TW201247704A - Half immunoglobulin binding proteins and uses thereof - Google Patents

Abstract

The invention provides compositions, methods, and kits related to half-Ig binding proteins that include a functional antibody binding site and a CH3 domain wherein the CH3 domain includes at least one mutation to inhibit CH3-CH3 dimerization.

Description

201247704 VI. INSTRUCTIONS: This application claims US Provisional Patent Applications Nos. 61/426, 2-7 and 61/539 13 of December 22, 2011 and September 26, 2011, respectively. The priority of the nickname. The entire contents of each of the aforementioned applications are hereby incorporated by reference. SEQUENCE LISTING This application contains a Sequence Listing filed in ASCII format via EFS-Web and incorporated herein by reference in its entirety. The Ascn replica was created in

On December 12, 2011, it was named 1 17813553 txt and the size was (10) 宄 (7). [Prior Art] A large amount of effort to study and modify the immunoglobulin amino acid sequence (eg, antibody amino acid sequence) and structure found in natural immunoglobulins to achieve desired characteristics reflects binding proteins (such as immunity). The importance of globulin as a therapeutic, diagnostic and research tool. The main point of view in the industry is that ideal therapeutic binding proteins (such as immunoglobulins) will have certain threshold characteristics including target specificity, biostability and bioavailability after administration to an individual, and sufficient target affinity for binding. Maximize the therapeutic effect. Unfortunately, the success of producing a binding protein therapeutic with all or even these minimal characteristics is limited. For example, a long antibody (such as IgG) exhibits the desired pharmacokinetics (e.g., a relatively long in vivo half-life) and good target binding affinity due to the presence of a 2-potential effect by the presence of two antigen-binding arms in a single antibody molecule. In no case, these full-length antibodies encounter problems due to their large molecular size (IV) 160877.doc 201247704. Furthermore, in some cases, full length antibodies may exhibit inappropriate agonism upon binding to a stimuli antigen, even though their corresponding (four) fragments function as: a resistance binding protein. See, for example, U.S. Patent No. M68,529, which is incorporated herein by reference. In some cases, this phenomenon may be attributed to the "cross-linking" effect of bivalent anti-It 1 "cross-linking" effect when the bivalent antibody binds to a cell surface receptor to promote the receptor dimerization leading to receptor activation. While monovalent binding proteins (such as immunoglobulins) are not expected to exhibit "crosslinking" effects, monovalent antibodies have hitherto been unsuitable as therapeutic agents due to certain limitations inherent in their structure/architecture. For example, the pharmacodynamic properties of a monovalent antibody in the form of a sputum are, for example, unstable in vivo and rapidly cleared upon administration. Furthermore, monovalent immunoglobulins generally have lower apparent binding affinities due to the absence of affinity binding compared to their multivalent counterparts. In general, the choice of a binding protein form for a therapeutic agent is subject to the undue limitation of acceptance of each alternative form. However, the full-length binding protein form = selected form in recent years may be due, at least in part, to its in vivo biostable nature. Monovalent binding proteins (such as immunoglobulins) are traded off, and biostability as a therapeutic factor may not be as important as bioavailability. For example, due in part to better tissue penetration than full length antibodies, a monovalent Fab antibody can be a preferred vehicle for delivery of a heterologous molecule, such as a toxin, to a target cell or tissue, a target cell. Or heterologous molecules in the tissue play a therapeutic role. See, for example, U.S. Patent No. 5,169,939, incorporated herein by reference. Attempts to develop monovalent antibodies 160877.doc 201247704 Other examples of use as therapeutic agents include monovalent environments that are critical to achieving therapeutic effects', such as where bivalent antibodies may induce target cells to undergo antigenic regulation as a problem, where this antigenicity Modulation may then provide a means to avoid target cells from cytotoxic agents, effector cells, and complement. Examples of such antibodies are described in Cobbold and Waldmann (1984) Nature 308.460-462, European Patent No. 〇i3 1424; Glennie and Stevenson (1982) Nature 295:712-714; Nielsen and Routledge (2002) Blood 100:4067- 4073; Stevenson et al., (1989) Anticancer Drug Des. 3(4): 219-230; Routledge et al., (1995) Transplant. 605347-853; Clark et al., (1989) Eur J. Immunol. 19: 381-388 'Bolt et al. (1993) Eur. J. Immunol. 23:403-411; Routledge et al., (1991) Eur. J. Immunol. 21:2717-2725; Staerz et al., (1985) Nature 314: 628-631; and U.S. Patent No. 5,968,509, the disclosures of each of each of It is worthwhile/intentional that this 荨 monovalent antibody fragment contains a functional dimeric F c sequence that is included for its therapeutic function (eg, complement-mediated T cell lysis) as needed for therapeutic function. This technique does not recognize the need or utility of incorporating an Fc region into a monovalent antibody for use as and/or development as a therapeutic agent. Difficulties in obtaining such antibodies emphasize the reluctance to include an Fc region in a monovalent antibody where the Fc region is not necessary for therapeutic function. Existing antibody manufacturing techniques do not provide an efficient means of obtaining a large number of fully purified heterodimers comprising a single antigen binding component (i.e., monovalent) and an Fc region. Efforts have been made to increase the in vivo stability of binding protein fragments with varying degrees of success. For example, a Fab fragment can be ligated to a stability moiety (160877.doc 201247704 such as polyethylene glycol) or other stabilizing molecule (such as a heterologous peptide). See for example

Dennis et al., (2002) J. Biol. Chem. 277:35035-35043; PCT Publication No. WO 01145746, each of which is incorporated herein by reference. The anti-c_Met monovalent molecule MetMAb with Fab-Fc/Fc structure is in Phase II clinical trials for non-small cell lung cancer. See PCT Publication No. WO2005063816, which is incorporated herein by reference. The Fc fragment can be ligated to the C-terminus of the light chain and then coupled to a fully heavy bond to achieve a monovalent binding to the antigen. See PCT Publication No. W020070105199, which is incorporated herein by reference. Can also be replaced by IgG4 backbone "(7)

