Dilutions of high titer phage stocks (typically 10.sup.6-fold) were
combined with 0.35 ml of a confluent E. coli strain XL culture and
20 .mu.g/ml tetracycline. The mixture was combined with 3.5 ml top
agar (0.7% Bacto-agar in Luria broth) and overlaid on an LB agar
plate (1.5% Bacto-agar in Luria broth). The plate was incubated 6-8
h at 37.degree. C. at which time a nitrocellulose filter (Whatman
82-mm diameter, 0.45 .mu.m pore size, GE Healthcare, Prod.
#10401116) was placed on top and the plate is incubated at
25.degree. C. for 12-15 h. The filter was removed, rinsed briefly
in PBS, and transferred to 5% M-P blocking solution for 2 h at
25.degree. C. with constant agitation.
Subsequently, the filter was cut into three sections: one to assess
light chain expression, one to assess heavy chain expression, and
one to assess antigen binding. Each section was transferred to the
primary detection reagent: goat anti-human kappa, HRP conjugate
(Southern Biotech, Prod. #2060-05, Lot # K3114-S506B) diluted
1000-fold in 5% M-P for detection of light chain, rat anti-HA, HRP
conjugate (Roche, Prod. #12013819001) diluted 1000-fold in 5% M-P
for detection of heavy chain, or biotinylated antigen at the
desired concentration in 5% M-P.
In order to label antigen with biotin, 500 .mu.g of human TL1A
(Fitzgerald, Prod. #30R-AT070, Lot # A13102302) was resuspended in
water to 1 mg/ml. Following suspension in water the protein was in
10 mM Tris, pH 8.5 with 75 mM arginine. The Tris and arginine were
removed by buffer exchange using a 7K MW cut-off, 5 ml Zeba spin
desalting column (Thermo Prod. #89891) that had been equilibrated
with 10 mM phosphate buffer, pH 8.0 with 65 mM NaCl. After
recovering the protein, it was immediately biotinylated by
combining it with EZ-Link Sulfo-NHS-LC-Biotin (Thermo Prod. #21327)
at a 5:1 molar at 25.degree. C. for 30 min. The reaction was
terminated by the addition of 750 mM arginine to achieve a final
concentration of 75 mM. The reaction was transferred to ice and
stored at 4.degree. C.
Filters were incubated 2 h at 25.degree. C. with constant
agitation, washed 5 times in PBS-0.05% Tween 20 (2 minutes each
wash with constant agitation). The filters probed with biotinylated
antigen were transferred to 10 ml of High Sensitivity Neutravidin,
HRP conjugate (Thermo Scientific, Prod. #31030) diluted 5000-fold
in 1% BSA in PBS and incubated for 1 h at 25.degree. C. The filters
were then washed 5 times in PBS-0.05% Tween 20 (2 minutes each wash
with constant agitation). All filters were developed with 1-Step
Ultra TMB-Blotting (Thermo Scientific, Prod. #37574).
Using this approach, expression of heavy chain (FIG. 1A) and light
chain (FIG. 1B) was demonstrated. Furthermore, when the filter was
probed with 8 nM biotinylated human TL1A staining was observed
(FIG. 1C) indicating the bacteria were expressing functional
Fab.
Characterization of Fab Binding by ELISA
The filter lift assay provides a qualitative assessment of antigen
binding. In order to compare the binding activity of chimeric
5C3D11 with the humanized construct 12835 in a more quantitative
fashion an ELISA was developed. A 96-well Costar-3366 plate was
coated with 50 .mu.l/well of 2 .mu.g/ml human TL1A (Fitzgerald,
Prod. #30R-AT070, Lot # A13102302) in PBS overnight at 4.degree. C.
The plate was rinsed once with PBS-T and blocked with 100
.mu.l/well of 1% BSA in PBS (1% BSA) for 1 h at 25.degree. C. Fab
samples were serially diluted 3-fold using 1% BSA and were
incubated for 1 h at 25.degree. C. (50 .mu.l/well). The plate was
washed three times with PBS-T and 50 .mu.l/well anti-human kappa,
HRP conjugate (Southern Biotech, Prod. #2060-05, Lot # K3114-5506B)
diluted 10,000-fold in 1% BSA was added for 1 h at 25.degree. C. In
certain assays (extended wash format), the plate was placed in
large volumes (up to 1L) of PBS-T and incubated, with mixing, for
2-5 hours prior to the addition of anti-human kappa, HRP conjugate.
The plate was washed three times with PBS-T, developed with 50
.mu.l/well 1-Step Ultra TMB-ELISA (Thermo Scientific, Prod. #34028,
Lot # SF2405221). The reaction was terminated by the addition of 2
N H.sub.2SO.sub.4 and the A650 and A450 were determined before and
after addition of H.sub.2SO.sub.4, respectively, using a Spectramax
plate reader. For measuring binding to murine TL1A the same
protocol was used, but the plates were coated with 2 .mu.g/ml
murine TL1A (BioLegend, Prod. #753004, Lot # B204691) and the Fab
samples were serially diluted 2-fold.
The binding activity of chimeric 5C3D11 Fab was compared to
humanized 12835 Fab (FIG. 2). Although the binding activities of
5C3D11 and humanized 12835 in an IgG format (bivalent) appeared
similar, it was observed that the binding activity of 12835 in the
Fab format (monovalent) was somewhat diminished compared to
chimeric Fab (FIG. 2). It is likely that this discrepancy reflects
differences in true affinity (monovalent format) versus similar
avidities (bivalent format). Using the monovalent assay format, the
chimeric Fab appears to be 2- to 3-fold higher affinity than
humanized 12835 Fab.
Capture Lift Assay
A nitrocellulose filter (Whatman 82-mm diameter, 0.45 .mu.m pore
size, GE Healthcare, Prod. #10401116) was floated on top of 10 ml
of 10 mg/ml goat anti-human kappa (Southern Biotech Prod. #2060-01)
for 2 h at 25.degree. C. The filter was submersed briefly before
removing and transferred to 10 ml of 5% M-P for 2 h at 25.degree.
C. The filter was removed from 5% M-P, rinsed briefly one time with
PBS, and was air-dried. Subsequently, the filter was processed in
the same way as the filter lift assay described above, with minor
modifications. Briefly, dilutions of high titer phage stocks
(typically 10.sup.6-fold) were combined with 0.35 ml of a confluent
E. coli strain XL culture and 20 .mu.g/ml tetracycline. The mixture
was combined with 3.5 ml top agar (0.7% Bacto-agar in Luria broth)
and overlaid on an LB agar plate (1.5% Bacto-agar in Luria broth).
The plate was incubated 6-8 h at 37.degree. C. at which time the
pre-treated nitrocellulose filter (described above) was placed on
top and the plate was incubated for 12-15 h at 25.degree. C. The
filter was removed, rinsed briefly in PBS, and transferred to
biotinylated antigen at the desired concentration in 5% M-P.
Filters were incubated 2 h at 25.degree. C. with constant
agitation, washed 5 times in PBS-0.05% Tween 20 (2 minutes each
wash with constant agitation) and were transferred to 10 ml of High
Sensitivity Neutravidin, HRP conjugate (Thermo Scientific, Prod.
#31030) diluted 5000-fold in 1% BSA in PBS and incubated for 1 h at
25.degree. C. The filters were then washed 5 times in PBS-0.05%
Tween 20 (2 minutes each wash with constant agitation). All filters
were developed with 1-Step Ultra TMB-Blotting (Thermo Scientific,
Prod. #37574). The developed filter as shown in FIG. 3 demonstrates
high sensitivity and avidity of 5C3D11 to TL1A.
Removal of Murine Framework Residues from 12835 to Identify
Multiple Active Humanized Clones, Including 18-7 and 21-3
Murine framework residues were removed using Kunkel mutagenesis
(Kunkel TA 1985. PNAS 82:488-492). Briefly, single strand M13
plasmid was isolated and primed for DNA replication with mutagenic
oligonucleotide(s) encoding the human instead of the murine
framework residue. After extension to complete the circle,
transformation of bacteria resulted in a mixture of wild type
(murine framework residue unmated) and mutated (human framework
residue) plasmids. Mutagenesis was performed at multiple sites
simultaneously to generate small combinatorial libraries containing
mixtures of clones containing various combinations of murine and
human framework mutations. Subsequently, the mixtures were plated
and screened by capture lift to identify the most active framework
combinations. Library clones were characterized by DNA
sequencing.
Fab was expressed in E. coli, quantitated by ELISA, and binding
activity was assessed by ELISA by titrating against immobilized
antigen. Expression of Fab, isolation of the periplasmic fraction,
quantitation of Fab expression and binding to antigen by ELISA were
all performed as described above.
Using this approach, multiple active clones containing differing
numbers of murine framework residues were identified. Examples of
active clones with varying numbers and positions of murine
framework residues are summarized in Table 3.
TABLE-US-00003 TABLE 3 Active humanized 5C3D11 constructs with
varying amounts of framework back mutations Mu FR Back Clone Light
Chain Heavy Chain Mutations 12835 V19 M21 P46 W47 V58 Y71 L20 T71
S93 9 1-3 V58I 8 5-2 T71R 8 22 Y71F 8 5-4 V19A M21L 7 1-4 P46L V58I
7 2-3 V19A M21L V58I 6 3-4 V19A M21L Y71F 6 5-1 V19A M21L T71R 6
7-4 V19A M21L P46L 6 26 V19A M21L L20V 6 9-1 V19A M21L Y71F T71R 5
13-2 V19A M21L T71R S93A 5 21 V19A M21L P46L V58I 5 27 V19A M21L
W47L L20V 5 10-1 V19A M21L P46L W47L T71R 4 19 V19A M21L P46L W47L
V58I 4 16-2 V19A M21L P46L W47L V58I T71R 3 17-2 V19A M21L P46L
W47L Y71F T71R 3 18-7 V19A M21L P46L W47L V58I Y71F T71R 2 19-5
V19A M21L P46L W47L L20V T71R 3 20-7 V19A M21L P46L W47L T71R S93A
3 21-3 V19A M21L P46L W47L L20V T71R S93A 2
Synthesis of CDR-Grafted Construct
Two of the clones identified, 18-7 and 21-3, contained only two
murine framework back mutations. The light chain of clone 18-7 did
not contain any murine framework residues while the heavy chain of
clone 21-3 did not contain any murine framework residues. Screening
of the framework combinatorial libraries did not identify a
CDR-grafted variant (no murine framework residues on both the heavy
and light chains). For comparison, the CDR-grafted variant was
synthesized using Kunkel mutagenesis and compared its binding
activity was characterized by ELISA. Although the CDR grafted
construct bound antigen the humanized 12835 variant consistently
displayed stronger binding to antigen (FIG. 4B).
After making the back mutations of murine framework residues as
indicated in Table 3, the heavy chain variable region frameworks
1-3 were identical to human germlines IGHV1-46*01, IGHV1-46*02, and
IGHV1-46*03 while the light chain variable region frameworks were
identical to human germline IGKV3-20*01.
In addition, different back-mutations were introduced into the
third framework of the heavy chain variable region such that the
new heavy chain variable regions were homologous to human germline
IGHV1-3*01 (see VH SEQ IDs 519, 521, 523, 525, 527, 529, 531, 533,
535, 537, 539, and 541). Collectively, clones containing these
back-mutations are referred to as alternative framework
variants.
Characterization of Humanized Variants in an Alternative ELISA
Format
Multiple Fab variants were characterized by ELISA, using an
alternative format that permits the rapid and direct comparison
(single-well determination with no dilution series required) of the
relative affinities of different Fab clones isolated from different
cultures, regardless of the relative expression levels of the
clones (Watkins et al. 1997, Analytical Biochemistry 253). This
assay enables a more quantitative comparison of the relative
binding strengths of the variants as the plate is saturated with
the different Fabs despite their different expression levels. Thus,
slight differences in binding profiles caused by variation in the
Fab quantitation assay are eliminated. Briefly, a 96-well
Costar-3366 plate was coated with 50 .mu.l/well of 2 .mu.g/ml goat
anti-human kappa (Southern Biotech Prod. #2060-01) for 2 h at
25.degree. C., washed once with PBS-0.05% Tween 20, and incubated
with 50 .mu.l/well of sample Fab for 2 h at 25.degree. C. The plate
was washed 4 times with PBS-0.05% Tween 20 and incubated with 50
.mu.l/well serial dilutions of biotinylated antigen for 2 h at
25.degree. C. Preparation of biotinylated antigen was described
above. The plate was washed 4 times with PBS-0.05% Tween 20 and
incubated with 50 .mu.l/well of high sensitivity Neutravidin, HRP
conjugate (Thermo Scientific, Prod. #31030) diluted 5000-fold in 1%
BSA in PBS for 1 h at 25.degree. C. The plate was washed three
times with PBS-T, developed with 50 .mu.l/well 1-Step Ultra
TMB-ELISA (Thermo Scientific, Prod. #34028, Lot # SF2405221). The
reaction was terminated by the addition of 2 N H.sub.2SO.sub.4 and
the A650 and A450 were determined before and after addition of
H.sub.2SO.sub.4, respectively, using a Spectramax plate reader.
The chimeric Fab binds antigen more strongly than the CDR grafted
Fab in the alternative ELISA format (FIG. 5, compare open circles
with closed circles). The humanized 12835 clone has slightly
diminished binding compared to the chimeric (compare open triangles
with open circles), followed by clone 18-7 and 21-3. The binding of
clone 21-3 was most similar to the CDR-grafted variant, suggesting
that one of the murine heavy chain framework back mutations may be
important for maintaining full binding activity with the parental
(wild-type) CDRs.
Example 2: Generation and Characterization of Anti-TL1A Antibodies
having Optimized CDRs
In order to identify CDR mutations that could restore and improve
the binding activity of the CDR grafted construct (fully human,
germline frameworks) each position of all six CDRs (LCDR1, LCDR2,
LCDR3, HCDR1, HCDR2, and HCDR3) was mutagenized individually by
Kunkel mutagenesis using degenerate oligonucleotides in which the
codon encoding the target amino acid was replaced with NNK.
Initially, one library was synthesized at each position of HCDR3
and LCDR3. These positional libraries, with a theoretical diversity
of 32 codons/20 amino acids/1 stop codon, were screened by capture
lift. In some cases, each position was screened by itself
(theoretical diversity of library equals 32) while in other cases
the positions of a particular CDR were pooled and screened as a CDR
library (theoretical diversity of library equals 32 times the
number of positions pooled; e.g. HCDR3 consists of 7 positions, so
the theoretical library size was 32.times.7 equals 224 members).
The libraries were screened at concentrations of biotinylated human
TL1A ranging from 15 to 1,000 pM. Positive plaques were picked and
sequenced. Fab was expressed, isolated from periplasmic fraction,
and characterized by ELISA, as described previously. Summaries of
the capture lift screening, DNA sequencing, and relative binding
activities by ELISA for some of the initial screens of LCDR3 and
HCDR3 are shown in Tables 4 and 5, respectively. The results of
more exhaustive capture lift screening of all six CDRs are
summarized in Tables 6 to 12.
In addition, LCDR3 and HCDR3 libraries constructed using an
alternative heavy chain variable region framework were made and
screened by capture lift (Tables 13 and 14). Screening CDR
libraries constructed on the alternative framework identified some
mutations that were identified on the VH1-46*01 framework, but also
identified novel mutations not previously identified (heavy chain
CDR3 L98S, V102H, V102F and light chain CDR3 S92A, S92F, and S92Y,
for example).
TABLE-US-00004 TABLE 4 Light Chain CDR3 Positional Scan (screened
with 200 pM antigen) Relative ELISA Activity Amino CL CL SEQS
Mutations (Strongest to Pos. Acid Plaques Hits Picks Acquired
(frequency) weakest binders) 1 Q89 374 19 12 10 H (3) Q = N > H,
S N (3) S (2) Q (2) 2 Q90 322 4 4 2 Q (2) 3 W91 234 0 6 (random) 1
S (1) W >>> S (inactive) 4 S92 212 44 12 11 E (4) D, E
> H, N, Q > S D (2) Q (2) N (1) V (1) H (1) 5 G93 168 22 12
11 S (3) G = A > D > Q, S A (1) D (1) Q (1) G (5) 6 N94 224 2
2 1 N (1) 7 P95 160 10 10 10 P (10) 8 R96 202 12 12 9 R (9) 9 T97
662 108 12 12 S (4) T > S
TABLE-US-00005 TABLE 5 Heavy Chain CDR3 Positional Scan (screened
with 500 pM antigen) Relative ELISA Activity Amino CL CL SEQS
Mutations (Strongest to Pos. Acid Plaques Hits Picks Acquired
(frequency) weakest binders) 1 S95 228 29 12 12 L (1) S
>>> L (inactive) S (11) 2 G96 272 6 6 5 A (1) G > A G
(4) 3 G97 188 7 7 7 G (7) 4 L98 192 23 12 10 M (2) L = M > A A
(2) L (6) 5 P99 396 58 12 11 P (11) 6 D101 165 2 2 1 E (1) D > E
7 V102 >300 57 12 12 M (5) M, K, Q, W > V = T K (2) R (1) S
(1) T (1) Q (1) W (1)
TABLE-US-00006 TABLE 6 Heavy Chain CDR1 Screening Summary (L8
template; 1-46*02 framework) SEQ [b- ID TL1A] 26 27 28 29 30 31 32
33 34 35 NO: CDR (pM) G F D I Q D T Y M H 9 SEQ 500 G F E I Q D T Y
M H 591 GFEIQD TYMH 500 G F D P Q D T Y M H 592 GFDPQD TYMH 500 G F
D V Q D T Y M H 593 GFDVQD TYMH 500 G F D I G D T Y M H 594 GFDIGD
TYMH 500 G F D I S D T Y M H 595 GFDISD TYMH 500 G F D I V D T Y M
H 596 GFDIVD TYMH 500 G F D I Q D A Y M H 597 GFDIQD AYMH 500 G F D
I Q D S Y M H 598 GFDIQD SYMH 500 G F D I Q D T F M H 599 GFDIQD
TFMH 500 G F D I Q D T Y I H 600 GFDIQD TYIH 150 G F D L Q D T Y M
H 601 GFDLQD TYMH 150 G F D P Q D T Y M H 592 GFDPQD TYMH 150 G F D
I S D T Y M H 595 GFDISD TYMH 150 G F D I Q D T Y I H 600 GFDIQD
TYIH 150 G F D I Q D T Y L H 602 GFDIQD TYLH
TABLE-US-00007 TABLE 7 Heavy Chain CDR2a Screening Summary (L8
template; 1-46*02 framework) SEQ [b- ID TL1A1 50 51 52 52a 53 54 55
56 57 58 NO: CDR (pM) R I D P A S G H T K 603 SEQ 200 R L D P A S G
H T K 604 RLDP ASGH TK 200 R I E P A S G H T K 605 RIEP ASGH TK 200
R I D P E S G H T K 606 RIDP ESGH TK 200 R I D P A S G H T K 603
RIDP ASGH TK 200 R I D P A G G H T K 607 RIDP AGGH TK 200 R I D P A
S A H T K 608 RIDP ASAH TK 200 R I D P A S G H I K 609 RIDP ASGH IK
200 R I D P A S G H L K 610 RIDP ASGH LK 200 R I D P A S G H V K
611 RIDP ASGH VK
TABLE-US-00008 TABLE 8 Heavy Chain CDR2b Screening Summary (various
templates; 1-46*02 framework) SEQ [b- ID TL1A] 59 60 61 62 63 64 65
NO: CDR Tem- (pM) Y D P K F Q V 612 SEQ plate 100 I D P K F Q V 613
IDPK 17V FQV 100 L D P K F Q V 614 LDPK 20L FQV 100 M D P K F Q V
615 MDPK 20EL FQV 100 S D P K F Q V 616 SDPK 20L FQV 100 T D P K F
Q V 617 TDPK 20L FQV 100 V D P K F Q V 618 VDPK 20EL FQV 100 Y I P
K F Q V 619 YIPK 20EL FQV 100 Y N P K F Q V 620 YNPK 17V, FQV 20EL
100 Y R P K F Q V 621 YRPK 17R FQV 100 Y S P K F Q V 622 YSPK 20EL
FQV 100 Y D P K F R V 623 YDPK 6EV FRV 100 Y D P K F Q A 624 YDPK
6EI, FQA 17L 100 Y D P K F Q D 625 YDPK 17L FQD 100 Y D P K F Q E
626 YDPK 20EL FQE 100 Y D P K F Q G 627 YDPK 17V FQG 100 Y D P K F
Q H 628 YDPK 20EL FQH 100 Y D P K F Q K 629 YDPK 17L, FQK 17I 100 Y
D P K F Q L 630 YDPK 17V FQL 100 Y D P K F Q M 631 YDPK 20EL FQM
100 Y D P K F Q N 632 YDPK 20EL FQN 100 Y D P K F Q P 633 YDPK 17L,
FQP 17I, 17V 100 Y D P K F Q R 634 YDPK 17L, FQR 17I 100 Y D P K F
Q S 635 YDPK 17L FQS 100 Y D P K F Q T 636 YDPK 17L, FQT 17I
TABLE-US-00009 TABLE 9 Heavy Chain CDR3 Screening Summary (L8
template; 1-46*02 framework) SEQ [b- ID TL1A] 95 96 97 98 99 101
102 NO: CDR (pM) S G G L P D V 15 SEQ 500 L G G L P D V 637 LGG LPD
V 500 S A G L P D V 638 SAG LPD V 500 S G G A P D V 639 SGG APD V
500 S G G M P D V 640 SGG MPD V 500 S G G L P E V 641 SGG LPE V 500
S G G L P D K 642 SGG LPD K 500 S G G L P D M 643 SGG LPD M 500 S G
G L P D Q 644 SGG LPD Q 500 S G G L P D R 645 SGG LPD R 500 S G G L
P D S 646 SGG LPD S 500 S G G L P D T 647 SGG LPD T 500 S G G L P D
W 648 SGG LPD W
TABLE-US-00010 TABLE 10 Light Chain CDR1 Screening Summary (L8
template; paired with heavy chain 1-46*02 framework) SEQ [b- ID
TL1A] 24 25 26 27 29 30 31 32 33 34 NO: CDR (pM) R A S S S V S Y M
Y 569 SEQ 150 G A S S S V S Y M Y 570 GAS SSV SYM Y 150 W A S S S V
S Y M Y 649 WAS SSV SYM Y 150 R A S S S V I Y M Y 650 RAS SSV IYM Y
150 R A S S S V S F M Y 651 RAS SSV SFM Y 150 R A S S S V S Y L Y
652 RAS SSV SYL Y 150 R A S S S V S Y M R 653 RAS SSV SYM R
TABLE-US-00011 TABLE 11 Light Chain CDR2 Screening Summary (L8
template; paired with heavy chain 1-46*02 framework) SEQ [b- ID
TL1A] 50 51 52 53 54 55 56 NO: CDR (pM) A T S N L A S 488 SEQ 150 A
K S N L A S 654 AKSN LAS 150 A T P N L A S 655 ATPN LAS 150 A T E N
L A S 656 ATEN LAS 150 A T S L L A S 657 ATSL LAS 150 A T S P L A S
658 ATSP LAS 150 A T S N L T S 659 ATSN LTS
TABLE-US-00012 TABLE 12 Light Chain CDR3 Screening Summary (various
templates; paired with heavy chain 1-46*02 framework) SEQ [b- ID
TL1A] 89 90 91 92 93 94 95 96 97 NO: CDR Tem- (pM) Q Q W S G N P R
T 24 SEQ plate 200 H Q W S G N P R T 660 HQW L8 SGN PRT 200 N Q W S
G N P R T 661 NQW L8 SGN PRT 200 S Q W S G N P R T 582 SQW L8 SGN
PRT 200 Q Q S S G N P R T 662 QQS L8 SGN PRT 200 Q Q W D G N P R T
663 QQW L8 DGN PRT 200 Q Q W E G N P R T 572 QQW L8 EGN PRT 200 Q Q
W H G N P R T 664 QQW L8 HGN PRT 200 Q Q W N G N P R T 665 QQW L8
NGN PRT 200 Q Q W Q G N P R T 573 QQW L8 QGN PRT 200 Q Q W V G N P
R T 666 QQW L8 VGN PRT 200 Q Q W S A N P R T 667 QQW L8 SAN PRT 200
Q Q W S D N P R T 668 QQW L8 SDN PRT 200 Q Q W S Q N P R T 669 QQW
L8 SQN PRT 200 Q Q W S S N P R T 670 QQW L8 SSN PRT 500 Q Q W S G N
P R S 583 QQW L8 SGN PRS 500 Q Q F S G N P R T 671 QQF 16 SGN PRT
500 Q Q H S G N P R T 672 QQH 46 SGN PRT 500 Q Q I S G N P R T 673
QQI 16 SGN PRT 500 Q Q P S G N P R T 674 QQP 16 SGN PRT 500 Q Q R S
G N P R T 675 QQR 46 SGN PRT 500 Q Q Y S G N P R T 676 QQY 46 SGN
PRT 500 Q Q W S G H P R T 677 QQW 16, SGH PRT 46 500 Q Q W S G L P
R T 678 QQW 46 SGL PRT 500 Q Q W S G Q P R T 679 QQW 46 SGQ PRT 500
Q Q W S G S P R T 86 QQW 16, SGS PRT 46 500 Q Q W S G T P R T 76
QQW 46 SGT PRT 500 Q Q W S G M P R T 680 QQW 16, SGM PRT 46 500 Q Q
W S G F P R T 80 QQW 46 SGF PRT 500 Q Q W S G K P R T 82 QQW 46 SGK
PRT 500 Q Q W S G R P R T 84 QQW 46 SGR PRT 1000 Q Q W S G D P R T
78 QQW L8 SGD PRT 1000 Q Q W S G T P R T 76 QQW L8 SGT PRT
TABLE-US-00013 TABLE 13 Heavy Chain CDR3 Screening Summary (L8mod
template; heavy chain 1-3*01 related framework) SEQ [b- ID TL1A] 95
96 97 98 99 101 102 NO: CDR (pM) S G G L P D V 15 SEQ 15 S G G L P
D H 681 SGGL PDH 15 S G G L P D R 645 SGGL PDR 15 S G G L P D F 682
SGGL PDF 15 S G G L P D V 15 SGGL PDV 15 S G G S P D V 683 SGGS
PDV
TABLE-US-00014 TABLE 14 Light Chain CDR3 Screening Summary (L8mod
template; paired with heavy chain 1-3*01 related framework) SEQ [b-
ID TL1A 89 90 91 92 93 94 95 96 97 NO: CDR (pM) Q Q W S G N P R T
24 SEQ 15 Q Q W V G N P R T 666 QQWVG NPRT 15 Q Q W A G N P R T 684
QQWAG NPRT 15 Q Q W Y G N P R T 685 QQWYG NPRT 15 Q Q W S G N P R T
24 QQWSG NPRT 15 Q Q W F G N P R T 686 QQWFG NPRT 15 Q Q W Q G N P
R T 573 QQWQG NPRT 15 Q Q W S Q N P R T 669 QQWSQ NPRT
Example 3. Generation and Characterization of Anti-TL1A Antibodies
having Optimized CDRs using a Mutated (S93) Heavy Chain
Clones containing the CDR-grafted heavy chain (CDR-grafted and
21-3) had lower binding activity than clones that contained the
murine back mutation (S) at heavy chain position 93. For this
reason, an additional HCDR3 library was constructed. The library
was constructed as described above except the degenerate
oligonucleotides for all positions were mixed prior to mutagenesis
and the library was synthesized and expressed as a pool, as opposed
to examining each position separately. Similar to the HCDR3
positional scanning performed on the L8 heavy chain backbone
(S93A), position 102 of HCDR3 yielded multiple mutations that
enhanced antigen binding in the capture lift format. Screening the
HCDR3 library based on heavy chain S93 template identified some
mutations that were identified on the heavy chain A93 template, but
also identified novel mutations not previously identified (heavy
chain CDR3 V102I and V102Y, for example). A summary of the capture
lift screening and DNA sequencing is shown in Table 15.
