Immunomodulators

Abstract

The present invention relates to immunomodulators. The present invention provides antibodies that specifically bind to PD-L1 and fusion molecules comprising PD-L1 binding proteins constructed using IL15 receptor binding domains, nucleic acid molecules encoding the antibodies or fusion molecules, and therapeutic compositions thereof. The agents inhibit PD-L1-mediated immunosuppression and enhance cell- and cytokine-mediated immunity for the treatment of neoplastic and infectious diseases.

Description

[0001] This application is a divisional application. The application date of the original application is January 15, 2015, the application number is 201580000119.1 (PCT / US2015 / 011657), and the name of the invention is “Immunomodulator”. Field of the Invention

[0002] The present invention provides a monoclonal antibody that specifically binds to PD-L1 and a bispecific fusion molecule comprising a PD-L1 binding protein constructed using IL15 and IL15 receptor αsushi domains, a nucleic acid molecule encoding the antibody and fusion molecule, and a therapeutic composition thereof. The reagent enhances the function of T cells and NK cells to enhance cell- and cytokine-mediated immunity for the treatment of various immune dysfunction-related disorders, including cancer and infectious diseases.

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 61 / 927,907, filed on January 15, 2014, the contents of which are incorporated herein by reference in their entirety. Background of the Invention

[0005] Programmed death 1 (PD-1) is a member of the CD28 receptor family (including CD28, CTLA-4, PD-1, ICOS and BTLA) (Freeman et al. (2000) J Exp Med 192: 1027-34; Latchman et al. (2001) Nat Immunol 2: 261-8). PD-1 is an inducible immunosuppressive receptor that is upregulated primarily on activated T cells and B cells during the progression of immunopathological conditions. The interaction of PD-1 with its ligand PD-L1 leads to the inhibition of TCR- and BCR-mediated proliferation and cytokine production and the induction of apoptosis of antigen-specific T cells through intrinsic PD-1-mediated negative signal transduction via immunoreceptor tyrosine-based inhibitory motifs (ITIMs) (Agata et al. (1996) Int. Immunol. 8: 765, Unkeless and Jin. (1997) Curr. Opin. Immunol. 9: 338-343, Okzaki et al. (2001) PNAS 98: 13866-71, Dong et al. (2002) Nat. Med. 8: 793-800). PD-L1 is a cell surface glycoprotein and the main ligand of PD-1. PD-L1 is also inducible on lymphoid and non-lymphoid peripheral tissues after cell activation. PD-L1 is upregulated in a variety of affected cell types (including cancer and stromal cells in addition to immune cells) and plays an active role in immunosuppression during disease progression (Iwai et al. (2002) PNAS 99: 12293-7, Ohigashi et al. (2005) Clin Cancer Res 11: 2947-53). Upregulation of PD-L1 has been associated with poor clinical outcomes in a variety of cancers and viral infections (Hofmeyer et al. (2011) J. BioMed. Biotech. 2011: 1-9, McDermott and Atkins. (2013) Cancer Med. 2: 662-73). Blockade of PD-1 or PD-L1 by antibodies promotes CD8 T cell infiltration, CTL activity, and increased presence of the Th1 cytokine IFN-γ in preclinical and clinical settings (Zhou et al. (2010) J. Immunol. 185: 5082-92, Nomi et al. (2007) Clin Cancer Res. 13: 2152-7, Flies et al. (2011) Yale J. Bio. Med. 48: 409-21, Zitvogel and Kroemer. (2012) Onco Immunol. 1: 1223-25). PD-L1 antibodies have been shown to be effective as immunomodulators when used as monotherapy or in combination with antibodies to other immunosuppressive molecules.

[0006] However, immunomodulatory interventions directed against immunosuppressive factors have only partially solved the problems associated with impaired immunity in cancer, infection, and other diseases. It is still highly desirable to utilize biotherapeutics to directly stimulate and amplify effector immune cells to enhance weakened innate and adaptive immune responses to more effective levels to control tumors and infections. Immunotherapy using cytokines (including interleukins, i.e., IL-2, IL-12, IL15, IL-21, and TNFα, GM-CSF, etc.) has been shown to be effective to some extent in the treatment of cancer and infection, but clinical results are often limited by systemic toxicity associated with high blood concentrations of cytokines required for efficacy and lack of specificity of targets in affected cells and tissues.

[0007] Among the cytokines evaluated, IL15 is recognized to be focused on stimulating effector and central memory CD8 T cells (composed of a subset of antigen-specific CD8 cells) to exert anti-tumor immunity, without modulating effects on other T cell populations. In addition, unlike IL-2, which activates Tregs, IL15 has been shown to have the ability to rescue T cells from apoptosis induced by Tregs and other immunosuppressive cells in addition to its ability to activate natural killer (NK) cells and effector and memory CD8 T cells (Van Belle et al. (2012) PLoS One 7: e45299, Obar and Lefrancois. (2010) J. Immunol. 185: 263-72, Pelletier and Girard. (2006) J Immunol 177: 100-108, Elpek et al. (2010) PNAS 107: 21647–21652).

[0008] Based on its ability to stimulate the proliferation of the murine T cell line CTLL-2, IL15 was identified as a γc cytokine in 1994 (Grabstein et al. (1994) Science 264: 965-8, Bamford et al. (1996) PNAS 93: 2897-902). Human IL15 shares approximately 97% and 96% amino acid sequence identity with simian and cynomolgus monkey IL15, respectively. Human and mouse IL15 have 73% homology and are comparable in activity to mouse cells. IL15 is a 12.5KD protein (114 amino acids) secreted by DC, macrophages and granulocytes as a 14-15kDa glycoprotein, and is also a member of the four cytokines containing an α-helical bundle (Anderson et al. (1995) J Biol Chem. 270: 29862-9, Steel et al. (2012) Trends Pharmacol. Sci. 33: 35-41). IL15 usually forms a complex with the IL15 receptor α expressed on APCs before binding to functional IL15 receptor β and γ units on T cells and NK cells. IL15 can be trans-presented to responder cells expressing CD122 and CD132 by cells expressing cytokines that are themselves bound to the membrane form of the receptor α chain (Dubois et al. (2002) Immunity 17: 537-47). The IL15 receptor αsushi domain (size 29.5 KD) is a critical component for complex formation with IL15 prior to proper engagement with receptors β and γ (Wei et al. (2001) J. Immunol. 167: 277-82). Complexes of IL15 with IL15Rα and IL15 / IL15Rαsushi binding domain fusion proteins have been reported to stimulate CD8 T cells and NK cells more efficiently in vitro and in vivo than IL15 alone (Mortier et al. (2005) J Biol Chem. 281: 1612-19, Stoklasek et al. (2006) J. Immunol. 177: 6072-80). IL15 also induces proliferation and differentiation of stimulated human B cells (Armitage et al. (1995) J Immunol. 154: 483-90). It has been proposed that IL15 primarily counteracts activation-induced cell death (AICD) by acting to prolong the survival of T lymphocytes (Marks-Konczalik et al. (2000) PNAS 97: 11445-50). IL15 has the specific ability to support NK cells and the maintenance of memory phenotype and antigen-specific memory CD8 T cells (Ma et al. (2006) Annu Rev Immunol. 24: 657-79).Therefore, among the most active cytokines in immune regulation, IL15 has the unique ability to mediate many important aspects of immunity against a variety of tumor types and viral infections (including HIV, HBV, HCV, LCMV, etc.) (Steel et al. (2012) Trends Pharmacol. Sci. 33: 35-41, Verbist and Klonowski, (2012) Cytokine. 59: 467-478). SUMMARY OF THE INVENTION

[0009] The present invention provides antibodies and binding proteins that bind to PD-L1. In certain embodiments of the present invention, the antibody binds to PD-L1 and blocks the interaction with PD-1. By blocking the interaction between PD-L1 and PD-1, such antibodies are used to reduce or inhibit immunosuppression.

[0010] In another aspect, the invention provides antibodies and binding proteins that specifically bind to PD-L1 and at least one other molecule. Examples of such embodiments include PD-L1 binding proteins that also bind to one or more other ligands and / or receptors, which binding proteins can be membrane-bound or soluble.

[0011] In another aspect, the present invention provides a molecule that promotes one or more immune responses by interacting with other ligands or receptors, such as a fusion protein that binds to PD-L1, in addition to weakening or inhibiting immunosuppression by binding to PD-L1. In an embodiment of the present invention, the molecule binds to PD-L1 on the target cell, and also stimulates a cell-mediated immune response, for example, by promoting the proliferation of T cells and / or NK cells. In an embodiment of the present invention, the molecule stimulates cells that respond to interleukins or interferons such as, but not limited to, IL2, IL7, IL15, and IL21. In an embodiment of the present invention, the molecule includes a sequence or domain that promotes IL15 stimulation of the IL15 receptor (IL15R). In an embodiment of the present invention, the molecule that promotes IL15R stimulation is part of an IL15Rα chain comprising a sushi domain. In an embodiment of the present invention, the molecule provides a sushi domain of an IL15Rα chain. In an embodiment of the present invention, the molecule provides a complex of IL15 with a sushi domain of an IL15Rα chain, which complex may be covalent or non-covalent. Experiments disclosed herein show that a single molecule containing a PD-L1 binding domain that blocks the binding of PD-L1 to PD-1 and an IL15R stimulatory domain promotes a better immune response than separate molecules used together. More specifically, molecules having an anti-PD-L1 antibody domain and a mixed domain comprising IL15 and IL15 α chain sushi domains are provided, which promote increased proliferation, Th1 cytokine release, and NK cell and T cell killing activity-related molecules compared to providing the domains in separate molecules.

[0012] In one embodiment, the invention provides an antibody or fragment that binds to PD-L1, comprising a heavy chain CDR-1H having the sequence X1YX2MX3 (SEQ ID NO: 328), wherein X1 is A, G, M, Q, S, Y or W, X2 is A, L, M, Q, R, S, V, W or Y, and X3 is A, F, L, M, S, T, V or Y; a heavy chain CDR-2H having SEQ ID NO: 243; and a heavy chain CDR-3H having the sequence of SEQ ID NO: 245. In certain such embodiments, the heavy chain CDR-1H has a sequence selected from the group consisting of SEQ ID NO: 241, SEQ ID NO: 264, SEQ ID NO: 266, SEQ ID NO: 268, SEQ ID NO: 270, SEQ ID NO: 272, SEQ ID NO: 274, SEQ ID NO: 276, SEQ ID NO: 278, SEQ ID NO: 280, SEQ ID NO: 282, SEQ ID NO: 284, SEQ ID NO: 286, SEQ ID NO: 288, SEQ ID NO: 290, SEQ ID NO: 292, SEQ ID NO: 294, SEQ ID NO: 296, SEQ ID NO: 298, SEQ ID NO: 300, SEQ ID NO: 302, SEQ ID NO: 304, SEQ ID NO: 306, SEQ ID NO: 308, SEQ ID NO: 310, and SEQ ID NO: 312. In such embodiments, the heavy chain variable domain is at least 80%, or at least 85%, or at least 90%, or at least 95% identical to SEQ ID NO: 246. The antibody may further comprise a light chain variable domain comprising a CDR-1L having SEQ ID NO: 247, a CDR-2L having SEQ ID NO: 248, and a CDR-3L having SEQ ID NO: 249. In some such embodiments, the light chain variable domain is at least 80%, or at least 85%, or at least 90%, or at least 95% identical to SEQ ID NO: 250. In another embodiment, the invention provides an antibody or fragment thereof that binds to PD-L1, wherein the light chain comprises a CDR-1L having SEQ ID NO: 247, a CDR-2L having SEQ ID NO: 248, and a CDR-3L having SEQ ID NO: 249.

[0013] The present invention also provides conjugates of the antibodies with, for example but not limited to, imaging agents, therapeutic agents or cytotoxic agents.

[0014] The present invention also provides a composition comprising the antibody and conjugate and a pharmaceutically acceptable carrier.

[0015] In another aspect, the present invention provides a fusion protein capable of binding to PDL1, which also stimulates an immune response mediated by, for example, T cells or NK cells. In an embodiment of the present invention, the fusion protein comprises a portion that binds to an IL15 receptor. In other embodiments, the fusion protein comprises a portion that binds to, for example, an interleukin receptor or an interferon receptor. In an embodiment of the present invention, the portion of the fusion protein that binds to PD-L1 is an antibody or a PD-L1 binding fragment thereof. In an embodiment of the present invention, the IL15 receptor binding portion is IL15, and its binding can be enhanced by the presence of the IL15Rαsushi domain in the fusion protein.

[0016] The present invention provides a method for inhibiting the interaction between PD1 and PD-L1 in a subject, the method comprising administering an effective amount of an antibody or fragment of the present invention. The present invention also provides a method for inhibiting immunosuppression mediated by PD-L1, the method comprising administering an effective amount of an antibody or fragment of the present invention, or a fusion protein of the present invention.