The main chain achieves a unit price combination. See PcT Publication No. WO 2007059782, which is incorporated herein by reference. The CH3 mediated dimerization of the IgG4 primary bond is extremely weak. Thus, there remains a significant need for improved binding protein forms, for example, as therapeutic or prophylactic agents, and methods of making and using such binding proteins. SUMMARY OF THE INVENTION The present invention provides a monovalent (as appropriate) binding protein comprising a non-dimeric immunoglobulin CH3 domain, which is referred to herein as a semi-immunoglobulin or a half. A binding protein of the invention binds to one or more specific antigens of interest and includes an Fc region for binding to an effector molecule. The binding proteins of the invention retain many of the antibodies, but are small in size, thereby providing altered pharmacokinetic and pharmacodynamic properties, including improved bioavailability due to smaller size and in antibody fragments (such as Fab fragments). The effect function is not lost. Furthermore, the binding proteins of the invention preferably do not promote the cross-linking observed in naturally occurring antibodies which can result in antigen clustering and inappropriate activity. 160877.doc 201247704 The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-X2, wherein: VD1 comprises a heavy chain antigen binding domain; XI comprises a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain, and a linker; N is 0 or 1; and X2 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit CH3 -CH3 dimerization, wherein the binding protein forms a functional antigen binding site. In certain embodiments, the VD1 is selected from the group consisting of: a heavy chain variable domain, a double heavy chain variable domain, a triple heavy chain variable domain, a light chain variable domain, a dual light chain variable domain, triple a light chain variable domain, a heavy chain variable domain and a light chain variable domain, a combination of two heavy chain variable domains and a light chain variable domain, a heavy chain variable domain and two light chain variable domains Combination, domain antibody, camelid antibody, scFv, receptor and backbone antigen binding protein. In certain embodiments, the binding protein further comprises a hinge (H) region between VD1 and X2. In certain embodiments, at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. In certain embodiments, at least one mutation is at a residue selected from the group consisting of C220, C226, C229, T366, L368, P395, F405, Y407, and K409 according to the Kabat nomenclature. In certain embodiments, the binding protein comprises a second polypeptide chain, wherein the second polypeptide chain comprises VD1-(X1)N, wherein 160877.doc 201247704 VD1 comprises a light chain antigen binding domain; X1 comprises a composition selected from the group consisting of The domains of the population: polypeptide, CL domain, CL-CH2 domain, (v) domain, CH2 domain, cm domain and (10) domain and linker; and N is 〇 or 1. In certain embodiments, the VD1 is selected from the group consisting of: a light chain variable domain, a dual light chain variable domain, a triple light chain variable domain, a heavy chain variable domain, a double heavy chain variable domain, a heavy bond variable domain, a combination of a heavy bond variable domain and a light bond variable domain, a combination of two heavy chain variable domains and a light chain variable domain, a heavy chain variable domain and two light bond variable domains Combination, camelid antibody, domain antibody 'scFv, receptor and backbone antigen binding protein. The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-X1-X2, wherein: VD1 comprises a first heavy chain variable domain; X1 comprises a domain polypeptide selected from the group consisting of: CH1 domain, CH2 a domain, a CH1 domain, and a CH2 domain, and a linker; and X2 comprises at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit CH3-CH3 dimerization, wherein the binding protein forms a functional antigenic binding Site. In certain embodiments, the binding protein further comprises a hinge region between VD1 and X2. In certain embodiments, at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. In certain embodiments 'at least one mutation that inhibits CH3-CH3 dimerization 160877.doc 201247704 at a residue selected from the group consisting of C220, C226, C229, T366, L368, P395 according to Kabat nomenclature F405, Y407 and K409 ° The present invention provides a binding protein comprising a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises VD1-X1-X2, wherein: VD1 comprises a first heavy bond variable Domain; XI comprises a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain, and a linker; and φ X2 comprises at least a portion of a CH3 domain; and wherein the second polypeptide chain comprises VD1-XI, wherein VD1 comprises a light chain variable domain; and XI comprises a light chain constant domain, a CH1 domain, a CH2 domain, a CH1 domain and a CH2 domain; wherein the binding protein comprises at least one mutation at the residue to inhibit CH3-CH3 Dimerization, and the binding protein forms a functional antigen binding site. In certain embodiments, the binding protein further comprises a φ hinge region between VD1 and X2. In certain embodiments, at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. And X. And K409 ° The present invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises 160877.doc 201247704 VD1-X1-X2, wherein: VD1 comprises a heavy chain variable domain; XI comprises a CH1 domain and a hinge region, wherein the hinge region Located at the C-terminus of the CH1 domain; and X2 comprises at least a portion of the CH3 domain; wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, C229, T366, L368, P395, F405, Y407 and K409, thereby inhibiting CH3-CH3 dimerization; and the binding protein forms a functional antigen binding site. The invention provides a binding protein comprising a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises VD1-X1-X2, wherein: VD1 comprises a heavy chain variable domain; XI comprises a CH1 domain and a hinge region Wherein the hinge region is at the C-terminus of the CH1 domain; and X2 comprises at least a portion of the CH3 domain; and wherein the second polypeptide chain comprises VD1-XI, wherein VD1 comprises a light chain variable domain; and XI comprises a light chain constant domain Wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, C229, T366, L368, P395, F405, Y407 and K409, thereby inhibiting CH3-CH3 dimerization; and the binding protein forms a functional antigen binding site 160877.doc -10- 201247704 The present invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2 ^-X3, wherein: VD1 comprises a first heavy chain antigen binding domain; XI is a linker; VD2 comprises a second heavy chain antigen binding domain; X2 comprises a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 Domain, CH1 domain and CH2 domain, light chain constant region, and linker; Sites are selected from 0 and 1; and X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit CH3-CH3 dimerization, wherein the binding protein forms a functional antigen binding site . In certain embodiments, each of VD1 and VD2 is selected from the group consisting of a heavy chain variable domain, a light chain variable domain, a domain antibody, a scFv, a receptor, and a framework antigen binding protein. In certain embodiments, the binding protein further comprises a hinge region between VD2 and X3. In certain embodiments, at least one mutation that inhibits CH3-CH3 dimerization is in the CH3/CH3 dimerization contact zone or in the hinge region. And X. And K409 ° In certain embodiments, the binding protein comprises a second polypeptide chain, wherein the second polypeptide chain comprises VD1-(X1)N-VD2-(X2)N, wherein 160877.doc 201247704 VD1 comprises the first a light chain antigen binding domain; XI is a linker; VD2 comprises a second light chain antigen binding domain; X2 comprises a domain selected from the group consisting of: a polypeptide, a light chain constant domain, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 Domain; and each N is independently selected from 0 and 1. In certain embodiments, VD1 and VD2 are each independently selected from the group consisting of a light chain variable domain, a heavy chain variable domain, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-X3, wherein: VD1 comprises a first heavy chain variable domain; XI is a linker; Independently selected from 0 and 1; VD2 comprises a second heavy chain variable domain; .X2 comprises a heavy chain constant 1 (CH1) domain, and X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein is selected from the group consisting of The residues of the population comprise at least one mutation to inhibit CH3-CH3 dimerization: T366, L368, P395, F405, Y407 and K409; wherein the binding protein forms a functional antigen binding site. In certain embodiments, the binding protein further comprises a more keypad between VD2 and X3. In certain embodiments, the binding protein comprises a mutation at a residue selected from the group consisting of: 157877.doc 12 201247704 to inhibit CH3-CH3 dimerization: C220, C226, and C229. The present invention provides a binding protein comprising a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises 乂01-(also 1; ^-乂02-(also 2)1^-乂3, wherein VD1 comprises a first heavy chain variable domain; XI is a linker; each N is independently selected from 〇 and 1; VD2 comprises a second heavy chain variable domain; X2 comprises a heavy chain constant l (CHl) domain; and X3 comprises a polypeptide having at least a portion of a CH3 domain; and wherein the second polypeptide chain comprises VD1-(X1)N-VD2-(X2)N, wherein VD1 comprises a first light chain variable domain; X1 is a linker; VD2 comprises a two light chain variable domain; X2 comprises a light chain constant domain; and each N is independently selected from the group consisting of 〇 and 1, wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 Polymerization: T366, L368, P395, F405, Y407, and K409; and wherein the binding protein forms a functional antigen binding site. In certain embodiments, the binding protein further comprises a 狡 chain region between VD2 and X3. In certain embodiments, the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, and 160877 .doc 13 201247704 C229. The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-VD3-(X3)N-X4, wherein: VD1 comprises first a heavy chain antigen binding domain; XI is a first linker; VD2 comprises a second heavy chain antigen binding domain; X2 is a second linker; VD3 comprises a third heavy chain antigen binding domain; X3 comprises a structure selected from the group consisting of Domain: polypeptide, CH1 domain, CH2 domain, CH1 domain and CH2 domain, light chain constant domain, and linker; each N is independently selected from 0 and 1; and X4 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein At least one mutation is included at the residue to inhibit CH3-CH3 dimerization, and wherein the binding protein forms a functional antigen binding site. In certain embodiments, the binding protein further comprises a hinge region between VD3 and X4. And R. In the embodiment, at least one mutation that inhibits CH3-CH3 dimerization is in CH In the 3/CH3 dimerization contact zone or in the hinge region. In certain embodiments, at least one mutation is at a residue selected from the group consisting of C220, C226, C229, 160877 according to the Kabat nomenclature. Doc 201247704 T366, L368, P395, F405, Y407 and K409. In certain embodiments, the binding protein further comprises a second polypeptide chain, wherein the second polypeptide chain comprises VD1-(X1)n-VD2-(X2 n-VD3-(X3)n, wherein VD1 comprises a first light chain antigen binding domain; XI is a first linker; VD2 comprises a second light chain antigen binding domain; X2 is a second linker; VD3 comprises a third lighter a strand antigen binding domain; X3 comprises a domain selected from the group consisting of a polypeptide, a light chain constant domain, a CH1 domain, a CH2 domain, and a CH1 domain and a CH2 domain; and each N is independently selected from 0 and 1. And R. The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-VD3-(X3)N-X4, wherein: VD1 comprises a first heavy chain variable domain; XI is the first linker; VD2 comprises the second heavy chain variable domain; X2 is the second linker; VD3 comprises the third heavy chain variable domain; each N is independently selected from 0 and 1; X3 comprises a heavy chain constant 1 (CH1) domain; and 160877.doc •15·201247704 X4 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization : T366, L368, P395, F405, Y407 and K409; and wherein the binding protein forms a functional antigen binding site. The invention provides a binding protein comprising a first and a second polypeptide chain, wherein the first polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-VD3-(X3)N-X4, wherein; VD1 Including a first heavy chain variable domain; XI is a first linker; VD2 comprises a second heavy chain variable domain; X2 is a second linker; VD3 comprises a third heavy chain variable domain; each N is independently selected from 0 And X; X3 comprises a heavy chain constant 1 (CH1) domain; and X4 comprises a polypeptide having at least a portion of the CH3 domain; and wherein the second polypeptide chain comprises VD1-(X1)n-VD2-(X2)n-VD3- (X3)n, wherein VD1 comprises a first light chain variable domain; XI is a first linker; VD2 comprises a second light chain variable domain; X2 is a second linker; VD3 comprises a third light chain variable domain; X 3 comprises a light chain 丨亘 localization; and 160877.doc -16 - 201247704 each N is independently selected from 0 and 1; wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3 -CH3 dimerization: Τ366, L368, Ρ395, F405, Υ407 and Κ409, and wherein the binding protein forms a functional antigen binding site. In certain embodiments, the binding protein further comprises a hinge region between VD3 and Χ4. In certain embodiments, the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, and C229. The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises a format selected from the group consisting of R-(X1)N-(VD1)N-(X2)N-X3, or (VD1)n-( X1)nR-(X2)n-X3, or (VD1)n-(X2)n-X3-(X1)nR, wherein: R comprises a receptor; XI is a linker; VD1 comprises a heavy chain antigen binding domain; X2 a domain comprising one or more selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain and a CH2 domain, a hinge region, and a linker; each N is independently selected from 0 and 1; and X3 comprises a polypeptide of at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit CH3-CH3 dimerization, and wherein the binding protein forms a functional antigen binding site 160877.doc -17-201247704 in some implementations In one embodiment, the VD2 is selected from the group consisting of a heavy chain variable domain, a light chain variable domain, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. In certain embodiments, at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. In certain embodiments, at least one mutation is at a residue selected from the group consisting of C220, C226, C229, T366, L368, P395, F405, Y407, and K409 according to the Kabat nomenclature. In certain embodiments, the binding protein further comprises a second polypeptide chain, wherein the second polypeptide chain comprises a format selected from the group consisting of: R-(X1)n-VD1-(X2)n, or VD1 -(X1)nR-(X2)n, or VD1-(X2)n-(X1)nR, wherein R comprises a receptor; XI is a linker; VD1 comprises a light chain antigen binding domain; X2 comprises a composition selected from the group consisting of The domains of the population: polypeptide, light chain constant domain, CH1 domain, CH2 domain, CH1 domain, and CH2 domain; and each N is independently selected from 0 and 1. In certain embodiments, the VD2 is selected from the group consisting of a light chain variable domain, a heavy chain variable domain, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. The invention provides a binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises R-(X1)N-VD1-(X2)N-X3, wherein; R comprises a receptor; 160877.doc • 18· 201247704 xi is a linker; Each N is independently selected from 〇 and 1; VD1 comprises a heavy bond variable domain; X2 comprises a heavy chain constant l (CH1) domain; and X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein X3 is selected from the group consisting of The residue contains at least one mutation to inhibit CH3-CH3 dimerization: T366, L368, P395, F405, Y407, and K409; φ where the binding protein forms a functional antigen binding site. The invention provides a binding protein comprising a first and a second polypeptide chain, wherein the first polypeptide chain comprises R-(X1)N-VD2-(X2)N-X3, wherein; R comprises a receptor; XI is a linkage Each N is independently selected from 〇 and 1; VD2 comprises a heavy chain variable domain; X2 comprises a heavy chain constant 1 (CH1) domain; and φ X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein X3 is selected from the group consisting of The residues of the population comprise at least one mutation to inhibit CH3-CH3 dimerization: T366, L368, P395, F405, Y407 and K409; and wherein the second polypeptide chain comprises R-(X1)N-VD1-( X2)N, wherein R is a receptor; XI is a linker; VD1 is a light bond variable domain; 160877.doc -19- 201247704 X2 is a light chain constant domain; and each N is independently selected from 0 and 1. Wherein the binding protein forms a functional antigen binding site. In certain embodiments, the binding protein further comprises a Qinling region between VD3 and X4. In certain embodiments, the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, and C229. In certain embodiments, at least one mutation that inhibits CH3-CH3 dimerization is at a residue selected from the group consisting of C220, C226, and C229. In certain embodiments, at least one mutation that inhibits CH3-CH3 dimerization is two mutations selected from the group consisting of C220S, C226S, and C229S. In certain embodiments, at least one mutation that inhibits CH3-CH3 dimerization is all three mutations C220S, C226S, and C229S. In certain embodiments, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85% of the binding protein population At least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% are not dimerized via the CH3 domain. In certain embodiments, a binding protein comprising at least one mutation that inhibits CH3-CH3 dimerization has altered biological activity compared to a corresponding binding protein that does not comprise at least one mutation that inhibits CH3-CH3 dimerization. 160877.doc -20- 201247704 In certain embodiments, the binding protein is an antagonist. In certain embodiments, the binding protein is an agonist. In certain embodiments, at least one Fc function of a binding protein that is not dimerized via the CH3 domain is altered compared to its corresponding binding protein that will dimerize. In certain embodiments, the FcRn binding efficiency in a binding protein that is not dimerized via the CH3 domain is altered compared to the corresponding binding protein that it will dimerize. In certain embodiments, the FcRn binding efficiency in a binding protein that is not dimerized via the CH3 domain is increased compared to the corresponding binding protein that it will dimerize. In certain embodiments, the FcRn binding potency in the binding protein that is not dimerized via the CH3 domain is reduced by the corresponding binding protein that will dimerize. In certain embodiments, the FcyR binding potency in a binding protein that is not dimerized via the CH3 domain is altered compared to the corresponding binding protein that it will dimerize. In certain embodiments, the FcyR binding potency in a binding protein that is not dimerized via the CH3 domain is increased compared to the corresponding binding protein that it will dimerize. In certain embodiments, the FcyR binding potency in a binding protein that is not dimerized via the CH3 domain is reduced compared to its corresponding binding protein that would dimerize. In certain embodiments, the ClqR binding potency in a binding protein that is not dimerized via the CH3 domain is altered compared to the corresponding binding protein that it will dimerize. In certain embodiments, the ClqR binding potency in a binding protein that is not dimerized via the CH3 domain is increased compared to its corresponding binding protein that will dimerize. 160877.doc -21 - 201247704 In certain embodiments, the ClqR binding potency in a binding protein that is not dimerized via the CH3 domain is reduced compared to its corresponding binding protein that would dimerize. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, T366F, T368F, P395A, F405R, Y407R, and K409D. In certain embodiments, the binding protein has a mutation at the following residues: C220S, C226S, C229S, T366F, T368F, P395A, F405A, Y407R, and K409D ° In certain embodiments, the binding protein has Mutations: C220S, C226S, C229S, P395A, F405R, Y407R and K409D « In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, P395A, F405A, Y407A and K409D. In certain embodiments, the binding protein has a mutation at the following residues: C220S, C226S, C229S, P395A, F405R, and Y407A. In certain embodiments, the binding protein has a mutation at the following residues: C220S, C226S, C229S, F405R, Y407A and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, P395A, Y407A, and K409D. In certain embodiments, the binding protein has at the following residues: C220S, C226S, C229S, P395A, F405R, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, P395A, and F405R. • 22·160877.doc 201247704 In certain embodiments, the binding protein has at the following residues: C220S, C226S, C229S, P395A, and Y407R. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, P395A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, F405R, and F407R. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, F405R, and K409D. φ In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, F407R, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, C229S, and P395A. In certain embodiments, the binding protein has residues C220S, C226S, C229S, and K405R » In certain embodiments, the binding protein has a mutation at the following residues: C220S, C226S, C229S, and F407R » φ in some implementations In the case, the binding protein has mutations at the following residues: C220S, C226S, C229S and K409D. In certain embodiments, the binding protein has mutations at the following residues: T366F, T368F, P395A, F405R, Y407R, and K409D. In certain embodiments, the binding protein has a mutation at the following residues: T366F 'T368F, P395A, F405A, Y407R, and K409D. In certain embodiments, the binding protein has a mutation at the following residues: P395A, F405R, Y407R, and K409D. 160877.doc -23- 201247704 In certain embodiments, the binding protein has a mutation at the following residues: P395A, F405A, Y407A, and K409D ° In certain embodiments, the binding protein has a mutation at the residue: P395A , F405R and Y407A. In certain embodiments, the binding protein has mutations at the following residues: F405R, Y407A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: P395A, Y407A, and K409D. In certain embodiments, the binding protein has a mutation at the following residues: P395A, F405R, and K409D. In certain embodiments, the binding protein has a mutation at the following residues: P395A and K405R. In certain embodiments, the binding protein has a mutation at the following residues: P395A and Y407R. In certain embodiments, the binding protein has a mutation at the following residues: P395A and K409D. In certain embodiments, the binding protein has a mutation at the following residues: F405R and F407R ° In certain embodiments, the binding protein has a mutation at the following residues: F405R and K409D. In certain embodiments, the binding protein has a mutation at the following residues: F407R and K409D. In certain embodiments, the binding protein has a mutation at the residue: P395A. 160877.doc •24- 201247704 In certain embodiments, the binding protein has a mutation at the following residue: K405R ° In certain embodiments, the binding protein has a mutation at the residue: F407R ° In certain embodiments In this case, the binding protein has a mutation at the following residue: K409D. In certain embodiments, the binding protein comprises a wild type hinge region sequence. In certain embodiments, the binding protein comprises a wild-type amino acid at at least one of C220, C226, and C229. In certain embodiments, the binding protein comprises a wild-type amino acid at at least two of C220, C226, and C229. In certain embodiments, the binding protein comprises a wild-type amino acid at C220, C226, and C229. In certain embodiments, the CH3 domain is at least 50% identical to the wild-type CH3 domain, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, At least 80%, at least 85%, at least 87.5%, at least 90%, at least 92%, at least 94% consistent, at least 96%, or at least 98%. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, T366F, T368F, P395A, F405R, Y407R, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, T366F, T368F, P395A, F405R, Y407R, and K409D. 160877.doc •25· 201247704 In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, T366F, T368F, P395A, F405A, Y407A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, T366F, T368F, P395A, F405A, Y407A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, F405R, Y407R, and K409D. In certain embodiments, the binding protein has a mutation at the following residues: C220S, C226S, P395A, F405R, Y407R, and K409D » In certain embodiments, the binding protein has a mutation at the following residues: C226S, C229S, P395A, F405A, Y407A and K409D. In certain embodiments, the binding protein has mutations at the following residues: C220S, C226S, P395A, F405A, Y407A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, F405R, and Y407A. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, F405R, Y407A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, Y407A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, F405R, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, and F405R. 160877.doc •26- 201247704 In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, and Y407R. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, P395A, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, F405R, and F407R. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, F405R, and K409D. φ In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, F407R, and K409D. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, and P395A. In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, and K405R. In certain embodiments, the binding protein has a mutation at the following residues: C226S, C229S, and F407R. φ In certain embodiments, the binding protein has mutations at the following residues: C226S, C229S, and K409D. In certain embodiments, the binding protein forms a functional antigen binding site for an antigen selected from the group consisting of: cell surface binding molecules, soluble molecules, cytokines, chemokines, enzymes, haptens, lipids, and body. In certain embodiments, the binding protein forms a functional antigen binding site for an antigen selected from the group consisting of: c-Met; Muc-1; 160877.doc -27- 201247704