TABLE-US-00015 TABLE 15 Capture lift screening of HCDR3 library
pool on heavy chain template with S93. Mutation Frequency V102K 10
V102M 7 V102Y 4 V102L 2 V102I 1 V102E 1 V102T 1
Example 4. Identification and Engineering of Potential Sequence
Liabilities
A structural homology model of the variable region of CDR-grafted
construct L8, based on known PDB antibody structures, was built
using Molecular Operating Environment (MOE) 2018.01 software
(Chemical Computing Group, Montreal Canada). The model and BioMOE
prediction algorithms were used to perform a sequence liability
assessment. In addition, variants 12835 and L8 were analyzed for
potential sequence liabilities based on known potentially labile
sequence motifs (Jarasch et al., J. Pharm. Sci, 104:1885-1898
(2015); Sydow et al., PLOS ONE 9:e100736 (2014), Vlasak and Ionescu
mnAbs 3:253-263 (2011)). Using these approaches, multiple residues
were identified as potentially labile, including: light chain M33,
W35, W47, W91 and N94; heavy chain D31T32, M34, D52P52a, M69 and
W103 (summarized in Table 17; FR designates framework).
Exhaustive capture lift screening at all potentially labile sites
located within CDRs was performed to identify amino acid
substitutions that can eliminate these residues while preserving
antigen binding. As an example, one screen was focused on light
chain CDR3 N94 (potential deamidation site). The initial capture
lift screen of LCDR3 positions 94 did not identify mutations that
displayed enhanced affinity relative to the wild-type sequence.
Therefore, in order to identify acceptable mutations to eliminate
these potentially labile residues the conditions of the initial
capture lift screen were altered. Specifically, instead of
screening with antigen at a concentration where the wild-type
sequence did not provide a signal the antigen concentration was
raised so that clones expressing the wild-type sequence were
visible on the lift. In this way, variants that bind with
affinities similar to the wild-type sequence, but eliminate the
problematic residue, can he identified.
For the LCDR3 position 94 library, approximately 3,000 clones were
plated and assayed by capture lift. The capture was screened using
1000 pM human TL1A and plaques displaying the eight darkest and six
lighter staining intensities were picked and sequenced. The results
are summarized in Table 16. As expected, two of the darkest
staining plaques expressed the wild-type residue, N94, However, the
other six dark staining plaques expressed T94, indicating that the
variant N94T largely preserves the binding affinity of the
wild-type sequence while eliminating the potential deamidation
site. In addition, five different sequences were identified from
the lighter staining plaques. These included D, F, K, R and S.
Although these alternative sequences may be somewhat lower in
affinity than the wild-type sequence, all may also serve as
replacements for N94 when combined with other, higher affinity
mutations identified elsewhere.
TABLE-US-00016 TABLE 16 Identification of Alternative Residues at
Potentially Labile Site LCDR3 N94 (screened with 1000 pM antigen)
Capture Lift Nucleotide Amino acid Clone Staining Sequence DNA
Sequence Protein L8 Dark AAT -- N -- L3-6-01, -04 Dark ACT T SEQ ID
76 L3-6-02, -07 Dark AAT -- N -- L3-6-03, -05, Dark ACG T SEQ ID 76
-06, -08 L3-6-09 Light GAT D SEQ ID 78 L3-6-10 Light TTT F SEQ ID
80 L3-6-11, -12 Light AAG K SEQ ID 82 L3-6-14 Light CGG R SEQ ID 84
L3-6-15 Light TCT S SEQ ID 86
TABLE-US-00017 TABLE 17 Summary of potentially labile residues and
active variants that eliminate the lability Residue Chain
(Location) Alternative(s) Comments Light M33 (CDR1) L W35 (FR2) W47
(FR2) I Changed to I to match human germline sequence W91 (CDR3)
N94 (CDR3) D, F, K, R, S, T; H, L, M, Q Heavy D31T32 (CDR1) A32 M34
(CDR1) I, L D52P52a (CDR2) E52 M69 (FR3) I69 Removed in all
variants based on alternative heavy chain framework VH1-3*01 W103
(FR4)
Example 5. Identification of Mutations that Confer Enhanced
Expression in E. coli
Certain mutations identified during the screening of CDR libraries
by capture lift did not always demonstrate enhanced binding in the
ELISA format, but consistently expressed soluble Fab in the
periplasmic space of bacteria at higher levels than other variants.
In particular, this phenomenon was observed at heavy chain CDR2
position V65 (V65G, V65T, and V65K) and light chain CDR1 position
R24 (R24G). These results were surprising because the capture lift
screening format is configured to minimize the impact of different
expression levels, while maximizing the impact of affinity on the
signal intensity. Consequently, these mutations were noted and
integrated into later combinatorial libraries that included
mutations that enhanced affinity in order to determine if the
mutations would confer expression and/or thermostability benefits
to candidates expressed as intact immunoglobulins in mammalian
expression systems.
Example 6. Generation and Characterization of Anti-TL1A Antibodies
having Combinatorial HCDR3 and LCDR3 Mutations
Based on the initial identification of beneficial mutations in both
HCDR3 and LCDR3 an additional library was synthesized, expressed,
and screened to identify combinations of independent mutations that
could further improve the binding affinity The library was
constructed by two site mutagenesis using oligonucleotides encoding
a subset of the mutations identified in the positional scanning.
Oligonucleotides encoding the wild-type residue were also included.
This combinatorial library contained 30 distinct variants:
wild-type (no mutations), 9 variants containing a single mutation
(redundant with variants identified in positional screen, as shown
in Tables 4 and 5), and 20 unique combinations. Capture lift
screening with 200 pal antigen identified 21 active clones. DNA
sequencing of the 21 clones identified certain combinations more
frequently than others (Table 18).
TABLE-US-00018 TABLE 18 Combinatorial library screening and DNA
sequence summary HCDR3 V102X M K Q W V LCDR3 D 4 4 0 0 0 S92X E 0 1
0 1 0 H 0 0 0 0 0 N 0 4 0 1 0 Q 1 0 1 0 0 S 0 3 0 1 0
Subsequently, multiple combinatorial libraries were synthesized,
expressed and screened (details below). in general, these libraries
combined mutations that were identified as improving affinity
(Examples 2 and 3) with mutations that altered potentially labile
residues (Example 4) and with mutations that potentially conferred
enhanced thermostability/expression (Example 5). The combinatorial
libraries were screened in multiple ELISA formats to identify
clones with the best attributes for further development (affinity,
selectivity, binding to membrane-associated TL1A, and
developability). Multiple variants with optimized and diverse CDR
sequences utilizing different VH germline templates were identified
as summarized in Tables 19 to 22.
TABLE-US-00019 TABLE 19 Heavy Chain CDRs on 1-46*02 Heavy chain
template HCDR2 (50- HCDR3 Clone SEQ HCDR1 SEQ 65), (1.sup.st P SEQ
(93- ID ID (26-35) ID is 52a ID 102) Start 553 GFDIQDT 554
RIDPASGHTKY 565 ARSGGL YMH DPKFQV PDV 34 553 GFDIQDT 555
RIEPASGHIKY 566 ARSGGL YMH DPKFQG PDW 2 553 GFDIQDT 556 RIEPASGHIKY
566 ARSGGL YMH SPKFQG PDW 52 553 GFDIQDT 556 RIEPASGHIKY 566 ARSGGL
YMH PSKFQG PDW 46 553 GFDIQDT 557 RIEPASGHVKY 566 ARSGGL YMH PSKFQV
PDW 47 553 GFDIQDT 558 RIEPASGHVKY 566 ARSGGL YMH PDKFQT PDW 14 553
GFDIQDT 559 RIDPASGHIKY 567 ARSGGL YMH DPKFQK PDM 16 553 GFDIQDT
560 RIDPASGHVKI 567 ARSGGL YMH DPKFQV PDM 17L 553 GFDIQDT 561
RIDPASGHLKY 567 ARSGGL YMH DPKFQV PDM 17L-1 553 GFDIQDT 562
RIDPASGHLKY 567 ARSGGL YMH DPKFQR PDM 23 553 GFDIQDT 563
RIDPASGHLKY 568 ARSGGL YMH DPKFQN PDK A1 553 GFDIQDT 563
RIDPASGHLKY 568 ARSGGL YMH PDKFQN PDK 53 553 GFDIQDT 564
RIEPASGHLKY 568 ARSGGL YMH DPKFQE PDK E1 553 GFDIQDT 564
RIEPASGHLKY 568 ARSGGL YMH DPKFQE PDK 484 DTYMH 485 PASGH 486
SGGLPD
TABLE-US-00020 TABLE 20 Light Chain CDRs on 3-20*01 Light chain
template Clone SEQ LCDR1 SEQ LCDR2 SEQ LCDR3 ID ID (24-33) ID
(50-56) ID (89-97) Start 569 RASSSVSYMY 488 ATSNLAS 571 QQWSGNPRT
34 569 RASSSVSYMY 488 ATSNLAS 572 QQWEGNPRT 2 569 RASSSVSYMY 488
ATSNLAS 572 QQWEGNPRT 52 570 GASSSVSYMY 488 ATSNLAS 572 QQWEGNPRT
46 569 RASSSVSYMY 488 ATSNLAS 572 QQWEGNPRT 47 569 RASSSVSYMY 488
ATSNLAS 572 QQWEGNPRT 14 569 RASSSVSYMY 488 ATSNLAS 573 QQWQGNPRT
16 569 RASSSVSYMY 488 ATSNLAS 573 QQWQGNPRT 17L 569 RASSSVSYMY 488
ATSNLAS 573 QQWQGNPRT 17L-1 569 RASSSVSYMY 488 ATSNLAS 573
QQWQGNPRT 23 569 RASSSVSYMY 488 ATSNLAS 572 QQWEGNPRT A1 570
GASSSVSYMY 488 ATSNLAS 572 QQWEGNPRT 53 569 RASSSVSYMY 488 ATSNLAS
572 QQWEGNPRT E1 570 GASSSVSYMY 488 ATSNLAS 572 QQWEGNPRT SEQ ID
NO: SEQ ID SEQ ID 487 NO: 488 NO: 489 ASSSVSYMY ATSNLAS GNPRT
TABLE-US-00021 TABLE 21 Heavy Chain CDRs on 1-3*01 Heavy chain
template HCDR1 HCDR2 (50- HCDR3 Clone SEQ (26- SEQ 65), (1.sup.st P
SEQ (93- ID ID 35) ID is 52a ID 102) Start 553 GFDIQ 554
RIDPASGHTKY 578 ARSGG DTYMH DPKFQV LPDV 3-17L 553 GFDIQ 574
RIDPASGHLKY 579 ARSGG V-A DTYMH DPKFQG LPDM 3-17L 553 GFDIQ 574
RIDPASGHLKY 579 ARSGG DTYMH DPKFQG LPDM L8mod 553 GFDIQ 575
RIDPASGHTKY 578 ARSGG DTYMH DPKFQG LPDV X-V 553 GFDIQ 554
RIDPASGHTKY 580 ARSGG DTYMH DPKFQV LPDF X 553 GFDIQ 554 RIDPASGHTKY
580 ARSGG DTYMH DPKFQV LPDF H3-1 553 GFDIQ 575 RIDPASGHTKY 581
ARSGG DTYMH DPKFQG LPDL XL3-6 553 GFDIQ 575 RIDPASGHTKY 580 ARSGG
DTYMH DPKFQG LPDF XL3- 553 GFDIQ 575 RIDPASGHTKY 580 ARSGG 10 DTYMH
DPKFQG LPDF XL3- 553 GFDIQ 575 RIDPASGHTKY 580 ARSGG 15 DTYMH
DPKFQG LPDF L3-13 553 GFDIQ 575 RIDPASGHTKY 580 ARSGG DTYMH DPKFQG
LPDF H2-2 553 GFDIQ 576 RIDPASGHSKY 580 ARSGG DTYMH DPKFQV LPDF
H2-5 553 GFDIQ 577 RIDPASGHYKY 580 ARSGG DTYMH DPKFQV LPDF
TABLE-US-00022 TABLE 22 Light Chain CDRs on 3-20*01 Light chain
template Clone SEQ LCDR1 SEQ LCDR2 SEQ LCDR3 ID ID (24-33) ID
(50-56) ID (89-97) Start 569 RASSSVSYMY 488 ATSNLAS 571 QQWSGNPRT
3-17L 569 RASSSVSYMY 488 ATSNLAS 573 QQWQGNPRT V-A 3-17L 569
RASSSVSYMY 488 ATSNLAS 573 QQWQGNPRT L8mod 569 RASSSVSYMY 488
ATSNLAS 571 QQWSGNPRT X-V 569 RASSSVSYMY 488 ATSNLAS 571 QQWSGNPRT
X 569 RASSSVSYMY 488 ATSNLAS 571 QQWSGNPRT H3-1 569 RASSSVSYMY 488
ATSNLAS 571 QQWSGNPRT XL3-6 569 RASSSVSYMY 488 ATSNLAS 582
SQWSGNPRT XL3- 569 RASSSVSYMY 488 ATSNLAS 583 QQWSGNPRS 10 XL3- 569
RASSSVSYMY 488 ATSNLAS 584 QQWSRNPRT 15 L3-13 569 RASSSVSYMY 488
ATSNLAS 585 QQWKGNPRT H2-2 569 RASSSVSYMY 488 ATSNLAS 571 QQWSGNPRT
H2-5 569 RASSSVSYMY 488 ATSNLAS 571 QQWSGNPRT
Fabs with the CDRs shown in Tables 19 to 22 were tested for binding
to human TL1A in multiple formats. First, human TL1A was
immobilized on the surface of an ELISA plate and soluble Fab
variants were titrated, as shown in FIGS. 8 and 9. Next, uniform
(saturating) quantities of soluble Fab variants were captured on
the surface of an ELISA plate and soluble, biotinylated human TL1A
was titrated as shown in FIGS. 10 to 13. In both ELISA formats all
the Fab variants bound human TL1A and displayed significantly
enhanced binding relative to the CDR-grafted variant, L8. In
addition, the variants all bound as well, or better than, variant
12835 while having no, or significantly fewer, murine
back-mutations in the frameworks. As a result these experiments
elucidated a set of anti-TLIA variable regions that exhibited both
high binding affinity and high homology to human germline Ig
sequences.
All the Fab variants were tested for binding to membrane-associated
human TL1A. For these studies a HEK293 cell line that had been
transfected with human TL1A was used. The 293 cells expressing
membrane bound human TL1A were maintained in DMEM containing
L-glutamine, glucose, sodium pyruvate, and phenol red (ThermoFisher
cat #11995-065) plus 10% fetal bovine serum, 1.times.
penicillin-streptomycin (Fisher cat #15140122) and 2 .mu.g/ml
puromycin (Gibco cat # A11138-03) in a 37.degree. C. incubator with
5% CO.sub.2. Three days prior to the assay, a T-75 flask was seeded
with 3.times.10.sup.6 cells so that the flask was 90-95% confluent
on the day of the assay. The media was aspirated and the cell
monolayer was gently washed with 5 ml PBS. Adherent cells were
removed by repeatedly pipetting 10 ml ice cold 1% BSA/PBS against
the monolayer. The cells were counted and 5.times.10.sup.5 were
aliquoted for each sample to be analyzed. The cells were collected
by centrifuging at 300.times.g at 4.degree. C. for 5 minutes and
the wash was discarded. The cells were resuspended in 100 .mu.l Fab
or IgG diluted in 1% BSA/PBS and placed on ice for 30 min. Next,
the cells were washed with 1 ml 1% BSA/PBS and collected by
centrifuging at 300.times.g at 4.degree. C. for 5 min. The wash was
discarded and 100 .mu.l secondary goat F(ab').sub.2 anti-human
kappa FITC (Southern Biotech cat #2062-02) or goat F(ab').sub.2
anti-human IgG PE (Southern Biotech cat #2043-09) conjugate,
diluted 1:200 in BSA/PBS was added. Cells were placed on ice for 30
min. Finally, the cells were washed with 1 ml BSA/PBS, collected by
centrifuging at 300.times.g at 4.degree. C. for 5 min. The wash was
removed and the cells were resuspended in 500 .mu.l 1% BSA/PBS. One
drop of Sytox AADvanced ReadyFlow Reagent (ThermoFisher cat #
R37173) was added per sample and the samples were analyzed on the
Attune NxT Flow Cytometer (ThermoFisher). All of the variants bound
membrane-associated human TL1A, as shown in FIG. 14.
Next, all the Fab variants were characterized for their selectivity
for human TL1A relative to other TNFSF members TRAIL, LIGHT and
Fas. Briefly, ELISA plates were coated overnight at 4.degree. C.
with 50 .mu.l/well antigen (Fas/TNFSF6, R&D Systems, cat. no.