[0017] The present invention also provides a method for stimulating an immune response against cells or tissues expressing PD-L1, the method comprising administering to a subject an effective amount of an antibody or fragment of the present invention, or a fusion protein of the present invention. In certain embodiments, the cells or tissues expressing PD-L1 are tumor cells or infected cells. <110> cardman enterprises co., ltd. <120> Immunomodulators <130> 15487 / 474WO1 <140> Unspecified <141> 2015-01-15 <150> US 61 / 927,907 <151> 2014-01-15 <160> 328 <170> PatentIn Version 3.5 <210> 1 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 1 Leu Tyr Trp Met His 1 5 <210> 2 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 2 Gly Phe Thr Phe Ser Leu Tyr 1 5 <210> 3 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 3 Ser Ile Tyr Ser Ser Gly Val Met Thr Phe Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 4 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 4 Tyr Ser Ser Gly Val Met 1 5 <210> 5 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 5 Gly Ser Arg Val Asp Ala Phe Asp Ile 1 5 <210> 6 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 6 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Leu Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Ser Ser Ser Gly Val Met Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Ser Arg Val Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 <210> 7 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 7 Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala 1 5 10 <210> 8 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 8 Gly Ala Ser Ala Arg Ala Ile 1 5 <210> 9 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 9 Gln Gln Ser Tyr Ser Ile Pro Ile Thr 1 5 <210> 10 <211> 108 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 10 Asp Ile Gln Met 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 Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ala Arg Ala Ile Gly Val Pro Asp Arg Phe Arg 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Ile Pro 85 90 95 Ile Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 11 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 11 Pro Tyr Pro Met Trp 1 5 <210> 12 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 12 Gly Phe Thr Phe Ser Pro Tyr 1 5 <210> 13 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 13 Ser Ile Ser Pro Ser Gly Gly Phe Thr Phe Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 14 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 14 Ser Pro Ser Gly Gly Phe 1 5 <210> 15 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 15 Asp Arg Val Ser Trp Asn Asp Tyr Thr Gly Met Asp Val 1 5 10 <210> 16 <211> 122 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 16 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Pro Tyr 20 25 30 Pro Met Trp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Pro Ser Gly Gly Phe Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Arg Val Ser Trp Asn Asp Tyr Thr Gly Met Asp Val Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 17 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 17 Arg Ala Ser Glu Ser Ile Gly Lys Phe Leu Asn 1 5 10 <210> 18 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 18 Ser Ala Ser Ser Leu Gln Ser 1 5 <210> 19 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 19 Gln Gln Ser Phe Asp Met Pro Ile Thr 1 5 <210> 20 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 20 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Ile Gly Lys Phe 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Thr Leu Leu Ile 35 40 45 His Ser Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Thr Thr Tyr Phe Cys Gln Gln Ser Phe Asp Met Pro Ile 85 90 95 Thr Phe Gly Gln Gly Thr Gln Leu Glu Ile Lys 100 105 <210> twenty one <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> twenty one A La Tyr Ile Met Gly 1 5 <210> twenty two <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> twenty two Gly Phe Thr Phe Ser Ala Tyr 1 5 <210> twenty three <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> twenty three Tyr Ile Ser Ser Ser Gly Gly Trp Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> twenty four <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> twenty four Ser Ser Ser Gly Gly Trp 1 5 <210> 25 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 25 Asp Gln Asp Gly Tyr Asn Asp Phe Asp Tyr 1 5 10 <210> 26 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 26 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Ile Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Ser Ser Ser Gly Gly Trp Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Asp Gln Asp Gly Tyr Asn Asp Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 27 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 27 Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu Asn 1 5 10 <210> 28 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 28 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 29 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 29 Gln Gln Ser Tyr Ser Thr Pro Arg Thr 1 5 <210> 30 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 30 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Arg 85 90 95 Thr Phe Gly His Gly Thr Lys Val Glu Ile Lys 100 105 <210> 31 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 31 Met Tyr Trp Met Ala 1 5 <210> 32 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 32 Gly Phe Thr Phe Ser Met Tyr 1 5 <210> 33 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 33 Tyr Ile Val Pro Ser Gly Gly Ile Thr Leu Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 34 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 34 Val Pro Ser Gly Gly Ile 1 5 <210> 35 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 35 Gln Val Arg Gly Gly Ala Phe Asp Ile 1 5 <210> 36 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 36 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Met Tyr 20 25 30 Trp Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Val Pro Ser Gly Gly Ile Thr Leu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Gln Val Arg Gly Gly Ala Phe Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 <210> 37 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 37 Gln Ala Ser His Asp Ile Ser Asn Tyr Leu Asn 1 5 10 <210> 38 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 38 Asp Ala Ser Asn Leu Glu Thr 1 5 <210> 39 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 39 Gln His Tyr Asp Asn Leu Pro Pro Ser 1 5 <210> 40 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 40 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Gln Ala Ser His Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln His Tyr Asp Asn Leu Pro Pro 85 90 95 Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 41 <211> 5 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 41 His Tyr Pro Met Val 1 5 <210> 42 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 42 Gly Phe Thr Phe Ser His Tyr 1 5 <210> 43 <211> 17 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 43 Trp Ile Gly Ser Ser Gly Gly Phe Thr Met Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 44 <211> 6 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 44 Gly Ser Ser Gly Gly Phe 1 5 <210> 45 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 45 Asp Ser Phe Glu Asp His Tyr Tyr Met Asp Val 1 5 10 <210> 46 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 46 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Tyr 20 25 30 Pro Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Trp Ile Gly Ser Ser Gly Gly Phe Thr Met Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Asp Ser Phe Glu Asp His Tyr Tyr Met Asp Val Trp Gly Lys 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 47 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 47 Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp 1 5 10 15 <210> 48 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 48 Leu Gly Ser Asn Arg Ala Ser 1 5 <210> 49 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 49 Met Gln Ala Leu Gln Thr Pro Leu Thr 1 5 <210> 50 <211> 112 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 50 Asp Ile Gln Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20 25 30 Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala 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 Val Gly Val Tyr Tyr Cys Met Gln Ala 85 90 95 Leu Gln Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 51 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 51 Glu Tyr Val Met Ile 1 5 <210> 52 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 52 Gly Phe Thr Phe Ser Glu Tyr 1 5 <210> 53 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 53 Ser Ile Ser Ser Ser Gly Gly Phe Thr Trp Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 54 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 54 Ser Ser Ser Gly Gly Phe 1 5 <210> 55 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 55 Ala Arg Ala Pro Val Gly Ala Asp Ala Phe Asp Ile 1 5 10 <210> 56 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 56 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Glu Tyr 20 25 30 Val Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Ser Ser Gly Gly Phe Thr Trp Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Arg Ala Pro Val Gly Ala Asp Ala Phe Asp Ile Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 57 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 57 Arg Ala Ser Gln Arg Ile Ser Ser Tyr Leu Asn 1 5 10 <210> 58 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 58 Asn Ala Ala Ser Leu Trp Asn 1 5 <210> 59 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 59 Gln Gln Ser His Ser Ser Pro Tyr Thr 1 5 <210> 60 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 60 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Arg Ile Ser Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Lys Leu Leu Ile 35 40 45 Asp Asn Ala Ala Ser Leu Trp Asn Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Thr Leu Met Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ser His Ser Ser Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Arg 100 105 <210> 61 <211> 5 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 61 Ser Tyr Glu Met His 1 5 <210> 62 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 62 Gly Phe Thr Phe Ser Ser Tyr 1 5 <210> 63 <211> 17 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 63 Gly Ile Trp Pro Ser Gly Gly Val Thr Leu Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 64 <211> 6 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 64 Trp Pro Ser Gly Gly Val 1 5 <210> 65 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 65 Phe Arg Thr Gln Pro Phe Asp Ile 1 5 <210> 66 <211> 117 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 66 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Trp Pro Ser Gly Gly Val Thr Leu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Phe Arg Thr Gln Pro Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105 110 Val Thr Val Ser Ser 115 <210> 67 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 67 Arg Ala Ser Gln Gly Ile Ser Asn Tyr Leu Ala 1 5 10 <210> 68 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 68 Ala Ala Ser Thr Leu Gln Ser 1 5 <210> 69 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 69 His Asn Tyr Asn Ser Ala Leu Thr 1 5 <210> 70 <211> 106 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 70 Asp Ile Gln Met Thr Gln Ser Pro Asp Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Val Ala Thr Tyr Phe Cys His Asn Tyr Asn Ser Ala Leu Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 71 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 71 Arg Tyr Gly Met Gly 1 5 <210> 72 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 72 Gly Phe Thr Phe Ser Arg Tyr 1 5 <210> 73 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 73 Ser Ile Ser Ser Ser Gly Gly Gln Thr Ser Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 74 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 74 Ser Ser Ser Gly Gly Gln 1 5 <210> 75 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 75 Gly Gly Leu Trp Phe Gly Leu Asp Pro 1 5 <210> 76 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 76 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Gly Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Ser Ser Gly Gly Gln Thr Ser Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Gly Leu Trp Phe Gly Leu Asp Pro Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 77 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 77 Arg Ala Ser Gln Gly Ile Ser Thr Trp Leu Ala 1 5 10 <210> 78 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 78 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 79 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 79 Gln Gln Ala Asn Ser Phe Pro Ser Thr 1 5 <210> 80 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 80 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Thr Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Ser 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 81 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 81 Lys Tyr Val Met His 1 5 <210> 82 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 82 Gly Phe Thr Phe Ser Lys Tyr 1 5 <210> 83 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 83 Tyr Ile Ser Ser Ser Gly Gly Phe Thr Val Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 84 <211> 6 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 84 Ser Ser Ser Gly Gly Phe 1 5 <210> 85 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 85 Val Phe Asp Ser Gly Asp Ala Phe Asp Ile 1 5 10 <210> 86 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 86 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Tyr 20 25 30 Val Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Ser Ser Ser Gly Gly Phe Thr Val Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Phe Asp Ser Gly Asp Ala Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 87 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 87 Lys Ser Ser Gln Ser Val Leu Tyr Ser Ser Asn Asn Asn Asn Tyr Leu 1 5 10 15 Ala <210> 88 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 88 Trp Ala Ser Thr Arg Glu Ser 1 5 <210> 89 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 89 Gln Gln Tyr Tyr Ser Thr Pro Pro Thr 1 5 <210> 90 <211> 113 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 90 Asp Ile Gln Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30 Ser Asn Asn Asn Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 100 105 110 Lys <210> 91 <211> 5 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 91 Trp Tyr Pro Met His 1 5 <210> 92 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 92 Gly Phe Thr Phe Ser Trp Tyr 1 5 <210> 93 <211> 17 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 93 Ser Ile Ser Ser Ser Gly Gly Phe Thr Met Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 94 <211> 6 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 94 Ser Ser Ser Gly Gly Phe 1 5 <210> 95 <211> 11 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 95 Glu Gly Gly Tyr Ser Tyr Gly Pro Gly Asp Tyr 1 5 10 <210> 96 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 96 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Trp Tyr 20 25 30 Pro Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Ser Ser Gly Gly Phe Thr Met Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Lys Glu Gly Gly Tyr Ser Tyr Gly Pro Gly Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 97 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 97 Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala 1 5 10 <210> 98 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 98 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 99 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 99 Gln Gln Ala Asn Ser Phe Pro Tyr Thr 1 5 <210> 100 <211> 107 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 100 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 101 <211> 5 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 101 Trp Tyr Pro Met Thr 1 5 <210> 102 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 102 Gly Phe Thr Phe Ser Trp Tyr 1 5 <210> 103 <211> 17 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 103 Ser Ile Gly Ser Ser Gly Gly Phe Thr Leu Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 104 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 104 Ile Gly Ser Ser Gly Gly Phe 1 5 <210> 105 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 105 Asp Val Trp Gly Ile Ala Ala Pro Tyr Phe Asp Tyr 1 5 10 <210> 106 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 106 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Trp Tyr 20 25 30 Pro Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Gly Ser Ser Ser Gly Gly Phe Thr Leu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Asp Val Trp Gly Ile Ala Ala Pro Tyr Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 107 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 107 Arg Ala Ser Gln Gly Ile Gly Ser Trp Leu Gly 1 5 10 <210> 108 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 108 Ala Ala Ser Asn Leu Gln Ser 1 5 <210> 109 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 109 Gln Gln Val Asn Asn Phe Pro Arg Ala 1 5 <210> 110 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 110 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Gly Ile Gly Ser Trp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly 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 Asp Phe Thr Leu Thr Ile Ser Asn Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Val Asn Asn Phe Pro Arg 85 90 95 Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 111 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 111 Trp Tyr Ile Met Ala 1 5 <210> 112 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 112 Gly Phe Thr Phe Ser Trp Tyr 1 5 <210> 113 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 113 Tyr Ile Ser Ser Ser Gly Gly Phe Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 114 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 114 Ser Ser Ser Gly Gly Phe 1 5 <210> 115 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 115 Ile Gly Gly Thr Asp Val Phe Asp Ile 1 5 <210> 116 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 116 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Trp Tyr 20 25 30 Ile Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Ser Ser Ser Gly Gly Phe Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ile Gly Gly Thr Asp Val Phe Asp Ile Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 117 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 117 Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala 1 5 10 <210> 118 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 118 Lys Ala Ser Ser Leu Glu Ser 1 5 <210> 119 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 119 Gln Gln Ser Tyr Ser Ser Pro Trp Thr 1 5 <210> 120 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 120 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 Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Ser Leu Leu Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Ser Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Val Lys 100 105 <210> 121 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 121 Trp Tyr Leu Met Lys 1 5 <210> 122 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 122 Gly Phe Thr Phe Ser Trp Tyr 1 5 <210> 123 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 123 Tyr Ile Gly Ser Ser Gly Gly Phe Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 124 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 124 Gly Ser Ser Gly Gly Phe 1 5 <210> 125 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 125 Glu Asp Asp Phe Gly Ala Met Asp Val 1 5 <210> 126 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 126 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Trp Tyr 20 25 30 Leu Met Lys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Gly Ser Ser Gly Gly Phe Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Glu Asp Asp Phe Gly Ala Met Asp Val Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser 115 <210> 127 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 127 Arg Ala Ser Gln Thr Val Ser Lys Tyr Phe Asn 1 5 10 <210> 128 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 128 Ala Thr Ser Thr Leu Gln Ser 1 5 <210> 129 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 129 Gln Gln Ser Tyr Thr Thr Pro Trp Thr 1 5 <210> 130 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 130 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Val Ser Lys Tyr 20 25 30 Phe Asn Trp Phe Gln Gln Lys Pro Gly Glu Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Thr Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Thr Thr Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 131 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 131 Ser Tyr Gln Met Gly 1 5 <210> 132 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 132 Gly Phe Thr Phe Ser Ser Tyr 1 5 <210> 133 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 133 Trp Ile Val Pro Ser Gly Gly Phe Thr His Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 134 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 134 Val Pro Ser Gly Gly Phe 1 5 <210> 135 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 135 Asp Ala Gly Tyr Arg Ser Gly Tyr Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val 1 5 10 15 <210> 136 <211> 125 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 136 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gln Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Trp Ile Val Pro Ser Gly Gly Phe Thr His Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Ala Gly Tyr Arg Ser Gly Tyr Tyr Tyr Tyr Tyr Gly Met 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 137 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 137 Arg Ala Ser Gln Ser Ile Ala Ser Tyr Leu Asn 1 5 10 <210> 138 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 138 Gly Ala Ser Ser Leu Gln Ser 1 5 <210> 139 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 139 Gln Gln Ser Tyr Gly Ile Ser Tyr Thr 1 5 <210> 140 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 140 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ala Ser Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Arg Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Gly Ile Ser Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 141 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 141 His Tyr Pro Met Ser 1 5 <210> 142 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 142 Gly Phe Thr Phe Ser His Tyr 1 5 <210> 143 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 143 Arg Ile Trp Ser Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 144 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 144 Ile Trp Ser Ser Gly Gly Asn 1 5 <210> 145 <211> 12 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 145 Gly Gly Tyr Phe Asp Trp Leu Tyr Pro His Asp Tyr 1 5 10 <210> 146 <211> 121 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 146 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Tyr 20 25 30 Pro Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Arg Ile Trp Ser Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Gly Tyr Phe Asp Trp Leu Tyr Pro His Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 147 <211> 11 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 147 Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ser 1 5 10 <210> 148 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 148 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 149 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 149 Gln Gln Gly Asn Ser Phe Pro Pro Thr 1 5 <210> 150 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 150 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Ala Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Glu Phe Thr Leu Thr Ile Ser Asn Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Asp Ile Lys 100 105 <210> 151 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 151 Arg Tyr Glu Met Leu 1 5 <210> 152 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 152 Gly Phe Thr Phe Ser Arg Tyr 1 5 <210> 153 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 153 Ser Ile Tyr Ser Ser Gly Gly Trp Thr Trp Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 154 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 154 Tyr Ser Ser Gly Gly Trp 1 5 <210> 155 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 155 His Ser Val Thr Gly Val Ala Phe Asp Tyr 1 5 10 <210> 156 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 156 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Glu Met Leu Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Ser Ser Gly Gly Trp Thr Trp Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg His Ser Val Thr Gly Val Ala Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 157 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 157 Arg Ala Ser Gln Asn Ile Asp Thr Tyr Leu Ser 1 5 10 <210> 158 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 158 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 159 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 159 Gln Gln Ser Tyr Tyr Thr Pro Thr 1 5 <210> 160 <211> 106 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 160 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asp Thr Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Tyr Thr Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 161 <211> 5 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 161 Pro Tyr Asp Met Gly 1 5 <210> 162 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 162 Gly Phe Thr Phe Ser Pro Tyr 1 5 <210> 163 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 163 Ser Ile Tyr Ser Ser Gly Gly Trp Thr Lys Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 164 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 164 Tyr Ser Ser Gly Gly Trp 1 5 <210> 165 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 165 Asp Asn Trp Asn Asp Gly Ala Phe Asp Val 1 5 10 <210> 166 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 166 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Pro Tyr 20 25 30 Asp Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Ser Ser Gly Gly Trp Thr Lys Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asn Trp Asn Asp Gly Ala Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 167 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 167 Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala 1 5 10 <210> 168 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 168 Ala Ala Ser Arg Leu Gln Thr 1 5 <210> 169 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 169 Gln Gln Ala Lys Thr Phe Pro Leu Thr 1 5 <210> 170 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 170 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 Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Phe Ala Ala Ser Arg Leu Gln Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ser Ala Thr Tyr Tyr Cys Gln Gln Ala Lys Thr Phe Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Arg Val Glu Ile Lys 100 105 <210> 171 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 171 Phe Tyr Asp Met Ser 1 5 <210> 172 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 172 Gly Phe Thr Phe Ser Phe Tyr 1 5 <210> 173 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 173 Ser Ile Val Pro Ser Gly Gly Trp Thr Phe Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 174 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 174 Val Pro Ser Gly Gly Trp 1 5 <210> 175 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 175 Asp Ser Trp Asn Asp Gly Ala Ser Asp Ile 1 5 10 <210> 176 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 176 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Phe Tyr 20 25 30 Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Val Pro Ser Gly Gly Trp Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Ser Trp Asn Asp Gly Ala Ser Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 <210> 177 <211> 11 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 177 Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala 1 5 10 <210> 178 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 178 Ala Ala Ser Ser Leu Gln Ser 1 5 <210> 179 <211> 9 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 179 Gln Gln Ala Asn Ser Phe Pro Phe Thr 1 5 <210> 180 <211> 107 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 180 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Phe Lys 100 105 <210> 181 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 181 Pro Tyr Gly Met Arg 1 5 <210> 182 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 182 Gly Phe Thr Phe Ser Pro Tyr 1 5 <210> 183 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 183 Ser Ile Ser Pro Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 184 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 184 Ser Pro Ser Gly Gly Asn 1 5 <210> 185 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 185 Arg Ala Ser His Ser Val Ser Ser Asp Tyr Leu Ala 1 5 10 <210> 186 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 186 Ile Arg Tyr Cys Gly Ser Ala Tyr Cys Tyr Thr Asp Ala Phe Asp Ile 1 5 10 15 <210> 187 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 187 Ser Gly Glu Lys Leu Gly Asp Arg Tyr Val Ser 1 5 10 <210> 188 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From the Fab Library <400> 188 His Asp Lys Lys Arg Pro Pro 1 5 <210> 189 <211> 9 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 189 Gln Ala Trp Asp Ser Pro Thr Glu Val 1 5 <210> 190 <211> 109 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 190 Gln Ser Ala Leu Thr Gln Ala Pro Ser Val Ser Val Ser Pro Gly Gln 1 5 10 15 Thr Thr Thr Ile Thr Cys Ser Gly Glu Lys Leu Gly Asp Arg Tyr Val 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Ile Leu Val Leu Tyr 35 40 45 His Asp Lys Lys Arg Pro Pro Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60 Asn Ser Gly Asp Thr Ala Thr Leu Thr Ile Thr Gly Thr His Thr Met 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Pro Thr Glu Val 85 90 95 Phe Gly Pro Gly Thr Lys Leu Thr Val Leu Ser Gln Pro 100 105 <210> 191 <211> 5 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 191 Met Tyr Asp Met Ala 1 5 <210> 192 <211> 7 <212> PRT <213> Artificial sequence <220> <223> From human antibody library <400> 192 Gly Phe Thr Phe Ser Met Tyr 1 5 <210> 193 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 193 Gly Ile Trp Pro Ser Gly Gly Pro Thr Met Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 194 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 194 Trp Pro Ser Gly Gly Pro 1 5 <210> 195 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 195 Gly Tyr Ser Tyr Gly Asp Ala Leu Asp Tyr 1 5 10 <210> 196 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 196 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Met Tyr 20 25 30 Asp Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Trp