CD28; CD40; CD19; CD3; TWEAK; TNFR; TREM-1 ABCF1; ACVR1; ACVR1B; ACVR2; ACVR2B ACVRL1; ADORA2A; Aggrecan; AGR2 AICDA; AIF1; AIG1; AKAP1; AKAP2; AMH; AMHR2 ANGPT1; ANGPT2; ANGPTL3; ANGPTL4; ANPEP APC; APOC1; AR; AZGP1 (zinc-a-glycoprotein); B7.1; B7.2 BAD; BAFF; BAG1; BAI1; BCL2; BCL6;

BLNK ; BLR1 (MDR15) ; BlyS ; BMP1 ; BMP2 ; BMP3B

(GDF10); BMP4; BMP6; BMP8; BMPR1A; BMPR1B; BMPR2; BPAG1 (plectin); BRCA1; C19orfl0 (IL27w); C3; C4A; C5; C5R1; CANT1; CASP1; CASP4; CAV1; CCBP2( D6/JAB61); CCL1 (l-309); CCL11 (eotaxin); CCL13 (MCP-4); CCL15 (MIP-ld); CCL16 (HCC-4); CCL17 (TARC) CCL18 (PARC); CCL19 (MIP-3b); CCL2 (MCP-1) MCAF; CCL20 (MIP-3a); CCL21 (MIP-2); SLC; and g lymphoid tissue factor (exodus)- 2; CCL22 (MDC/STC-1); CCL23 φ (MPIF-1); CCL24 (MPIF-2/eosinophil chemoattractant-2); CCL25 (TECK); CCL26 (eosinophil chemotactic factor) -3); CCL27 (CTACK/ILC); CCL28; CCL3 (MIP-la); CCL4 (MIP-lb); CCL5 (RANTES); CCL7 (MCP-3); CCL8 (mcp-2); CCNA1; CCNA2; CCND1; CCNE1; CCNE2; CCR1(CKR1/HM145); CCR2(mcp-1RB/RA); CCR3(CKR3/ CMKBR3); CCR4; CCR5(CMKBR5/ChemRl3); CCR6 160877.doc • 28 - 201247704 (CMKBR6/CKR -L3/STRL22/DRY6) ; CCR7 (CKR7/EBI 1); CCR8 (CMKBR8/TE R1/CKR-L1); CCR9 (GPR-9-6); CCRL1 (VSHK1); CCRL2 (L-CCR); CD164; CD19; CD1C; CD20; CD200; CD-22; CD24; CD28; CD3; CD37; CD38; CD3E; CD3G; CD3Z; CD4; CD40; CD40L; CD44; CD45RB; CD52; CD69; CD72; CD74; CD79A; CD79B; CD8; CD80; CD81; CD83; CD86; CDH1 (E-cadherin (E- Cadherin ) ) ; CDH10 ; CDH12 ; CDH13 ; CDH18 ;