126-FL/CF; TRAIL/TNFSF10, R&D Systems, cat. no. 375-TL/CF;
LIGHT/TNFSF14, R&D Systems, cat. no. 664-LI/CF) at 1 .mu.g/ml
in PBS. The plate was washed 3 times with PBS-T and blocked with
100 .mu.l of 1% BSA/PBS. The block was discarded and the Fab
variants or control antibodies (Fas/TNFSF6, R&D Systems, cat.
no. AF126; TRAIL/TNFSF10, R&D Systems, cat. no. AF375;
LIGHT/TNFSF14, R&D Systems, cat. no. AF664) were titrated in 50
.mu.l 1% BSA/PBS and incubated for 1 h at 25.degree. C. The plate
was washed 3 times with PBS-T and secondary HRP-conjugated antibody
(diluted 5,000-fold in 1% BSA/PBS) was added for 1 h at 25.degree.
C. The plate was washed 3 times with PBS-T and developed. As shown
in FIGS. 15 to 20 none of the variants displayed detectable binding
to the related family members, indicating the selectivity for human
TL1A versus other TNFSF family members was preserved while
engineering higher affinity using human germline framework
templates.
Example 7: Characterization of Select Humanized Variants Expressed
on Different IgG Constant Regions
The light and heavy chain variable regions of clones 14, 17L, 23,
34, 47, and 53, from Tables 19 and 20 above, were cloned onto kappa
light chain constant region, and either a modified IgG1 or an IgG2
heavy chain backbone, respectively. The modified IgG1 backbone and
IgG2 were selected to reduce potential effector function of the
antibodies. Transient expression and purification characteristics
are shown in Table 23 below. For these variants, all expressed
better as modified IgG1 than as IgG2. Furthermore, the yields
obtained were consistent with the observation made regarding the
impact of certain mutations on expression in bacteria (see Example
5). Specifically, the highest expressing variants 14, 34, and 47
all contained mutations at heavy chain CDR2 V65G, V65T or V65K
while the lowest expressing variants 17L, 23, and 53 did not.
In general, the binding of all variants to human TL1A in both
formats (modified IgG1 and IgG2) was preserved, as assessed by
ELISA binding to antigen coated plates (FIG. 21) and by ELISA
capture of soluble, biotinylated antigen (FIG. 22). In addition,
binding was to membrane-associated human TL1A was preserved for all
variants, with the exception of variant 53 when expressed as an
IgG2 (FIG. 23). Finally, selectivity for human TL1A versus other
TNFSF members was maintained, as none of the clones showed
appreciable binding to TNFSF family members Fas, TRAIL, or LIGHT
(FIG. 24).
TABLE-US-00023 TABLE 23 Expression of Select CDR Variants as human
IgG1 (modified) and IgG2 IgG1 Yield IgG1 IgG2 Yield Clone (mg)
Purity (mg) IgG2 Purity 34 5.9 90% 3.1 90% 47 5.5 95% 3.2 95% 14
6.1 90% 2.6 95% 17L 0.15 90% 0.03 <80% 23 3.0 80% 0.6 80% 53 1.8
95% 0.1 85%
Example 8: Characterization of Potency and Species Selectivity in
Whole Blood Assay
The neutralizing activity and potency of the variants described
herein expressed as IgG1 (modified) and IgG2 was tested in a human
whole blood assay using healthy donors. This assay is a
modification of Cassatella et al., "Soluble TNF-like cytokine
(TL1A) production by immune complexes stimulated monocytes in
rheumatoid arthritis" J Immunol. 2007 Jun. 1;178(11):7325-33; and
measures the production of IFN-.gamma. under conditions where TL1A
and its receptor DR3 are upregulated and activated. In this assay,
both soluble and membrane-associated TL1A are produced. Results
from this assay have been shown to correlate with in vivo outcomes
in a mouse model of colitis. See Takedatsu, "TL1A (TNFSF15)
regulates the development of chronic colitis by modulating both
T-helper 1 and T-helper 17 activation. Gastroenterology. 2008 Aug;
135(2): 552-567.
Briefly, 96-well plates are coated with human gamma globulin in PBS
overnight at 4.degree. C., washed with PBS, and incubated with
anti-human IgG (Fc fragment specific) for at least 1 hour at
25.degree. C. to generate immune complex (IC). Just prior to use,
the plates are washed three times with PBS. Collected blood samples
are treated with IL-12 and IL-18, antibody is titrated in the
samples, and the samples are added to the plates and placed at
37.degree. C. for 24 hours. Next, 100 .mu.l of PBS/5% BSA was added
to each well and mixed. Plates were centrifuged at 500 g for 5
minutes and then .about.150 .mu.l of diluted plasma was collected
for IFN-.gamma. measurement. PBS is added to make collection of
a-cellular plasma easier from the plate since the blood percentage
is high (95%). All samples are diluted to ensure that values are
within the linear range of the standard curve. All variants,
regardless of the format of IgG (modified IgG1 or IgG2), displayed
potent inhibition of IFN-.gamma. production (Table 24). For
comparison, the typical IC50 values for murine parental antibody
5C3D11 and humanized 12835 are 1.38.+-.0.95 nM (donor n 32 16) and
9.28.+-.10.71 nM (donor n=4), respectively.
TABLE-US-00024 TABLE 24 Potency in Human Whole Blood Assay IgG Mean
IC50 +/- SD Clone Subclass (nM) # Donors 34 G1 0.18 .+-. 0.05 6 34
G2 0.16 .+-. 0.06 3 47 G1 1.26 .+-. 0.40 6 47 G2 1.01 .+-. 0.16 3
14 G1 0.19 .+-. 0.06 6 14 G2 0.35 .+-. 0.13 3 23 G1 0.41 .+-. 0.10
6 23 G2 0.33 .+-. 0.12 3 53 G1 0.39 .+-. 0.06 6
Next, the samples were evaluated in the same assay, using blood
obtained from cynomolgus monkeys, in order to evaluate the
cross-reactivity of the optimized, humanized variants with
cynomolgus TLIA. The assay was performed similar to the assay that
utilized human whole blood, except the variants were tested at a
single concentration (10 nM), rather than performing a full
titration. The variants all inhibited IFN-.gamma. production,
though variants 47 and 53 did not inhibit to the extent of variants
14, 23, or 34 and murine 5C3D11 (FIG. 25). These data demonstrate
that the optimized humanized variants preserved cross-reactivity
with cynomolgus TL 1A.
Neutralizing TL1A antibodies were also formatted as effectorless
IgG1 (as shown in SEQ ID NO: 542) molecules, expressed and purified
from CHO cells, and tested in a potency assay using human (FIG.
26A-C) and cynomolgus monkey (FIG. 27A-C) whole blood as described
above. Results are summarized below in Table 25.
TABLE-US-00025 TABLE 25 Potency (IC50, nM) of CHO Expressed
Variants in Human and Cynomolgus Whole Blood Assay Cynomolgus Whole
Human Whole Blood Blood Clone Mean SD Donors Mean SD Donors 14 0.30
0.15 3 0.28 0.04 3 23 0.64 0.21 3 0.72 0.16 3 34 0.24 0.03 3 0.21
0.06 3 53 0.59 0.11 3 1.32 0.23 3 1D1 0.27 0.12 3 5C3D11 0.18 0.07
3
These experiments establish that all variants are active and
potent, with variants 14 and 34 typically displaying the greatest
potency with human blood. All variants are potent using cynomolgus
blood. Variants 14, 23 and 34 display similar potency to human and
cynomolgus TL1A, while variant 53 displays .about.2-fold greater
potency towards human TL1A.
Example 9: Competition Assays
A binding competition assay using surface plasmon resonance (SPR)
is performed to evaluate whether a test anti-TL1A antibody binds to
the same region on TL1A as any anti-TL1A antibody described
herein.
The reference antibody is directly immobilized via amine coupling
onto a carboxymethylated dextran sensor chip surface (CMS) using a
Biacore 2000 or 3000 instrument. Recombinant soluble human TL1A or
murine TL1A diluted to 10 nM in 8.1 mM Na.sub.2HPO.sub.4, 1.47 mM
KF.sub.2PO.sub.4, pH 7.2, 237 mM NaCI, 2.7 mM KCI, 3.4 mM EDTA and
0.01% Tween 20 (PBS-NET) is injected for about 1 minute at a flow
rate of 10 RI/minute to achieve binding levels on the immobilized
antibody of at least 100 response units (RU). The reference
antibody is then injected at 30 nM for 5 minutes in order to
saturate all of the potential binding sites on the TL1A. A repeat
injection of the reference antibody is performed to confirm this
saturation. Next, the test antibody in PBS-NET or PBS-NET alone as
a control is injected at 30 nM for 5 minutes. If the test antibody
binds to the TL1A saturated with the first antibody, this indicates
that the test antibody binds to a non-competing region on TL1A as
compared to the reference antibody. If the test antibody does not
bind to the saturated TL1A, this indicates that the two antibodies
bind to the same region or compete with binding to TL1A. This
strategy may be repeated with the test antibody immobilized and the
reference antibody injected after the test antibody is bound with
TL1A. Each cycle may be repeated. At the end of each cycle, the
immobilized antibody surface is regenerated either by a 30-second
pulse of 3M MgCl.sub.2 or by 0.1% TFA followed by two consecutive
15-second pulses of PBS-NET. All injections are performed at
25.degree. C. at a collection rate of 10 Hz. All sensorgrams are
double referenced by using both a control surface and buffer
injections.
Another binding competition assay using SPR is performed to
evaluate whether a test anti-TL1A antibody binds to the same region
on TL1A as any anti-TL1A antibody described herein. The reference
antibody is immobilized to the SPR chip via amine coupled at three
or four different densities across the array. The TL1A protein is
injected in an increasing concentration series to estimate kinetic
parameters and the appropriate concentration for injections during
the competition binning experiment. Once the optimal antigen
concentration for the binning experiment is determined,
regeneration conditions (typically a brief low pH injection) are
evaluated to establish the optimal conditions for regeneration
between cycles of the binning assay.
Binning is performed using a pre-mix approach, where a moderate
concentration of TL1A is injected over the array, either by itself,
or pre-complexed to the test antibody at saturating antibody
concentrations (e.g., 30-50 .mu.g/mL). The assay may be performed
such that the test antibody is immobilized and the reference
antibody is pre-complexed to the TL1A. Clones that bind to unique
regions from the immobilized antibody provide an increase in
signal, while competitive clones will decrease the antigen binding
signal. The competition assay is run so that all clones are tested
as both ligands and analytes.
Example 10: Comparison of 5C3D11 Binding to other Anti-TL1A
Antibodies
Two epitope binning studies were performed to compare the epitope
recognized by 5C3D11 and 12835 to the epitopes recognized by other
TL1A antibodies, including 1D1, 1681, 1B4, and 1A9, as shown in
Table 26.
TABLE-US-00026 TABLE 26 Antibody variable region sequences used for
epitope binning studies Anti- body Heavy Chain Light Chain 5C3D11
SEQ ID NO 3 SEQ ID NO 6 12835 SEQ ID NO 26 SEQ ID NO 28 1D1
QVQLVQSGAEVKKPGASVKVS EIVLTQSPATLSLSPGERA CKASGYDFTYYGISWVRQAPG
TLSCRASQSVSSYLAWYQQ QGLEWMGWISTYNGNTHYARM KPGQAPRLLIYDASNRATG
LQGRVTMTTDTSTRTAYMELR IPARFSGSGSGTDFTLTIS SLRSDDTAVYYCARENYYGSG
SLEPEDFAVYYCQQRSNWP AYRGGMDVWGQGTTVTVSS WTFGQGTKVEIK (SEQ ID NO:
687) (SEQ ID NO: 691) 1681 EVQLLESGGGLVQPGKSLRLS
DIQMTQSPSTLSASVGDRV CAVSGFTFSTYGMNWVRQAPG TITCRASQTISSWLAWYQQ
KGLEWVSSISGTGRTTYHADS TPEKAPKLLIYAASNLQSG VQGRFTVSRDNSKNILYLQMN
VPSRFSGSGSGTEFTLTIS SLRADDTAVYFCTKERGDYYY SLQPDDFATYYCQQYHRSW
GVFDYWGQGTLVTVSS TFGQGTKVEIT (SEQ ID NO: 688) (SEQ ID NO: 692) 1B4
QVTLKESGPALVKPTQTLTLT DIQLTQSPSFLSASVGDRV CTFSGFSLSTSNMGVVWIRQP
TITCSASSSVNYMHWYQQK PGKALEWLAHILWDDREYSNP PGKAPKLLIYSTSNLASGV
ALKSRLTISKDTSKNQVVLTM PSRFSGSGSGTEFTLTISS TNMDPVDTATYYCARMSRNYY
LQPEDFATYYCHQWNNYGT GSSYVMDYWGQGTLVTVSS FGQGTKVEIKR (SEQ ID NO:
689) (SEQ ID NO: 693) 1A9 QIQLVQSGPELKKPGETVKIS DVLMTQTPLSLPVSLGDQA
CKASGYTFTTYGMSWVKQAPG SISCRSSQNIVHSDGNTYL KGLKWMGWMNTYSGVTTYADD
EWYLQKPGQSPKLLIYKVS FKGRFAFSLETSASTAYMQID NRFSGVPDRFSGSGSGTDF
NLKNEDTATYFCAREGYVFDD TLKISRVEAEDLGIYYCFQ YYATDYWGQGTSVTVSS
GSHVPLTFGAGTKLELK (SEQ ID NO: 690) (SEQ ID NO: 694)
To minimize avidity effects a planar carboxymethyldextran surface
sensor chip was used (Xantec Prod. # SPMXCMDP) in the first study
while a HC30M sensor chip was used (Xantec Prod. # SPMXHC30M) in
the second study. The running buffer for continuous flow
micro-spotting was HBS-EP+ and the flow rate was 65 .mu.l/min. The
chip was activated with 7 min of 18 mM EDC and 4.5 nM sulfo-NHS in
100 mM MES, pH 5.5. Antibodies were then immobilized for 15 min in
two replicate prints. Antibodies were diluted to 10 .mu.g/ml in 10
mM acetate, pH 4.5. The antibodies were titrated in a 3-fold serial
dilution three places across the plate, establishing a
concentration series of different spot densities. Each antibody was
spotted 8 times--twice at each of four dilutions. This created a
10.times.8 array. The remaining active groups were neutralized with
a 7 min quench using 1 M ethanolamine, pH 8.5. Because the antigen
is a homotrimeric protein and IgG is bivalent the epitope binning
was performed using pre-mix conditions.
First epitope binning study--TL1A was prepared at a final
concentration of 50 nM (3.3 .mu.g/ml) and mixed with 333 nM (50
.mu.g/ml) analyte (solution phase antibody) or with running buffer
(control). For samples in IgG format, 50 .mu.g/ml is 333 nM while
for samples in Fab format 50 .mu.g/ml is 1 .mu.M. Mixed samples
were injected for 5 min over the array and regenerated for 30 sec
after every cycle using a 4:1 mixture of Pierce IgG Elution buffer
and 5 M NaCl (1 M final concentration).
Second epitope binning study--TL1A was prepared at a final
concentration of 50 nM (3.3 .mu.g/ml) and mixed with 1 .mu.M (150
.mu.g/ml) IgG or 2 .mu.M (200 .mu.g/ml) Fab analyte (solution phase
antibody) or with running buffer (control). The antibody samples
were serially diluted 2-fold seven times (7.8 nM final for IgG, 15
nM final for Fab). Mixed samples were injected for 5 min over the
array and regenerated for 30 sec after every cycle using a 4:1
mixture of Pierce IgG Elution buffer and 5 M NaCl (1 M final
concentration).
In epitope binning studies a clear signal (sandwich) was observed
with immobilized 5C3D11 and 12835 with all of the control
antibodies tested as analyte (Table 27, top two rows). These
results demonstrate that 5C3D11 and 12835 can bind TL1A
simultaneously with the other antibodies and thus, recognize a
distinct epitope.
TABLE-US-00027 TABLE 27 Summary of ability of antibodies to form
sandwich (Yes) Analyte Epitope Bin Ligand 5C3D11 12835 1D1 1B4 1681
1A9 1 5C3D11 No No Yes Yes Yes Yes 1 12835 No No Yes Yes Yes Yes 2
1D1 Yes Yes No No Yes Yes 2 1B4 Yes Yes No No Yes Yes 3 1681 Yes
Yes Yes Yes No Yes 4 1A9 Yes Yes Yes Yes Yes No Yes indicates that
the antibodies are able to simultaneously bind TL1A target
Example 11: In Vivo Assessment of Anti-TL1A Efficacy
The efficacy of anti-TL1A antibodies in animal models of colitis is
performed. Anti-TL1A antibodies are tested in rodent models of
acute colitis induced by intrarectal administration of di- or
tri-nitrobenzenesulfonic acid (D/TNBS) or oxazolone, and chronic
colitis induced by administration of DSS in drinking water or
transfer of CD45RB.sup.hi T cells. DNBS and oxazolone induce
localized ulceration and inflammation. DSS administration induces
robust generalized inflammation of the intestinal tract
characterized by erosive lesions and inflammatory infiltrate.
Symptoms of all these models usually include diarrhea, occult
blood, weight loss and occasionally rectal prolapse. In a
prophylactic model, antibody treatment begins at the start of
administration of the colitis-inducing compound. In a therapeutic
model, antibody treatment begins several days after commencement of
induction. The effect of the treatment on weight, stool consistency
and occult blood, as well as microscopic effects on epithelial
integrity and degree of inflammatory infiltrate is determined.
Daily clinical scoring is performed based on stool consistency and
presence of occult blood giving a disease activity index (DAI)
score.
Example 12: Phase 1 Clinical Trial
A phase 1 clinical trial is performed to evaluate the safety,
tolerability, pharmacokinetics and pharmacodynamics of an anti-TL1A
antibody provided herein in subjects having Crohn's Disease.
Single ascending dose (SAD) arms: Subjects in each group (subjects
are grouped based on the presence or absence of a risk variant)
receive either a single dose of the antibody or a placebo.
Exemplary doses are 1, 3, 10, 30, 100, 300, 600 and 800 mg of
antibody, or between 5 to 30 milligrams per kilogram. Safety
monitoring and PK assessments are performed for a predetermined
time. Based on evaluation of the PK data, and if the antibody is
deemed to be well tolerated, dose escalation occurs, either within
the same groups or a further group of healthy subjects. Dose
escalation continues until the maximum dose has been attained
unless predefined maximum exposure is reached or intolerable side
effects become apparent.
Multiple ascending dose (MAD) arms: Subjects in each group
(subjects are grouped based on the presence or absence of a risk
variant) receive multiple doses of the antibody or a placebo. The
dose levels and dosing intervals are selected as those that are
predicted to be safe from the SAD data. Dose levels and dosing
frequency are chosen to achieve therapeutic drug levels within the
systemic circulation that are maintained at steady state for
several days to allow appropriate safety parameters to be
monitored. Samples are collected and analyzed to determination PK
profiles.
Inclusion Criteria: Healthy subjects of non-childbearing potential
between the ages of 18 and 55 years. Healthy is defined as no
clinically relevant abnormalities identified by a detailed medical
history, full physical examination, including blood pressure and
pulse rate measurement, 12 lead ECG and clinical laboratory tests.
Female subjects of non-childbearing potential must meet at least
one of the following criteria: (1) achieved postmenopausal status,
defined as: cessation of regular menses for at least 12 consecutive
months with no alternative pathological or physiological cause; and
have a serum follicle stimulating hormone (FSH) level within the
laboratory's reference range for postmenopausal females; (2) have
undergone a documented hysterectomy and/or bilateral oophorectomy;
(3) have medically confirmed ovarian failure. All other female
subjects (including females with tubal ligations and females that
do NOT have a documented hysterectomy, bilateral oophorectomy
and/or ovarian failure) will be considered to be of childbearing
potential. Body Mass Index (BMI) of 17.5 to 30.5 kg/m2; and a total
body weight >50 kg (110 lbs). Evidence of a personally signed
and dated informed consent document indicating that the subject (or
a legal representative) has been informed of all pertinent aspects
of the study.
Two groups of healthy subjects are selected: subjects having a risk
variant whose presence is associated with an increase in
susceptibility to Crohn's Disease, and subjects lacking the risk
variant.
Exclusion Criteria: Evidence or history of clinically significant
hematological, renal, endocrine, pulmonary, gastrointestinal,
cardiovascular, hepatic, psychiatric, neurologic, or allergic
disease (including drug allergies, but excluding untreated,
asymptomatic, seasonal allergies at time of dosing). Subjects with
a history of or current positive results for any of the following
serological tests: Hepatitis B surface antigen (HBsAg), Hepatitis B
core antibody (HBcAb), anti-Hepatitis C antibody (HCV Ab) or human
immunodeficiency virus (HIV). Subjects with a history of allergic
or anaphylactic reaction to a therapeutic drug. Treatment with an
investigational drug within 30 days (or as determined by the local
requirement, whichever is longer) or 5 half-lives or 180 days for
biologics preceding the first dose of study medication. Pregnant
females; breastfeeding females; and females of childbearing
potential.
Primary Outcome Measures: Incidence of dose limiting or
intolerability treatment related adverse events (AEs) [Time Frame:
12 weeks]. Incidence, severity and causal relationship of treatment
emergent AEs (TEAEs) and withdrawals due to treatment emergent
adverse events [Time Frame: 12 weeks]. Incidence and magnitude of
abnormal laboratory findings [Time Frame: 12 weeks]. Abnormal and
clinically relevant changes in vital signs, blood pressure (BP) and
electrocardiogram (ECG) parameters [Time Frame: 12 weeks].
Secondary Outcome Measures: Single Ascending Dose: Maximum Observed
Plasma Concentration (C.sub.max) [Time Frame: 12 weeks]. Single
Ascending Dose: Time to Reach Maximum Observed Plasma Concentration
(T.sub.max) [Time Frame: 12 weeks]. Single Ascending Dose: Area
under the plasma concentration-time profile from time zero to 14
days (AUC14 days) [Time Frame: 12 weeks]. Single Ascending Dose:
Area under the plasma concentration-time profile from time zero
extrapolated to infinite time (AUC.sub.inf) [Time Frame: 12 weeks].