Pro Ser Gly Gly Pro Thr Met Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Gly Gly Tyr Ser Tyr Gly Asp Ala Leu Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 197 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 197 Ser Gly Thr Ser Ser Asn Ile Gly Arg Asn Tyr Val Ser 1 5 10 <210> 198 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 198 Asp Asp Arg Asn Arg Pro Ser 1 5 <210> 199 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 199 Gly Thr Trp Asp Thr Ser Leu Ser Val Val Val 1 5 10 <210> 200 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 200 Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Val Thr Ile Ser Cys Ser Gly Thr Ser Ser Ser Asn Ile Gly Arg Asn 20 25 30 Tyr Val Ser Trp Tyr Arg His Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Asp Asp Arg Asn Arg Pro Ser Gly Ile Val Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Thr Ser Leu 85 90 95 Ser Val Val Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu Ser Gln 100 105 110 Pro <210> 201 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 201 Glu Tyr Arg Met Ile 1 5 <210> 202 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 202 Gly Phe Thr Phe Ser Glu Tyr 1 5 <210> 203 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 203 Gly Ile Tyr Pro Ser Gly Gly Trp Thr Asn Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 204 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 204 Tyr Pro Ser Gly Gly Trp 1 5 <210> 205 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 205 Ile Gly Gly Ala Asn Ala Phe Asp Ile 1 5 <210> 206 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 206 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Glu Tyr 20 25 30 Arg Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Tyr Pro Ser Gly Gly Trp Thr Asn Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ile Gly Gly Ala Asn Ala Phe Asp Ile Trp Gly Gln Gly Thr 100 105 110 Met Val Thr Val Ser Ser 115 <210> 207 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 207 Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr Asn Tyr Val Ser 1 5 10 <210> 208 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 208 Glu Val Ser Asn Arg Pro Ser 1 5 <210> 209 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 209 Ser Ser Tyr Thr Ser Gly Ser Thr Arg Tyr Val 1 5 10 <210> 210 <211> 114 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 210 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Glu Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Gly 85 90 95 Ser Thr Arg Tyr Val Phe Gly Pro Gly Thr Lys Val Thr Val Leu Gly 100 105 110 Gln Pro <210> 211 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 211 Lys Tyr Arg Met Ala 1 5 <210> 212 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 212 Gly Phe Thr Phe Ser Lys Tyr 1 5 <210> 213 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 213 Tyr Ile Tyr Pro Ser Gly Gly Phe Thr Phe Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 214 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 214 Tyr Pro Ser Gly Gly Phe 1 5 <210> 215 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 215 Asp Ile Gly Gln Trp Leu Phe Val Phe Asp Tyr 1 5 10 <210> 216 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 216 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Tyr 20 25 30 Arg Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Tyr Pro Ser Gly Gly Phe Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Asp Ile Gly Gln Trp Leu Phe Val Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 217 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 217 Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser 1 5 10 <210> 218 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 218 Asp Val Ser Asn Arg Pro Ser 1 5 <210> 219 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 219 Ser Ser Tyr Thr Asn Thr Ile Thr Val Val 1 5 10 <210> 220 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 220 Gln Ser Glu Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Asn Thr 85 90 95 Ile Thr Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 110 Pro <210> 221 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 221 Lys Tyr Asp Met Tyr 1 5 <210> 222 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 222 Gly Phe Thr Phe Ser Lys Tyr 1 5 <210> 223 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 223 Gly Ile Trp Pro Ser Gly Gly Leu Thr Met Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 224 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 224 Trp Pro Ser Gly Gly Leu 1 5 <210> 225 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 225 Asp Gly Val Val Gly Gly Ser Tyr Ala Phe Asp Ile 1 5 10 <210> 226 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 226 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Tyr 20 25 30 Asp Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Trp Pro Ser Gly Gly Leu Thr Met Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Asp Gly Val Val Gly Gly Ser Tyr Ala Phe Asp Ile Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 227 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 227 Ser Gly Ala Ser Ser Asn Leu Gly Arg Asn Thr Val Asn 1 5 10 <210> 228 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 228 Thr Asn Asp His Arg Pro Ser 1 5 <210> 229 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 229 Ala Ala Trp Asp His Ser Leu Asn Gly Tyr Val 1 5 10 <210> 230 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Human antibody library <400> 230 Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Ser Gly Ala Ser Ser Asn Leu Gly Arg Asn 20 25 30 Thr Val Asn Trp Tyr Gln Gln Val Pro Gly Ser Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Thr Asn Asp His Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Arg Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln 65 70 75 80 Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp His Ser Leu 85 90 95 Asn Gly Tyr Val Phe Gly Pro Gly Thr Lys Val Thr Val Leu Ser Gln 100 105 110 Pro <210> 231 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 231 Ile Tyr Ser Met Asn 1 5 <210> 232 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 232 Gly Phe Thr Phe Ser Ile Tyr 1 5 <210> 233 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 233 Val Ile Tyr Pro Ser Gly Gly Phe Thr His Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 234 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 234 Tyr Pro Ser Gly Gly Phe 1 5 <210> 235 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 235 Gly Arg Lys Thr Thr Val Thr Asp Phe Asp Tyr 1 5 10 <210> 236 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> Human Antibody Library <400> 236 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ile Tyr 20 25 30 Ser Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Tyr Pro Ser Gly Gly Phe Thr His Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Arg Lys Thr Thr Val Thr Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 237 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 237 Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser 1 5 10 <210> 238 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 238 Glu Val Ser His Arg Pro Ser 1 5 <210> 239 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 239 Ser Ser Tyr Thr Ser Gly Ser Thr Arg Val 1 5 10 <210> 240 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 240 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Val 35 40 45 Met Ile Tyr Glu Val Ser His Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Gly 85 90 95 Ser Thr Arg Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gln 100 105 110 Pro <210> 241 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 241 Met Tyr Met Met Met 1 5 <210> 242 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 242 Gly Phe Thr Phe Ser Met Tyr 1 5 <210> 243 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 243 Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 244 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 244 Tyr Pro Ser Gly Gly Ile 1 5 <210> 245 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 245 Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr 1 5 10 <210> 246 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 246 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Met Tyr 20 25 30 Met Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 247 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 247 Thr Gly Thr Ser Ser Asp Val Gly Ala Tyr Asn Tyr Val Ser 1 5 10 <210> 248 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 248 Asp Val Ser Asn Arg Pro Ser 1 5 <210> 249 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 249 Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val 1 5 10 <210> 250 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 250 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ala Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95 Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly Gln 100 105 110 Pro <210> 251 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 251 Ala Tyr Arg Met Val 1 5 <210> 252 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 252 Gly Phe Thr Phe Ser Ala Tyr 1 5 <210> 253 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 253 Arg Ile Tyr Pro Ser Gly Gly Phe Thr Phe Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 254 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 254 Tyr Pro Ser Gly Gly Phe 1 5 <210> 255 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 255 Ala Leu Gly Pro Leu Ser Pro Leu Asp Ser 1 5 10 <210> 256 <211> 119 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 256 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Arg Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Arg Ile Tyr Pro Ser Gly Gly Phe Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Leu Gly Pro Leu Ser Pro Leu Asp Ser Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 257 <211> 14 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 257 Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser 1 5 10 <210> 258 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 258 Glu Val Ser Asn Arg Pro Ser 1 5 <210> 259 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 259 Ser Ser Tyr Arg Ser Gly Asn Thr Leu Val 1 5 10 <210> 260 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Human Antibody Library <400> 260 Gln Ser Glu Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Glu Val Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Gly Asp Tyr Tyr Cys Ser Ser Tyr Arg Ser Gly 85 90 95 Asn Thr Leu Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gln 100 105 110 Pro <210> 261 <211> 209 <212> PRT <213> Artificial sequence <220> <223> SD15 fusion protein <220> <221> Domain <222> (1)..(61) <223> Human ILL-15Ra Sushi domain (NM-002189) <220> <221> Domain <222> (62)..(75) <223> IRD-11exone3 <220> <221> Domain <222> (76)..(95) <223> Linker <220> <221> Domain <222> (96)..(209) <223> Human IL-15 (NM-000585-3) <400> 261 Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 1 5 10 15 Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 20 25 30 Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala 35 40 45 Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 50 55 60 Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro Ser Gly Gly Ser Gly 65 70 75 80 Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Gln Asn 85 90 95 Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln 100 105 110 Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro 115 120 125 Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val 130 135 140 Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn 145 150 155 160 Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr 165 170 175 Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys 180 185 190 Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr 195 200 205 Ser <210> 262 <211> 659 <212> PRT <213> Artificial Sequence <220> <223> Fusion Protein <400> 262 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Met Tyr 20 25 30 Met Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Ser Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val 450 455 460 Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly 465 470 475 480 Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn 485 490 495 Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile 500 505 510 Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro Ser Gly Gly 515 520 525 Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu 530 535 540 Gln Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu 545 550 555 560 Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val 565 570 575 His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu 580 585 590 Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val 595 600 605 Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn 610 615 620 Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn 625 630 635 640 Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile 645 650 655 Asn Thr Ser <210> 263 <211> 659 <212> PRT <213> artificial sequence <220> <223> fusion protein <400> 263 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Met Tyr 20 25 30 Met Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu 405 410 415 Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr 420 425 430 Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro 435 440 445 Ala Pro Pro Ser Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Leu Gln Asn Trp Val Asn Val Ile Ser Asp Leu 465 470 475 480 Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu 485 490 495 Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys 500 505 510 Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala 515 520 525 Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser 530 535 540 Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu 545 550 555 560 Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His 565 570 575 Ile Val Gln Met Phe Ile Asn Thr Ser Phe Phe Leu Tyr Ser Lys Leu 580 585 590 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 595 600 605 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 610 615 620 Leu Ser Pro Gly Ser Cys Pro Pro Pro Met Ser Val Glu His Ala Asp 625 630 635 640 Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys 645 650 655 Asn Ser Gly <210> 264 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 264 Ala Tyr Ala Met Ala 1 5 <210> 265 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 265 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Ala Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 266 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 266 Ala Tyr Arg Met Phe 1 5 <210> 267 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 267 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Arg Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 268 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 268 Ala Tyr Leu Met Val 1 5 <210> 269 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 269 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Leu Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 270 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 270 Ala Tyr Val Met Phe 1 5 <210> 271 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 271 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Val Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 272 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 272 Ala Tyr Val Met Ser 1 5 <210> 273 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 273 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Val Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 274 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 274 Gly Tyr Leu Met Val 1 5 <210> 275 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 275 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30 Leu Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 276 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 276 Gly Tyr Gln Met Leu 1 5 <210> 277 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 277 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30 Gln Met Leu Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 278 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 278 Gly Tyr Ser Met Phe 1 5 <210> 279 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 279 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30 Ser Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 280 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 280 Gly Tyr Trp Met Ala 1 5 <210> 281 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 281 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gly Tyr 20 25 30 Trp Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 282 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 282 Gln Tyr Leu Met Tyr 1 5 <210> 283 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 283 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gln Tyr 20 25 30 Leu Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 284 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 284 Gln Tyr Val Met Phe 1 5 <210> 285 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 285 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gln Tyr 20 25 30 Val Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 286 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 286 Gln Tyr Tyr Met Tyr 1 5 <210> 287 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 287 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Gln Tyr 20 25 30 Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 288 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 288 Ser Tyr Leu Met Ser 1 5 <210> 289 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 289 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Leu Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 290 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 290 Ser Tyr Leu Met Val 1 5 <210> 291 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 291 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Leu Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 292 <211> 12 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 292 Ser Tyr Leu Met Thr Thr Cys Cys Leu Asp His Lys 1 5 10 <210> 293 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 293 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Leu Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 294 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 294 Ser Tyr Gln Met Val 1 5 <210> 295 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 295 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gln Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 296 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 296 See Tyr See Met Ala 1 5 <210> 297 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 297 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ser Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 298 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 298 Ser Tyr Val Met Phe 1 5 <210> 299 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 299 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Val Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 300 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 300 Watch Tyr Val Met Watch 1 5 <210> 301 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 301 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Val Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 302 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 302 See Tyr Val Met Tyr 1 5 <210> 303 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 303 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Val Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 304 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 304 Ser Tyr Tyr Met Phe 1 5 <210> 305 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 305 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Tyr Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 306 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 306 Watch Tyr Tyr Met Val 1 5 <210> 307 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 307 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Tyr Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 308 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 308 Tyr Tyr Sees With Val 1 5 <210> 309 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 309 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Tyr Tyr 20 25 30 Ser Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 310 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 310 Trp Tyr Leu Met Ala 1 5 <210> 311 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 311 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Trp Tyr 20 25 30 Leu Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 312 <211> 5 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 312 Trp Tyr Gln Met Ser 1 5 <210> 313 <211> 120 <212> PRT <213> artificial sequence <220> <223> synthetic <400> 313 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Trp Tyr 20 25 30 Gln Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 314 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 314 Ser Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly 1 5 10 15 Gly Ser Leu Gln 20 <210> 315 <211> 198 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(61) <223> Human IL-15Ra Sushi domain (NM-002189) <220> <221> Domain <222> (62)..(64) <223> IRD-11exone3 <220> <221> Domain <222> (65)..(84) <223> Connector 1 <220> <221> Domain <222> (85)..(198) <223> Human IL-15 (NM-000585-3) <400> 315 Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 1 5 10 15 Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 20 25 30 Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala 35 40 45 Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 50 55 60 Ser Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly 65 70 75 80 Gly Ser Leu Gln Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile 85 90 95 Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu 100 105 110 Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu 115 120 125 Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His 130 135 140 Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser 145 150 155 160 Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu 165 170 175 Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln 180 185 190 Met Phe Ile Asn Thr Ser 195 <210> 316 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 316 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 Gly Gly Gly Gly Ser Gly Gly Gly Gly 20 25 <210> 317 <211> 203 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(61) <223> Human ILL-15Ra Sushi domain (NM-002189) <220> <221> Domain <222> (62)..(64) <223> Human IL-15Ra Sushi domain (NM-002189) IRD-11exone3 <220> <221> Domain <222> (65)..(89) <223> Connector 2 <220> <221> Domain <222> (90)..(203) <223> Human IL-15 (NM-000585-3) <400> 317 Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 1 5 10 15 Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 20 25 30 Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala 35 40 45 Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 50 55 60 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 65 70 75 80 Gly Gly Gly Gly Ser Gly Gly Gly Gly Asn Trp Val Asn Val Ile Ser 85 90 95 Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala 100 105 110 Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala 115 120 125 Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly 130 135 140 Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn 145 150 155 160 Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu 165 170 175 Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe 180 185 190 Val His Ile Val Gln Met Phe Ile Asn Thr Ser 195 200 <210> 318 <211> 30 <212> PRT <213> Artificial sequence <220> <223> Synthetic <400> 318 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 20 25 30 <210> 319 <211> 208 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(61) <223> Human ILL-15Ra Sushi domain (NM-002189) <220> <221> Domain <222> (62)..(64) <223> IRD-11exone3 <220> <221> Domain <222> (65)..(94) <223> Connector 3 <220> <221> Domain <222> (95)..(208) <223> Human IL-15 (NM-000585-3) <400> 319 Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 1 5 10 15 Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 20 25 30 Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala 35 40 45 Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 50 55 60 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 65 70 75 80 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Asn Trp 85 90 95 Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser 100 105 110 Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser 115 120 125 Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile 130 135 140 Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu 145 150 155 160 Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu 165 170 175 Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu 180 185 190 Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser 195 200 205 <210> 320 <211> 40 <212> PRT <213> Artificial sequence <220> <223> Synthetic <400> 320 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 20 25 30 Gly Gly Gly Ser Gly Gly Gly Gly 35 40 <210> 321 <211> 218 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(61) <223> Human ILL-15Ra Sushi domain (NM-002189) <220> <221> Domain <222> (62)..(64) <223> IRD-11exone3 <220> <221> Domain <222> (65)..(104) <223> Linker 4 <220> <221> Domain <222> (105)..(218) <223> Human IL-15 (NM-000585-3) <400> 321 Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 1 5 10 15 Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 20 25 30 Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala 35 40 45 Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 50 55 60 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 65 70 75 80 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 85 90 95 Gly Gly Gly Ser Gly Gly Gly Gly Asn Trp Val Asn Val Ile Ser Asp 100 105 110 Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr 115 120 125 Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met 130 135 140 Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp 145 150 155 160 Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn 165 170 175 Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys 180 185 190 Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val 195 200 205 His Ile Val Gln Met Phe Ile Asn Thr Ser 210 215 <210> 322 <211> 50 <212> PRT <213> Artificial Sequence <220> <223> Synthetic <400> 322 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 20 25 30 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 35 40 45 Gly Gly 50 <210> 323 <211> 228 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(61) <223> Human ILL-15Ra Sushi domain (NM-002189) <220> <221> Domain <222> (62)..(64) <223> IRD-11exone3 <220> <221> Domain <222> (65)..(114) <223> Connector 5 <220> <221> Domain <222> (115)..(228) <223> Human IL-15 (NM-000585-3) <400> 323 Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser 1 5 10 15 Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys 20 25 30 Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala 35 40 45 Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp 50 55 60 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 65 70 75 80 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 85 90 95 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 100 105 110 Gly Gly Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp 115 120 125 Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp 130 135 140 Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu 145 150 155 160 Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr 165 170 175 Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly 180 185 190 Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys 195 200 205 Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe 210 215 220 Ile Asn Thr Ser 225 <210> 324 <211> 18 <212> PRT <213> Artificial sequence <220> <223> Synthetic <400> 324 Ser Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly 1 5 10 15 Gly Ser <210> 325 <211> 646 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(120) <223> tccLD7-102 heavy chain variable domain <220> <221> Domain <222> (121)..(449) <223> IgG1 CH1-CH2-CH3 domain <220> <221> Domain <222> (450)..(646) <223> deSD15-20 <400> 325 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ala Tyr 20 25 30 Arg Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Ser Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val 450 455 460 Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly 465 470 475 480 Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn 485 490 495 Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile 500 505 510 Arg Asp Ser Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Gly 515 520 525 Gly Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile 530 535 540 Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu 545 550 555 560 Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu 565 570 575 Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His 580 585 590 Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser 595 600 605 Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu 610 615 620 Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln 625 630 635 640 Met Phe Ile Asn Thr Ser 645 <210> 326 <211> 216 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(113) <223> V L <220> <221> Domain <222> (114)..(216) <223> CL <400> 326 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ala Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95 Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly Gln 100 105 110 Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 327 <211> 215 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Domain <222> (1)..(113) <223> V L <220> <221> Domain <222> (114)..(215) <223> CL w / missing C-terminal serine <400> 327 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ala Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95 Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly Gln 100 105 110 Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys 210 215 <210> 328 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Synthetic <220> <221> Variants <222> (1) <223> Residues are A, G, M, Q, S, Y, or W <220> <221> Variants <222> (3) <223> Residue is A, L, M, Q, R, S, V, W or Y <220> <221> Variants <222> (5) <223> Residue is A, F, L, M, S, T, V or Y <400> 328 Xaa Tyr Xaa Met Xaa 1 5 BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1Depicted are the binding of antibodies tccR3λF8, tccR3κA11, tccR3λH4, tctR3κA8, sR3λD7, and R2κA6 to human hPDL1-Fc (upper left panel), blocking of hPDL1 binding to hPD1 (lower left panel), binding to mouse mPDL1-Fc (upper right panel), and blocking of mPDL1 binding to hPD1 (lower right panel).