CDH19 ; CDH8 ; CDH5 ; CDH7 ; CDH8 ; CDH9 ; CDK2 ; CDK3 ; CDK4 ; CDK5 ; CDK6 ; CDK7 ; CDK9 ; CEBPB; CER1; CHGA; CHGB; Chitinase; CHST10; CKLFSF2; CKLFSF3; CKLFSF4; CKLFSF5; CKLFSF6; CKLFSF7; CKLFSF8; CLDN3; CLDN7 (cludin_7); CLN3; CLU ( Clusterin); CMKLR1; CMKOR1 (RDCl); CNR1; COL18A1; COL1A1; COL4A3; COL6A1; CR2; CRP; CSF1 (M-CSF); CSF2 (GM-CSF); CSF3 (GCSF); CTLA4; CTNNBl (p-sodium cyclin); CTSB (cathepsin B); CX3CL1 (SCYD1); CX3CR1 (V28); CXCL1 (GRO1); CXCL10 (IP-10); CXCLll (I- TAC/IP-9); CXCL12 (SDF1); CXCL13; CXCL14; CXCL16; CXCL2(GR02); CXCL3(GR03); CXCL5(ENA-78/LIX); CXCL6(GCP-2); CXCL9(MIG); .doc -29- 201247704 CXCR3(GPR9/CKR-L2) ; CXCR4 ; CXCR6(TYMSTR/STRL33/ Bonzo) ; CYB5 ; CYC1 CYSLTR1 , DAB2IP , DES , DKFZp451J0118 , DNCL1 , DPP4 , E2F1 , ECGF1 , EDG1 , EFNA1 , EFNA3 , EFNB2 , EGF , EGFR , ELAC2 , ENG , ENOl , EN02 , EN03 , EPHB4 , EPO , ERBB2 ( Her-2 ) ; ERK1 , ERK8 , ESR1 , ESR2 , F3 ( TF ) , FADD , FasL , FASN , FCER1A , FCER2 , FCGR3A , FGF1 , FGF1 , FGF12 , FGF11 , FGF12 , FGF12B , FGF13 , FGF14 , FGF16 , FGF17 , FGF18 ; FGF19; FGF2 (bFGF); FGF20; FGF21; FGF22; FGF23; FGF3 (int-2); FGF4 (HST); FGF5; FGF6 (HST-2); FGF7 (KGF); FGF8; FGF9; FGFR3; FIGF (VEGFD) FIL1 (EPSILON); FIL1 (ZETA); FLJ12584; FLJ25530; FLRT1 (fibronectin); FLT1; FOS; FOSL1 (FRA-1); FY (DARC); GABRP (GABAa); GAGEB1; GAGEC1 GALNAC4S-6ST , GATA3 , GDF5 , GFI1 , GGT1 , GM-CSF , GNAS1 ;

GNRH1; GPR2 (CCR10); GPR31; GPR44; GPR81 (FKSG80); GRCCIO (CIO); GRP; GSN (Gelsolin); GSTP1; HAVCR2; HDAC4; HDAC5; HDAC7A; HDAC9; HGF; HIF1A; HIP1 Histamine and histamine receptors; HLA-A; HLA-DRA; HM74; HMOX1; HUMCYT2A; ICEBERG; ICOSL; ID2; IFN-a; IFNA1, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNB1, IFNy, IFNW1 IGBP1 , IGF1 , IGF1R , IGF2 , IGFBP2 , IGFBP3 , IGFBP6 , IL-1 , I60877.doc -30- 201247704 IL10 , IL10RA , IL10RB , IL11 , IL11RA , IL-12 , IL12A , IL12B , IL12RB1 , IL12RB2 , IL13 ; IL13RA1 , IL13RA2 , IL14 , IL15 , IL15RA , IL16 , IL17 , IL17B , IL17C , IL17R , IL18 , IL18BP , IL18R1 , IL18RAP ;

IL1F1 , IL1F5 , IL1F6 , IL1F7 , IL1F8 , IL1F9 , IL1F1 , IL1F1 , IL1F1 , IL1HY1 , IL1R1 , IL1R2 , IL1RAP1 , IL1RL1 , IL1RAPL2 , IL1RL1 , IL1RL2 , IL1RN , IL2 , IL20 , IL20RA , IL21R , IL22 , IL22R , IL22R2 ; IL23 , IL24 , IL25 , IL26 , IL27 , IL28A , IL28B , IL29 , IL2RA , IL2RB , IL2RG , IL3 , IL30 , IL3RA , IL4 , IL4R , IL5 , IL5RA , IL6 , IL6R , IL6ST ( glycoprotein 130 ) , IL7 , IL7R IL8; IL8RA; IL8RB; IL8RB; IL9; IL9R; ILK; INHA; INHBA; INSL3; INSL4; IRAKI; IRAK2; ITGA1; ITGA2: ITGA3; ITGA6 (a6 integrin); ITGAV; ITGB3; ITGB4 (b4 integrin) JAG1; JAK1; JAK3; JUN; K6HF; KAI1; KDR; KITLG; KLF5 (GC box BP); KLF6; KLK10; KLK12; KLK13; KLK14; KLK15; KLK3; KLK4; KLK5; KLK6; KLK9; KRT1; KRT19( Keratin 19 (Keratin 19); KRT2A; KRTHB6 (hair-specific type II keratin); LAMA5; LEP (leptin); Lingo-p75; Lingo-Troy; LPS; LTA (TNF-b); LTB; LTB4R (GPR16); LTB4R2; LTBR; MACMARCKS; MAG or Omgp; MAP2K7 (c-Jun); MDK; MIB1; midkine; MIF; MIP-2; MKI67 (Ki -67); MMP2; MMP9; MS4A1; MSMB; MT3 (metal sulfur 160877.doc -31 - 201247704 protein-Ill (metallothionectin-III)); MTSS1; MUC1 (mucin); MYC; MYD88; NCK2; Neurocan; NFKB1; NFKB2; NGFB (NGF); NGFR; NgR-Lingo; NgR-Nogo66 (Nogo); NgR-p75; NgR-Troy; NME1 (NM23A); NOX5; NPPB; NROB1; NR0B2 NR1D1 , NR1D2 , NR1H2 , NR1H3 , NR1H4 , NRII2 , NRII3 , NR2C1 , NR2C2 , NR2E1 , NR2E3 , NR2F1 , NR2F2 , NR2F6 , NR3C1 , NR3C2 , NR4A1 , NR4A2 ;

NR5A3 , NR5A1 , NR5A2 , NR6A1 , NRP1 , NRP2 , NT5E , NTN4 , ODZ1 , OPRD1 , P2RX7 , PAP , PARTI , PATE , PAWR , PCA3 , PCNA , PDGFA , PDGFB , PECAM1 , PF4 ( CXCL4 ) , PGF , PGR , phosphoric acid Phosphatase , PIAS2 , PIK3CG , PLAU ( uPA ) , PLG , PLXDC1 , PPBP ( CXCL7 ) , PPID , PR1 , PRKCQ , PRKD1 , PRL , PROC , PROK2 , PSAP , PSCA , PTAFR ;

PTEN; PTGS2 (COX-2); PTN; RAC2 (p21 Rac2); RARB; RGS1; RGS13; RGS3; RNF110 (ZNF 144); R0B02; SI00A2; SCGB1D2 (lipophilin B); SCGB2A1 ( Mammaglobin 2; SCGB2A2 (mammon globin 1); SCYE1 (endothelial mononuclear cell cytokine); SDF2; SERPINA1; SERPINA3; SERPINB5 (maspin); SERPINE1 (PAI- 1); SERPINF1; SHBG; SLA2; SLC2A2; SLC33A1; SLC43A1; SLIT2; SPP1; SPRRlB(Sprl); ST6GAL1; STAB1; STAT6; STEAP; 160877.doc -32- 201247704

STEAP2; TB4R2; TBX21; TCP10; TDGF1; TEK; TGFA; TGFB1; TGFB111; TGFB2; TGFB3; TGFBI; TGFBR1; TGFBR2; TGFBR3; TH1L; THBS1 (thrombospondin-l); THBS2; TIE (Tie-1); TIMP3; tissue factor; TLR10; TLR2; TLR3; TLR4; TLR5; TLR6; TLR7; TLR8; TLR9; TNF; TNF-a; TNFAIP2 (B94); TNFAIP3; TNFRSF11A; TNFRSF1A; TNFRSF1B TNFRSF21; TNFRSF5; TNFRSF6 (Fas); TNFRSF7; TNFRSF8; TNFRSF9; TNFSF10 (TRAIL); TNFSF11 (TRANCE); TNFSF 12 (AP03L); TNFSF 13 (April); TNFSF13B; TNFSF14 (HVEM-L); TNFSF 15 ( VESF); TNFSF18; TNFSF4 (OX40 Ligand); TNFSF5 (CD40 Ligand); TNFSF6 (FasL); TNFSF7 (CD27 Ligand); TNFSF8 (CD30 Ligand); TNFSF9 (4-1BB Ligand) TOLLIP; Toll-like receptor; TOP2A (topoisomerase Iia); TP53; TPM1; TPM2; TRADD; TRAF1; TRAF2; TRAF3; TRAF4; TRAF5; TRAF6; TREM1; TREM2; TRPC6; TSLP; TWEAK; GF; VEGFB; VEGFC; versican; VHL C5; VLA-4; XCL1 (lymphotactin); XCL2 (SCM-lb); XCR1 (GPR5/CCXCR1); YY1; And ZFPM2. And Rs. , 120, 122, 124, 126, 128, 130, 132, 138, 160, 162, 164, 166, 168, 170, 172, 160877.doc -33- 201247704 174, 176, 178, 180, 182, 184, 186 , 188 , 190 , 192 , 194 ' 196 ' 198 ' 200, 202 and 204 » In certain embodiments, the light chain variable domain comprises an amino acid sequence selected from the group consisting of: 3 £(^1 〇>10:4, 28, 39, 41, 79, 81-83, 85, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 161, 163, 165, 167 , 169 , 171 , 173 , 175 , 177 , 179 , 181 , 183 , 185 , 187 , 189 , 191 , 193 , 195 , 197 , 199 , 201 & 203 ° In certain embodiments, the heavy chain R or The body comprises amino acid sequences SEQ ID NOs: 84, 206 and 207 selected from the group consisting of: In certain embodiments, the R or receptor package of the light chain The amino acid sequences of the following composition are selected to be SEQ ID NOs: 84, 206 and 207. In certain embodiments, the binding protein is capable of binding one or more targets. In certain embodiments, one or more targets Is selected from the group consisting of c-Met; CD-28; CD-3; CD-19; ABCF1; ACVR1; ACVR1B; ACVR2; ACVR2B; ACVRL1; ADORA2A; aggrecan; AGR2; AICDA; AIF1; AIG1 AKAP1 , AKAP2 , AMH , AMHR2 , ANGPT1 , ANGPT2 , ANGPTL3 , ANGPTL4 , ANPEP , APC , APOC1 , AR , AZGP1 ( zinc - a-glycoprotein ) , B7.1 , B7.2 , BAD , BAFF , BAG1 , BAI1 BCL2; BCL6; BDNF; BLNK; BLR1 (MDR15); BlyS; BMP1; BMP2; BMP3B (GDF10); BMP4; BMP6; BMP8; BMPR1A; BMPR1B; BMPR2; BPAG1 (retinin); BRCA1; C19orflO (IL27w); C3; C4A; C5; C5R1; CANT1; 160877.doc • 34- 201247704