Single Ascending Dose: Area under the plasma concentration-time
profile from time zero to the time of last quantifiable
concentration (AUC.sub.last) [Time Frame: 12 weeks]. Single
Ascending Dose: Dose normalized maximum plasma concentration
(C.sub.max[dn]) [Time Frame: 12 weeks]. Single Ascending Dose: Dose
normalized area under the plasma concentration-time profile from
time zero extrapolated to infinite time (AUC.sub.inf[dn]) [Time
Frame: 12 weeks]. Single Ascending Dose: Dose normalized area under
the plasma concentration-time profile from time zero to the time of
last quantifiable concentration (AUC.sub.last[dn]) [Time Frame: 12
weeks]. Single Ascending Dose: Plasma Decay Half-Life (t1/2) [Time
Frame: 12 weeks]. Plasma decay half-life is the time measured for
the plasma concentration to decrease by one half. Single Ascending
Dose: Mean residence time (MRT) [Time Frame: 12 weeks]. Single
Ascending Dose: Volume of Distribution at Steady State (V.sub.ss)
[Time Frame: 6 weeks]. Volume of distribution is defined as
theoretical volume in which the total amount of drug would need to
be uniformly distributed to produce the desired blood concentration
of a drug. Steady state volume of distribution (V.sub.ss) is the
apparent volume of distribution at steady-state. Single Ascending
Dose: Systemic Clearance (CL) [Time Frame: 6]. CL is a quantitative
measure of the rate at which a drug substance is removed from the
body.
Multiple Ascending Dose First Dose: Maximum Observed Plasma
Concentration (C.sub.max) [Time Frame: 12 weeks]. Multiple
Ascending Dose First Dose: Time to Reach Maximum Observed Plasma
Concentration (T.sub.max) [Time Frame: 12 weeks]. Multiple
Ascending Dose First Dose: Area under the plasma concentration-time
profile from time zero to time .tau., the dosing interval where
.tau.=2 weeks (AUC.sub.t) [Time Frame: 12 weeks]. Multiple
Ascending Dose First Dose: Dose normalized maximum plasma
concentration (C.sub.max[dn]) [Time Frame: 12 weeks]. Multiple
Ascending Dose First Dose: Dose normalized Area under the plasma
concentration-time profile from time zero to time .tau., the dosing
interval where .tau.=2 weeks (AUC.sub..tau.[dn]) [Time Frame: 12
weeks]. Plasma Decay Half-Life (t1/2) [Time Frame: 12 weeks].
Plasma decay half-life is the time measured for the plasma
concentration to decrease by one half. Multiple Ascending Dose
First Dose: Mean residence time (MRT) [Time Frame: 12 weeks].
Apparent Volume of Distribution (Vz/F) [Time Frame: 12 weeks].
Volume of distribution is defined as theoretical volume in which
the total amount of drug would need to be uniformly distributed to
produce the desired plasma concentration of a drug. Apparent volume
of distribution after oral dose (Vz/F) is influenced by the
fraction absorbed. Multiple Ascending Dose First Dose: Volume of
Distribution at Steady State (V.sub.ss) [Time Frame: 12 weeks].
Volume of distribution is defined as theoretical volume in which
the total amount of drug would need to be uniformly distributed to
produce the desired blood concentration of a drug. Steady state
volume of distribution (V.sub.ss) is the apparent volume of
distribution at steady-state. Multiple Ascending Dose First Dose:
Apparent Oral Clearance (CL/F) [Time Frame: 12 weeks]. Clearance of
a drug is a measure of the rate at which a drug is metabolized or
eliminated by normal biological processes. Clearance obtained after
oral dose (apparent oral clearance) is influenced by the fraction
of the dose absorbed. Clearance is estimated from population
pharmacokinetic (PK) modeling. Drug clearance is a quantitative
measure of the rate at which a drug substance is removed from the
blood. Multiple Ascending Dose First Dose: Systemic Clearance (CL)
[Time Frame: 12 weeks]. CL is a quantitative measure of the rate at
which a drug substance is removed from the body.
Multiple Ascending Dose Multiple Dose: Maximum Observed Plasma
Concentration (C.sub.max) [Time Frame: 12 weeks]. Multiple
Ascending Dose Multiple Dose: Time to Reach Maximum Observed Plasma
Concentration (T.sub.max) [Time Frame: 12 weeks]. Multiple
Ascending Dose Multiple Dose: Area under the plasma
concentration-time profile from time zero to time .tau., the dosing
interval where .tau.=2 weeks (AUCT) [Time Frame: 12 weeks].
Multiple Ascending Dose Multiple Dose: Dose normalized maximum
plasma concentration (Cmax[dn]) [Time Frame: 12 weeks]. Multiple
Ascending Dose Multiple Dose: Dose normalized Area under the plasma
concentration-time profile from time zero to time .tau., the dosing
interval where .tau.=2 weeks (AUCT [dn]) [Time Frame: 12 weeks].
Multiple Ascending Dose Multiple Dose: Plasma Decay Half-Life
(t1/2) [Time Frame: 12 weeks]. Plasma decay half-life is the time
measured for the plasma concentration to decrease by one half
Multiple Ascending Dose Multiple Dose: Apparent Volume of
Distribution (Vz/F) [Time Frame: 12 weeks]. Volume of distribution
is defined as theoretical volume in which the total amount of drug
would need to be uniformly distributed to produce the desired
plasma concentration of a drug. Apparent volume of distribution
after oral dose (Vz/F) is influenced by the fraction absorbed.
Multiple Ascending Dose Multiple Dose: Volume of Distribution at
Steady State (V.sub.ss) [Time Frame: 12 weeks]. Volume of
distribution is defined as theoretical volume in which the total
amount of drug would need to be uniformly distributed to produce
the desired blood concentration of a drug. Steady state volume of
distribution (V.sub.ss) is the apparent volume of distribution at
steady-state.
Multiple Ascending Dose Multiple Dose: Apparent Oral Clearance
(CL/F) [Time Frame: 12 weeks ]. Clearance of a drug is a measure of
the rate at which a drug is metabolized or eliminated by normal
biological processes. Clearance obtained after oral dose (apparent
oral clearance) is influenced by the fraction of the dose absorbed.
Clearance was estimated from population pharmacokinetic (PK)
modeling. Drug clearance is a quantitative measure of the rate at
which a drug substance is removed from the blood. Multiple
Ascending Dose Multiple Dose: Systemic Clearance (CL) [Time Frame:
12 weeks]. CL is a quantitative measure of the rate at which a drug
substance is removed from the body. Multiple Ascending Dose
Multiple Dose: Minimum Observed Plasma Trough Concentration
(C.sub.min) [Time Frame: 12 weeks]. Multiple Ascending Dose
Multiple Dose: Average concentration at steady state (Cav) [Time
Frame: 12 weeks]. Multiple Ascending Dose Multiple Dose: Observed
accumulation ratio (Rac) [Time Frame: 12 weeks]. Multiple Ascending
Dose Multiple Dose: Peak to trough fluctuation (PTF) [Time Frame:
12 weeks]. Multiple Ascending Dose Additional Parameter: estimate
of bioavailability (F) for subcutaneous administration at the
corresponding intravenous dose [Time Frame: 12 weeks].
Immunogenicity for both Single Ascending Dose and Multiple
Ascending Dose: Development of anti-drug antibodies (ADA) [Time
Frame: 12 weeks].
Example 13: Phase 1b Clinical Trial
A phase 1b open label clinical trial is performed to evaluate
efficacy of an anti-TL1A antibody provided herein on patients
having a risk variant associated with Crohn's Disease.
Arms: 10 patients positive for a risk variant whose presence is
associated with an increase in susceptibility to Crohn's Disease
are administered the antibody. 5-10 patients negative for the risk
variant are administered the antibody. Patients are monitored in
real-time. Central ready of endoscopy and biopsy is employed, with
readers blinded to point of time of treatment and endpoints.
Inclusion Criteria: Two groups of subjects are selected: subjects
having a risk variant whose presence is associated with an increase
in susceptibility to Crohn's Disease, and subjects lacking the risk
variant.
Primary Outcome Measures: Simple Endoscopic Score for Crohn's
Disease (SESCD), Crohn's Disease Activity Index (CDAI), and Patient
Reported Outcome (PRO). If risk variant positive group shows 50%
reduction from baseline, a Phase 2a clinical trial is
performed.
Inclusion Criteria: PRO entry criteria: Abdominal pain score of 2
or more and/or stool frequency score of 4 or more. Primary outcome
would be pain core of 0 or 1 and stool frequency score of 3 or less
with no worsening from baseline. Endoscopy entry criteria: SESCD
ileum only entry at score of 4 and 6 if colon is involved. Primary
endoscopic outcome is 40-50% delta of mean SESCD.
Example 14: Phase 2a Clinical Trial
A phase 2a clinical trial is performed to evaluate the efficacy of
an anti-TL1A antibody provided herein in subjects having Crohn's
Disease.
Arms: 40 patients per arm (antibody and placebo arms) are treated
with antibody or placebo for 12 weeks. An interim analysis is
performed after 20 patients from each group are treated at the
highest dose to look for a 40-50% delta between placebo and treated
group in primary outcome (50% reduction from baseline in SESCD,
CDAI, and PRO).
Primary Outcome Measures: Simple Endoscopic Score for Crohn's
Disease (SESCD), Crohn's Disease Activity Index (CDAI), and Patient
Reported Outcome (PRO).
Inclusion Criteria: PRO entry criteria: Abdominal pain score of 2
or more and/or stool frequency score of 4 or more. Primary outcome
would be pain core of 0 or 1 and stool frequency score of 3 or less
with no worsening from baseline. Endoscopy entry criteria: SESCD
ileum only entry at score of 4 and 6 if colon is involved. Primary
endoscopic outcome is 40-50% delta of mean SESCD.
Various embodiments are described above in the Detailed
Description. While these descriptions directly describe the above
embodiments, it is understood that those skilled in the art may
conceive modifications and/or variations to the specific
embodiments shown and described herein. Any such modifications or
variations that fall within the purview of this description are
intended to be included therein as well. Unless specifically noted,
it is the intention of the inventors that the words and phrases in
the specification and claims be given the ordinary and accustomed
meanings to those of ordinary skill in the applicable art(s).
The foregoing description of various embodiments known to the
applicant at this time of filing the application has been presented
and is intended for the purposes of illustration and description.
The present description is not intended to be exhaustive nor
limited to the precise form disclosed and many modifications and
variations are possible in the light of the above teachings. The
embodiments described serve to explain principles and practical
applications, and to enable others skilled in the art to utilize
the various embodiments, optionally with various modifications, as
are suited to the particular use contemplated. Therefore, it is
intended that the disclosure not be limited to the particular
embodiments disclosed.
While particular embodiments have been shown and described, it will
be obvious to those skilled in the art that, based upon the
teachings herein, changes and modifications may be made without
departing from this disclosure and its broader aspects and,
therefore, the appended claims are to encompass within their scope
all such changes and modifications as are within the true spirit
and scope of this disclosure. It will be understood by those within
the art that, in general, terms used herein are generally intended
as "open" terms (e.g., the term "including" should be interpreted
as "including but not limited to," the term "having" should be
interpreted as "having at least," the term "includes" should be
interpreted as "includes but is not limited to," etc.).
TABLE-US-00028 SEQUENCES SEQ ID NO Description Sequence 1 murine
mAb 5C3D11
gaagttcagctgcaacagtctggcgccgagctggttaagcctggcgcttctgtgaagctga heavy
chain variable
gctgtaccgcctctggcttcgacatccaagacacctacatgcactgggtcaagcagaggcc
region tgagcagggactcgagtggatcggcagaattgatcctgccagcggccacaccaaatacga
ccccaagttccaagtgaaggccaccatcaccaccgacaccagcagcaataccgcctacct
gcagctgagcagcctgacctctgaagataccgccgtgtactactgcagcagatctggcgg
actgcccgatgtttggggagccggaacaaccgtgacagtgtccagc 2 murine mAb 5C3D11
gaggttcaacttcaacaatcgggggccgagctggttaagcccggcgcttctgtaaaattgtc
heavy chain variable
ttgcactgcctctgggtttgacatccaagatacatatatgcattgggtgaaacagcgtcccga
region--codon
gcagggcttggagtggattggacgtattgaccccgcctctgggcacacgaaatatgatc- ct
optimized for E. coli
aagttccaggttaaagcgactatcacaacggacacctccagcaatacggcttatttacagtta
tcctcgctgacctctgaggatactgcagtgtactactgctctcgctctggtggtctgccagac
gtgtggggtgcaggaactacagttactgtgtcttca 3 murine mAb 5C3D11
EVQLQQSGAELVKPGASVKLSCTASGFDIQDTYMHWVK heavy chain variable
QRPEQGLEWIGRIDPASGHTKYDPKFQVKATITTDTSSNT region--amino acid
AYLQLSSLTSEDTAVYYCSRSGGLPDVWGAGTTVTVSS 4 murine mAb 5C3D11
Caaattgtgctgtctcagagccccgccatcctgagtgcttctccaggcgagaaagtgacca light
chain variable
tgacctgcagagccagcagcagcgtgtcctacatgtactggtatcagcagaagcccggca region
gcagccccaagccttggatctacgccacaagcaatctggccagcggcgtgcccgatagat
tttctggctctggcagcggcaccagctacagcctgacaatctctagagtggaagccgagga
tgccgccacctactactgtcaacagtggagcggcaaccccagaacctttggcggaggcac
caagctggaaatcaag 5 murine mAb 5C3D11
Caaatcgtcctgtcacagtccccggcgatcctttctgcttcaccaggagagaaggtaaccat
light chain variable
gacatgtcgcgcctcttcctcagtttcttacatgtactggtaccagcagaaaccaggatcatct-
region--codon
cccaaaccctggatctacgctacatcaaaccttgcatctggcgtgccagaccgtttttc- agg
optimized for E. coli
gtcgggctcggggacttcctattcattaaccatttctcgcgtagaagcggaagacgccgcca
cgtattattgtcagcagtggtcaggaaatccgcgcacattcggaggcggaacgaaattgga
gatcaaa 6 murine mAb 5C3D11
QIVLSQSPAILSASPGEKVTMTCRASSSVSYMYWYQQKPG light chain variable
SSPKPWIYATSNLASGVPDRFSGSGSGTSYSLTISRVEAED region--amino acid
AATYYCQQWSGNPRTFGGGTKLEIK 7 5C3D11 HCDR1
ggcttcgacatccaagacacctacatgcac 8 5C3D11 HCDR1--codon
gggtttgacatccaagatacatatatgcat optimized for E. coli 9 5C3D11
HCDR1--amino GFDIQDTYMH acid 10 5C3D11 HCDR2
agaattgatcctgccagcggccacaccaaatacgaccccaagttccaagtg 11 5C3D11
HCDR2--codon cgtattgaccccgcctctgggcacacgaaatatgatcctaagttccaggtt-
optimized for E. coli 554 5C3D11 HCDR2--amino RIDPASGHTKYDPKFQV
acid 13 5C3D11 HCDR3 Tctggcggactgcccgatgtt 14 5C3D11 HCDR3--codon
Tctggtggtctgccagacgtg optimized for E. coli 15 5C3D11 HCDR3--amino
SGGLPDV acid 16 5C3D11 LCDR1 agagccagcagcagcgtgtcctacatgtac 17
5C3D11 LCDR1--codon cgcgcctcttcctcagtttcttacatgtac optimized for E.
coli 569 5C3D11 LCDR1--amino RASSSVSYMY acid 19 5C3D11 LCDR2
Gccacaagcaatctggccagc 20 5C3D11 LCDR2--codon Gctacatcaaaccttgcatct
optimized for E. coli 488 5C3D11 LCDR2--amino ATSNLAS acid 22
5C3D11 LCDR3 caacagtggagcggcaaccccagaacc 23 5C3D11 LCDR3
cagcagtggtcaggaaatccgcgcaca codon optimized for E. coli 24 5C3D11
LCDR3--amino QQWSGNPRT acid 25 12835 (humanized
caagtacaattagtccagtcgggtgccgaggtaaaaaaacctggagcatccgta- aaactgt
5C3D11 heavy chain
cttgcaaagcatcggggtttgacatccaggacacctacatgcactgggtgcgtcaagctcc
variable region)--codon
aggacagggattagagtggatgggtcgcatcgaccccgcgagcggacac- acgaaatac
optimized for
gaccctaaatttcaagtacgtgtcacgatgactaccgacactagtacgagcactgttta- tatg
E. coli
gaattgtcctcgttacgctcagaggatacggcagtctattattgcagccgttccggaggctta
cccgacgtctggggacagggaactactgtaacagtcagtagt 26 12835 (humanized
QVQLVQSGAEVKKPGASVKLSCKASGFDIQDTYMHWVR 5C3D11 heavy chain
QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTTDTSTS variable region)--amino
TVYMELSSLRSEDTAVYYCSRSGGLPDVWGQGTTVTVSS acid 27 12835 (humanized
Gagattgtgttaacgcaatcaccggggactttatcgctgtcgccgggggagcgc- gttacaa
5C3D11 light chain
tgtcttgtcgcgcttcctcttcggtttcatacatgtattggtatcaacaaaaaccgggacaggct
variable region)--codon
ccacgcccctggatttacgctactagcaatttggcctcgggcgttcccg- accgcttcagcgg
optimized for
gtcagggagcggcaccgattacacgttgaccatctctcgtctggaacctgaagacttcg- cg E.