[0019] Figure 2 Depicted are the binding of antibodies sR3λD7, tccR3κB7, tccR3κA4, tccR3λF8, tccR3λD7, tccR3λH4, and tccR3κD9 to human hPDL1-Fc (upper left panel), blocking of hPDL1 binding to hPD1 (lower left panel), binding to mouse mPDL1-Fc (upper right panel), and blocking of mPDL1 binding to hPD1 (lower right panel).

[0020] Figure 3 Depicted are the binding of antibodies tccR3κF8, tccR3κD9, tccR3λD7, tccR3λD7sR3κF10, sR3λD7, and tccR3λF8 to human hPDL1-Fc (upper left panel), blocking of hPDL1 binding to hPD1 (lower left panel), binding to mouse mPDL1-Fc (upper right panel), and blocking of mPDL1 binding to hPD1 (lower right panel).

[0021] Figure 4 Antibodies R2κA6, sR3λD7, tccR3λD7, tccR3κB7, and tccR3κH4 are depicted for binding to human hPDL1-Fc (upper left panel), blocking of hPDL1 binding to hPD1 (lower left panel), binding to mouse mPDL1-Fc (upper right panel), and blocking of mPDL1 binding to hPD1 (lower right panel).

[0022] Figure 5 Binding of antibodies sR3λD7, tctR3κA8, tccR3κA11, tccR3λD7, tccR3κD9, tccR3λF8, tccR3κF8, tccR3κF10, tccR3λH4, tccR3κB7, and tccR3κA4 to PDLI-293 cells (top) and MDS-MB-231 (bottom) cells are depicted.

[0023] Figure 6 Anti-PD-L1 antibodies are depicted binding to (A) human monocyte-derived dendritic cells, (B) MDA-MB-231 human cancer line cells expressing PD-L1, and (C) mouse cell line expressing PD-L1 B16-F10.