CASP1; CASP4; CAV1; CCBP2(D6/JAB61); CCL1(1-309); CCL11 (eosinophil chemokine); CCL13 (MCP-4); CCL15 (MIP-ld); CCL16 (HCC-4) CCL17(TARC); CCL18 (PARC); CCL19(MIP-3b); CCL2(MCP-1); MCAF; CCL20 (MIP-3a); CCL21(MIP-2); SLC; non-lymphoid tissue chemokine (exodus)-2; CCL22 (MDC/STC-1); CCL23 (MPIF-1); CCL24 (MPIF-2/eosinophil chemoattractant-2); CCL25 (TECK); CCL26 (eosinophils) Chemokine-3); CCL27 (CTACK/ILC); CCL28; CCL3 (MIP-la); CCL4 (MIP-lb); CCL5 (RANTES); CCL7 (MCP-3); CCL8 (mcp-2); CCNA1 CCNA1 ; CCND1 ; CCNE1 ; CCNE2 ; CCR1 (CKR1/HM145) ; CCR2 ( mcp - lRB / RA ) ; CCR3 ( CKR3 / CMKBR3 ) ; CCR4 ; CCR5 ( CMKBR 5 / ChemR 13 ) ; CCR 6 ( CMKBR6 / CKR - L3 / STRL 22 /DRY6); CCR7(CKR7/EBI1); CCR8(CMKBR8/TER1/CKR-L1); CCR9(GPR-9-6); CCRL1 (VSHK1); CCRL2(L-CCR); CD164; CD19; CD1C; CD20 CD200 ; CD24 ; CD28 ; CD3 ; CD37 ; CD3 ; CD3 ; CD3 ; CD3Z ; CD4 ; CD40 ; CD40L ; CD44 ; CD45 RB ; CD52 ; CD69 ; CD79 ; CD8 ; CD80 ; CD81 ; CD83 ; CD86 ; CDH1 ( E - # 5 mucin ) ; CDH7 ; CDH8 ; CDK9 ; CDK2 ; CDK3 ; CDK4 ; CDK5 ; CDK6 ; CDK7 ; CDK9 ; CER1; CHGA; 160877.doc -35- 201247704 CHGB; chitinase; CHST10; CKLFSF2; CKLFSF3; CKLFSF4; CKLFSF5; CKLFSF6; CKLFSF7; CKLFSF8; CLDN3; CLDN7 (Claudin-7); CLN3; CLU CMKOR1; CMKOR1 (RDCl); CNR1; COL18A1; COL1A1; COL4A3; COL6A1; CR2; CRP; CSF1(M-csi); CSF2(GM-CSF); CSF3(GCSF); CTLA4; CTNNB1 (b-sole hydrocarbon )); CTSB (Cathepsin B); CX3CL1 (SCYD1); CX3CR1 (V28); CXCL1 (GRO1); CXCL10 (IP-10); CXCL11 (I-TAC/IP-9); CXCL12 (SDF1); CXCL13; CXCL14; CXCL16; CXCL2(GR02) ; CXCL3(GR03) ; CXCL5(ENA -78/LIX); CXCL6(GCP-2); CXCL9(MIG); CXCR3(GPR9/CKR-L2); CXCR4; CXCR6(TYMSTR/STRL33/Bonzo); CYB5; CYC1; CYSLTR1; DAB2IP; DES; DKFZp451 JO DNCL1 , DPP4 , E2F1 , ECGFI , EDG1 , EFNA1 , EFNA3 , EFNB2 , EGF , EGFR , ELAC2 1 ENG , ENOl , EN02 , EN03 , EPHB4 , EPO , ERBB2 ( Her-2 ) , EREG , ERK8 , ESR1 , ESR2 F3 (TF) , FADD , FasL , FASN , FCER1A , FCER2 , FCGR3A , FGF , FGF1 ( aFGF ) , FGF10 , FGF11 , FGF12 , FGF12B , FGF13 , FGF14 , FGF16 , FGF17 , FGF18 , FGF19 , FGF2 ( bFGF ) ; FGF21; FGF21; FGF23; FGF3(int-2); FGF4(HST); FGF5; FGF6(HST-2); FGF7(KGF); FGF8; FGF9; FGFR3; FIGF(VEGFD); FIL1 (EPSILON); FIL1 (ZETA); FLJ12584; FLJ25530; FLRT1 (Fiber binding protein); FLT1; 160877.doc -36- 201247704

FOS; FOSL1 (FRA-1); FY (DARC); GABRP (GABAa); GAGEB1; GAGEC1; GALNAC4S-6ST; GATA3; GDF5; GFI1; GGT1; GM-CSF; GNAS1; GNRH1; GPR2 (CCR10); GPR31; GPR44; GPR81 (FKSG80); GRCC10 (CIO); GRP; GSN (plycosin); GSTP1; HAVCR2; HDAC4; HDAC5; HDAC7A; HDAC9; HGF; HIF1A; HIP1; histamine and histamine receptor; HLA-A HLA-DRA , HM74 , HMOX1 , HUMCYT2A , ICEBERG , ICOSL , ID2 , IFN - a , IFNA1 , IFNA2 , IFNA4 , IFNA5 , IFNA6 , IFNA1 , IFNB1 , IFNy , IFNW1 , IGBP1 , IGF1 , IGF1R , IGF2 , IGFBP2 , IGFBP3 IGFBP6 , IL-1 , IL10 , IL10RA , IL10RB , IL11 , IL11RA , IL-12 , IL12A , IL12B , IL12RB1 , IL12RB2 , IL13 , IL13RA1 , IL13RA2 , IL14 , IL15 , IL15RA , IL16 , IL17 , IL17B , IL17C , IL17R IL18 , r IL18BP , IL18R1 , IL18RAP , IL19 , ILIA , IL1B , IL1F10 , IL1F5 , IL1F6 , IL1F7 , IL1F8 , IL1F9 , IL1HY1 , IL1R1 , IL1R2 , IL1RAP IL1RAPL1 , IL1RL1 , IL1RL1 , IL1RL2 , IL1RN , IL2 , IL20 , IL20RA , IL21R , IL22 , IL22R , IL22RA2 , IL23 , IL24 , IL25 , IL26 , IL27 , IL28A , IL28B , IL29 , IL2RA , IL2RB , IL2RG , IL3 , IL30 IL3RA , IL4 , IL4R , IL5 , IL5RA , IL6 , IL6R , IL6ST ( glycoprotein 130 ) , IL7 , IL7R , IL8 , IL8RA , IL8RB , IL8RB , IL9 , IL9R , ILK , INHA , INHBA , INSL3 , INSL4 , IRAKI ; IRAK2; 160877.doc -37- 201247704 ITGAl; ITGA2; ITGA3; ITGA6 (a6 integrin); ITGAV; ITGB3; ITGB4 (P4 integrin); JAG1; JAK1; JAK3; JUN; K6HF; KAI1; KDR; KITLG; KLF5 (GC box BP); KLFK; KLK10; KLK12; KLK13; KLK14; KLK15; KLK3; KLK4; KLK5; KLK6; KLK9; KRT1; KRT19 (keratin 19); KRT2A; KRTHB6 (hair-specific type II keratin) LAMA5; LEP (alkaline); Lingo-p75; Lingo-Troy; LPS; LTA (TNF-b); LTB; LTB4R (GPR16); LTB4R2; LTBR; MACMARCKS; MAG or Omgp; MAP2K7 (c-Jun); MDK ; MIB1 ; medium factor MIF; MIP-2; MKI67 (Ki-67); MMP2; MMP9; MS4A1; MSMB; MT3 (metallothionein-III); MTSS1; MUC1 (mucin); MYC; MYD88; NCK2; neuroglycan; NFKB1 NFKB2; NGFB(NGF); NGFR; NgR-Lingo; NgR-Nogo66 (Nogo); NgR-p75; NgR-Troy; NME1 (NM23A); NOX5; NPPB; NR0B1; NR0B2; NR1D1; NR1D2; NR1H2; NR1H3; NR1H4 ; NRII2 ;

NR23; NR2C1; NR2C2; NR2E1; NR2E3; NR2F1 NR2F2; NR2F6; NR3C1; NR3C2; NR4A1;

NR5A3; NR5A1; NR5A2; NR6A1; NRP1; NRP2 NT5E; NTN4; ODZ1; OPRD1; P2RX7; PAP;

PATE; PAWR; PCA3; PCNA; PDGFA; PDGFB PECAM1; PF4 (CXCL4); PGF; PGR; phosphoproteoglycan PIAS2; PIK3CG; PLAU (uPA); PLG; PLXDC1 PPBP (CXCL7); PPID; PR1; PRKCQ; PRKD1 ; PRL 160877.doc • 38 - 201247704