coli gtctattactgtcaacaatggtcgggaaatccccgtacatttggcggagggacgaagttgga
aattaaa 28 12835 (humanized
EIVLTQSPGTLSLSPGERVTMSCRASSSVSYMYWYQQKPG 5C3D11 light chain
QAPRPWIYATSNLASGVPDRFSGSGSGTDYTLTISRLEPED variable region)--amino
FAVYYCQQWSGNPRTFGGGTKLEIK acid 29 12835 HCDR1--codon
gggtttgacatccaggacacctacatgcac optimized for E. coli 30 12835
HCDR2--codon cgcatcgaccccgcgagcggacacacgaaatacgaccctaaatttcaagta
optimized for E. coli 31 12835 HCDR3--codon tccggaggcttacccgacgtc
optimized for E. coli 32 12835 LCDR1--codon
cgcgcttcctcttcggtttcatacatgtattggtat optimized for E. coli 33 12835
LCDR2--codon gctactagcaatttggcctcg optimized for E. coli 34 12835
LCDR3--codon caacaatggtcgggaaatccccgtaca optimized for E. coli 35
18-7 (CDR-grafted
caagtacaattagtccagtcgggtgccgaggtaaaaaaacctggagcatccgt- aaaactgt
light chain) heavy
cttgcaaagcatcggggtttgacatccaggacacctacatgcactgggtgcgtcaagctcc chain
variable region
aggacagggattagagtggatgggtcgcatcgaccccgcgagcggacacacgaaatac
gaccctaaatttcaagtacgtgtcacgatgactcgtgacactagtacgagcactgtttatatg
gaattgtcctcgttacgctcagaggatacggcagtctattattgcagccgttccggaggctta
cccgacgtctggggacagggaactactgtaacagtcagtagt 36 18-7 (CDR-grafted
QVQLVQSGAEVKKPGASVKLSCKASGFDIQDTYMHWVR light chain) heavy
QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDTST chain variable
STVYMELSSLRSEDTAVYYCSRSGGLPDVWGQGTTVTVS region--amino acid S 37
18-7 (CDR-grafted
gagattgtgttaacgcaatcaccggggactttatcgctgtcgccgggggagcg- cgcgaca
light chain) light
ctgtcttgtcgcgcttcctcttcggtttcatacatgtattggtatcaacaaaaaccgggacagg
chain variable region
ctccacgcctgctgatttacgctactagcaatttggcctcgggcatccccgaccgcttcagc
gggtcagggagcggcaccgattttacgttgaccatctctcgtctggaacctgaagacttcgc
ggtctattactgtcaacaatggtcgggaaatccccgtacatttggcggagggacgaagttgg
aaattaaa 38 18-7 (CDR-grafted
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG light chain) light
QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF chain variable
AVYYCQQWSGNPRTFGGGTKLEIK region--amino acid 39 21-3 (CDR-grafted
caagtacaattagtccagtcgggtgccgaggtaaaaaaacctggagcatccgt- aaaagtct
heavy chain) heavy
cttgcaaagcatcggggtttgacatccaggacacctacatgcactgggtgcgtcaagctcc chain
variable region
aggacagggattagagtggatgggtcgcatcgaccccgcgagcggacacacgaaatac
gaccctaaatttcaagtacgtgtcacgatgactcgtgacactagtacgagcactgtttatatg
gaattgtcctcgttacgctcagaggatacggcagtctattattgcgcacgttccggaggctta
cccgacgtctggggacagggaactactgtaacagtcagtagt 40 21-3 (CDR-grafted
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVR heavy chain) heavy
QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDTST chain variable
STVYMELSSLRSEDTAVYYCARSGGLPDVWGQGTTVTVS region--amino acid S 41
21-3 (CDR-grafted
gagattgtgttaacgcaatcaccggggactttatcgctgtcgccgggggagcg- cgcgaca
heavy chain) light
ctgtcttgtcgcgcttcctcttcggtttcatacatgtattggtatcaacaaaaaccgggacagg
chain variable region
ctccacgcctgctgatttacgctactagcaatttggcctcgggcgttcccgaccgcttcagcg
ggtcagggagcggcaccgattacacgttgaccatctctcgtctggaacctgaagacttcgc
ggtctattactgtcaacaatggtcgggaaatccccgtacatttggcggagggacgaagttgg
aaattaaa 42 21-3 (CDR-grafted
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG heavy chain) light
QAPRLLIYATSNLASGVPDRFSGSGSGTDYTLTISRLEPED chain variable
FAVYYCQQWSGNPRTFGGGTKLEIK region--amino acid 43 21-3 V102K (CDR-
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVR grafted heavy chain)
QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDTST heavy chain variable
STVYMELSSLRSEDTAVYYCARSGGLPDKWGQGTTVTVS region--amino acid S 44
21-3 V102M (CDR- QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVR grafted
heavy chain) QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDTST heavy chain
variable STVYMELSSLRSEDTAVYYCARSGGLPDMWGQGTTVTV region--amino acid
SS 45 21-3 V102Q (CDR- QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVR
grafted heavy chain) QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDTST heavy
chain variable STVYMELSSLRSEDTAVYYCARSGGLPDQWGQGTTVTVS
region--amino acid S 46 21-3 V102W (CDR-
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVR grafted heavy chain)
QAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDTST heavy chain variable
STVYMELSSLRSEDTAVYYCARSGGLPDWWGQGTTVTV region--amino acid SS 47
18-7 S92D (CDR- EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG grafted
light chain) QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF light chain
variable AVYYCQQWDGNPRTFGGGTKLEIK region--amino acid
48 18-7 S92E (CDR- EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG grafted
light chain) QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF light chain
variable AVYYCQQWEGNPRTFGGGTKLEIK region--amino acid 49 18-7 S92H
(CDR- EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG grafted light chain)
QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF light chain variable
AVYYCQQWHGNPRTFGGGTKLEIK region--amino acid 50 18-7 S92N (CDR-
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG grafted light chain)
QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF light chain variable
AVYYCQQWNGNPRTFGGGTKLEIK region--amino acid 51 18-7 S92Q (CDR-
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG grafted light chain)
QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF light chain variable
AVYYCQQWQGNPRTFGGGTKLEIK region--amino acid 52 21-3 CDRv (CDR-
QVQLVQSGAEVKKPGASVKVSCKASGFX.sub.1X.sub.2X.sub.3DTX.sub.4X.sub.5H
grafted heavy chain) WVRQAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTR heavy
chain variable DTSTSTVYMELSSLRSEDTAVYYCARSGGX.sub.6PDX.sub.7WGQGT
region--amino acid TVTVSS X.sub.1 = D or E X.sub.2 = I, P, or V
X.sub.3 = G, Q, S, or V X.sub.4 = F or Y X.sub.5 = I or M X.sub.6 =
L or M X.sub.7 = E, I, K, L, M, Q, T, V, W, or Y 53 18-7 CDRv (CDR-
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPG grafted light chain)
QAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTISRLEPEDF light chain variable
AVYYCX.sub.1QWX.sub.2X.sub.3X.sub.4PRTFGGGTKLEIK region--amino acid
X.sub.1 = Q or N X.sub.2 = D, E, H, N, Q, or S X.sub.3 = A or G
X.sub.4 = D, F, K, N, R, S, or T 54 21-3 CDRv (heavy
QVQLVQSGAEVKKPGASVKVSCKASGFX.sub.1X.sub.2X.sub.3DTX.sub.4X.sub.5H
chain contains murine WVRQAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTR S93)
heavy chain DTSTSTVYMELSSLRSEDTAVYYCSRSGGX.sub.6PDX.sub.7WGQGT
variable region--amino TVTVSS acid X.sub.1 = D or E X.sub.2 = I, P,
or V X.sub.3 = G, Q, S, or V X.sub.4 = F or Y X.sub.5 = I or M
X.sub.6 = L or M X.sub.7 = E, I, K, L, M, Q, T, V, W, or Y 76
QQWSGTPRT 78 QQWSGDPRT 80 QQWSGFPRT 82 QQWSGKPRT 84 QQWSGRPRT 86
QQWSGSPRT (L8; VL) SEQ ID NO: 490
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (L8; VH) SEQ ID NO: 491
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQVRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDVWGQGTIVIVSS (Clone 34; VL) SEQ ID
NO: 492
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 34; VH) SEQ ID NO: 493
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHIKYDPKFQGRVIMIRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDWWGQGTIVIVSS (Clone 2; VL) SEQ ID
NO: 494
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 2; VH) SEQ ID NO: 495
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHIKYSPKFQGRVIMIRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDWWGQGTIVIVSS (Clone 52; VL) SEQ ID
NO: 496
EIVLTQSPGTLSLSPGERATLSCGASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 52; VH) SEQ ID NO: 497
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHIKYSPKFQGRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDWWGQGTTVTVSS (Clone 46; VL) SEQ ID
NO: 498
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 46; VH) SEQ ID NO: 499
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHVKYSPKFQVRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDWWGQGTTVTVSS (Clone 47; VL) SEQ ID
NO: 500
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 47; VH) SEQ ID NO: 501
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHVKYDPKFQTRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDWWGQGTTVTVSS (Clone 14; VL) SEQ ID
NO: 502
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWQGNP RTFGGGTKLEIK (Clone 14; VH) SEQ ID NO: 503
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHiKYDPKFQkRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDMWGQGTTVTVSS (Clone 16L; VL) SEQ ID
NO: 504
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWQGNP RTFGGGTKLEIK (Clone 16L; VH) SEQ ID NO: 505
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHvKiDPKFQVRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDMWGQGTTVTVSS (Clone 17L; VL) SEQ ID
NO: 506
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWQGNP RTFGGGTKLEIK (Clone 17L; VH) SEQ ID NO: 507
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHLKYDPKFQVRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDMWGQGTTVTVSS (Clone 17L-1; VL) SEQ
ID NO: 508
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWQGNP RTFGGGTKLEIK (Clone 17L-1; VH) SEQ ID NO: 509
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHLKYDPKFQRRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDMWGQGTTVTVSS (Clone 23; VL) SEQ ID
NO: 510
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 23; VH) SEQ ID NO: 511
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHLKYDPKFQNRVTMTRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDKWGQGTTVTVSS (Clone A1; VL) SEQ ID
NO: 512
EIVLTQSPGTLSLSPGERATLSCGASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone A1; VH) SEQ ID NO: 513
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHLKYDPKFQNRVIMIRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDKWGQGTIVIVSS (Clone 53; VL) SEQ ID
NO: 514
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone 53; VH) SEQ ID NO: 515
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHLKYDPKFQERVIMIRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDKWGQGTIVIVSS (Clone E1; VL) SEQ ID
NO: 516
EIVLTQSPGTLSLSPGERATLSCGASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWEGNP RTFGGGTKLEIK (Clone E1; VH) SEQ ID NO: 517
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIEPASGHLKYDPKFQERVIMIRDT-
STSTVYMELSSLRSEDTAVY YCARSGGLPDKWGQGTIVIVSS (Clone 3-17L V-A; VL)
SEQ ID NO: 518
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWQGNP RTFGGGTKLEIK (Clone 3-17L V-A; VH) SEQ ID NO:
519
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHLKYDPKFQGRVTITRDT-
SASTAYMELSSLRSEDTAVY YCARSGGLPDMWGQGTIVIVSS (Clone 3-17L; VL) SEQ
ID NO: 520
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWQGNP RTFGGGTKLEIK (Clone 3-17L; VH) SEQ ID NO: 521
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHLKYDPKFQGRVTITRDT-
SASTVYMELSSLRSEDTAVY YCARSGGLPDMWGQGTIVIVSS (Clone L8mod; VL) SEQ
ID NO: 522
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (Clone L8mod; VH) SEQ ID NO: 523
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDVWGQGTIVIVSS (Clone X-V; VL) SEQ ID
NO: 524
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (Clone X-V; VH) SEQ ID NO: 525
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQVRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone X; VL) SEQ ID
NO: 526
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (Clone X; VH) SEQ ID NO: 527
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone H3-1; VL) SEQ ID
NO: 528
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (Clone H3-1; VH) SEQ ID NO: 529
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDLWGQGTIVIVSS (Clone XL3-6; VL) SEQ
ID NO: 530
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCSQWSGNP RTFGGGTKLEIK (Clone XL3-6; VH) SEQ ID NO: 531
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone XL3-10; VL) SEQ
ID NO: 532
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RSFGGGTKLEIK (Clone XL3-10; VH) SEQ ID NO: 533
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone XL3-15; VL) SEQ
ID NO: 534
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSRNP RTFGGGTKLEIK (Clone XL3-15; VH) SEQ ID NO: 535
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone L3-13; VL) SEQ
ID NO: 536
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWKGNP RTFGGGTKLEIK (Clone L3-13; VH) SEQ ID NO: 537
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHTKYDPKFQGRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone H2-2; VL) SEQ ID
NO: 538
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (Clone H2-2; VH) SEQ ID NO: 539
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHSKYDPKFQVRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS (Clone H2-5; VL) SEQ ID
NO: 540
EIVLTQSPGTLSLSPGERATLSCRASSSVSYMYWYQQKPGQAPRLLIYATSNLASGIPDRFSGSGSGTDFTLTI-
SRLEPEDFAVYYCQQWSGNP RTFGGGTKLEIK (Clone H2-5; VH) SEQ ID NO: 541
QVQLVQSGAEVKKPGASVKVSCKASGFDIQDTYMHWVRQAPGQGLEWMGRIDPASGHYKYDPKFQVRATITTDT-
SASTAYLQLSSLRSEDTAVY YCARSGGLPDFWGQGTIVIVSS modified G1 SEQ ID NO:
542
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS-
SLGTQTYICNVNHKPSNTKV
DKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH-
NAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAV-
EWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK G2 constant
domains SEQ ID NO: 543
ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS-
NFGTQTYTCNVDHKPSNTKV
DKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKT-
KPREEQFNSTFRVVSVLTVV
HQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWES-
NGQPENNYKTTPPMLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Kappa constant domain SEQ ID NO: 544
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL-
SKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC (L8 HFR1) SEQ ID NO: 545
QVQLVQSGAEVKKPGASVKVSCKAS (L8 HFR2) SEQ ID NO: 546 WVRQAPGQGLEWMG
(L8 HFR3) SEQ ID NO: 547 RVTMTRDTSTSTVYMELSSLRSEDTAVYYC (L8 HFR4)
SEQ ID NO: 548 WGQGTTVTVSS (L8 LFR1) SEQ ID NO: 549
EIVLTQSPGTLSLSPGERATLSC (L8 LFR2) SEQ ID NO: 550 WYQQKPGQAPRLLIY
(L8 LFR3) SEQ ID NO: 551 GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC (L8 LFR4)
SEQ ID NO: 552 FGGGTKLEIK (VH FR3) SEQ ID NO: 586
RATITTDTSASTAYLQLSSLRSEDTAVYYC (VH FR3) SEQ ID NO: 587
RVTITRDTSASTVYMELSSLRSEDTAVYYC (VH FR3) SEQ ID NO: 588
RVTITRDTSASTAYMELSSLRSEDTAVYYC
SEQUENCE LISTINGS
1
SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 694 <210>
SEQ ID NO 1 <211> LENGTH: 348 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polynucleotide <400> SEQUENCE: 1 gaagttcagc
tgcaacagtc tggcgccgag ctggttaagc ctggcgcttc tgtgaagctg 60
agctgtaccg cctctggctt cgacatccaa gacacctaca tgcactgggt caagcagagg
120 cctgagcagg gactcgagtg gatcggcaga attgatcctg ccagcggcca
caccaaatac 180 gaccccaagt tccaagtgaa ggccaccatc accaccgaca
ccagcagcaa taccgcctac 240 ctgcagctga gcagcctgac ctctgaagat
accgccgtgt actactgcag cagatctggc 300 ggactgcccg atgtttgggg
agccggaaca accgtgacag tgtccagc 348 <210> SEQ ID NO 2
<211> LENGTH: 348 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polynucleotide <400> SEQUENCE: 2 gaggttcaac ttcaacaatc
gggggccgag ctggttaagc ccggcgcttc tgtaaaattg 60 tcttgcactg
cctctgggtt tgacatccaa gatacatata tgcattgggt gaaacagcgt 120
cccgagcagg gcttggagtg gattggacgt attgaccccg cctctgggca cacgaaatat
180 gatcctaagt tccaggttaa agcgactatc acaacggaca cctccagcaa
tacggcttat 240 ttacagttat cctcgctgac ctctgaggat actgcagtgt
actactgctc tcgctctggt 300 ggtctgccag acgtgtgggg tgcaggaact
acagttactg tgtcttca 348 <210> SEQ ID NO 3 <211> LENGTH:
116 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE: 3
Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5
10 15 Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asp Ile Gln Asp
Thr 20 25 30 Tyr Met His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu
Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys
Tyr Asp Pro Lys Phe 50 55 60 Gln Val Lys Ala Thr Ile Thr Thr Asp
Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Thr
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Gly
Leu Pro Asp Val Trp Gly Ala Gly Thr Thr Val 100 105 110 Thr Val Ser
Ser 115 <210> SEQ ID NO 4 <211> LENGTH: 318 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic polynucleotide <400> SEQUENCE: 4
caaattgtgc tgtctcagag ccccgccatc ctgagtgctt ctccaggcga gaaagtgacc
60 atgacctgca gagccagcag cagcgtgtcc tacatgtact ggtatcagca
gaagcccggc 120 agcagcccca agccttggat ctacgccaca agcaatctgg
ccagcggcgt gcccgataga 180 ttttctggct ctggcagcgg caccagctac
agcctgacaa tctctagagt ggaagccgag 240 gatgccgcca cctactactg
tcaacagtgg agcggcaacc ccagaacctt tggcggaggc 300 accaagctgg aaatcaag
318 <210> SEQ ID NO 5 <211> LENGTH: 318 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic polynucleotide <400> SEQUENCE: 5
caaatcgtcc tgtcacagtc cccggcgatc ctttctgctt caccaggaga gaaggtaacc
60 atgacatgtc gcgcctcttc ctcagtttct tacatgtact ggtaccagca
gaaaccagga 120 tcatctccca aaccctggat ctacgctaca tcaaaccttg
catctggcgt gccagaccgt 180 ttttcagggt cgggctcggg gacttcctat
tcattaacca tttctcgcgt agaagcggaa 240 gacgccgcca cgtattattg
tcagcagtgg tcaggaaatc cgcgcacatt cggaggcgga 300 acgaaattgg agatcaaa
318 <210> SEQ ID NO 6 <211> LENGTH: 106 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic polypeptide <400> SEQUENCE: 6 Gln Ile Val
Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu
Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25
30 Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser
Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg
Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp
Ser Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu
Ile Lys 100 105 <210> SEQ ID NO 7 <211> LENGTH: 30
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic oligonucleotide <400>
SEQUENCE: 7 ggcttcgaca tccaagacac ctacatgcac 30 <210> SEQ ID
NO 8 <211> LENGTH: 30 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 8 gggtttgaca tccaagatac
atatatgcat 30 <210> SEQ ID NO 9 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 9 Gly
Phe Asp Ile Gln Asp Thr Tyr Met His 1 5 10 <210> SEQ ID NO 10
<211> LENGTH: 51 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 10 agaattgatc ctgccagcgg
ccacaccaaa tacgacccca agttccaagt g 51 <210> SEQ ID NO 11
<211> LENGTH: 51 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 11 cgtattgacc ccgcctctgg
gcacacgaaa tatgatccta agttccaggt t 51 <210> SEQ ID NO 12
<211> LENGTH: 17 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
peptide
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (2)..(2) <223> OTHER INFORMATION: Ile or Leu
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (3)..(3) <223> OTHER INFORMATION: Asp or Glu
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (5)..(5) <223> OTHER INFORMATION: Ala or Glu
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (6)..(6) <223> OTHER INFORMATION: Gly or Ser
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (7)..(7) <223> OTHER INFORMATION: Ala or Gly
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (9)..(9) <223> OTHER INFORMATION: Ile, Leu, Thr, or
Val <220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (11)..(11) <223> OTHER INFORMATION: Ile, Leu, Met,
Ser, Thr, Val, or Tyr <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (12)..(12) <223> OTHER
INFORMATION: Asp, Ile, Asn, Arg, or Ser <220> FEATURE:
<221> NAME/KEY: MOD_RES <222> LOCATION: (16)..(16)
<223> OTHER INFORMATION: Gln or Arg <220> FEATURE:
<221> NAME/KEY: MOD_RES <222> LOCATION: (17)..(17)
<223> OTHER INFORMATION: Ala, Asp, Glu, Gly, His, Lys, Leu,
Met, Asn, Pro, Arg, Ser, Thr, or Val <400> SEQUENCE: 12 Arg
Xaa Xaa Pro Xaa Xaa Xaa His Xaa Lys Xaa Xaa Pro Lys Phe Xaa 1 5 10
15 Xaa <210> SEQ ID NO 13 <211> LENGTH: 21 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic oligonucleotide <400> SEQUENCE: 13
tctggcggac tgcccgatgt t 21 <210> SEQ ID NO 14 <211>
LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic oligonucleotide
<400> SEQUENCE: 14 tctggtggtc tgccagacgt g 21 <210> SEQ
ID NO 15 <211> LENGTH: 7 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 15 Ser Gly Gly Leu Pro Asp Val 1 5
<210> SEQ ID NO 16 <211> LENGTH: 30 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic oligonucleotide <400> SEQUENCE: 16 agagccagca
gcagcgtgtc ctacatgtac 30 <210> SEQ ID NO 17 <211>
LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic oligonucleotide
<400> SEQUENCE: 17 cgcgcctctt cctcagtttc ttacatgtac 30
<210> SEQ ID NO 18 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (1)..(1) <223> OTHER
INFORMATION: Gly, Arg, or Trp <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (7)..(7) <223> OTHER
INFORMATION: Ile or Ser <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (8)..(8) <223> OTHER
INFORMATION: Phe or Tyr <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (9)..(9) <223> OTHER
INFORMATION: Leu or Met <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (10)..(10) <223> OTHER
INFORMATION: Arg or Tyr <400> SEQUENCE: 18 Xaa Ala Ser Ser
Ser Val Xaa Xaa Xaa Xaa 1 5 10 <210> SEQ ID NO 19 <211>
LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic oligonucleotide
<400> SEQUENCE: 19 gccacaagca atctggccag c 21 <210> SEQ
ID NO 20 <211> LENGTH: 21 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 20 gctacatcaa accttgcatc t 21
<210> SEQ ID NO 21 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (2)..(2) <223> OTHER
INFORMATION: Lys or Thr <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (3)..(3) <223> OTHER
INFORMATION: Glu, Pro, or Ser <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (4)..(4) <223> OTHER
INFORMATION: Leu, Asn, or Pro <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (6)..(6) <223> OTHER
INFORMATION: Ala or Thr <400> SEQUENCE: 21 Ala Xaa Xaa Xaa
Leu Xaa Ser 1 5 <210> SEQ ID NO 22 <211> LENGTH: 27
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic oligonucleotide <400>
SEQUENCE: 22 caacagtgga gcggcaaccc cagaacc 27 <210> SEQ ID NO
23 <211> LENGTH: 27 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 23 cagcagtggt caggaaatcc
gcgcaca 27 <210> SEQ ID NO 24 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 24 Gln Gln Trp Ser Gly Asn Pro Arg
Thr 1 5 <210> SEQ ID NO 25 <211> LENGTH: 348
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polynucleotide <400> SEQUENCE:
25 caagtacaat tagtccagtc gggtgccgag gtaaaaaaac ctggagcatc
cgtaaaactg 60 tcttgcaaag catcggggtt tgacatccag gacacctaca
tgcactgggt gcgtcaagct 120 ccaggacagg gattagagtg gatgggtcgc
atcgaccccg cgagcggaca cacgaaatac 180 gaccctaaat ttcaagtacg
tgtcacgatg actaccgaca ctagtacgag cactgtttat 240 atggaattgt
cctcgttacg ctcagaggat acggcagtct attattgcag ccgttccgga 300
ggcttacccg acgtctgggg acagggaact actgtaacag tcagtagt 348
<210> SEQ ID NO 26 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 26 Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys
Leu Ser Cys Lys Ala Ser Gly Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Val Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Gly Leu Pro Asp Val Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 27 <211> LENGTH: 318 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polynucleotide <400> SEQUENCE: 27 gagattgtgt
taacgcaatc accggggact ttatcgctgt cgccggggga gcgcgttaca 60
atgtcttgtc gcgcttcctc ttcggtttca tacatgtatt ggtatcaaca aaaaccggga
120 caggctccac gcccctggat ttacgctact agcaatttgg cctcgggcgt
tcccgaccgc 180 ttcagcgggt cagggagcgg caccgattac acgttgacca
tctctcgtct ggaacctgaa 240 gacttcgcgg tctattactg tcaacaatgg
tcgggaaatc cccgtacatt tggcggaggg 300 acgaagttgg aaattaaa 318
<210> SEQ ID NO 28 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 28 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Val
Thr Met Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Trp Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 29 <211> LENGTH: 30 <212>
TYPE: DNA <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic oligonucleotide <400> SEQUENCE: 29