[0024] Figure 7 Shown are the functional blocking activities of anti-PD-L1 antibodies measured by: (A) increase in CD4 proliferation when activated by aCD3 and PD-L1Fc-coated beads, (B) increase in cytokine secretion in SEB-activated human PBMCs, and (C) increase in CD4 proliferation in mixed lymphocyte reactions with mo-DCs.

[0025] Figure 8 Shown are the activation of CD4 and CD8 when both anti-PDL1 antibody and IL15 were present in (A) mixed lymphocyte reaction with mo-DCs and (B) CD8 stimulation by αCD3 and PD-L1Fc-coated beads.

[0026] Fig. 9 The anti-PD-L1-sushi domain-IL15 (referred to as anti-PDL1-SD15) fusion protein is shown to retain binding to PD-L1 as measured by (A) solid phase ELISA and (B) binding to CD4 activated by aCD3-coated beads.

[0027] Fig.10 It is shown that PBMCs cultured in vitro with anti-PD-L1-SD15 fusion protein resulted in increased NK cell numbers (A), increased CD8 cell numbers (B), and activation status (C) as measured by the percentage of granzyme B. No effect on CD4 cells was observed (D).

[0028] Fig.11 Anti-PD-L1-SD15 fusion protein is shown to activate CD8 similarly to IL15 when added to in vitro CD8 stimulation in the presence of aCD3-coated beads (A). However, in the presence of aCD3 and PD-L1Fc-coated beads, anti-PD-L1-SD15 fusion protein can increase CD8 proliferation more than 5-fold when compared to IL15 (B). cD7-SD15neg is an anti-PD-L1 cD7 with non-functional IL15 used as a negative control.

[0029] Fig.12Activation of CD8 in the presence of PD-L1Fc on antigen presenting cells is shown. Anti-PD-L1-SD15 fusion protein cD7-SD15 stimulates CD8 at significantly lower concentrations, as measured by (A) an increase in the percentage of granzyme B-positive CD8 and (B) an increase in total cytokine secretion. Compared to IL15, the highest level of CD8 activation was increased in CD8 activated by aCD3 and PD-L1Fc with the addition of cD7-SD15. (C) Data on CD8 proliferation in the presence of anti-PD-L1 antibodies and free IL15 (dashed line) overlap with data on CD8 proliferation in the presence of anti-PD-L1-SD15 fusion protein (straight line).

[0030] FIG. 13 provides V values ​​of anti-PD-L1 antibodies. H ( Figure 13A1-3 ) and V L ( Figure 13B1-3 ) chain. H Sequences, boxed regions indicate CDRs. For CDR-1H, Chothia CDRs are in italics, and Kabat CDRs are underlined. For CDR-2H, Kabat CDRs are coextensive with the boxed sequences and Chothia CDRs in italics. L Sequences, boxed regions indicate Kabat / Chothia CDRs.

[0031] Fig.14 Shown are the amino acid sequences of SD15 (SEQ ID NO: 261), tccλD7HC-SD15 (SEQ ID NO: 262), and a LALA mutant of tccλD7HC-SD15 (SEQ ID NO: 263) comprising the IL15Rα sushi domain and IL15, wherein the LALA mutant contains two adjacent leucines (Leu) at positions in the heavy chain constant region that are important for FcγRI. 234 and Leu 235 ) is substituted with alanine.

[0032] Fig.15 Cytotoxicity of an anti-PD-L1-SD15 fusion protein (CD7SD15) compared to the PD-L1 binding portion of the molecule alone (cD7) and a fusion protein containing binding domains specific for KLH and IL15 domains (KLHSD15) is shown. Human CD8 T cells and MAD-MB-231 tumor cells were co-cultured for 7 days in IMDM supplemented with 10% FBS. Tumor cell killing activity was assessed by measuring the number of dead tumor cells stained with Viability Dye eFluor 780 in FACS.

[0033] Fig.16 The anti-PD-L1-SD15 fusion protein was shown to prolong the survival of mice with tumors expressing PD-L1. 5 Balb / c mice were injected intravenously with CT26 colon tumor cells. After 24 hours, mice received intraperitoneal administration of anti-PD-L1 antibody cD7 (purple line: 75ug / dose), anti-PD-L1-SD15 fusion protein cD7-SD15 (green line: 75ug / dose, blue line: 25ug / dose) or sD7-SD15 (gray line: 75ug / dose, red line: 25ug / dose) twice a week in the first week, and then once a week for the rest of the treatment course. Mice in the control group received an equal volume of saline or normal IgG solution. Survival was measured using a Kaplan-Meier plot.

[0034] Fig.17 Binding of the two affinity matured anti-PDL1 antibodies to soluble human PDL1, soluble mouse PDL1, and soluble rat PDL1 and no binding to human PDL2 were shown.

[0035] Fig.18 Blockade of human PD1 to human PDL1 (upper panel) and mouse PD1 to mouse PDL1 (lower panel) by two affinity matured anti-PDL1 antibodies compared to their parental tccλD7 antibody.

[0036] Fig.19 It was shown that two affinity matured variants of the anti-PD-L1 antibody tccλD7 had higher binding activity to human MDA-MB-231 tumor cells expressing PD-L1 as measured by flow cytometry.

[0037] Fig. 20 Affinity matured variants of the anti-PD-L1 antibody tccλD7 showing enhanced potency in promoting the production of the Th1 cytokines IL2 (upper panel) and IFNγ (lower panel).

[0038] Fig.21 The binding of the fusion protein of the present invention to MDA-MB-231 tumor cells expressing PD-L1 is shown.

[0039] Fig. 22 The protein of the present invention was shown to have stimulatory activity on IL15-responsive human megakaryoblastic leukemia cells.

[0040] Fig.23 The hPD-L1 binding (upper panel) and ligand blocking (lower panel) activities of the fusion proteins of the present invention are shown.

[0041] Fig.24 The results of size exclusion chromatography of the fusion protein of the present invention are shown.

[0042] Fig.25 The serum stability of the fusion protein of the present invention is shown.

[0043] Details

[0044] The interaction of PD-1 and PD-L1 on immune cells inhibits the proliferation and cytokine production performed by immune cells. PD-L1 is also inducible and upregulated in various tissues (including cancer). In short, PD-1 and PD-L1 play a role in immunosuppression. The present invention provides novel antibodies or antigen-binding fragments of such antibodies that bind to PD-L1 and block the interaction with PD-1. In an embodiment of the present invention, the antibody weakens or inhibits immunosuppression.

[0045] The novel antibodies of the present invention are shown in Table 1 and the attached sequence table, which show the amino acid sequences of heavy chain and light chain CDRs (identified according to the identification system of Kabat and Chothia) and complete heavy chain and light chain variable regions. The first two heavy chain CDRs are identified according to the common system of Kabat and Chothia, which provides different but overlapping CDR positions. The comparison of many heavy chains and light chains shows significant similarities between many CDR sequences. Therefore, it is expected that many CDRs can be mixed and matched between sequences.

[0046] The antibody may have one or more amino acid substitutions, deletions, insertions and / or additions. In certain embodiments, the antibody comprises one of the above-mentioned heavy chain variable domains and one of the above-mentioned light chain variable domains. In certain embodiments, the PD-L1 antibody or its binding fragment comprises one or more CDRs or one or more variable domains having an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identical to the CDR and variable domain sequences shown in Table 1.

[0047] "Identity" is the number or percentage of identical positions shared by two amino acid or nucleic acid sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap. "Substantially identical" refers to amino acid sequences that differ only in conservative amino acid substitutions, such as substitution of one amino acid for another of the same kind (e.g., substitution of valine for glycine, substitution of arginine for lysine, etc.), or in one or more non-conservative substitutions, deletions or insertions at positions in the amino acid sequence that do not destroy the function of the protein. In some cases, amino acid substitutions are produced by selecting substitutions that do not differ significantly in their effects on maintaining: (a) the structure of the peptide backbone in the substituted region, (b) the charge or hydrophobicity of the molecule at the target site; or (c) the bulk of the side chain. For example, naturally occurring residues can be divided into the following groups based on the properties of their side chains: (1) hydrophobic amino acids (leucine, methionine, alanine, valine, leucine, and isoleucine); (2) neutral hydrophilic amino acids (cysteine, serine, and threonine); (3) acidic amino acids (aspartic acid and glutamic acid); (4) basic amino acids (asparagine, glutamine, histidine, lysine, and arginine); (5) amino acids that affect chain orientation (glycine and proline); and (6) aromatic amino acids (tryptophan, tyrosine, and phenylalanine). Substitutions made within these groups can be considered conservative substitutions. Examples of substitutions include, but are not limited to, valine for alanine, lysine for arginine, glutamine for asparagine, glutamic acid for aspartic acid, serine for cysteine, asparagine for glutamine, aspartic acid for glutamic acid, proline for glycine, arginine for histidine, leucine for isoleucine, isoleucine for leucine, arginine for lysine, leucine for methionine, leucine for phenylalanine, glycine for proline, threonine for serine, serine for threonine, tyrosine for tryptophan, phenylalanine for tyrosine, and / or leucine for valine.

[0048] Preferably, the amino acid sequence has at least 80%, or at least 85%, or at least 90%, or at least 95% identity with the amino acid sequence disclosed herein. Methods and computer programs for determining sequence similarity are publicly available, including but not limited to the GCG program package (Devereux et al., Nucleic Acids Research 12: 387, 1984), BLASTP, BLASTN, FASTA (Altschul et al., J. Mol. Biol. 215: 403 (1990) and the ALIGN program (version 2.0). The well-known Smith-Waterman algorithm can also be used to determine similarity. The BLAST program is publicly available from NCBI and other sources (BLAST Manual, Altschul, et al., NCBI NLM NIH, Bethesda, Md. 20894; BLAST at http: / / www.ncbi.nlm.nih.gov / blast / 2.0). These methods account for various substitutions, deletions, and other modifications when comparing sequences. Conservative substitutions generally include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.

[0049] The antibodies of the present invention also include those whose binding characteristics have been improved by direct mutation, affinity maturation, phage display or chain shuffling. Affinity and specificity can be modified or improved by mutating CDRs and screening for antigen binding sites with desired characteristics. CDRs are mutated in a variety of ways. One way is to randomize single residues or combinations of residues so that all 20 amino acids are found at specific positions in a group of otherwise identical antigen binding sites. Alternatively, mutations are induced within the scope of CDR residues by error-prone PCR methods (see, e.g., Hawkins et al., J. Mol. Biol., 226: 889-896 (1992)). For example, phage display vectors containing heavy and light chain variable region genes can be amplified in a mutator strain of Escherichia coli (E. coli) (see, e.g., Low et al., J. Mol. Biol., 250: 359-368 (1996)). These mutagenesis methods are illustrative of many methods known to those skilled in the art.

[0050] In order to minimize immunogenicity, antibodies comprising human constant domain sequences are preferred. The antibody can be any immunoglobulin class such as IgG, IgM, IgA, IgD or IgE and its subclass, or a member of any of the immunoglobulin classes and its subclass can be combined. Antibody classes can also be selected to optimize the effector functions (e.g., complement dependent cellular toxicity (CDC) and antibody dependent cellular toxicity (ADCC)) of natural antibodies.

[0051] Certain embodiments of the present invention include the use of PD-L1 binding antibody fragments. Fv is the smallest fragment containing a complete heavy chain variable domain and a light chain variable domain, including all 6 hypervariable loops (CDRs). Since there are no constant domains, the variable domains are non-covalently associated. H With V L The linker that the domains associate to form the antigen binding site connects the heavy and light chains into a single polypeptide chain ("single-chain Fv" or "scFv"). In an embodiment of the invention, the linker is (Gly-Gly-Gly-Gly-Ser)3. Since scFv fragments do not have the constant domains of intact antibodies, they are much smaller than intact antibodies. ScFv fragments also do not contain the normal heavy chain constant domains that interact with other biological molecules that may not be desirable in certain embodiments.

[0052] You can also use V H 、V L and optionally C L , C H 1 or other constant domains. The monovalent fragment of the antibody produced by papain digestion is called Fab and does not have a heavy chain hinge region. The fragment produced by pepsin digestion (called F (ab ') 2) retains the heavy chain hinge and is bivalent. Such fragments can also be recombinantly produced. Many other useful antigen-binding antibody fragments are known in the art and include but are not limited to diabodies, triabodies, tetrabodies, single domain antibodies and other monovalent and multivalent forms.

[0053] The present invention also provides multivalent antigen binding proteins, which may be in the form of (but not limited to) antibodies, antigen binding fragments thereof, and proteins comprising all or part of the antigen binding portion of antibodies. Multivalent antigen binding proteins may be monospecific, bispecific, or multispecific. The term specificity refers to the number of different types of antigenic determinants to which a particular molecule can bind. If an immunoglobulin molecule binds to only one type of antigenic determinant, the immunoglobulin molecule is monospecific. If an immunoglobulin molecule binds to different types of antigenic determinants, the immunoglobulin molecule is multispecific.

[0054] In an embodiment of the present invention, the PD-L1 binding protein has at least about 10 2 M -1 s -1 , at least about 10 3 M -1 s -1 , at least about 10 4 M -1 s -1 , at least about 10 5 M -1 s -1 or at least about 10 6 M -1 s -1 In one embodiment, the PD-L1 binding protein has an association rate constant (Kon) of 10 2 M -1 s -1 Up to 10 3 M -1 s -1 , 10 3 M -1 s -1 Up to 10 4 M -1 s -1 , 10 4 M -1 s -1 Up to 10 5 M -1 s -1 or 10 5 M -1 s -1 Up to 10 6 M -1 s -1 The association rate constant (Kon) of , as measured by surface plasmon resonance.