PROC; PROK2; PSAP; PSCA; PTAFR; PTEN; PTGS2 (COX-2); PTN; RAC2 (p21 Rac2); RARB; RGS1; RGS13; RGS3; RNF110(ZNF 144); R0B02; SI00A2; SCGB1D2 (lipophile) B); SCGB2A1 (mammon globin 2); SCGB2A2 (mammon globin 1); SCYE1 (endothelial mononuclear cell cytokine); SDF2; SERPINA1; SERPINA3; SERPINB5 (mammography); SERPINEl (PAI-l) SELPINF1 , SHBG , SLA2 , SLC2A2 , SLC33A1 , SLC43A1 , SLIT2 , SPP1 , SPRR1B ( Sprl ) , ST6GAL1 , STAB1 , STAT6 , STEAP , STEAP2 , TB4R2 , TBX21 , TCP10 , TDGF1 , TEK , TGFA , TGFB1 , TGFB1 , TGFB2 ; TGFB3; TGFBR1; TGFBR1; TGFBR2; TGFBR3; TH1L; THBS1 (platelet-reactive protein-1); THBS2; THBS4; ΤΗΡΟ; TIE(Tie-1); TIMP3; tissue factor; TLR10; TLR2; TLR3; TLR4; TLR5; TLR6 TLR7; TLR8; TLR9; TNF; TNF-a; TNFAIP2 (B94); TNFAIP3; TNFRSF11A; TNFRSF1A; TNFRSF1B; TNFRSF21; TNFRSF5; TNFRSF6 (Fas); TNFRSF7; TNFRSF TNFSFIO (TRAIL); TNFSF11 (TRANCE); TNFSF12 (AP03L); TNFSF13 (April); TNFSF13B; TNFSF14 (HVEM-L); TNFSF15 (VEGI); TNFSF1.8; TNFSF4 (OX40 ligand); TNFSF5 (CD40 ligand); TNFSF6 (FasL); TNFSF7 (CD27 ligand); TNFSF8 (CD30 ligand); TNFSF9 (4-1BB ligand); TOLLIP; Toll-like receptor; TOP2A (topologically different) Structured enzyme Iia); TP53; TPM1; TPM2; TRADD; TRAF1; TRAF2; 160877.doc-39-201247704 TRAF3; TRAF4; TRAF5; TRAF6; TREM1; TREM2; TRPC6; TSLP; TWEAK; VEGF; VEGFB; VEGFC; Proteoglycan; VHL C5; VLA-4; XCL1 (lymphocyte chemotactic factor); XCL2 (SCM-lb); XCR1 (GPR5/CCXCR1); YY1; and ZFPM2 ° In certain embodiments, the binding protein is capable of binding Two targets, wherein the two targets are selected from the group consisting of c-Met and CD-28; c-Met and CD-3; c-Met and CD-19; CD-28 and CD-3; -28 and CD-19; CD-3 and CD-19; CD138 and CD20; CD138 and CD40; CD20 and CD3; CD38 and CD138; CD38 and CD20; CD38 and CD40; CD40 and CD20; CD19 and CD20; CD-8 and IL-6; PDL-1 and CTLA-4; CTLA-4 and BTN02; CSPGs and RGM A; IGF1 and IGF2; IGF1/2 and Erb2B; IL-12 and IL -18; IL-12 and TWEAK; IL-13 and ADAM8; IL-13 and CL25; IL-13 and IL-Ιβ; IL-13 and IL-25; IL-13 and IL-4; IL-13 and IL IL-13 and IL-9; IL-13 and LHR agonists; IL-13 and MDC; IL-13 and MIF; IL-13 and PED2; IL-13 and SPRR2a; IL-13 and SPRR2b; IL-13 and TARC; IL-13 φ and TGF-β; lL-1-α and IL-Ιβ; MAG and RGM A; NgR and RGM A; Nog〇A and RGM A; OMGp and RGM A; RGM A and RGM B; Te38 and TNF-α; TNF-α and IL-12; TNF-α and IL-12p40; TNF-α and IL-13; TNF-α and IL-15; TNF-α and IL-17; -α and IL-18; TNF-α and IL-Ιβ; TNF-α and IL-23; TNF-α and MIF; TNF-α and PEG2; TNF-α and PGE4; TNF-α and VEGF; EGFR; TNF-α and RANK ligands; TNF-α and Blys; 160877.doc •40- 201247704 TNF-α and GP130; TNF-α and CD-22; and TNFa and CTLA-4. In certain embodiments, the binding protein is capable of modulating the biological function of one or more targets. In certain embodiments, the binding protein is capable of neutralizing one or more targets. In certain embodiments, one or more are 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10" In certain embodiments, the cytokine is selected from the group consisting of: Lymphocytes, mononuclear hormones and peptide hormones. φ In certain embodiments, the cytokines are IL-la and IL-Ιβ. In certain embodiments, the cytokines are TNF-a and IL-13. In certain embodiments, the cytokines are IL-12 and IL-18. In certain embodiments, the chemokine is selected from the group consisting of: CCR2, CCR5, and CXCL-13. In certain embodiments, the cell surface protein is integrins. In certain embodiments, the cell surface protein line is selected from the group consisting of: CD-20 and CD3. # In certain embodiments, the enzyme is selected from the group consisting of kinases and proteases. In certain embodiments, the subject is selected from the group consisting of a lymphokinin receptor, a mononuclear hormone receptor, and a polypeptide hormone receptor. In certain embodiments, the linker is selected from the group consisting of: ASTKGPSVFPLAP (SEQ ID NO: 46); ASTKGP (SEQ ID NO: 48); TVAAPSVFIFPP (SEQ ID NO: 50); TVAAP (SEQ ID NO: 52) ; AKTTPKLEEGEFSEAR (SEQ ID NO: 160877.doc •41 · 201247704

AKTTPKLEEGEFSEARV (SEQ ID NO: 95); AKTTPKLGG (SEQ ID NO: 96); SAKTTPKLGG (SEQ ID NO: 97); SAKTTP (SEQ ID NO: 98); RADAAP (SEQ ID NO: 99); RADAAPTVS ( SEQ ID NO: 100); RADAAAAGGPGS (SEQ ID NO: 101); RADAAAA(G4S)4 (SEQ ID NO: 102);

SAKTTPKLEEGEFSEARV (SEQ ID NO: 103); ADAAP (SEQ ID NO: 104); ADAAPTVSIFPP (SEQ ID NO: 105); QPKAAP (SEQ ID NO: 106); QPKAAPSVTLFPP (SEQ ID NO: 107); AKTTPP (SEQ ID NO) : AKTTPPSVTPLAP (SEQ ID NO: 109); AKTTAP (SEQ ID NO: 110); AKTTAPSVYPLAP (SEQ ID NO: 111); GGGGSGGGGSGGGGS (SEQ ID NO: 112); GENKVEYAPALMALS (SEQ ID NO: 113); GPAKELTPLKEAKVS (SEQ ID NO: 114); GHEAAA VMQVQYPAS (SEQ ID NO: 115); TVAAPSVFIFPPTVAA

PSVFIFPP (SEQ ID NO: 116); and ASTKGPSVFPLAPAST KGPSVFPLAP (SEQ ID NO: 117). In certain embodiments, as measured by surface plasmon resonance, the binding protein _ has one or more targets having an association rate constant (Kon) selected from the group consisting of: at least about ΙΟ2 Μ _, 丨; At least about 3 Μ···1; at least about 104 Μ···1; at least about 105 Μ···1; and at least about Μ6 Μ,·1. In certain embodiments, the binding protein has one or more targets having a dissociation rate constant (Koff) selected from the group consisting of: up to about 103 ΜΛ·1; up to about 104, as measured by surface plasmon resonance. Mi·1; up to about 105 M-Y1; and up to about Μ6 Μ,·1. 160877.doc •42- 201247704

In certain embodiments, the binding protein has one or more targets having a dissociation constant (Kd) selected from the group consisting of: up to about ι · 6 μ; up to about 1 〇 7 up to about 10 〜 up to about 1 〇 - 9Μ; at most about ι〇υ about 10·11 Μ; and up to about 10-12 Μ. The invention provides a binding protein comprising a binding protein as described herein in combination with an agent comprising a population selected from the group consisting of an immunoadhesive molecule, a developer, a therapeutic agent, and a cytotoxic agent. The agent is selected from the group consisting of: a radioactive label, a glory label, a luminescent label, a bioluminescent label, a magnetic label, and biotin. In some embodiments, the developer is selected Sexual markers of the group consisting of free: 3H, 14r, 35

C Ύ 99 Ι77ι

Tc 111

Lu, 166Ho and (1) Sm

In , 125I , u In certain embodiments, the agent is a cytotoxic agent or an cytotoxic agent: an antimetabolite, a base: a therapeutic agent of a second group of sheep (4) an alkylating agent 'antibiotics, growth factors

... anti-angiogenic agents, anti-mitotic agents, anthracyclines, toxins and cell death agents. In some embodiments, the crystal is released in certain embodiments. , 'Ό σ protein is a crystal binding protein. The crystallized binding protein crystals are drug-free drug-free and have a higher in vivo half-life than the binding protein. In certain embodiments, the crystal-bound egg-like counterpart has a higher in vivo half-life. In certain embodiments, the invention provides methods of crystallizing a binding protein that retain biological activity. In certain embodiments, the binding protein is cultured in a medium in a medium sufficient to produce a binding protein according to 160877.doc • 43· 201247704

The method is produced wherein the host cell comprises a vector and the vector comprises a nucleic acid encoding a protein of a. ° Q The present invention provides a pharmaceutical composition. In some embodiments, the medical group

A binding protein and a pharmaceutically acceptable carrier are provided herein. Q In certain embodiments, the pharmaceutical composition further comprises at least one agent. In some embodiments, the other agent is selected from the group consisting of: a second therapeutic agent; a developer, a cytotoxic agent; an angiogenesis inhibitor; a kinase inhibitor; a costimulatory molecular blocker; an adhesion molecule Broken agent; anti-cytokine antibody or functional fragment thereof; methotrexate; cyclosporin; rapaniycin; FK506; detectable label or reporter; TNF antagonist; Rheumatoid; muscle relaxant; anesthetic; non-steroidal anti-inflammatory drugs (NSAID); analgesics; anesthetics; sedatives; local anesthetics; neuromuscular blockers; antimicrobial agents; antipsoriatic drugs; corticosteroids; anabolic steroids; Immunization 'immunoglobulin; immunosuppressant; growth hormone; hormone replacement drug; radiopharmaceutical; 3H; 14C; 35S; 90Y; 99Tc; 丨丨丨In; 丨251; 131T. 177τ .