gggtttgaca tccaggacac ctacatgcac 30 <210> SEQ ID NO 30
<211> LENGTH: 51 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 30 cgcatcgacc ccgcgagcgg
acacacgaaa tacgacccta aatttcaagt a 51 <210> SEQ ID NO 31
<211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 31 tccggaggct tacccgacgt c 21
<210> SEQ ID NO 32 <211> LENGTH: 36 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic oligonucleotide <400> SEQUENCE: 32 cgcgcttcct
cttcggtttc atacatgtat tggtat 36 <210> SEQ ID NO 33
<211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
oligonucleotide <400> SEQUENCE: 33 gctactagca atttggcctc g 21
<210> SEQ ID NO 34 <211> LENGTH: 27 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic oligonucleotide <400> SEQUENCE: 34 caacaatggt
cgggaaatcc ccgtaca 27 <210> SEQ ID NO 35 <211> LENGTH:
348 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polynucleotide <400> SEQUENCE:
35 caagtacaat tagtccagtc gggtgccgag gtaaaaaaac ctggagcatc
cgtaaaactg 60 tcttgcaaag catcggggtt tgacatccag gacacctaca
tgcactgggt gcgtcaagct 120 ccaggacagg gattagagtg gatgggtcgc
atcgaccccg cgagcggaca cacgaaatac 180 gaccctaaat ttcaagtacg
tgtcacgatg actcgtgaca ctagtacgag cactgtttat 240 atggaattgt
cctcgttacg ctcagaggat acggcagtct attattgcag ccgttccgga 300
ggcttacccg acgtctgggg acagggaact actgtaacag tcagtagt 348
<210> SEQ ID NO 36 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 36 Gln Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys
Leu Ser Cys Lys Ala Ser Gly Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro
Lys Phe 50 55 60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr
Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Gly Leu Pro Asp
Val Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115
<210> SEQ ID NO 37 <211> LENGTH: 318 <212> TYPE:
DNA <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polynucleotide <400> SEQUENCE: 37 gagattgtgt
taacgcaatc accggggact ttatcgctgt cgccggggga gcgcgcgaca 60
ctgtcttgtc gcgcttcctc ttcggtttca tacatgtatt ggtatcaaca aaaaccggga
120 caggctccac gcctgctgat ttacgctact agcaatttgg cctcgggcat
ccccgaccgc 180 ttcagcgggt cagggagcgg caccgatttt acgttgacca
tctctcgtct ggaacctgaa 240 gacttcgcgg tctattactg tcaacaatgg
tcgggaaatc cccgtacatt tggcggaggg 300 acgaagttgg aaattaaa 318
<210> SEQ ID NO 38 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 38 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 39 <211> LENGTH: 348
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polynucleotide <400> SEQUENCE:
39 caagtacaat tagtccagtc gggtgccgag gtaaaaaaac ctggagcatc
cgtaaaagtc 60 tcttgcaaag catcggggtt tgacatccag gacacctaca
tgcactgggt gcgtcaagct 120 ccaggacagg gattagagtg gatgggtcgc
atcgaccccg cgagcggaca cacgaaatac 180 gaccctaaat ttcaagtacg
tgtcacgatg actcgtgaca ctagtacgag cactgtttat 240 atggaattgt
cctcgttacg ctcagaggat acggcagtct attattgcgc acgttccgga 300
ggcttacccg acgtctgggg acagggaact actgtaacag tcagtagt 348
<210> SEQ ID NO 40 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 40 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Val Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 41 <211> LENGTH: 318 <212> TYPE: DNA
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polynucleotide <400> SEQUENCE: 41 gagattgtgt
taacgcaatc accggggact ttatcgctgt cgccggggga gcgcgcgaca 60
ctgtcttgtc gcgcttcctc ttcggtttca tacatgtatt ggtatcaaca aaaaccggga
120 caggctccac gcctgctgat ttacgctact agcaatttgg cctcgggcgt
tcccgaccgc 180 ttcagcgggt cagggagcgg caccgattac acgttgacca
tctctcgtct ggaacctgaa 240 gacttcgcgg tctattactg tcaacaatgg
tcgggaaatc cccgtacatt tggcggaggg 300 acgaagttgg aaattaaa 318
<210> SEQ ID NO 42 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 42 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 43 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE: 43
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 Phe Asp Ile Gln Asp
Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys
Tyr Asp Pro Lys Phe 50 55 60 Gln Val Arg Val Thr Met Thr Arg Asp
Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly
Leu Pro Asp Lys Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser
Ser 115 <210> SEQ ID NO 44 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE: 44
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 Phe Asp Ile Gln Asp
Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys
Tyr Asp Pro Lys Phe 50 55 60
Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65
70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Met Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 45 <211> LENGTH: 116 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 45 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Gln Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 46 <211> LENGTH: 116 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 46 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Trp Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 47 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 47 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Asp Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 48 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 48 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 49 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE: 49
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5
10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr
Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp
Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys
Gln Gln Trp His Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr
Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 50 <211>
LENGTH: 106 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polypeptide
<400> SEQUENCE: 50 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr
Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg
Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys
Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn
Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Asn Gly Asn Pro Arg Thr 85
90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210>
SEQ ID NO 51 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 51 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Gln Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 52
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (28)..(28) <223> OTHER INFORMATION: Asp
or Glu <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (29)..(29) <223> OTHER INFORMATION:
Ile, Pro, or Val <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (30)..(30) <223> OTHER INFORMATION:
Gly, Gln, Ser, or Val <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (33)..(33) <223> OTHER
INFORMATION: Phe or Tyr <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (34)..(34) <223> OTHER
INFORMATION: Ile or Met <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (102)..(102) <223> OTHER
INFORMATION: Leu or Met <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (105)..(105) <223> OTHER
INFORMATION: Glu, Ile, Lys, Leu, Met, Gln, Thr, Val, Trp, or Tyr
<400> SEQUENCE: 52 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 Phe Xaa Xaa Xaa Asp Thr 20 25 30 Xaa Xaa His Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp
Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Val
Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Ser Gly Gly Xaa Pro Asp Xaa Trp Gly Gln Gly Thr Thr
Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID NO 53
<211> LENGTH: 106 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (88)..(88) <223> OTHER INFORMATION: Gln
or Asn <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (91)..(91) <223> OTHER INFORMATION:
Asp, Glu, His, Asn, Gln, or Ser <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (92)..(92) <223>
OTHER INFORMATION: Ala or Gly <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (93)..(93) <223>
OTHER INFORMATION: Asp, Phe, Lys, Asn, Arg, Ser, or Thr <400>
SEQUENCE: 53 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu
Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser
Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln
Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser
Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala
Val Tyr Tyr Cys Xaa Gln Trp Xaa Xaa Xaa Pro Arg Thr 85 90 95 Phe
Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO
54 <211> LENGTH: 116 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (28)..(28) <223> OTHER INFORMATION: Asp
or Glu <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (29)..(29) <223> OTHER INFORMATION:
Ile, Pro, or Val <220> FEATURE: <221> NAME/KEY: MOD_RES
<222> LOCATION: (30)..(30) <223> OTHER INFORMATION:
Gly, Gln, Ser, or Val <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (33)..(33) <223> OTHER
INFORMATION: Phe or Tyr <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (34)..(34) <223> OTHER
INFORMATION: Ile or Met <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (102)..(102) <223> OTHER
INFORMATION: Leu or Met <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (105)..(105) <223> OTHER
INFORMATION: Glu, Ile, Lys, Leu, Met, Gln, Thr, Val, Trp, or Tyr
<400> SEQUENCE: 54 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 Phe Xaa Xaa Xaa Asp Thr 20 25 30 Xaa Xaa His Trp Val Arg
Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp
Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Val
Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ser Arg Ser Gly Gly Xaa Pro Asp Xaa Trp Gly Gln Gly Thr Thr
Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID NO 55
<400> SEQUENCE: 55 000 <210> SEQ ID NO 56 <400>
SEQUENCE: 56 000 <210> SEQ ID NO 57 <400> SEQUENCE: 57
000 <210> SEQ ID NO 58 <400> SEQUENCE: 58 000
<210> SEQ ID NO 59 <400> SEQUENCE: 59 000 <210>
SEQ ID NO 60 <400> SEQUENCE: 60 000 <210> SEQ ID NO 61
<400> SEQUENCE: 61 000 <210> SEQ ID NO 62 <400>
SEQUENCE: 62 000 <210> SEQ ID NO 63 <400> SEQUENCE: 63
000 <210> SEQ ID NO 64 <400> SEQUENCE: 64 000
<210> SEQ ID NO 65
<400> SEQUENCE: 65 000 <210> SEQ ID NO 66 <400>
SEQUENCE: 66 000 <210> SEQ ID NO 67 <400> SEQUENCE: 67
000 <210> SEQ ID NO 68 <400> SEQUENCE: 68 000
<210> SEQ ID NO 69 <400> SEQUENCE: 69 000 <210>
SEQ ID NO 70 <400> SEQUENCE: 70 000 <210> SEQ ID NO 71
<400> SEQUENCE: 71 000 <210> SEQ ID NO 72 <400>
SEQUENCE: 72 000 <210> SEQ ID NO 73 <400> SEQUENCE: 73
000 <210> SEQ ID NO 74 <400> SEQUENCE: 74 000
<210> SEQ ID NO 75 <400> SEQUENCE: 75 000 <210>
SEQ ID NO 76 <211> LENGTH: 9 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 76 Gln Gln Trp Ser Gly Thr
Pro Arg Thr 1 5 <210> SEQ ID NO 77 <400> SEQUENCE: 77
000 <210> SEQ ID NO 78 <211> LENGTH: 9 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 78 Gln Gln Trp
Ser Gly Asp Pro Arg Thr 1 5 <210> SEQ ID NO 79 <400>
SEQUENCE: 79 000 <210> SEQ ID NO 80 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 80 Gln
Gln Trp Ser Gly Phe Pro Arg Thr 1 5 <210> SEQ ID NO 81
<400> SEQUENCE: 81 000 <210> SEQ ID NO 82 <211>
LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 82 Gln Gln Trp Ser Gly Lys Pro Arg Thr 1 5 <210>
SEQ ID NO 83 <400> SEQUENCE: 83 000 <210> SEQ ID NO 84
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 84 Gln Gln Trp Ser Gly Arg Pro Arg Thr 1 5
<210> SEQ ID NO 85 <400> SEQUENCE: 85 000 <210>
SEQ ID NO 86 <211> LENGTH: 9 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 86 Gln Gln Trp Ser Gly Ser
Pro Arg Thr 1 5 <210> SEQ ID NO 87 <400> SEQUENCE: 87
000 <210> SEQ ID NO 88 <400> SEQUENCE: 88 000
<210> SEQ ID NO 89 <400> SEQUENCE: 89 000 <210>
SEQ ID NO 90 <400> SEQUENCE: 90 000 <210> SEQ ID NO 91
<400> SEQUENCE: 91 000 <210> SEQ ID NO 92 <400>
SEQUENCE: 92 000 <210> SEQ ID NO 93 <400> SEQUENCE: 93
000 <210> SEQ ID NO 94
<400> SEQUENCE: 94 000 <210> SEQ ID NO 95 <400>
SEQUENCE: 95 000 <210> SEQ ID NO 96 <400> SEQUENCE: 96
000 <210> SEQ ID NO 97 <400> SEQUENCE: 97 000
<210> SEQ ID NO 98 <400> SEQUENCE: 98 000 <210>
SEQ ID NO 99 <400> SEQUENCE: 99 000 <210> SEQ ID NO 100
<400> SEQUENCE: 100 000 <210> SEQ ID NO 101 <400>
SEQUENCE: 101 000 <210> SEQ ID NO 102 <400> SEQUENCE:
102 000 <210> SEQ ID NO 103 <400> SEQUENCE: 103 000
<210> SEQ ID NO 104 <400> SEQUENCE: 104 000 <210>
SEQ ID NO 105 <400> SEQUENCE: 105 000 <210> SEQ ID NO
106 <400> SEQUENCE: 106 000 <210> SEQ ID NO 107
<400> SEQUENCE: 107 000 <210> SEQ ID NO 108 <400>
SEQUENCE: 108 000 <210> SEQ ID NO 109 <400> SEQUENCE:
109 000 <210> SEQ ID NO 110 <400> SEQUENCE: 110 000
<210> SEQ ID NO 111 <400> SEQUENCE: 111 000 <210>
SEQ ID NO 112 <400> SEQUENCE: 112 000 <210> SEQ ID NO
113 <400> SEQUENCE: 113 000 <210> SEQ ID NO 114
<400> SEQUENCE: 114 000 <210> SEQ ID NO 115 <400>
SEQUENCE: 115 000 <210> SEQ ID NO 116 <400> SEQUENCE:
116 000 <210> SEQ ID NO 117 <400> SEQUENCE: 117 000
<210> SEQ ID NO 118 <400> SEQUENCE: 118 000 <210>
SEQ ID NO 119 <400> SEQUENCE: 119 000 <210> SEQ ID NO
120 <400> SEQUENCE: 120 000 <210> SEQ ID NO 121
<400> SEQUENCE: 121 000 <210> SEQ ID NO 122 <400>
SEQUENCE: 122 000 <210> SEQ ID NO 123 <400> SEQUENCE:
123 000 <210> SEQ ID NO 124 <400> SEQUENCE: 124 000
<210> SEQ ID NO 125 <400> SEQUENCE: 125 000 <210>
SEQ ID NO 126 <400> SEQUENCE: 126 000 <210> SEQ ID NO
127 <400> SEQUENCE: 127 000 <210> SEQ ID NO 128
<400> SEQUENCE: 128 000 <210> SEQ ID NO 129 <400>
SEQUENCE: 129 000
<210> SEQ ID NO 130 <400> SEQUENCE: 130 000 <210>
SEQ ID NO 131 <400> SEQUENCE: 131 000 <210> SEQ ID NO
132 <400> SEQUENCE: 132 000 <210> SEQ ID NO 133
<400> SEQUENCE: 133 000 <210> SEQ ID NO 134 <400>
SEQUENCE: 134 000 <210> SEQ ID NO 135 <400> SEQUENCE:
135 000 <210> SEQ ID NO 136 <400> SEQUENCE: 136 000
<210> SEQ ID NO 137 <400> SEQUENCE: 137 000 <210>
SEQ ID NO 138 <400> SEQUENCE: 138 000 <210> SEQ ID NO
139 <400> SEQUENCE: 139 000 <210> SEQ ID NO 140
<400> SEQUENCE: 140 000 <210> SEQ ID NO 141 <400>
SEQUENCE: 141 000 <210> SEQ ID NO 142 <400> SEQUENCE:
142 000 <210> SEQ ID NO 143 <400> SEQUENCE: 143 000
<210> SEQ ID NO 144 <400> SEQUENCE: 144 000 <210>
SEQ ID NO 145 <400> SEQUENCE: 145 000 <210> SEQ ID NO
146 <400> SEQUENCE: 146 000 <210> SEQ ID NO 147
<400> SEQUENCE: 147 000 <210> SEQ ID NO 148 <400>
SEQUENCE: 148 000 <210> SEQ ID NO 149 <400> SEQUENCE:
149 000 <210> SEQ ID NO 150 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <220> FEATURE:
<221> NAME/KEY: MOD_RES <222> LOCATION: (3)..(3)
<223> OTHER INFORMATION: Asp or Glu <220> FEATURE:
<221> NAME/KEY: MOD_RES <222> LOCATION: (4)..(4)
<223> OTHER INFORMATION: Ile, Leu, Pro, or Val <220>
FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION:
(5)..(5) <223> OTHER INFORMATION: Gly, Gln, Ser, or Val
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (7)..(7) <223> OTHER INFORMATION: Ala, Ser, or Thr
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (8)..(8) <223> OTHER INFORMATION: Phe or Tyr
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (9)..(9) <223> OTHER INFORMATION: Ile, Leu, or Met
<400> SEQUENCE: 150 Gly Phe Xaa Xaa Xaa Asp Xaa Xaa Xaa His 1
5 10 <210> SEQ ID NO 151 <400> SEQUENCE: 151 000
<210> SEQ ID NO 152 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (1)..(1) <223> OTHER
INFORMATION: Leu or Ser <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (2)..(2) <223> OTHER
INFORMATION: Ala or Gly <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (4)..(4) <223> OTHER
INFORMATION: Ala, Leu, or Met <220> FEATURE: <221>
NAME/KEY: MOD_RES <222> LOCATION: (6)..(6) <223> OTHER
INFORMATION: Asp or Glu <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (7)..(7) <223> OTHER
INFORMATION: Lys, Met, Gln, Arg, Ser, Thr, Val, or Trp <400>
SEQUENCE: 152 Xaa Xaa Gly Xaa Pro Xaa Xaa 1 5 <210> SEQ ID NO
153 <400> SEQUENCE: 153 000 <210> SEQ ID NO 154
<400> SEQUENCE: 154 000 <210> SEQ ID NO 155 <211>
LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <220>
FEATURE: <221> NAME/KEY: MOD_RES <222> LOCATION:
(1)..(1) <223> OTHER INFORMATION: His, Asn, Gln, or Ser
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (3)..(3) <223> OTHER INFORMATION: Phe, His, Ile,
Pro, Arg, Ser, Trp, or Tyr
<220> FEATURE: <221> NAME/KEY: MOD_RES <222>
LOCATION: (4)..(4) <223> OTHER INFORMATION: Asp, Glu, His,
Asn, Gln, Ser, or Val <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (5)..(5) <223> OTHER
INFORMATION: Ala, Asp, Gly, Gln, or Ser <220> FEATURE:
<221> NAME/KEY: MOD_RES <222> LOCATION: (6)..(6)
<223> OTHER INFORMATION: Asp, Phe, His, Lys, Leu, Met, Asn,
Gln, Arg, Ser, or Thr <220> FEATURE: <221> NAME/KEY:
MOD_RES <222> LOCATION: (9)..(9) <223> OTHER
INFORMATION: Ser or Thr <400> SEQUENCE: 155 Xaa Gln Xaa Xaa
Xaa Xaa Pro Arg Xaa 1 5 <210> SEQ ID NO 156 <400>
SEQUENCE: 156 000 <210> SEQ ID NO 157 <400> SEQUENCE:
157 000 <210> SEQ ID NO 158 <400> SEQUENCE: 158 000
<210> SEQ ID NO 159 <400> SEQUENCE: 159 000 <210>
SEQ ID NO 160 <400> SEQUENCE: 160 000 <210> SEQ ID NO
161 <400> SEQUENCE: 161 000 <210> SEQ ID NO 162
<400> SEQUENCE: 162 000 <210> SEQ ID NO 163 <400>
SEQUENCE: 163 000 <210> SEQ ID NO 164 <400> SEQUENCE:
164 000 <210> SEQ ID NO 165 <400> SEQUENCE: 165 000
<210> SEQ ID NO 166 <400> SEQUENCE: 166 000 <210>
SEQ ID NO 167 <400> SEQUENCE: 167 000 <210> SEQ ID NO
168 <400> SEQUENCE: 168 000 <210> SEQ ID NO 169
<400> SEQUENCE: 169 000 <210> SEQ ID NO 170 <400>
SEQUENCE: 170 000 <210> SEQ ID NO 171 <400> SEQUENCE:
171 000 <210> SEQ ID NO 172 <400> SEQUENCE: 172 000
<210> SEQ ID NO 173 <400> SEQUENCE: 173 000 <210>
SEQ ID NO 174 <400> SEQUENCE: 174 000 <210> SEQ ID NO
175 <400> SEQUENCE: 175 000 <210> SEQ ID NO 176
<400> SEQUENCE: 176 000 <210> SEQ ID NO 177 <400>
SEQUENCE: 177 000 <210> SEQ ID NO 178 <400> SEQUENCE:
178 000 <210> SEQ ID NO 179 <400> SEQUENCE: 179 000
<210> SEQ ID NO 180 <400> SEQUENCE: 180 000 <210>
SEQ ID NO 181 <400> SEQUENCE: 181 000 <210> SEQ ID NO
182 <400> SEQUENCE: 182 000 <210> SEQ ID NO 183
<400> SEQUENCE: 183 000 <210> SEQ ID NO 184 <400>
SEQUENCE: 184 000 <210> SEQ ID NO 185 <400> SEQUENCE:
185 000 <210> SEQ ID NO 186 <400> SEQUENCE: 186 000
<210> SEQ ID NO 187 <400> SEQUENCE: 187 000 <210>
SEQ ID NO 188
<400> SEQUENCE: 188 000 <210> SEQ ID NO 189 <400>
SEQUENCE: 189 000 <210> SEQ ID NO 190 <400> SEQUENCE:
190 000 <210> SEQ ID NO 191 <400> SEQUENCE: 191 000
<210> SEQ ID NO 192 <400> SEQUENCE: 192 000 <210>
SEQ ID NO 193 <400> SEQUENCE: 193 000 <210> SEQ ID NO
194 <400> SEQUENCE: 194 000 <210> SEQ ID NO 195
<400> SEQUENCE: 195 000 <210> SEQ ID NO 196 <400>
SEQUENCE: 196 000 <210> SEQ ID NO 197 <400> SEQUENCE:
197 000 <210> SEQ ID NO 198 <400> SEQUENCE: 198 000
<210> SEQ ID NO 199 <400> SEQUENCE: 199 000 <210>
SEQ ID NO 200 <400> SEQUENCE: 200 000 <210> SEQ ID NO
201 <400> SEQUENCE: 201 000 <210> SEQ ID NO 202
<400> SEQUENCE: 202 000 <210> SEQ ID NO 203 <400>
SEQUENCE: 203 000 <210> SEQ ID NO 204 <400> SEQUENCE:
204 000 <210> SEQ ID NO 205 <400> SEQUENCE: 205 000
<210> SEQ ID NO 206 <400> SEQUENCE: 206 000 <210>
SEQ ID NO 207 <400> SEQUENCE: 207 000 <210> SEQ ID NO
208 <400> SEQUENCE: 208 000 <210> SEQ ID NO 209
<400> SEQUENCE: 209 000 <210> SEQ ID NO 210 <400>
SEQUENCE: 210 000 <210> SEQ ID NO 211 <400> SEQUENCE:
211 000 <210> SEQ ID NO 212 <400> SEQUENCE: 212 000
<210> SEQ ID NO 213 <400> SEQUENCE: 213 000 <210>
SEQ ID NO 214 <400> SEQUENCE: 214 000 <210> SEQ ID NO
215 <400> SEQUENCE: 215 000 <210> SEQ ID NO 216
<400> SEQUENCE: 216 000 <210> SEQ ID NO 217 <400>
SEQUENCE: 217 000 <210> SEQ ID NO 218 <400> SEQUENCE:
218 000 <210> SEQ ID NO 219 <400> SEQUENCE: 219 000
<210> SEQ ID NO 220 <400> SEQUENCE: 220 000 <210>
SEQ ID NO 221 <400> SEQUENCE: 221 000 <210> SEQ ID NO
222 <400> SEQUENCE: 222 000 <210> SEQ ID NO 223
<400> SEQUENCE: 223 000 <210> SEQ ID NO 224
<400> SEQUENCE: 224 000 <210> SEQ ID NO 225 <400>
SEQUENCE: 225 000 <210> SEQ ID NO 226 <400> SEQUENCE:
226 000 <210> SEQ ID NO 227 <400> SEQUENCE: 227 000
<210> SEQ ID NO 228 <400> SEQUENCE: 228 000 <210>
SEQ ID NO 229 <400> SEQUENCE: 229 000 <210> SEQ ID NO
230 <400> SEQUENCE: 230 000 <210> SEQ ID NO 231
<400> SEQUENCE: 231 000 <210> SEQ ID NO 232 <400>
SEQUENCE: 232 000 <210> SEQ ID NO 233 <400> SEQUENCE:
233 000 <210> SEQ ID NO 234 <400> SEQUENCE: 234 000
<210> SEQ ID NO 235 <400> SEQUENCE: 235 000 <210>
SEQ ID NO 236 <400> SEQUENCE: 236 000 <210> SEQ ID NO
237 <400> SEQUENCE: 237 000 <210> SEQ ID NO 238
<400> SEQUENCE: 238 000 <210> SEQ ID NO 239 <400>
SEQUENCE: 239 000 <210> SEQ ID NO 240 <400> SEQUENCE:
240 000 <210> SEQ ID NO 241 <400> SEQUENCE: 241 000
<210> SEQ ID NO 242 <400> SEQUENCE: 242 000 <210>
SEQ ID NO 243 <400> SEQUENCE: 243 000 <210> SEQ ID NO
244 <400> SEQUENCE: 244 000 <210> SEQ ID NO 245
<400> SEQUENCE: 245 000 <210> SEQ ID NO 246 <400>
SEQUENCE: 246 000 <210> SEQ ID NO 247 <400> SEQUENCE:
247 000 <210> SEQ ID NO 248 <400> SEQUENCE: 248 000
<210> SEQ ID NO 249 <400> SEQUENCE: 249 000 <210>
SEQ ID NO 250 <400> SEQUENCE: 250 000 <210> SEQ ID NO
251 <400> SEQUENCE: 251 000 <210> SEQ ID NO 252
<400> SEQUENCE: 252 000 <210> SEQ ID NO 253 <400>
SEQUENCE: 253 000 <210> SEQ ID NO 254 <400> SEQUENCE:
254 000 <210> SEQ ID NO 255 <400> SEQUENCE: 255 000
<210> SEQ ID NO 256 <400> SEQUENCE: 256 000 <210>
SEQ ID NO 257 <400> SEQUENCE: 257 000 <210> SEQ ID NO
258 <400> SEQUENCE: 258 000 <210> SEQ ID NO 259
<400> SEQUENCE: 259 000
<210> SEQ ID NO 260 <400> SEQUENCE: 260 000 <210>
SEQ ID NO 261 <400> SEQUENCE: 261 000 <210> SEQ ID NO
262 <400> SEQUENCE: 262 000 <210> SEQ ID NO 263
<400> SEQUENCE: 263 000 <210> SEQ ID NO 264 <400>
SEQUENCE: 264 000 <210> SEQ ID NO 265 <400> SEQUENCE:
265 000 <210> SEQ ID NO 266 <400> SEQUENCE: 266 000
<210> SEQ ID NO 267 <400> SEQUENCE: 267 000 <210>
SEQ ID NO 268 <400> SEQUENCE: 268 000 <210> SEQ ID NO
269 <400> SEQUENCE: 269 000 <210> SEQ ID NO 270
<400> SEQUENCE: 270 000 <210> SEQ ID NO 271 <400>
SEQUENCE: 271 000 <210> SEQ ID NO 272 <400> SEQUENCE:
272 000 <210> SEQ ID NO 273 <400> SEQUENCE: 273 000
<210> SEQ ID NO 274 <400> SEQUENCE: 274 000 <210>
SEQ ID NO 275 <400> SEQUENCE: 275 000 <210> SEQ ID NO
276 <400> SEQUENCE: 276 000 <210> SEQ ID NO 277
<400> SEQUENCE: 277 000 <210> SEQ ID NO 278 <400>
SEQUENCE: 278 000 <210> SEQ ID NO 279 <400> SEQUENCE:
279 000 <210> SEQ ID NO 280 <400> SEQUENCE: 280 000
<210> SEQ ID NO 281 <400> SEQUENCE: 281 000 <210>
SEQ ID NO 282 <400> SEQUENCE: 282 000 <210> SEQ ID NO
283 <400> SEQUENCE: 283 000 <210> SEQ ID NO 284
<400> SEQUENCE: 284 000 <210> SEQ ID NO 285 <400>
SEQUENCE: 285 000 <210> SEQ ID NO 286 <400> SEQUENCE:
286 000 <210> SEQ ID NO 287 <400> SEQUENCE: 287 000
<210> SEQ ID NO 288 <400> SEQUENCE: 288 000 <210>
SEQ ID NO 289 <400> SEQUENCE: 289 000 <210> SEQ ID NO
290 <400> SEQUENCE: 290 000 <210> SEQ ID NO 291
<400> SEQUENCE: 291 000 <210> SEQ ID NO 292 <400>
SEQUENCE: 292 000 <210> SEQ ID NO 293 <400> SEQUENCE:
293 000 <210> SEQ ID NO 294 <400> SEQUENCE: 294 000
<210> SEQ ID NO 295 <400> SEQUENCE: 295 000
<210> SEQ ID NO 296 <400> SEQUENCE: 296 000 <210>
SEQ ID NO 297 <400> SEQUENCE: 297 000 <210> SEQ ID NO
298 <400> SEQUENCE: 298 000 <210> SEQ ID NO 299
<400> SEQUENCE: 299 000 <210> SEQ ID NO 300 <400>
SEQUENCE: 300 000 <210> SEQ ID NO 301 <400> SEQUENCE:
301 000 <210> SEQ ID NO 302 <400> SEQUENCE: 302 000
<210> SEQ ID NO 303 <400> SEQUENCE: 303 000 <210>
SEQ ID NO 304 <400> SEQUENCE: 304 000 <210> SEQ ID NO
305 <400> SEQUENCE: 305 000 <210> SEQ ID NO 306
<400> SEQUENCE: 306 000 <210> SEQ ID NO 307 <400>
SEQUENCE: 307 000 <210> SEQ ID NO 308 <400> SEQUENCE:
308 000 <210> SEQ ID NO 309 <400> SEQUENCE: 309 000
<210> SEQ ID NO 310 <400> SEQUENCE: 310 000 <210>
SEQ ID NO 311 <400> SEQUENCE: 311 000 <210> SEQ ID NO
312 <400> SEQUENCE: 312 000 <210> SEQ ID NO 313
<400> SEQUENCE: 313 000 <210> SEQ ID NO 314 <400>