[0055] In another embodiment, the PD-L1 binding protein has at most about 10 -3 s -1 , up to about 10 -4 s -1 , up to about 10 -5 s -1 or at most about 10 -6 s -1 In one embodiment, the PD-L1 binding protein has a dissociation rate constant (Koff) of 10 -3 s -1 Up to 10 -4 s -1 , 10 -4 s-1 Up to 10 -5 s -1 or 10 -5 s -1 Up to 10 -6 s -1 The dissociation rate constant (Koff) of , as measured by surface plasmon resonance.

[0056] In another embodiment, the PD-L1 binding protein has at most about 10 -7 M, up to about 10 -8 M, up to about 10 -9 M, up to about 10 -10 M, up to about 10 -11 M, up to about 10 -12 M or at most about 10 -13 The dissociation constant (K D In one embodiment, the binding protein has 10 -7 M to 10 -8 M, 10 -8 M to 10 -9 M, 10 -9 M to 10 -10 M, 10 -10 M to 10 -11 M, 10 -11 M to 10 -12 M or 10 -12 M to 10 -13 The dissociation constant (K) of M to its target D ).

[0057] The binding proteins described herein may be conjugates that also include an imaging agent, a therapeutic agent, or a cytotoxic agent. In one embodiment, the imaging agent is a radioactive label, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, or biotin. In another embodiment, the radioactive label is: 3 H. 14 C. 35 S. 90 Y. 99 Tc, 111 In, 125 I. 131 I. 177 Lu or 153 In yet another embodiment, the therapeutic or cytotoxic agent is an antimetabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, a toxin, or an apoptotic agent. As discussed below, immunostimulatory cytokines are particularly important.

[0058] The present invention also provides molecules that bind to PD-L1 to inhibit immunosuppression, which also promote immune responses by interacting with other ligands or receptors. As exemplified herein, such a molecule combines the PD-L1 binding domain of an antibody with a domain that stimulates the function of NK cells or T cells. Such a stimulatory domain can be, but is not limited to, a stimulatory domain that binds and stimulates receptors that respond to interleukins or interferons such as, but not limited to, IL2, IL7, IL15, and IL21. The stimulatory domain exemplified herein is a mixed domain comprising a sushi domain of the IL15Rα chain of IL15 connected to IL15 by a linker (e.g., SEQ ID NO: 261). An example of a complete molecule is shown in SEQ ID NO: 262. An almost identical molecule modified with 2 amino acid substitutions in the region between the antibody domain and the IL15R stimulatory domain to inhibit proteolysis of the region is shown in SEQ ID NO: 263. As demonstrated herein, molecules comprising a PD-L1 binding domain that inhibits immunosuppression and a second domain that promotes an immune response provide enhanced immune cell activity compared to two different molecules that provide said functions alone.

[0059] As exemplified herein, the PD-L1 binding portion of the molecule is an antigen binding domain of an antibody. Several novel antibody heavy chain and light chain variable domains and antibodies comprising them are provided. According to the present invention, the PD-L1 binding protein can be any agent that binds to PD-L1 and blocks immunosuppression. These agents include anti-PD-L1 antibodies and fragments, not limited to those novel antibodies disclosed herein, as well as peptides and proteins derived from PD1, natural ligands of PD-L1.

[0060] As disclosed herein, the PD-L1 binding domain is connected to a domain that stimulates NK and T cell activity. The domain comprises IL15 and is connected to it by a flexible linker, and the "sushi" domain is from the alpha chain of the IL15 receptor. The sushi domain binds IL15 with high affinity, and the complex of IL15 and the sushi domain is particularly active for stimulating NK and T cell proliferation. It is particularly noteworthy that, as shown in the Examples, treatment with an agent combining a PD-L1 binding domain in the same molecule as the IL15 stimulatory domain is more effective than combined treatment using the PD-L1 binding domain and the IL15 stimulatory domain as separate molecules.

[0061] Thus, in certain embodiments, the invention encompasses hybrid molecules comprising a domain that binds to PD-L1 and blocks binding to PD1 and a domain that stimulates IL15R, thereby stimulating the proliferation of immune cells. As exemplified, the IL15R stimulatory domain comprises a sushi domain linked to the IL15Rα chain of IL15 via a flexible linker similar to those used for single chain Fv molecules (i.e., containing 15-20 amino acids, primarily serine and glycine). In practice, there are other methods that can be used, which may be preferred, for example, for manufacturing procedures. In addition, one recognizes the domain structure and, thus, the modular aspects and other properties of the disclosed hybrid proteins. For example, the linker that links the sushi domain to IL15 is used to express the hybrid protein as one polypeptide, but may likewise be replaced by other reagents, linkers, or cross-linking agents. Alternatively, the high affinity of IL15 for the sushi-containing portion of the IL15Rα chain suggests that the sushi domain may form a stable complex with IL15 that is not necessarily covalent. Similarly, while the exemplified proteins comprise complete antibody constant regions, other antigen-binding fragments of PD-L1 binding antibodies may also be sufficient.

[0062] Thus, the present invention provides a PD-L1 binding domain connected to an IL15R stimulatory domain, the IL15R stimulatory domain comprising a sushi domain of an IL15R alpha chain or a variant thereof and IL15 or a variant thereof. In certain embodiments, the variant may have 80%, 85%, 87%, 90%, 91%, 92%, 93%, 94% or 95% identity to a sequence disclosed herein. In one embodiment, the sushi domain of the IL15R alpha chain forms a covalent complex with IL15. In another embodiment, the sushi domain of the IL15R alpha chain and IL15 form a non-covalent complex. The PD-L1 binding domain may comprise 1, 2, 3, 4, 5 or 6 CDRs, or heavy and / or light chain variable domains, of an antibody thereof disclosed herein, or an antigen-binding fragment thereof, or a variant thereof, such as a variant having 80%, 85%, 87%, 90%, 91%, 92%, 93%, 94% or 95% identity thereto, or a PD-L1 antibody or antigen-binding fragment thereof known in the art that blocks binding to PD-1.

[0063] It should be understood that the anti-PD-L1 antibodies and hybrid proteins of the present invention, when used in mammals for prophylactic or therapeutic purposes, will be administered in the form of a composition that additionally comprises a pharmaceutically acceptable carrier. Suitable pharmaceutically acceptable carriers include, for example, one or more of water, saline, phosphate-buffered saline, glucose, glycerol, sucrose, polysorbate, ethanol, etc. and combinations thereof. Pharmaceutically acceptable carriers may also contain small amounts of auxiliary substances, such as wetting agents or emulsifiers, preservatives or buffers, which enhance the shelf life or efficacy of the antibody.

[0064] In the methods of the present invention, a therapeutically effective amount of an antibody of the hybrid protein of the present invention is administered to a mammal in need thereof. As used herein, the term "administering" refers to delivering the antibodies and fusion proteins of the present invention to a mammal by any method that can achieve the desired result. They can be administered, for example, intravenously or intramuscularly. Although the exemplary antibodies of the present invention are particularly useful for administration to humans, they can also be administered to other mammals. As used herein, the term "mammal" is intended to include, but is not limited to, humans, laboratory animals, household pets, and farm animals. "Therapeutically effective amount" refers to an antibody of the present invention that is an effective amount in producing a desired therapeutic effect, such as inhibiting kinase activity, when administered to a mammal.

[0065] The antibodies and hybrid proteins of the present invention are used to inhibit tumors and other neoplastic diseases, and to treat other pathological conditions associated with immunosuppression. Tumors that can be treated include primary tumors, metastatic tumors, and refractory tumors. Refractory tumors include tumors that do not respond or are resistant to treatment with a single chemotherapeutic agent, a single antibody, radiation alone, or a combination thereof. Refractory tumors also include tumors that appear to be suppressed by treatment with such agents, but recur up to 5 years, sometimes 10 years or more after discontinuation of treatment. The antibodies are effective for treating vascularized tumors and tumors that are not vascularized or not yet significantly vascularized.

[0066] Examples of solid tumors that can be treated accordingly include breast cancer, lung cancer, colorectal cancer, pancreatic cancer, gliomas and lymphomas. Some examples of such tumors include epidermoid tumors, squamous tumors such as head and neck tumors, colorectal tumors, prostate tumors, breast tumors, lung tumors (including small cell and non-small cell tumors), pancreatic tumors, thyroid tumors, ovarian tumors and liver tumors. Other examples include Kaposi's sarcoma, central nervous system tumors, neuroblastomas, capillary hemangioblastomas, meningiomas and brain metastases, melanomas, gastrointestinal and renal cancers and sarcomas, rhabdomyosarcomas, glioblastomas, preferably glioblastoma multiforme and leiomyosarcomas. Examples of vascularized skin cancers for which the antagonists of the present invention are effective include squamous cell carcinomas, basal cell carcinomas and skin cancers that can be treated by inhibiting the growth of malignant keratinocytes such as human malignant keratinocytes.

[0067] Examples of non-solid tumors include leukemias, multiple myeloma, and lymphomas that are unresponsive to cytokines such as IL15. Some examples of leukemias include acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), erythrocytic leukemia, or monocytic leukemia. Some examples of lymphomas include Hodgkin's and non-Hodgkin's lymphomas.

[0068] The PD-L1 antibodies and immune cell stimulatory hybrid proteins of the present invention are also used for the treatment of viral infections. PD-1 expression on T cells is associated with viral load in patients infected with HIV and HCV, and PD-1 expression has been identified as a marker of exhausted virus-specific CD8+ T cells. For example, PD-1 + CD8 + T cells show impaired effector function and PD-1-related T cell exhaustion that can be restored by blocking PD-1 / PD-L1 interactions. This leads to the recovery of virus-specific CD8+T cell-mediated immunity, indicating that interruption of PD-1 signaling using antagonistic antibodies can restore T cell effector function. Immunotherapy based on the blockade of PD-1 / PD-L1 leads not only to the collapse of T cell tolerance to tumor antigens, but also provides a strategy for reactivating virus-specific effector T cells and eradicating pathogens in chronic viral infections. Therefore, the antibodies and hybrid proteins of the present invention can be used to treat chronic viral infections, including but not limited to HCV and HIV, and lymphocytic choriomeningitis virus (LCMV).

[0069] The antibodies and hybrid proteins of the present invention can be advantageously administered to a subject in need thereof together with a second agent. For example, in some embodiments, the antibodies or hybrid proteins of the present invention are administered to a subject together with an anti-tumor agent. In some embodiments, the antibodies or hybrid proteins of the present invention are administered to a subject together with a second angiogenesis inhibitor. In some embodiments, the antibodies or hybrid proteins of the present invention are administered together with an anti-inflammatory agent or an immunosuppressant.

[0070] Antineoplastic agents include cytotoxic chemotherapeutic agents, targeted small molecules and biomolecules, and radiation. Non-limiting examples of chemotherapeutic agents include cisplatin, dacarbazine (DTIC), dactinomycin, irinotecan, mechlorethamine (nitrogen mustard), mustard), streptozocin, cyclophosphamide, carmustine (BCNU), lomustine (CCNU), doxorubicin (Adriamycin), daunorubicin, procarbazine, mitomycin, cytarabine, etoposide, methotrexate, 5-fluorouracil, vinblastine, vincristine, bleomycin, paclitaxel (Taxol), docetaxel (Taxotere), aldesleukin, asparaginase, busulfan, carboplatin, cladribine, dacarbazine, floxuridine, fludarabine, hydroxyurea, ifosfamide, interferon alpha, leuprolide, megestrol acetate, melphalan, mercaptopurine, plicamycin, mitotane, pegaspargase, pentostatin, pipobroman, plicamycin, streptozocin, tamoxifen, teniposide, testolactone, thioguanine, thiotepa, uracil mustard, vinorelbine, chlorambucil, paclitaxel, and combinations thereof.

[0071] Targeted small molecules and biomolecules include, but are not limited to, inhibitors of components of signal transduction pathways, such as modulators of tyrosine kinases and receptor tyrosine kinase inhibitors, and agents that bind to tumor-specific antigens. Non-limiting examples of growth factor receptors involved in tumorigenesis are receptors for platelet-derived growth factor (PDGFR), insulin-like growth factor (IGFR), nerve growth factor (NGFR), and fibroblast growth factor (FGFR), and receptors of the epidermal growth factor receptor family, including EGFR (erbB1), HER2 (erbB2), erbB3, and erbB4.

[0072] EGFR antagonists include antibodies that bind to EGFR or EGFR ligands and inhibit ligand binding and / or receptor activation. For example, the agent can block the formation of receptor dimers or heterodimers with other EGFR family members. The ligands of EGFR include, for example, EGF, TGF-α amphiregulin, heparin-binding EGF (HB-EGF) and βregullulin. EGFR antagonists can bind to the extracellular portion of EGFR from the outside (which may or may not inhibit the binding of the ligand), or bind to the tyrosine kinase domain internally. EGFR antagonists also include, for example, agents that inhibit EGFR-dependent signal transduction by inhibiting the function of components of the EGFR signal transduction pathway. Examples of EGFR antagonists that bind to EGFR include, but are not limited to, biomolecules, such as antibodies (and their functional equivalents) that are specific for EGFR, and small molecules, such as synthetic kinase inhibitors that act directly on the cytoplasmic domain of EGFR.

[0073] Small molecule and biologic inhibitors include inhibitors of epidermal growth factor receptor (EGFR), including gefitinib, erlotinib and cetuximab, inhibitors of HER2 (e.g., trastuzumab, trastuzumab emtansine (trastuzumab-DM1; T-DM1) and pertuzumab), anti-VEGF antibodies and fragments (e.g., bevacizumab), antibodies that inhibit CD20 (e.g., rituximab, ibritumomab tiuxetan), anti-VEGFR antibodies (e.g., ramucirumab (IMC-1121B), IMC-1C11 and CDP791), anti-PDGFR antibodies and imatinib. Small molecule kinase inhibitors may be specific for a particular tyrosine kinase or be inhibitors of two or more kinases. For example, the compound N-(3,4-dichloro-2-fluorophenyl)-7-({[(3aR,6aS)-2-methyloctahydrocyclopenta[c]pyrrol-5-yl]methyl}oxy)-6-(methyloxy)quinazolin-4-amine (also known as XL647, EXEL-7647, and KD-019) is an in vitro inhibitor of several receptor tyrosine kinases (RTKs), including EGFR, EphB4, KDR (VEGFR), Flt4 (VEGFR3), and ErbB2, and is also an inhibitor of SRC kinases, which are involved in pathways that lead to tumor non-responsiveness to certain TKIs. In an embodiment of the invention, treatment of a subject in need thereof comprises administration of a rho-kinase inhibitor of Formula I and administration of KD-019.