Lu * Ho ; 153Sm ; fluorescent label; luminescent label; bioluminescent label; magnetic label; biotin; antidepressant; antipsychotic; stimulant; asthma drug; beta agonist; inhaled steroid; adrenaline or similar ; cytokines and cytokine antagonists. The invention provides a pharmaceutical composition comprising a binding protein complex provided herein and a pharmaceutically acceptable carrier. 160877.doc -44 - 201247704 The invention provides a nucleic acid encoding a polypeptide of the invention as provided herein. The invention provides expression constructs comprising a nucleic acid encoding a polypeptide of the invention as provided herein. The invention provides a cell comprising a representation construct having a nucleic acid encoding a polypeptide of the invention as provided herein. The invention provides the use of a binding protein of the invention. In one embodiment, the binding proteins provided herein are used to prepare a medicament. In certain embodiments, the medicament is for treating a disease or condition selected from the group consisting of: arthritis, osteoarthritis, juvenile chronic arthritis, septic arthritis, Lyme arthritis, Psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic lupus erythematosus, Crohn's disease, ulcerative colitis, inflammatory bowel disease, insulin-dependent diabetes, sputum Gonaditis, asthma, allergic diseases, psoriasis, inflammatory scleroderma, graft versus host disease, organ transplant rejection', acute or chronic immune diseases associated with organ transplantation, sarcoidosis, atherosclerosis , disseminated intracoronary coagulation, Kawasaki, s disease, Grave's disease, renal syndrome, chronic fatigue syndrome, Wegener's granulomatosis, Heng Bao _ Si Chien Henoch-Schoenlein purpurea, renal microscopic vasculitis, chronic active hepatitis, uveitis, septic shock, toxic shock syndrome , sepsis syndrome, cachexia, infectious diseases, parasitic diseases, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington's chorea, Parkins〇n, s disease Alzheimer's disease, stroke, original 160877.doc -45- 201247704 biliary cirrhosis, hemolytic anemia, malignant disease, heart failure, myocardial infarction, Addison's disease, sporadic I Sporadic polyglandular deficiency type I and type II polyglandular secretion deficiency, Schmidt's syndrome, adult (acute) respiratory distress syndrome, alopecia, plaque alopecia, seronegative Arthropathy, arthropathy, Reiter's disease, psoriatic arthropathy, ulcerative colitis, joint disease, intestinal synovitis, chlamydia, yersinia, and salmonella (salmonella) related arthropathy, spondyloarthropathy, atherosclerosis/arteriosclerosis, atopic hypersensitivity, autoimmune Blisters, pemphigus vulgaris, pebbicular pemphigus, pemphigoid, linear IgA disease, autoimmune; gluteal anemia, C〇〇mbs positive haemolytic anaemia, acquired pernicious anemia , adolescent pernicious anemia, myalgia, free radical disease, chronic skin mucosal candidiasis, giant cell arteritis, primary sclerosing hepatitis, unexplained autoimmune hepatitis, acquired immunity Lack of disease syndrome, acquired immunodeficiency related diseases, hepatitis B, hepatitis C, general variability immunodeficiency _ lack of (common variability hypogammaglobulinemia), dilated cardiomyopathy, female infertility' _ nest Functional failure, _ nest premature aging, fibrotic lung disease, cause

/polymyositis-associated lung disease, ~|曰"negative lung disease, rheumatoid arthritis, phase-related lupus erythematosus-related lung disease, dermatomyositis-related lung disease (Sj6gren, s 160877.doc •46- 201247704 Dose-associated lung disease associated with ankylosing spondylitis, disease, vasculitis, diffuse lung disease, haemosiderosis-related lung disease, drug-induced interstitial lung disease, fibrosis, radiofibrosis, Obstructive bronchiolitis, chronic eosinophilia

Spherical pneumonia, lymphocytic infiltrating lung disease, post-infection interstitial lung disease, gouty arthritis, autoimmune hepatitis, type 1 autoimmune hepatitis (typical autoimmune or lupus-like hepatitis), type 2 Autoimmune hepatitis (anti-lkm antibody hepatitis)' autoimmune-mediated hypoglycemia, type B tamsin resistance with acanthosis nigricans, parathyroid dysfunction, acute immune diseases associated with organ transplantation, and organ transplantation Related chronic immune diseases, osteoarthritis, primary sclerosing cholangitis, type 1 psoriasis, type 2 psoriasis, idiopathic leukopenia, autoimmune neutropenia, N〇s type Kidney disease, spheroid nephritis, renal microscopic vasculitis, lyme disease, discoid lupus erythematosus, idiopathic or NOS male infertility, sperm autoimmune, multiple sclerosis (all subtypes) ), sympathetic ophthalmia, pulmonary hypertension secondary to connective tissue disease, G〇〇dpasture, s syndrome, pulmonary manifestations of nodular polyarteritis, acute rheumatic fever, rheumatoid spondylitis Stiller Still's disease, systemic sclerosis, Hugh's syndrome, Takayasu's disease/arteritis, autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmune thyroid disease, thyroid Hyperthyroidism, goiter autoimmune 〖Gynecological dysfunction (Hashimoto's disease), atrophic autoimmune thyroid dysfunction, primary mucinous edema, lens-like uveitis, primary Vasculitis, leukoplakia acute liver disease, chronic liver 160877.doc •47- 201247704 Alcoholic cirrhosis, alcohol-induced liver injury, bile stagnation, characteristic liver disease, drug-induced hepatitis, non-alcoholic steatosis hepatitis, Allergies and asthma, group B streptococcus (GBS) infection, mental disorders (such as depression and schizophrenia), Th2 and Th1 type mediated diseases, acute and chronic pain (different forms of pain), and lung cancer , breast cancer, stomach cancer, bladder cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer and rectal cancer and hematopoietic malignancies ( Leukemia and lymphoma), no beta lipoproteinemia, hand and foot cyanosis, acute and chronic parasitic or infection processes, acute leukemia, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute or chronic bacterial infection, acute Pancreatitis, acute renal failure, adenocarcinoma, atrial ectopic pulsation, aids dementia syndrome, alcohol-induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergic rhinitis, allograft rejection, α-1 _Antitrypsin deficiency, amyotrophic lateral sclerosis, poor jk, angina pectoris, anterior horn cell degeneration, anti-ed3 therapy, anti-squamous syndrome, anti-receptor allergic reaction, aortic and peripheral aneurysm, aortic dissection, Arterial hypertension, atherosclerosis, arteriovenous sinus, ataxia, atrial fibrillation (sustained or paroxysmal), atrial flutter, atrioventricular block, B-cell lymphoma, bone graft rejection 'Bone marrow transplantation (BMT) rejection, bundle branch block, Burkiu, s lymphoma, burns, arrhythmia, cardiac arrest syndrome Group, cardiac tumor, cardiomyopathy, cardiopulmonary bypass inflammatory response, cartilage transplant rejection, cerebellar cortical degeneration, cerebellar disorder, turbulent or multifocal atrial tachycardia, chemotherapy-related disorders, chronic myeloid leukemia (CML) , chronic alcoholism, chronic inflammatory disease, chronic lymphocytic leukemia 160877.doc -48- 201247704 (CLL), chronic obstructive pulmonary disease (c〇PD), chronic salicylic acidosis, colorectal cancer, congestive heart failure , conjunctivitis, contact dermatitis, pulmonary heart disease, coronary artery disease, Creutzfeldt_Jak〇b disease, culture-negative sepsis, cystic fibrosis, cytokine therapy-related disorders, boxer dementia, off Cycling disease, dengue hemorrhagic fever, dermatitis, dermatological conditions, diabetes (diabete/diabetes mellitus), diabetic arteriosclerosis, Diffuse Lewy body disease, dilated congestion φ cardiomyopathy, basal ganglia disease, middle-aged Down syndrome (Down's

Syndrome in middle age), drug-induced dyskinesia induced by drugs that block CNS dopamine receptors, drug sensitivity, genital warts, cerebrospinal gamma, endocarditis, endocrine disease, epiglottis, EB virus infection (epstein_ Barr virus infection), acromegaly, extrapyramidal and cerebellar disorders, familial hemophagocytic histiocytoma, fetal thymic transplant rejection, Friedreich's ataxia, functional periphery Arterial disease, fungal sepsis, gas gangrene, gastric ulcer, renal glomerulonephritis, graft rejection of any organ or tissue, gram negative sepsis, Gram-positive sepsis, intracellular organisms Body-induced granuloma, hairy cell leukemia, Hallerrorden-Spatz disease, hashimoto's thyroiditis, hay fever, heart transplant rejection, hemochromatosis, Hemodialysis, hemolytic uremic syndrome / thrombolytic thrombocytopenic purpura, hemorrhage, hepatitis (A), His bundle of heart rhythm (His bundle arrythmias), HIV infection/HIV neuropathy, Hodgkin's 160877.doc •49-201247704 disease, hyperactive exercise disorder, allergic reaction, hypersensitivity pneumonitis, high-gold pressure, slow-motion exercise disorder , hypothalamic-pituitary-adrenal axis assessment, idiopathic Addison's disease, idiopathic pulmonary fibrosis, antibody-mediated cytotoxicity, debilitation, infant spinal muscular atrophy, aortic inflammation, influenza VIII, Ionizing radiation exposure, iridocyclitis/uveitis/optic neuritis, ischemia-reperfusion injury, ischemic stroke, juvenile rheumatoid arthritis, adolescent spinal muscular atrophy, Kaposi's sarcoma (Kap〇 Si, s sarcoma), kidney transplant rejection, Legionnaire's disease (legi〇neUa), leishmaniasis, leprosy, corticosal system disease, fat edema, liver transplant rejection, lymphedema, malaria, Malignant lymphoma, malignant histiocytosis, malignant melanoma, meningitis, meningococcalemia, metabolic/idiopathic disease, migraine, mitochondrial multilineage Disease, mixed connective tissue disease, monoclonal immunoglobulin Ah disease, multiple myeloma, multiple system degeneration (Mance, on behalf of # _ because Thomas, Shideerge and Machado - Joseph (Mencel Dejerine_Th〇mas ShiDrager and