SEQUENCE: 314 000 <210> SEQ ID NO 315 <400> SEQUENCE:
315 000 <210> SEQ ID NO 316 <400> SEQUENCE: 316 000
<210> SEQ ID NO 317 <400> SEQUENCE: 317 000 <210>
SEQ ID NO 318 <400> SEQUENCE: 318 000 <210> SEQ ID NO
319 <400> SEQUENCE: 319 000 <210> SEQ ID NO 320
<400> SEQUENCE: 320 000 <210> SEQ ID NO 321 <400>
SEQUENCE: 321 000 <210> SEQ ID NO 322 <400> SEQUENCE:
322 000 <210> SEQ ID NO 323 <400> SEQUENCE: 323 000
<210> SEQ ID NO 324 <400> SEQUENCE: 324 000 <210>
SEQ ID NO 325 <400> SEQUENCE: 325 000 <210> SEQ ID NO
326 <400> SEQUENCE: 326 000 <210> SEQ ID NO 327
<400> SEQUENCE: 327 000 <210> SEQ ID NO 328 <400>
SEQUENCE: 328 000 <210> SEQ ID NO 329 <400> SEQUENCE:
329 000 <210> SEQ ID NO 330 <400> SEQUENCE: 330 000
<210> SEQ ID NO 331 <400> SEQUENCE: 331 000
<210> SEQ ID NO 332 <400> SEQUENCE: 332 000 <210>
SEQ ID NO 333 <400> SEQUENCE: 333 000 <210> SEQ ID NO
334 <400> SEQUENCE: 334 000 <210> SEQ ID NO 335
<400> SEQUENCE: 335 000 <210> SEQ ID NO 336 <400>
SEQUENCE: 336 000 <210> SEQ ID NO 337 <400> SEQUENCE:
337 000 <210> SEQ ID NO 338 <400> SEQUENCE: 338 000
<210> SEQ ID NO 339 <400> SEQUENCE: 339 000 <210>
SEQ ID NO 340 <400> SEQUENCE: 340 000 <210> SEQ ID NO
341 <400> SEQUENCE: 341 000 <210> SEQ ID NO 342
<400> SEQUENCE: 342 000 <210> SEQ ID NO 343 <400>
SEQUENCE: 343 000 <210> SEQ ID NO 344 <400> SEQUENCE:
344 000 <210> SEQ ID NO 345 <400> SEQUENCE: 345 000
<210> SEQ ID NO 346 <400> SEQUENCE: 346 000 <210>
SEQ ID NO 347 <400> SEQUENCE: 347 000 <210> SEQ ID NO
348 <400> SEQUENCE: 348 000 <210> SEQ ID NO 349
<400> SEQUENCE: 349 000 <210> SEQ ID NO 350 <400>
SEQUENCE: 350 000 <210> SEQ ID NO 351 <400> SEQUENCE:
351 000 <210> SEQ ID NO 352 <400> SEQUENCE: 352 000
<210> SEQ ID NO 353 <400> SEQUENCE: 353 000 <210>
SEQ ID NO 354 <400> SEQUENCE: 354 000 <210> SEQ ID NO
355 <400> SEQUENCE: 355 000 <210> SEQ ID NO 356
<400> SEQUENCE: 356 000 <210> SEQ ID NO 357 <400>
SEQUENCE: 357 000 <210> SEQ ID NO 358 <400> SEQUENCE:
358 000 <210> SEQ ID NO 359 <400> SEQUENCE: 359 000
<210> SEQ ID NO 360 <400> SEQUENCE: 360 000 <210>
SEQ ID NO 361 <400> SEQUENCE: 361 000 <210> SEQ ID NO
362 <400> SEQUENCE: 362 000 <210> SEQ ID NO 363
<400> SEQUENCE: 363 000 <210> SEQ ID NO 364 <400>
SEQUENCE: 364 000 <210> SEQ ID NO 365 <400> SEQUENCE:
365 000 <210> SEQ ID NO 366 <400> SEQUENCE: 366 000
<210> SEQ ID NO 367 <400> SEQUENCE: 367
000 <210> SEQ ID NO 368 <400> SEQUENCE: 368 000
<210> SEQ ID NO 369 <400> SEQUENCE: 369 000 <210>
SEQ ID NO 370 <400> SEQUENCE: 370 000 <210> SEQ ID NO
371 <400> SEQUENCE: 371 000 <210> SEQ ID NO 372
<400> SEQUENCE: 372 000 <210> SEQ ID NO 373 <400>
SEQUENCE: 373 000 <210> SEQ ID NO 374 <400> SEQUENCE:
374 000 <210> SEQ ID NO 375 <400> SEQUENCE: 375 000
<210> SEQ ID NO 376 <400> SEQUENCE: 376 000 <210>
SEQ ID NO 377 <400> SEQUENCE: 377 000 <210> SEQ ID NO
378 <400> SEQUENCE: 378 000 <210> SEQ ID NO 379
<400> SEQUENCE: 379 000 <210> SEQ ID NO 380 <400>
SEQUENCE: 380 000 <210> SEQ ID NO 381 <400> SEQUENCE:
381 000 <210> SEQ ID NO 382 <400> SEQUENCE: 382 000
<210> SEQ ID NO 383 <400> SEQUENCE: 383 000 <210>
SEQ ID NO 384 <400> SEQUENCE: 384 000 <210> SEQ ID NO
385 <400> SEQUENCE: 385 000 <210> SEQ ID NO 386
<400> SEQUENCE: 386 000 <210> SEQ ID NO 387 <400>
SEQUENCE: 387 000 <210> SEQ ID NO 388 <400> SEQUENCE:
388 000 <210> SEQ ID NO 389 <400> SEQUENCE: 389 000
<210> SEQ ID NO 390 <400> SEQUENCE: 390 000 <210>
SEQ ID NO 391 <400> SEQUENCE: 391 000 <210> SEQ ID NO
392 <400> SEQUENCE: 392 000 <210> SEQ ID NO 393
<400> SEQUENCE: 393 000 <210> SEQ ID NO 394 <400>
SEQUENCE: 394 000 <210> SEQ ID NO 395 <400> SEQUENCE:
395 000 <210> SEQ ID NO 396 <400> SEQUENCE: 396 000
<210> SEQ ID NO 397 <400> SEQUENCE: 397 000 <210>
SEQ ID NO 398 <400> SEQUENCE: 398 000 <210> SEQ ID NO
399 <400> SEQUENCE: 399 000 <210> SEQ ID NO 400
<400> SEQUENCE: 400 000 <210> SEQ ID NO 401 <400>
SEQUENCE: 401 000 <210> SEQ ID NO 402 <400> SEQUENCE:
402 000 <210> SEQ ID NO 403 <400> SEQUENCE: 403
000 <210> SEQ ID NO 404 <400> SEQUENCE: 404 000
<210> SEQ ID NO 405 <400> SEQUENCE: 405 000 <210>
SEQ ID NO 406 <400> SEQUENCE: 406 000 <210> SEQ ID NO
407 <400> SEQUENCE: 407 000 <210> SEQ ID NO 408
<400> SEQUENCE: 408 000 <210> SEQ ID NO 409 <400>
SEQUENCE: 409 000 <210> SEQ ID NO 410 <400> SEQUENCE:
410 000 <210> SEQ ID NO 411 <400> SEQUENCE: 411 000
<210> SEQ ID NO 412 <400> SEQUENCE: 412 000 <210>
SEQ ID NO 413 <400> SEQUENCE: 413 000 <210> SEQ ID NO
414 <400> SEQUENCE: 414 000 <210> SEQ ID NO 415
<400> SEQUENCE: 415 000 <210> SEQ ID NO 416 <400>
SEQUENCE: 416 000 <210> SEQ ID NO 417 <400> SEQUENCE:
417 000 <210> SEQ ID NO 418 <400> SEQUENCE: 418 000
<210> SEQ ID NO 419 <400> SEQUENCE: 419 000 <210>
SEQ ID NO 420 <400> SEQUENCE: 420 000 <210> SEQ ID NO
421 <400> SEQUENCE: 421 000 <210> SEQ ID NO 422
<400> SEQUENCE: 422 000 <210> SEQ ID NO 423 <400>
SEQUENCE: 423 000 <210> SEQ ID NO 424 <400> SEQUENCE:
424 000 <210> SEQ ID NO 425 <400> SEQUENCE: 425 000
<210> SEQ ID NO 426 <400> SEQUENCE: 426 000 <210>
SEQ ID NO 427 <400> SEQUENCE: 427 000 <210> SEQ ID NO
428 <400> SEQUENCE: 428 000 <210> SEQ ID NO 429
<400> SEQUENCE: 429 000 <210> SEQ ID NO 430 <400>
SEQUENCE: 430 000 <210> SEQ ID NO 431 <400> SEQUENCE:
431 000 <210> SEQ ID NO 432 <400> SEQUENCE: 432 000
<210> SEQ ID NO 433 <400> SEQUENCE: 433 000 <210>
SEQ ID NO 434 <400> SEQUENCE: 434 000 <210> SEQ ID NO
435 <400> SEQUENCE: 435 000 <210> SEQ ID NO 436
<400> SEQUENCE: 436 000 <210> SEQ ID NO 437 <400>
SEQUENCE: 437 000 <210> SEQ ID NO 438 <400> SEQUENCE:
438 000 <210> SEQ ID NO 439
<400> SEQUENCE: 439 000 <210> SEQ ID NO 440 <400>
SEQUENCE: 440 000 <210> SEQ ID NO 441 <400> SEQUENCE:
441 000 <210> SEQ ID NO 442 <400> SEQUENCE: 442 000
<210> SEQ ID NO 443 <400> SEQUENCE: 443 000 <210>
SEQ ID NO 444 <400> SEQUENCE: 444 000 <210> SEQ ID NO
445 <400> SEQUENCE: 445 000 <210> SEQ ID NO 446
<400> SEQUENCE: 446 000 <210> SEQ ID NO 447 <400>
SEQUENCE: 447 000 <210> SEQ ID NO 448 <400> SEQUENCE:
448 000 <210> SEQ ID NO 449 <400> SEQUENCE: 449 000
<210> SEQ ID NO 450 <400> SEQUENCE: 450 000 <210>
SEQ ID NO 451 <400> SEQUENCE: 451 000 <210> SEQ ID NO
452 <400> SEQUENCE: 452 000 <210> SEQ ID NO 453
<400> SEQUENCE: 453 000 <210> SEQ ID NO 454 <400>
SEQUENCE: 454 000 <210> SEQ ID NO 455 <400> SEQUENCE:
455 000 <210> SEQ ID NO 456 <400> SEQUENCE: 456 000
<210> SEQ ID NO 457 <400> SEQUENCE: 457 000 <210>
SEQ ID NO 458 <400> SEQUENCE: 458 000 <210> SEQ ID NO
459 <400> SEQUENCE: 459 000 <210> SEQ ID NO 460
<400> SEQUENCE: 460 000 <210> SEQ ID NO 461 <400>
SEQUENCE: 461 000 <210> SEQ ID NO 462 <400> SEQUENCE:
462 000 <210> SEQ ID NO 463 <400> SEQUENCE: 463 000
<210> SEQ ID NO 464 <400> SEQUENCE: 464 000 <210>
SEQ ID NO 465 <400> SEQUENCE: 465 000 <210> SEQ ID NO
466 <400> SEQUENCE: 466 000 <210> SEQ ID NO 467
<400> SEQUENCE: 467 000 <210> SEQ ID NO 468 <400>
SEQUENCE: 468 000 <210> SEQ ID NO 469 <400> SEQUENCE:
469 000 <210> SEQ ID NO 470 <400> SEQUENCE: 470 000
<210> SEQ ID NO 471 <400> SEQUENCE: 471 000 <210>
SEQ ID NO 472 <400> SEQUENCE: 472 000 <210> SEQ ID NO
473 <400> SEQUENCE: 473 000 <210> SEQ ID NO 474
<400> SEQUENCE: 474 000 <210> SEQ ID NO 475
<400> SEQUENCE: 475 000 <210> SEQ ID NO 476 <400>
SEQUENCE: 476 000 <210> SEQ ID NO 477 <400> SEQUENCE:
477 000 <210> SEQ ID NO 478 <400> SEQUENCE: 478 000
<210> SEQ ID NO 479 <400> SEQUENCE: 479 000 <210>
SEQ ID NO 480 <400> SEQUENCE: 480 000 <210> SEQ ID NO
481 <400> SEQUENCE: 481 000 <210> SEQ ID NO 482
<400> SEQUENCE: 482 000 <210> SEQ ID NO 483 <400>
SEQUENCE: 483 000 <210> SEQ ID NO 484 <211> LENGTH: 5
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 484
Asp Thr Tyr Met His 1 5 <210> SEQ ID NO 485 <211>
LENGTH: 5 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 485 Pro Ala Ser Gly His 1 5 <210> SEQ ID NO 486
<211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 486 Ser Gly Gly Leu Pro Asp 1 5 <210>
SEQ ID NO 487 <211> LENGTH: 9 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 487 Ala Ser Ser Ser Val Ser
Tyr Met Tyr 1 5 <210> SEQ ID NO 488 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 488
Ala Thr Ser Asn Leu Ala Ser 1 5 <210> SEQ ID NO 489
<211> LENGTH: 5 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 489 Gly Asn Pro Arg Thr 1 5 <210> SEQ
ID NO 490 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 490 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 491 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 491 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Val Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 492 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 492 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile
Lys
100 105 <210> SEQ ID NO 493 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
493 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Glu Pro Ala Ser Gly His Ile
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Arg
Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Trp Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 494 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
494 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Trp Glu Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 495
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 495 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Glu Pro Ala Ser Gly His Ile Lys Tyr Ser Pro Lys Phe 50 55
60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Trp Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 496 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 496 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Gly Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 497 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 497 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Glu Pro Ala Ser Gly His Ile Lys Tyr Ser Pro Lys Phe
50 55 60 Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Trp Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 498 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 498 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 499 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
499 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Glu Pro Ala Ser Gly His Val
Lys Tyr Ser Pro Lys Phe 50 55 60 Gln Val Arg Val Thr Met Thr Arg
Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Trp Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 500 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 500 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 501 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
501 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Glu Pro Ala Ser Gly His Val
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Thr Arg Val Thr Met Thr Arg
Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Trp Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 502 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
502 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Trp Gln Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 503
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 503 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Ile Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Lys Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Met Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 504 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 504 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Gln Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 505 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 505 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Val Lys Ile Asp Pro Lys Phe
50 55 60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Met Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 506 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 506 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Gln Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 507 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
507 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 Phe Asp Ile Gln
Asp Thr 20 25 30
Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35
40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro Lys
Phe 50 55 60 Gln Val Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser
Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Met
Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115
<210> SEQ ID NO 508 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 508 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Gln Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 509 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
509 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Leu
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Arg Arg Val Thr Met Thr Arg
Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Met Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 510 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
510 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Trp Glu Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 511
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 511 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Asn Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr
65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Lys Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 512 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 512 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Gly Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 513 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 513 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Asn Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Lys Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 514 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 514 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 515 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 515 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Glu Pro Ala Ser Gly His Leu Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Glu Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Lys Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 516 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 516 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Gly Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Glu Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 517 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
517 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Glu Pro Ala Ser Gly His Leu
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Glu Arg Val Thr Met Thr Arg
Asp Thr Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Lys Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 518 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
518 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Trp Gln Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 519
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 519 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Met Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 520 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 520 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Gln Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 521 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 521 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr
Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Met Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115
<210> SEQ ID NO 522 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 522 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 523 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
523 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Gly Arg Ala Thr Ile Thr Thr
Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Val Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 524 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
524 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Trp Ser Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 525
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 525 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Val Arg Ala Thr Ile Thr Thr Asp Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80 Leu Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Phe Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 526 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 526 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 527 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 527 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Gly Arg Ala Thr Ile Thr Thr Asp Thr Ser Ala Ser Thr
Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Phe Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 528 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 528 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 529 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
529
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 Phe Asp Ile Gln Asp
Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys
Tyr Asp Pro Lys Phe 50 55 60 Gln Gly Arg Ala Thr Ile Thr Thr Asp
Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly
Leu Pro Asp Leu Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser
Ser 115 <210> SEQ ID NO 530 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
530 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Ser Gln Trp Ser Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 531
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 531 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Gly Arg Ala Thr Ile Thr Thr Asp Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80 Leu Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Phe Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 532 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 532 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly Asn Pro
Arg Ser 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 533 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 533 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Gly Arg Ala Thr Ile Thr Thr Asp Thr Ser Ala Ser Thr
Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Phe Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 534 <211> LENGTH: 106 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 534 Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40
45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Arg
Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 <210> SEQ ID NO 535 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
535 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Gly Arg Ala Thr Ile Thr Thr
Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Phe Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 536 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
536 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr
35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser
Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg
Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp
Lys Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu
Ile Lys 100 105 <210> SEQ ID NO 537 <211> LENGTH: 116
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
537 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 Phe Asp Ile Gln
Asp Thr 20 25 30 Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ala Ser Gly His Thr
Lys Tyr Asp Pro Lys Phe 50 55 60 Gln Gly Arg Ala Thr Ile Thr Thr
Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu
Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly
Gly Leu Pro Asp Phe Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val
Ser Ser 115 <210> SEQ ID NO 538 <211> LENGTH: 106
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
538 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Ser Ser Val Ser
Tyr Met 20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Ile Pro
Asp Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu Pro Glu 65 70 75 80 Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Trp Ser Gly Asn Pro Arg Thr 85 90 95 Phe Gly Gly Gly
Thr Lys Leu Glu Ile Lys 100 105 <210> SEQ ID NO 539
<211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 539 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Arg Ile Asp Pro Ala Ser Gly His Ser Lys Tyr Asp Pro Lys Phe 50 55
60 Gln Val Arg Ala Thr Ile Thr Thr Asp Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80 Leu Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Phe Trp Gly Gln
Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210> SEQ ID
NO 540 <211> LENGTH: 106 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 540 Glu Ile Val Leu Thr Gln Ser
Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu
Ser Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Tyr
Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr 35 40 45 Ala
Thr Ser Asn Leu Ala Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55
60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro Glu
65 70 75 80 Asp Phe Ala Val Tyr Tyr Cys Gln Gln Trp Ser Gly Asn Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105
<210> SEQ ID NO 541 <211> LENGTH: 116 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 541 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 Phe Asp Ile Gln Asp Thr 20 25 30 Tyr
Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40
45 Gly Arg Ile Asp Pro Ala Ser Gly His Tyr Lys Tyr Asp Pro Lys Phe
50 55 60 Gln Val Arg Ala Thr Ile Thr Thr Asp Thr Ser Ala Ser Thr
Ala Tyr 65 70 75 80 Leu Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Leu Pro Asp Phe Trp
Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <210>
SEQ ID NO 542 <211> LENGTH: 330 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 542 Ala Ser Thr Lys Gly
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe
Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40
45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Ala Ala Gly Ala
Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170
175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser
Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275
280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330 <210> SEQ ID NO 543 <211> LENGTH: 326
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
543 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro
Ser Ser Asn Phe Gly Thr Gln Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp
His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Thr Val Glu Arg
Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 100 105 110 Pro Val
Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 115 120 125
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 130
135 140 Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
Gly 145 150 155 160 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
Glu Gln Phe Asn 165 170 175 Ser Thr Phe Arg Val Val Ser Val Leu Thr
Val Val His Gln Asp Trp 180 185 190 Leu Asn Gly Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Gly Leu Pro 195 200 205 Ala Pro Ile Glu Lys Thr
Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 210 215 220 Pro Gln Val Tyr
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 225 230 235 240 Gln
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 245 250
255 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
260 265 270 Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys 275 280 285 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys 290 295 300 Ser Val Met His Glu Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu 305 310 315 320 Ser Leu Ser Pro Gly Lys 325