[0074] Dasatinib (BMS-354825; Bristol-Myers Squibb, New York) is another orally bioavailable ATP site competitive Src inhibitor. Dasatinib also targets Bcr-Abl (FDA approved for patients with chronic myeloid leukemia (CML) or Philadelphia chromosome positive (Ph+) acute lymphoblastic leukemia (ALL)) as well as c-Kit, PDGFR, C-FMS, EphA2 and SFK. Two other oral tyrosine kinase inhibitors of Src and Bcr-Abl are bosutinib (SKI-606) and saratinib (AZD0530).

[0075] In an embodiment of the present invention, the PD-L1 antibody or conjugate of the present invention is combined with an antiviral agent for the treatment of chronic viral infection. For example, for HCV, the following agents can be used. HCV protease inhibitors include, but are not limited to, boceprevir, telaprevir (VX-950), ITMN-191, SCH-900518, TMC-435, BI-201335, MK-7009, VX-500, VX-813, BMS790052, BMS650032 and VBY376. HCV nonstructural protein 4B (NS4B) inhibitors include, but are not limited to, clemizole and other NS4B-RNA binding inhibitors, including but not limited to benzimidazole RBI (B-RBI) and indazole RBI (I-RBI). HCV nonstructural protein 5A (NS5A) inhibitors include, but are not limited to, BMS-790052, A-689, A-831, EDP239, GS5885, and PP1461. Inhibitors of HCV polymerase (NS5B) include, but are not limited to, nucleoside analogs (e.g., valopicitabine, R1479, R1626, R7128), nucleotide analogs (e.g., IDX184, PSI-7851, PSI-7977, and non-nucleoside analogs (e.g., feribovir, HCV-796, VCH-759, VCH-916, ANA598, VCH-222 (VX-222), BI-207127, MK-3281, ABT-072, ABT-333, GS9190, BMS791325). In addition, ribavirin or ribavirin analogs such as viramidine (ICN 3142), mizoribine, merimepodib (VX-497), mycophenolate mofetil and mycophenolate mofetil may be used.

[0076] In certain embodiments, once a day, once every 1 day, once every 2 days, once every 3 days, once a week, twice a week, three times a week, or once every 2 weeks, the antibody or hybrid protein of the present invention of a dosage is applied to the subject. In other embodiments, once a day, once every 2 days, once every 3 days, once a week, or once every 2 weeks, the compound or composition of 2, 3 or 4 dosages is applied to the subject. In some embodiments, the dosage of the compound or composition is applied for 2 days, 3 days, 5 days, 7 days, 14 days or 21 days. In certain embodiments, the dosage of a compound or composition is applied for 1 month, 1.5 months, 2 months, 2.5 months, 3 months, 4 months, 5 months, 6 months or more months.

[0077] The methods of administration include, but are not limited to, parenteral, intradermal, intravitreal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, oral, sublingual, intranasal, intracerebral, intravaginal, transdermal, transmucosal, rectal, by inhalation, or topical administration (particularly to the ear, nose, eye or skin). The mode of administration is at the discretion of the physician. In most cases, administration will result in the release of the compound into the blood. For the treatment of eye diseases, intravitreal administration of biologics is preferred.

[0078] In specific embodiments, it may be desirable to administer the compound locally. This may be achieved, for example (but not by way of limitation), by local infusion, local administration, by injection, by catheter, or by implant (the implant being a porous, non-porous or gelatinous material, including a membrane, such as a silicone rubber membrane, or a fiber). In this case, administration may selectively target local tissues without releasing the compound in large quantities into the bloodstream.

[0079] Pulmonary administration may also be employed, for example, by use of an inhaler or nebulizer, and formulation with an aerosolizing agent, or by infusion of a fluorocarbon or synthetic lung surfactant. In certain embodiments, the compound is formulated as a suppository with conventional binders and vehicles such as triglycerides.

[0080] In another embodiment, the compound is delivered in a vesicle, particularly a liposome (see Langer, 1990, Science 249: 1527-1533; Treat et al., Liposomes in the Therapy of Infectious Disease and Bacterial infection, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez Berestein, supra, pp. 317-327; see generally above).

[0081] In another embodiment, the compound is delivered in a controlled release system (see, e.g., Goodson, Medical Applications of Controlled Release, supra, Vol. 2, pp. 115-138 (1984)). Examples of controlled release systems are discussed in the review by Langer, Science 249: 1527-1533, which examples of controlled release can be used. In one embodiment, a pump can be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14: 201; Buchwald et al., 1980, Surgery 88: 507; Saudek et al., 1989, N. Engl. J. Med. 321: 574). In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, 1983, J. Macromol. Sci. Rev. Macromol. Chem. 23:61; see also Levy et al., 1985, Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989, J. Neurosurg. 71:105).

[0082] The above administration scheme is provided for illustrative purposes only and should not be considered limiting. It will be readily understood by those of ordinary skill in the art that all dosages are within the scope of the present invention.

[0083] It is understood and anticipated that the principles of the invention disclosed herein may be made by one skilled in the art and that such modifications are intended to be included within the scope of the present invention.

[0084] Throughout this application, various publications are cited. These publications are hereby incorporated into this application in their entirety by reference to more fully describe the state of the prior art in the field to which the present invention belongs. The following examples further illustrate the present invention, but should not be construed as limiting the scope of the present invention in any way. Example

[0085] Mixed lymphocyte reaction:

[0086] CD14-positive monocytes were isolated from whole blood by negative selection using the RosetteSep human monocyte enrichment kit (StemCell technologies). Immature monocyte-derived dendritic cells (mo-DC) were produced by culturing CD14-positive cells in IMDM supplemented with 10% FBS for 6 to 7 days with 150ng / mL GM-CSF and 50ng / mL IL-4. CD4-positive cells were negatively isolated from whole blood using the RosetteSep human CD4 enrichment kit (StemCell technologies). Mo-DC and CD4-positive cells from different donors were then co-cultured at a ratio of 1:10 mo-DC to CD4 cells, respectively. In order to evaluate the blocking function of anti-PDL1 antibodies, increasing amounts of anti-PDL1 antibodies were added at the beginning of co-culture. In some cases, increasing amounts of IL15 were also added at the beginning of co-culture. On the 6th or 7th day, supernatants were collected for measurement of secreted IL-2 and IFNγ by ELISA. The number of CD4 cells and the expression of the proliferation marker Ki67 were evaluated by flow cytometry.

[0087] Activation of PBMCs:

[0088] PBMCs were isolated from whole blood using Histopaque-1077 (Sigma), cultured in IMDM supplemented with 10% FBS, and subsequently activated for 3 to 7 days with SEB (0.1 ug / mL), PHA (1 ug / mL), or anti-CD3 clone HiT3a (1 ug / mL, eBioscience). The binding of anti-PDL1 antibodies or anti-PDL1-SD15 fusion proteins was evaluated in activated PBMCs by flow cytometry after 3 days. Functional evaluation of anti-PDL1 antibodies was performed by adding increasing amounts of anti-PDL1 antibodies during PBMC activation with SEB. On the 2nd or 3rd day, supernatants were collected to measure IL-2 and IFNγ. In the case of anti-PDL1-SD15 fusion proteins, PBMCs were cultured in the presence of anti-PDL1-SD15 or anti-PDL1 antibodies and without other activation. On the 6th day, cells were collected and the number of CD8 and granzyme B, CD8, perforin, and CD4 cells was evaluated by flow cytometry.

[0089] Activation of CD4 and CD8 cells:

[0090] CD4 and CD8 positive cells were negatively separated from whole blood using RosetteSep enrichment kit (StemCell technologies). CD4 cells were activated by anti-CD3 or anti-CD3 and PDL1Fc coated beads in IMDM, 10% FBS in the presence of anti-PDL1 antibodies. On the 5th day, supernatants were collected for IFNγ measurement by ELISA, and the expression of Ki67, a proliferation marker of cells, was evaluated using flow cytometry. CD8 cells were activated by anti-CD3 coated beads and IL15 or anti-PDL1-SD15 fusion proteins. In some cases, anti-CD3 coated beads were replaced with anti-CD3 and PDL1Fc. On the 6th or 7th day, supernatants were collected to measure the secretion of IFNγ and TNFα by ELISA. The cells were collected to measure the activation of CD8 using flow cytometry by granzyme B and perforin markers.

[0091] Nomenclature for Antibody-Fusion Proteins:

[0092] In experiments utilizing anti-PD-L1 IL15 fusion proteins, shorter names of the fusion proteins are identified in the figure legends. Fusion protein tcclD7HC-SD15 is identified as cD7-SD15 in the figure legends. Fusion protein tcclF8HC-SD15 is identified as F8-SD15 in the figure legends.

[0093] Specific high-affinity antibodies against PD-L1 from phage display libraries

[0094] Obtaining anti-PD-L1 antibodies with high affinity using phage display libraries. In one approach, phage Fabs amplified from the Dyax library were panned against recombinant human PDL1-Fc (PDL1 ECD and human Fc fusion protein Q9NZQ7) or mouse PDL1-Fc (Q9EP73) immobilized on immunotubes for 3 rounds. ELISA-positive clones from rounds 2 (R2) and 3 (R3) were sequenced.

[0095] In the second approach, phage Fabs amplified from the Dyax library were subjected to the first round of panning against recombinant human PDL1-Fc (PDL1 ECD and human Fc fusion protein, Q9NZQ7), followed by the second round of panning against activated T cells. For the third round, either the activated T cells or recombinant human PDL1-Fc were used for panning. Clones that could bind to both soluble PDL1-Fc and cell-expressed PDL1-Fc were sequenced. The V H and V L The variable domain sequences are shown in Figure 13 and rows 1-26 of Table 1.

[0096] Unique clones were converted to IgG for further characterization. The variable domains were inserted into the Dyax expression vector pBhl. Wild-type CH1-CH2-CH3 domains and mutant CH1-CH2-CH3 (L234A and L235A, also referred to herein as LALA mutants) were prepared in IgG format.

[0097]

[0098]

[0099] These antibodies were validated for PD-L1 by solid phase ELISA ( Figure 1-4 )) and HEK-293 cells ( Figure 5 ). Blockade of PD-1:PD-L1 interaction in the presence of these antibodies was determined by solid phase ELISA and by 293-HEK cells expressing PD-L1. Biacore was used to calculate the affinity constant for each antibody.

[0100]

[0101]

[0102] Immature monocyte-derived dendritic cells ( Figure 6 A), breast cancer cell line MDA-MB-231 expressing human PD-L1 ( Figure 6 B) Tumor line B16-F10 cells expressing mouse PD-L1 ( Figure 6 C) and binding to activated human CD4 and CD8 T cells, validating the binding of these antibodies on native cells expressing PD-L1.

[0103] Functionally active anti-PD-L1 antibodies block PD-1PD-L1 interaction and increase T cell proliferation and activation

[0104] The function of high affinity binding anti-PD-L1 antibodies to block PD-1PD-L1 interactions and increase T cell proliferation was evaluated. Negatively purified CD4 T cells were activated in vitro with beads coated with αCD3 or αCD3 and PD-L1Fc in the presence of anti-PD-L1 antibodies. CD4 cells stimulated with beads coated with αCD3 and PD-L1Fc showed lower proliferation and IFNγ and IL-2 secretion compared to CD4 cells stimulated with beads coated with αCD3 alone. Adding functionally active anti-PD-L1 antibodies to CD4 cultures stimulated with beads coated with αCD3 and PD-L1Fc increased CD4 proliferation (measured by the percentage of total CD4 number or proliferation marker Ki67) ( Figure 7A). Addition of a functionally active blocking anti-PD-L1 antibody as well as αCD3 and PD-L1 Fc-coated beads to CD4 cultures also increased cytokine secretion by CD4 (measured by ELISA of IFNγ and IL-2 accumulated in the supernatant).

[0105] When PBMCs isolated from whole blood were stimulated with the superantigen Staphylococcus enterotoxin B (SEB) in the presence of anti-PD-L1 blocking antibodies, an increase in cytokine secretion was observed. Supernatants of PBMCs (previously frozen) cultured with SEB for 48 hours were collected, and IFNγ and IL-2 were measured by ELISA. When compared to controls to which no antibody was added, no increase in the number of T cells was observed in cultures with several anti-PD-L1 antibodies, but a significant increase in the levels of IFNγ and IL-2 was observed ( Figure 7 B).

[0106] Additionally, increases in CD4 proliferation and activation were observed in mixed lymphocyte reactions (MLRs) of CD4 T cells and mo-DC cultured in the presence of anti-PD-L1 blocking antibodies. Several anti-PD-L1 antibodies increased CD4 proliferation in MLRs when compared to cultures to which no antibody was added ( Figure 7 C) These antibodies also increased the secretion of IFNγ and IL-2 as assessed by ELISA.

[0107] IL15 enhances the effects of anti-PD-L1 antibodies on T cell proliferation and activation in vitro

[0108] When compared to cultures of CD4 and mo-DC with anti-PD-L1 antibody alone, MLR of CD4 T cells and mo-DC in the presence of anti-PD-L1 blocking antibody and cytokine IL15 resulted in CD4 cell proliferation ( Figure 8 A), IFNγ and IL-2 secretion. IL15 was added in these assays at equimolar concentrations to anti-PD-L1 antibody. At lower anti-PD-L1 antibody and IL15 concentrations (0.5 nM, Figure 8 A), a certain synergistic effect on CD4 proliferation was observed.

[0109] Negatively purified CD8 from whole blood stimulated with aCD3 and PD-L1Fc coated beads also responded to IL15 in a dose-dependent manner. Adding IL15 to the culture of CD8 with aCD3 and PD-L1Fc coated beads and anti-PD-L1 antibody resulted in a great increase in CD8 proliferation ( Figure 8 B).

[0110] Anti-PD-L1-IL15 fusion protein targets IL15 to antigen-presenting cells expressing PD-L1 and increases proliferation and activation of responding CD8 cells

[0111] Anti-PD-L1 antibodies and IL15 fusion proteins were constructed by connecting the Fc domain of the antibody to the sushi domain of IL15R and the IL15 molecule itself. A fusion of IL15Rαsushi domain, IRD-11exone3, linker and IL15 (designated "SD15") is provided as SEQ ID NO: 261. SD15 was attached to the c-terminus of the heavy chain of conventional IgG. A fusion protein with IL15Rαsushi domain, IRD-11exone3, linker and IL15 was attached to the tccλD7 variable domain and IgG1C H 1-C H 2-C H 3 heavy chain c-terminus of the variable domain (SEQ ID NO: 262). The construct also contained a K to S substitution at the end of the IgG1 heavy chain (1) to eliminate the possibility of "GK" cleavage; (2) to add a cloning site (BamHI) to the vector.

[0112] The light chain is that of a conventional antibody. The light chain and the fused heavy chain with or without the LALA mutant were inserted into the Dyax pBh1 vector for expression.

[0113] This fusion molecule was named anti-PD-L1-sushi domain-IL15 or anti-PD-L1-SD15. A different form of the fusion protein in which IL15 was linked to Fc instead of the sushi domain was also constructed, and since this fusion protein did not have IL15 functional activity, we used this protein as a negative control in some assays (called anti-PD-L1-SD15neg).

[0114] When the binding of anti-PD-L1-SD15 fusion protein was compared with anti-PD-L1 antibody in a solid phase PD-L1 Fc binding ELISA assay, no significant changes were observed ( Fig. 9 A). When the binding of anti-PD-L1-SD15 proteins was compared to their respective original anti-PD-L1 antibodies, some changes in binding affinity to activated CD4 cells expressing PD-L1 were observed ( Fig. 9 B) Anti-PD-L1-SD15 proteins have lower affinity for cells expressing PD-L1 when compared to their respective anti-PD-L1 antibodies; although there may be differences in the binding of the secondary antibody to bound anti-PD-L1-SD15 versus bound anti-PD-L1 on the cell surface.

[0115] To evaluate the function of IL15 of anti-PD-L1-SD15 fusion protein, PBMCs isolated from whole blood were cultured in the presence of anti-PD-L1-SD15 fusion protein or IL15. No other stimulation was added to the culture. Similar to IL15, anti-PD-L1-SD15 fusion protein increased the number of NK cells ( Fig.10 A) Increase CD8 proliferation ( Fig.10 B) and activation (measured by the percentage of granzyme B-positive CD8, Fig.10 C). No significant increase in CD4 counts was observed for all cultures ( Fig.10 D).

[0116] To evaluate the activity of anti-PD-L1-SD15 on CD8, these fusion proteins were added to CD8 cultures in the presence of αCD3 or αCD3 and PDL1Fc-coated beads. When PDL1Fc was present on antigen presenting cells (in this case αCD3 and PDL1Fc-coated beads), anti-PD-L1-SD15 significantly increased the proliferation of CD8 ( Fig.11 A, no PD-L1Fc relative Fig.11 B, with PD-L1 Fc on beads). In addition, a significant increase in CD8 activation was observed. cD7-SD15 reduced the effective dose required to activate CD8, as evidenced by an increase in the percentage of granzyme B-positive CD8 cells ( Fig.12 A) and IFNγ secretion increased by about 10-fold ( Fig.12 cD7-SD15 also increased the maximum level of CD8 activation when compared to IL15 ( Fig.12 A and B). When compared to the addition of anti-PD-L1 antibody + free IL15, anti-PD-L1-SD15 fusion protein increased CD8 proliferation to a higher level than the combination added alone ( Fig.12 C). These properties of anti-PD-L1-SD15 fusion proteins will be beneficial in the context of immunotherapy, as lower doses of anti-PD-L1-SD15 fusion proteins can be used to achieve higher levels of CD8 activation and proliferation. The highly amplified response of CD8 to anti-PD-L1-SD15 fusion proteins in situations where antigen presenting cells express PD-L1 will be advantageous in achieving selective CD8 activation.

[0117] Anti-PD-L1-IL15 fusion protein cytotoxicity

[0118] To determine whether anti-PD-L1-SD15 fusion protein would enhance IL15-induced CD8 T cell cytotoxicity against PD-L1-expressing tumor cells, CD8 T cells were co-cultured with MAD-MB-231 tumor cells expressing human PD-L1 for 7 days in the presence of anti-PD-L1-SD15 fusion protein or anti-KLH-SD15, which do not have binding activity to tumor cells expressing PD-L1, before measuring tumor cell death. Human CD8 T cells and tumor cells were co-cultured in IMDM supplemented with 10% FBS for 7 days. Tumor cell killing activity was assessed by measuring the number of dead tumor cells stained with ViabilityDye eFluor780 in FACS. Anti-PD-L1-SD15 fusion protein significantly enhanced CD8 T cell-mediated cytotoxicity of MDA-MB-231 compared to treatment with anti-KLH-SD15 in co-culture ( Fig.15 In addition, the PD-L1-SD15 fusion protein cD7-SD15 significantly increased the survival rate of mice with tumor cells expressing PD-L1 in a tumor model of mice injected intravenously with mouse CT26 colon tumor cells, compared with mice treated with vehicle or PD-L1-SD15 fusion protein sD7-SD15 (which has no binding activity with mouse PD-L1) ( Fig.16 ). These results suggest that targeting IL15-stimulated immune effector cells to PD-L1-overexpressing tumor sites via a bifunctional anti-PD-L1-SD15 fusion protein has the advantage of enhancing anti-tumor immunity while minimizing side effects. This type of bifunctional antibody-cytokine fusion protein has the potential to serve as a novel immunomodulatory therapeutic agent to achieve greater anti-tumor efficacy in the control of tumor progression.

[0119] Affinity Maturation

[0120] Variants of the tccλD7 heavy chain were generated by introducing amino acid substitutions at the three methionine positions of CDR-1H and screening for enhanced affinity. More specifically, variants containing approximately 1 x 10 8A library of variants of CDRH1 of tccλD7. The library was panned against recombinant human PDL1-Fc (PDL1ECD and human Fc fusion protein, Q9NZQ7) or mouse PDL1-Fc (Q9EP73) immobilized on immunotubes for 4 rounds. ELISA positive clones from rounds 3 and 4 were sequenced. Unique clones were compared by competitive ELISA. Table 4 shows the amino acid substitutions observed in the 25 variants obtained from the screening, with SEQ ID NOs representing affinity matured CDR-1H sequences and heavy chain variable regions containing the CDRs. The amino acid sequences of these variants are also listed in the sequence table indicated in Table 1.

[0121]

[0122]

[0123] As described elsewhere herein, two variants, tccD7_#114 and tccD7_#102 (also referred to herein as tccD7_#1 and tccD7_#2, respectively) were converted to IgG and also to an IgG format containing two Leu-Ala substitutions in the hinge region to reduce ADCC. The antibodies were expressed and purified for further characterization. For both affinity matured variants, enhanced binding to soluble PDL1 is shown in Fig.17 middle. Fig.18 The two variants block the binding of human PD1 to human PDL1 (upper figure) and block the binding of mouse PD1 to mouse PDL1 (lower figure). The variants also show higher binding activity to MDA-MB-231 cells than the parent ( Fig.19 ).

[0124] The affinity-matured variants were tested for their ability to promote the production of the Th1 cytokines IL2 and IFNγ. PBMCs isolated from whole blood were stimulated with superantigen Staphylococcus enterotoxin B (SEB, 0.1 ug / mL) in the presence of anti-PD-L1 antibodies. Supernatants of PBMCs cultured with SEB for 7 days were collected, and IFNγ and IL-2 were measured by ELISA. When compared to cD7, a significant increase in the levels of IFNγ and IL-2 was observed in cultures of variants with anti-PD-L1 antibodies cD7#1 and #2 ( Fig. 20 ).

[0125] Several fusion protein variants comprising a PD-L1 binding domain, an IL15Rαsushi domain, and IL15 were constructed. Certain constructs included a linker between the IL15Rαsushi domain and the IL15 portion. In one construct, 11 amino acids of exon 3 present in the c-terminus of the IL15 receptor αsushi domain were replaced with a "GS" linker of varying lengths. The GS linker contains SGGSGGGGSGGGSGGGGS (SEQ ID NO: 324; 18 amino acids), SGGSGGGGSGGGSGGGGSLQ (SEQ ID NO: 314; 20 amino acids), SGGGGSGGGGSGGGGSGGGGSGGGG (SEQ ID NO: 316; 25 amino acids), SGGGGSGGGGSGGGGSGGGGSGGGGSGGGG (SEQ ID NO: 318; 30 amino acids), SGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGG (SEQ ID NO: 320; 40 amino acids), and SGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS in constructs having SEQ ID NOs: 325, 315, 317, 319, 321, and 323, respectively. GGGGSGGGG (SEQ ID NO: 322; 50 amino acids).

[0126] The fusion proteins were transiently or stably expressed in HEK293 cells and purified by protein A column chromatography following the manufacturer's instructions. In certain embodiments, to stabilize the association between the Ig heavy chain and light chain constant domains of the anti-PD-L1 portion of the molecule, the C-terminal serine of the lambda light chain is deleted, referred to herein by the name "ds".

[0127] Fusion proteins containing the affinity-matured variant #102 of tccλD7 with a sushi domain and IL15 (SEQ ID NO: 325) were tested for binding to MDA-MB-231 by flow cytometry. All fusion proteins showed enhanced binding compared to the fusion proteins containing tccλD7 ( Fig.21 ). The fusion protein containing the affinity-matured variant #102 of tccλD7 was also confirmed to have stimulatory activity against IL15-responsive human megakaryoblastic leukemia cells. The cells were cultured with the anti-PD-L1-SD15 fusion protein in RPMI 1640 supplemented with 10% FBS and 20% conditioned medium from human bladder cancer 5637 cells for 48 hours. Luminescent cell viability assay measures cell proliferation as relative luminescence units (RLU) ( Fig. 22 ).

[0128] Analysis using size exclusion chromatography showed less than 5% aggregation of the expressed fusion protein ( Fig.24 ) and improved serum stability ( Fig.25 ).

Claims

1. A fusion protein comprising (i) comprising a first domain of a PD-L1 antibody, wherein the antibody comprises (1) an Fc domain and (2) a PD-L1 binding domain, wherein the PD-L1 binding domain comprises: (a) the heavy chain CDR-1H shown in SEQ ID NO:241, the heavy chain CDR-2H shown in SEQ ID NO:243, and the heavy chain CDR-3H shown in SEQ ID NO:245, and the light chain CDR-1L shown in SEQ ID NO:247, the light chain CDR-2L shown in SEQ ID NO:248, and the light chain CDR-3L shown in SEQ ID NO:249; or (b) the heavy chain CDR-1H set forth in SEQ ID NO:266, the heavy chain CDR-2H set forth in SEQ ID NO:243, and the heavy chain CDR-3H set forth in SEQ ID NO:245, and the light chain CDR-1L set forth in SEQ ID NO:247, the light chain CDR-2L set forth in SEQ ID NO:248, and the light chain CDR-3L set forth in SEQ ID NO:249; wherein the antibody or antigen-binding fragment of (b) is a variant of (a) with improved affinity for PD-L1, which is obtained by substituting three methionine residues in SEQ ID NO:241 to produce SEQ ID NO:266; (ii) a second domain that binds to an IL-15 receptor ("IL-15R"), wherein the second domain comprises IL-15, or an amino acid sequence that is at least 95% identical to IL-15, or an IL-15R binding fragment thereof, and (iii) IL15 receptor αsushi domain; wherein ((iii) is linked to the C-terminus of the Fc domain, and wherein (ii) is connected to (iii) via a flexible linker.

2. The fusion protein according to claim 1, comprising SEQ ID NO: 315, SEQ ID NO: 317, SEQ ID NO: 319, SEQ ID NO: 321, SEQ ID NO: 323 or SEQ ID NO:

261. The fusion protein according to claim 1 , wherein the flexible linker comprises 15-20 amino acids, which are mainly serine and glycine. The fusion protein according to claim 1 , comprising SEQ ID NO:

261.

5. The fusion protein according to any one of claims 1 to 4, wherein the PD-L1 antibody or antigen-binding fragment thereof comprises the heavy chain variable domain shown in SEQ ID NO: 267 and / or the light chain variable domain shown in SEQ ID NO:

250.

6. The fusion protein according to any one of claims 1 to 4, wherein the PD-L1 antibody or antigen-binding fragment thereof comprises a heavy chain variable domain shown in SEQ ID NO: 246 and / or a light chain variable domain shown in SEQ ID NO:

250.

7. A conjugate of the fusion protein according to any one of claims 1 to 6, further comprising an imaging agent, a therapeutic agent or a cytotoxic agent.

8. An isolated nucleic acid encoding the fusion protein according to any one of claims 1 to 6.

9. A nucleic acid vector comprising the nucleic acid according to claim 8.

10. A prokaryotic or eukaryotic host cell comprising the nucleic acid according to claim 8.

11. A composition comprising the fusion protein according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

12. Use of the fusion protein according to any one of claims 1 to 6 in the preparation of a medicament for treating solid tumors.

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