<210> SEQ ID NO 544 <211> LENGTH: 107 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 544 Arg Thr Val Ala Ala
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys
Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40
45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp
Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
Cys 100 105 <210> SEQ ID NO 545 <211> LENGTH: 25
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 545
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 20 25 <210> SEQ ID
NO 546 <211> LENGTH: 14 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 546 Trp Val Arg Gln Ala Pro Gly Gln
Gly Leu Glu Trp Met Gly 1 5 10 <210> SEQ ID NO 547
<211> LENGTH: 30 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 547 Arg Val Thr Met Thr Arg Asp
Thr Ser Thr Ser Thr Val Tyr Met Glu 1 5 10 15 Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 20 25 30 <210> SEQ ID NO
548 <211> LENGTH: 11 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 548 Trp Gly Gln Gly Thr Thr Val Thr
Val Ser Ser 1 5 10 <210> SEQ ID NO 549 <211> LENGTH: 23
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 549
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5
10 15 Glu Arg Ala Thr Leu Ser Cys 20 <210> SEQ ID NO 550
<211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 550 Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
Arg Leu Leu Ile Tyr 1 5 10 15 <210> SEQ ID NO 551 <211>
LENGTH: 32 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polypeptide
<400> SEQUENCE: 551 Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly
Ser Gly Thr Asp Phe Thr 1 5 10 15 Leu Thr Ile Ser Arg Leu Glu Pro
Glu Asp Phe Ala Val Tyr Tyr Cys 20 25 30 <210> SEQ ID NO 552
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 552 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 1
5 10
<210> SEQ ID NO 553 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 553 Gly Phe Asp Ile Gln Asp
Thr Tyr Met His 1 5 10 <210> SEQ ID NO 554 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 554 Arg Ile Asp Pro Ala Ser Gly His Thr Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Val <210> SEQ ID NO 555 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 555 Arg Ile Glu Pro Ala Ser Gly His Ile Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Gly <210> SEQ ID NO 556 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 556 Arg Ile Glu Pro Ala Ser Gly His Ile Lys Tyr Ser Pro
Lys Phe Gln 1 5 10 15 Gly <210> SEQ ID NO 557 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 557 Arg Ile Glu Pro Ala Ser Gly His Val Lys Tyr Ser Pro
Lys Phe Gln 1 5 10 15 Val <210> SEQ ID NO 558 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 558 Arg Ile Glu Pro Ala Ser Gly His Val Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Thr <210> SEQ ID NO 559 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 559 Arg Ile Asp Pro Ala Ser Gly His Ile Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Lys <210> SEQ ID NO 560 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 560 Arg Ile Asp Pro Ala Ser Gly His Val Lys Ile Asp Pro
Lys Phe Gln 1 5 10 15 Val <210> SEQ ID NO 561 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 561 Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Val <210> SEQ ID NO 562 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 562 Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Arg <210> SEQ ID NO 563 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 563 Arg Ile Asp Pro Ala Ser Gly His Leu Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Asn <210> SEQ ID NO 564 <211>
LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 564 Arg Ile Glu Pro Ala Ser Gly His Leu Lys Tyr Asp Pro
Lys Phe Gln 1 5 10 15 Glu <210> SEQ ID NO 565 <211>
LENGTH: 9 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 565 Ala Arg Ser Gly Gly Leu Pro Asp Val 1 5 <210>
SEQ ID NO 566 <211> LENGTH: 9 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 566 Ala Arg Ser Gly Gly Leu
Pro Asp Trp 1 5 <210> SEQ ID NO 567 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 567
Ala Arg Ser Gly Gly Leu Pro Asp Met 1 5 <210> SEQ ID NO 568
<211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 568
Ala Arg Ser Gly Gly Leu Pro Asp Lys 1 5 <210> SEQ ID NO 569
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 569 Arg Ala Ser Ser Ser Val Ser Tyr Met Tyr 1
5 10 <210> SEQ ID NO 570 <211> LENGTH: 10 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 570 Gly Ala Ser
Ser Ser Val Ser Tyr Met Tyr 1 5 10 <210> SEQ ID NO 571
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 571 Gln Gln Trp Ser Gly Asn Pro Arg Thr 1 5
<210> SEQ ID NO 572 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 572 Gln Gln Trp Glu Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 573 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 573
Gln Gln Trp Gln Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 574
<211> LENGTH: 17 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 574 Arg Ile Asp Pro Ala Ser Gly His Leu Lys
Tyr Asp Pro Lys Phe Gln 1 5 10 15 Gly <210> SEQ ID NO 575
<211> LENGTH: 17 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 575 Arg Ile Asp Pro Ala Ser Gly His Thr Lys
Tyr Asp Pro Lys Phe Gln 1 5 10 15 Gly <210> SEQ ID NO 576
<211> LENGTH: 17 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 576 Arg Ile Asp Pro Ala Ser Gly His Ser Lys
Tyr Asp Pro Lys Phe Gln 1 5 10 15 Val <210> SEQ ID NO 577
<211> LENGTH: 17 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 577 Arg Ile Asp Pro Ala Ser Gly His Tyr Lys
Tyr Asp Pro Lys Phe Gln 1 5 10 15 Val <210> SEQ ID NO 578
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 578 Ala Arg Ser Gly Gly Leu Pro Asp Val 1 5
<210> SEQ ID NO 579 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 579 Ala Arg Ser Gly Gly Leu
Pro Asp Met 1 5 <210> SEQ ID NO 580 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 580
Ala Arg Ser Gly Gly Leu Pro Asp Phe 1 5 <210> SEQ ID NO 581
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 581 Ala Arg Ser Gly Gly Leu Pro Asp Leu 1 5
<210> SEQ ID NO 582 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 582 Ser Gln Trp Ser Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 583 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 583
Gln Gln Trp Ser Gly Asn Pro Arg Ser 1 5 <210> SEQ ID NO 584
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 584 Gln Gln Trp Ser Arg Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 585 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 585
Gln Gln Trp Lys Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 586
<211> LENGTH: 30 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 586 Arg Ala Thr Ile Thr Thr Asp
Thr Ser Ala Ser Thr Ala Tyr Leu Gln 1 5 10 15 Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 20 25 30 <210> SEQ ID NO
587 <211> LENGTH: 30 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 587 Arg Val Thr Ile Thr Arg Asp
Thr Ser Ala Ser Thr Val Tyr Met Glu 1 5 10 15 Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 20 25 30 <210> SEQ ID NO
588 <211> LENGTH: 30 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 588 Arg Val Thr Ile Thr Arg Asp
Thr Ser Ala Ser Thr Ala Tyr Met Glu 1 5 10 15 Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 20 25 30 <210> SEQ ID NO
589 <211> LENGTH: 6 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 589 His His His His His His 1 5
<210> SEQ ID NO 590 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 590 Ser Arg Ser Gly Gly Leu
Pro Asp Val 1 5 <210> SEQ ID NO 591 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 591
Gly Phe Glu Ile Gln Asp Thr Tyr Met His 1 5 10 <210> SEQ ID
NO 592 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 592 Gly Phe Asp Pro Gln Asp Thr Tyr
Met His 1 5 10 <210> SEQ ID NO 593 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 593
Gly Phe Asp Val Gln Asp Thr Tyr Met His 1 5 10 <210> SEQ ID
NO 594 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 594 Gly Phe Asp Ile Gly Asp Thr Tyr
Met His 1 5 10 <210> SEQ ID NO 595 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 595
Gly Phe Asp Ile Ser Asp Thr Tyr Met His 1 5 10 <210> SEQ ID
NO 596 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 596 Gly Phe Asp Ile Val Asp Thr Tyr
Met His 1 5 10 <210> SEQ ID NO 597 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 597
Gly Phe Asp Ile Gln Asp Ala Tyr Met His 1 5 10 <210> SEQ ID
NO 598 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 598 Gly Phe Asp Ile Gln Asp Ser Tyr
Met His 1 5 10 <210> SEQ ID NO 599 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 599
Gly Phe Asp Ile Gln Asp Thr Phe Met His 1 5 10 <210> SEQ ID
NO 600 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence:
Synthetic
peptide <400> SEQUENCE: 600 Gly Phe Asp Ile Gln Asp Thr Tyr
Ile His 1 5 10 <210> SEQ ID NO 601 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 601
Gly Phe Asp Leu Gln Asp Thr Tyr Met His 1 5 10 <210> SEQ ID
NO 602 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 602 Gly Phe Asp Ile Gln Asp Thr Tyr
Leu His 1 5 10 <210> SEQ ID NO 603 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 603
Arg Ile Asp Pro Ala Ser Gly His Thr Lys 1 5 10 <210> SEQ ID
NO 604 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 604 Arg Leu Asp Pro Ala Ser Gly His
Thr Lys 1 5 10 <210> SEQ ID NO 605 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 605
Arg Ile Glu Pro Ala Ser Gly His Thr Lys 1 5 10 <210> SEQ ID
NO 606 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 606 Arg Ile Asp Pro Glu Ser Gly His
Thr Lys 1 5 10 <210> SEQ ID NO 607 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 607
Arg Ile Asp Pro Ala Gly Gly His Thr Lys 1 5 10 <210> SEQ ID
NO 608 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 608 Arg Ile Asp Pro Ala Ser Ala His
Thr Lys 1 5 10 <210> SEQ ID NO 609 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 609
Arg Ile Asp Pro Ala Ser Gly His Ile Lys 1 5 10 <210> SEQ ID
NO 610 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 610 Arg Ile Asp Pro Ala Ser Gly His
Leu Lys 1 5 10 <210> SEQ ID NO 611 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 611
Arg Ile Asp Pro Ala Ser Gly His Val Lys 1 5 10 <210> SEQ ID
NO 612 <211> LENGTH: 7 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 612 Tyr Asp Pro Lys Phe Gln Val 1 5
<210> SEQ ID NO 613 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 613 Ile Asp Pro Lys Phe Gln
Val 1 5 <210> SEQ ID NO 614 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 614 Leu Asp Pro
Lys Phe Gln Val 1 5 <210> SEQ ID NO 615 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 615
Met Asp Pro Lys Phe Gln Val 1 5 <210> SEQ ID NO 616
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 616 Ser Asp Pro Lys Phe Gln Val 1 5
<210> SEQ ID NO 617 <211> LENGTH: 7 <212> TYPE:
PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 617 Thr Asp Pro Lys Phe Gln
Val 1 5 <210> SEQ ID NO 618 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 618 Val Asp Pro
Lys Phe Gln Val 1 5 <210> SEQ ID NO 619 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 619
Tyr Ile Pro Lys Phe Gln Val 1 5 <210> SEQ ID NO 620
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 620 Tyr Asn Pro Lys Phe Gln Val 1 5
<210> SEQ ID NO 621 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 621 Tyr Arg Pro Lys Phe Gln
Val 1 5 <210> SEQ ID NO 622 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 622 Tyr Ser Pro
Lys Phe Gln Val 1 5 <210> SEQ ID NO 623 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 623
Tyr Asp Pro Lys Phe Arg Val 1 5 <210> SEQ ID NO 624
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 624 Tyr Asp Pro Lys Phe Gln Ala 1 5
<210> SEQ ID NO 625 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 625 Tyr Asp Pro Lys Phe Gln
Asp 1 5 <210> SEQ ID NO 626 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 626 Tyr Asp Pro
Lys Phe Gln Glu 1 5 <210> SEQ ID NO 627 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 627
Tyr Asp Pro Lys Phe Gln Gly 1 5 <210> SEQ ID NO 628
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 628 Tyr Asp Pro Lys Phe Gln His 1 5
<210> SEQ ID NO 629 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 629 Tyr Asp Pro Lys Phe Gln
Lys 1 5 <210> SEQ ID NO 630 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 630 Tyr Asp Pro
Lys Phe Gln Leu 1 5 <210> SEQ ID NO 631 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 631
Tyr Asp Pro Lys Phe Gln Met 1 5 <210> SEQ ID NO 632
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 632 Tyr Asp Pro Lys Phe Gln Asn 1 5
<210> SEQ ID NO 633 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 633 Tyr Asp Pro Lys Phe Gln
Pro 1 5
<210> SEQ ID NO 634 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 634 Tyr Asp Pro Lys Phe Gln
Arg 1 5 <210> SEQ ID NO 635 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 635 Tyr Asp Pro
Lys Phe Gln Ser 1 5 <210> SEQ ID NO 636 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 636
Tyr Asp Pro Lys Phe Gln Thr 1 5 <210> SEQ ID NO 637
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 637 Leu Gly Gly Leu Pro Asp Val 1 5
<210> SEQ ID NO 638 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 638 Ser Ala Gly Leu Pro Asp
Val 1 5 <210> SEQ ID NO 639 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 639 Ser Gly Gly
Ala Pro Asp Val 1 5 <210> SEQ ID NO 640 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 640
Ser Gly Gly Met Pro Asp Val 1 5 <210> SEQ ID NO 641
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 641 Ser Gly Gly Leu Pro Glu Val 1 5
<210> SEQ ID NO 642 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 642 Ser Gly Gly Leu Pro Asp
Lys 1 5 <210> SEQ ID NO 643 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 643 Ser Gly Gly
Leu Pro Asp Met 1 5 <210> SEQ ID NO 644 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 644
Ser Gly Gly Leu Pro Asp Gln 1 5 <210> SEQ ID NO 645
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 645 Ser Gly Gly Leu Pro Asp Arg 1 5
<210> SEQ ID NO 646 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 646 Ser Gly Gly Leu Pro Asp
Ser 1 5 <210> SEQ ID NO 647 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 647 Ser Gly Gly
Leu Pro Asp Thr 1 5 <210> SEQ ID NO 648 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 648
Ser Gly Gly Leu Pro Asp Trp 1 5 <210> SEQ ID NO 649
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 649 Trp Ala Ser Ser Ser Val Ser Tyr Met Tyr 1
5 10 <210> SEQ ID NO 650 <211> LENGTH: 10 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 650
Arg Ala Ser Ser Ser Val Ile Tyr Met Tyr 1 5 10 <210> SEQ ID
NO 651 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 651 Arg Ala Ser Ser Ser Val Ser Phe
Met Tyr 1 5 10 <210> SEQ ID NO 652 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 652
Arg Ala Ser Ser Ser Val Ser Tyr Leu Tyr 1 5 10 <210> SEQ ID
NO 653 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 653 Arg Ala Ser Ser Ser Val Ser Tyr
Met Arg 1 5 10 <210> SEQ ID NO 654 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 654
Ala Lys Ser Asn Leu Ala Ser 1 5 <210> SEQ ID NO 655
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 655 Ala Thr Pro Asn Leu Ala Ser 1 5
<210> SEQ ID NO 656 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 656 Ala Thr Glu Asn Leu Ala
Ser 1 5 <210> SEQ ID NO 657 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 657 Ala Thr Ser
Leu Leu Ala Ser 1 5 <210> SEQ ID NO 658 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 658
Ala Thr Ser Pro Leu Ala Ser 1 5 <210> SEQ ID NO 659
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 659 Ala Thr Ser Asn Leu Thr Ser 1 5
<210> SEQ ID NO 660 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 660 His Gln Trp Ser Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 661 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 661
Asn Gln Trp Ser Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 662
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 662 Gln Gln Ser Ser Gly Asn Pro Arg Thr 1 5
<210> SEQ ID NO 663 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 663 Gln Gln Trp Asp Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 664 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 664
Gln Gln Trp His Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 665
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 665 Gln Gln Trp Asn Gly Asn Pro Arg Thr 1 5
<210> SEQ ID NO 666 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 666 Gln Gln Trp Val Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 667 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 667 Gln Gln Trp Ser Ala Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 668 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 668
Gln Gln Trp Ser Asp Asn Pro Arg Thr 1 5 <210> SEQ ID NO 669
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 669 Gln Gln Trp Ser Gln Asn Pro Arg Thr 1 5
<210> SEQ ID NO 670 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 670 Gln Gln Trp Ser Ser Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 671 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 671
Gln Gln Phe Ser Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 672
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 672 Gln Gln His Ser Gly Asn Pro Arg Thr 1 5
<210> SEQ ID NO 673 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 673 Gln Gln Ile Ser Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 674 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 674
Gln Gln Pro Ser Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 675
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 675 Gln Gln Arg Ser Gly Asn Pro Arg Thr 1 5
<210> SEQ ID NO 676 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 676 Gln Gln Tyr Ser Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 677 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 677
Gln Gln Trp Ser Gly His Pro Arg Thr 1 5 <210> SEQ ID NO 678
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 678 Gln Gln Trp Ser Gly Leu Pro Arg Thr 1 5
<210> SEQ ID NO 679 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 679 Gln Gln Trp Ser Gly Gln
Pro Arg Thr 1 5 <210> SEQ ID NO 680 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 680
Gln Gln Trp Ser Gly Met Pro Arg Thr 1 5 <210> SEQ ID NO 681
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 681 Ser Gly Gly Leu Pro Asp His 1 5
<210> SEQ ID NO 682 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 682 Ser Gly Gly Leu Pro Asp
Phe 1 5 <210> SEQ ID NO 683 <211> LENGTH: 7 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 683 Ser Gly Gly
Ser Pro Asp Val 1 5 <210> SEQ ID NO 684 <211> LENGTH:
9
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 684
Gln Gln Trp Ala Gly Asn Pro Arg Thr 1 5 <210> SEQ ID NO 685
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 685 Gln Gln Trp Tyr Gly Asn Pro Arg Thr 1 5
<210> SEQ ID NO 686 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 686 Gln Gln Trp Phe Gly Asn
Pro Arg Thr 1 5 <210> SEQ ID NO 687 <211> LENGTH: 124
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
687 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 Asp Phe Thr
Tyr Tyr 20 25 30 Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly
Leu Glu Trp Met 35 40 45 Gly Trp Ile Ser Thr Tyr Asn Gly Asn Thr
His Tyr Ala Arg Met Leu 50 55 60 Gln Gly Arg Val Thr Met Thr Thr
Asp Thr Ser Thr Arg 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 Glu Asn
Tyr Tyr Gly Ser Gly Ala Tyr Arg Gly Gly Met Asp 100 105 110 Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> SEQ ID
NO 688 <211> LENGTH: 121 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 688 Glu Val Gln Leu Leu Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Lys 1 5 10 15 Ser Leu Arg Leu Ser
Cys Ala Val Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Gly Met Asn
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser
Ser Ile Ser Gly Thr Gly Arg Thr Thr Tyr His Ala Asp Ser Val 50 55
60 Gln Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Ile Leu Tyr
65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr
Phe Cys 85 90 95 Thr Lys Glu Arg Gly Asp Tyr Tyr Tyr Gly Val Phe
Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115
120 <210> SEQ ID NO 689 <211> LENGTH: 124 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic polypeptide <400> SEQUENCE: 689 Gln Val
Thr Leu Lys Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15
Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser 20
25 30 Asn Met Gly Val Val Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu
Glu 35 40 45 Trp Leu Ala His Ile Leu Trp Asp Asp Arg Glu Tyr Ser
Asn Pro Ala 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr
Ser Lys Asn Gln Val 65 70 75 80 Val Leu Thr Met Thr Asn Met Asp Pro
Val Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Met Ser Arg Asn
Tyr Tyr Gly Ser Ser Tyr Val Met Asp 100 105 110 Tyr Trp Gly Gln Gly
Thr Leu Val Thr Val Ser Ser 115 120 <210> SEQ ID NO 690
<211> LENGTH: 122 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 690 Gln Ile Gln Leu Val 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 Thr Tyr 20 25 30 Gly Met Ser
Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly
Trp Met Asn Thr Tyr Ser Gly Val Thr Thr Tyr Ala Asp Asp Phe 50 55
60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr
65 70 75 80 Met Gln Ile Asp Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr
Phe Cys 85 90 95 Ala Arg Glu Gly Tyr Val Phe Asp Asp Tyr Tyr Ala
Thr Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Ser Val Thr Val Ser Ser
115 120 <210> SEQ ID NO 691 <211> LENGTH: 107
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
691 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser
Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile
Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr
Tyr Cys Gln Gln Arg Ser Asn Trp Pro Trp 85 90 95 Thr Phe Gly Gln
Gly Thr Lys Val Glu Ile Lys 100 105 <210> SEQ ID NO 692
<211> LENGTH: 106 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic
polypeptide <400> SEQUENCE: 692 Asp Ile Gln Met Thr Gln Ser
Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile
Thr Cys Arg Ala Ser Gln Thr Ile Ser Ser Trp 20 25 30 Leu Ala Trp
Tyr Gln Gln Thr Pro Glu Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr
Ala Ala Ser Asn Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr His Arg Ser
Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Thr 100
105
<210> SEQ ID NO 693 <211> LENGTH: 106 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 693 Asp Ile Gln Leu Thr
Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val
Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Asn Tyr Met 20 25 30 His
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40
45 Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys His Gln Trp Asn Asn
Tyr Gly Thr Phe 85 90 95 Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105 <210> SEQ ID NO 694 <211> LENGTH: 112
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
694 Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly
1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val
His Ser 20 25 30 Asp Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys
Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn
Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser
Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu
Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro
Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110
* * * * *
References