Machado-Joseph)), myasthenia gravis, mycobacterium avium intracellulare, mycobacterium tuberculosis, myelodysplastic syndrome, myocardial infarction, myocardial ischemic condition, nasopharyngeal carcinoma, neonatal Chronic lung disease, nephritis, nephropathy, neurodegenerative disease, spastic neuromuscular atrophy, neutropenic fever, non-Hodgkin's lymphoma (n〇n_h〇dgkins iymphoma), abdominal aorta and its branch blockage, Obstructive arterial disease, okt3 therapy, testicular/episositis, testicular/vasectomy, organ enlargement, osteoporosis, pancreatic transplant rejection, pancreatic cancer, paraneoplastic 160877.doc -50· 201247704 syndrome /Malignant hypercalcemia, parathyroid transplant rejection, pelvic inflammatory disease, perennial rhinitis, pericardial disease, peripheral atherosclerotic disease, peripheral vascular disease, peritonitis, pernicious anemia, Pneumocystis carinii pneumonia (pneumocystis carinii Pneumonia), pneumonia, POEMS syndrome (polyneuropathy, organ enlargement, endocrine disease, gamma globulin disease) Skin Change Syndrome), Post-Perfusion Syndrome, Post-Pump Syndrome, Post-Cleavage Symptoms, Pre-eclampsia, Progressive Nuclear Paralysis, Primary Pulmonary Hypertension, Radiotherapy, Raynaud's phenomenon (Raynaud, S phen〇men ()n) φ and disease, Raynoud's disease, Refsiim's disease, regular stenosis qRS tachycardia, renal vascular hypertension, reperfusion injury, restrictive cardiomyopathy, sarcoma , scleroderma, senile chorea, senile dementia (Senile Dementia 〇f Lewy body type), seronegative joint disease, shock, sickle cell anemia, skin allograft rejection, skin variability syndrome, small bowel transplantation Rejection, solid tumors, specific arrhythmia, spinal ataxia, spinocerebellar degeneration, streptococcal myositis, cerebellar structural lesions, subacute • sclerosing panencephalitis, fainting, cardiovascular syphilis, systemic allergy, Systemic inflammatory response syndrome, systemic adolescent rheumatoid arthritis, T-cell or FAB ALL, capillary expansion Thromboangiitis obliterans, thrombocytopenia, poisoning, transplantation, trauma/bleeding, m-type allergic reaction, type IV allergy, unstable angina pectoris, uremia, septicemia, urticaria, valvular heart disease, varicose veins , vasculitis, venous disease, venous thrombosis 'ventricular fibrillation, viral and fungal infections, viral encephalitis/aseptic meningeal hernia, virus-associated hemophagocytic syndrome' Weini 160877.doc •51· 201247704 克-科Xercke syndrome (Wernicke-Korsakoff Syn (jr〇me), Wilson's disease or any organ or tissue xenograft rejection" The invention is further illustrated by the following detailed description and circular. [Embodiment] The present invention relates to monovalent and optionally multispecific binding proteins which bind one or more antigens or targets (e.g., receptor ligands). In particular, the present invention relates to recombinant expression vectors and host cells, referred to herein as semi-immunoglobulins (half (4) binding proteins and pharmaceutical compositions thereof, and nucleic acids used to prepare such +Ig. A method of detecting a particular antigen in vitro or in vivo using a binding protein of the invention is encompassed. Unless otherwise defined herein, scientific and technical terms used in connection with the invention will have the meaning commonly understood by the ordinary skill of the art. Fan Ming should be a clear U. In the event of any potential ambiguity, the definition provided in this article is superior to any dictionary or foreign definition. In addition, unless the context requires otherwise, the singular term should include plural and plural, and should include In the singular case, unless otherwise stated, the use of "or" means "and/or". In addition, the use of the term "including (4) (4)" (and other forms such as "Eludes" and "ineluded") There is a limit to J. Life, unless otherwise specified, such as "to = or "component j term covers a unit And the components and components of the above-mentioned sub-units. Unless otherwise defined by the context, all values herein are understood to be modified by the term "about." The amount of change will depend on the particular value, but is generally considered Within the standard deviation of I60877.doc •52· 201247704, “about” can be understood as changes of up to 〇%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 〇1% or 〇〇1%; or mean +/- up to 2 or up to 3 standard deviations. The ranges provided herein are to be understood to include all values within the range, or ranges within the range Or any subset of values. For example, 1-10 should be understood to include any range or subset of i, 2, 3, 4, 5, 6, 7, 8, 9 and 10' or their values, and a fractional value when appropriate. Similarly, the scope of the "at most" specific value form is reasonable.

The solution is a value including 0 to the top of the range; and "less than" is understood to include the value from 0 to 0. In general, the nomenclature and techniques used in cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are well known and commonly employed in the art. Nomenclature and technology. Unless otherwise stated, the methods and techniques of the present invention are generally performed in accordance with conventional methods well known in the art and methods of the present invention as generally described and discussed in the entire specification. . Enzymatic reactions and purification techniques are performed as commonly practiced in the art or as described herein, according to the manufacturer's instructions. The methods used in analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein, as well as laboratory materials and techniques (4) are well known and commonly used in the art, and (4) and techniques. Standard techniques for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. In order to make the invention easier to understand, the selected terms are defined as follows. The term "half-heart, "semi-Ig molecule" or "semi-Ig binding protein" is used interchangeably to refer to the smallest structure having an antigen-binding domain (eg, 160877.doc -53-201247704 heavy-bond antigen-binding domain) at the c-terminus. An immunoglobulin-based binding protein conjugated to at least a portion of the N-terminus of the immunoglobulin cH3 domain, wherein the ch3 domain preferably includes one or more mutations in the CH3/CH3 contact region to inhibit ch3_ch3: polymerization. Half 18 may be referred to herein as a "binding protein." The semi-fourth functional antigen binding site 'this binding site can be provided by a single heavy chain antigen binding chain' or form a functional antigen binding site by complementary pairing with a light bond antigen binding domain including a heavy bond antigen binding domain Click to provide. In addition to the minimal structure of the heavy chain antigen binding domain joined to at least a portion of the CH3 domain, the semi-Ig molecules of the invention may also include other domains. For the sake of simplicity, the antigen present in the peptide comprising the CH3 domain, the binding domain is referred to as the heavy bond antigen binding domain, although the antigen binding domain need not be derived from the antibody heavy chain. Exemplary embodiments of the half include, but are not limited to: VD1-(X1)n-X2; VD1-(X1)n-VD2-(X2)n-X3; and VD1-(X1)n-VD2-(X2) n-VD3-(X3)n-X4; In the exemplary embodiment 'each VD (or VDH) line is independently selected from the group consisting of a heavy chain variable domain, a double heavy chain variable domain, a triple heavy chain variable domain, and a light Key variable domain, double light chain variable domain, triple light chain variable domain, combination of heavy chain variable domain and light bond variable domain, combination of two heavy bond variable domains and light bond variable domains, heavy A combination of a chain variable domain and two light bond variable domains, a domain antibody, a Lucifer antibody, a scFv, a receptor, and a backbone antigen binding protein. As used herein, a combination of one or more heavy chain variable domains and one or more light bond variable domains is understood to be a covalent bond (eg, by a peptide bond) either directly or via a non-variable domain peptide sequence ( Such as a linker sequence) in any order (eg heavy-light-key; light-key; heavy 160877.doc-54-201247704 chain-heavy chain-light chain; heavy chain-light chain_heavy chain; light chain- Heavy chain_heavy chain; heavy chain_light chain-light chain; light chain-heavy chain-light chain; light chain_light chain-heavy chain) Two or more domains bonded to each other. In certain embodiments, the linker is sufficiently long to allow for complementary pairing between the light and heavy chains. In certain embodiments, the length of the linker is insufficient to allow for complementary pairing between the light and heavy bonds. In certain embodiments, 'unrelated to the length of the linker, the light bond does not match the heavy chain and does not form a complementary pair or the light and heavy chains can form a complementary pair without being joined by the peptide linker. For simplicity, a VD herein may be referred to as a variable domain in the context of an exemplary embodiment of a binding protein peptide provided herein, but in the context of a peptide comprising a heavy chain antigen binding domain, it is understood to include as provided herein. Heavy chain variable domain, double heavy chain variable domain, triple heavy chain variable domain, light chain variable domain, double light chain variable domain, triple light chain variable domain, heavy chain variable domain and light chain variable Combination of domains, combination of two heavy chain variable domains and light chain variable domains, combination of heavy chain variable domains and two light chain variable domains, domain antibodies, camelid antibodies, scFv, receptors and backbone antigens Binding protein. In certain embodiments, each VD (or VDH) is independently selected from the group consisting of a heavy chain variable domain, a light chain variable domain, a domain antibody, a camelid antibody, a scFv, a receptor, and a backbone antigen binding protein. . In an exemplary embodiment, the identity of each X depends on its position in the semi-Ig binding protein. The most C-terminal X (for example, X2 in the first embodiment, X3 in the second embodiment, and χ4 in the third embodiment) may include a polypeptide having at least a portion of the CH3 domain in the CH3/CH3 contact region. The residue within the residue has at least one mutation that inhibits the dimerization of CH3-CH3. The second-end C-terminal enthalpy (for example, XI in the first embodiment, Χ2 in the second embodiment, and X3 in the third embodiment 160877.doc • 55-201247704) may include a polypeptide, a CH1 domain, a CH2 domain, a CH1, and CH2 domain, or linker. In some embodiments, the second and last C-ends X further comprise a sequence of hinge regions that can be a linker. In some embodiments, the hinge region is preferably between the CH1 and CH2 domains when both the CH1 and CH2 domains are present. When there is a CH1 and CH3 domain and no CH2 domain exists, the hinge region is preferably between the CH1 and CH3 domains. When there are CH2 and CH3 domains and no CH1 domain exists, the hinge region is preferably located at the N-terminus of the CH2 domain. When there are other defects (e.g., XI in the second embodiment and Χ1 & Χ2 in the third embodiment), it includes a linker sequence.

Each Ν is independently selected from 〇 and 丨. In certain embodiments, the half provided herein includes "addition of other sequences (eg, linker sequences, functional sequences) in a peptide comprising a heavy chain antigen binding domain. In certain embodiments, the semi-Ig binding proteins provided herein are only Included above are the domains raised in peptides comprising a heavy chain antigen binding domain. In certain embodiments, χ does not include a CH1 domain and/or a CH2 domain. In certain embodiments, the binding protein does not include a CH1 domain and / or (: 112 domain. Furthermore, the semi-Ig binding protein may comprise a squamous bond comprising at least a light chain antigen binding domain. For simplicity, the CH3 domain is not included and preferably does not include c.

The antigen-binding domain present in the peptide of the domain or C Η 2 domain is called the light chain antigen-binding domain. The domain does not need to be derived from the antibody light chain. A light chain antigen binding domain is understood to include a light bond variable domain, a dual light chain variable domain, a triple light bond variable domain, a heavy chain variable domain, a double heavy chain variable domain, a triple heavy chain variable domain heavy bond. Combination of variable domain and light bond variable domains, combination of two heavy bond variable domains and light chain variable domains, combination of heavy chain variable domains with two light chain variable domains, camelid antibody, white. As used herein, a combination of a domain antibody, a scFv, a receptor, and a backbone antigen binding one or more heavy chain variable domains and one or more light chains can be understood as being by non- Covalent bonds (eg, hydrogen bonds, electrostatic interactions); or covalent bonds (eg, by peptide bonds) either directly or via a non-variable domain peptide sequence 诸如j (such as a linker sequence) in any order (eg, heavy chain-light Chain; light chain - heavy chain; heavy chain - heavy chain - light chain; heavy chain - light chain - heavy chain; light chain - heavy chain - heavy chain; heavy chain - light chain - light chain; Light chain; light chain - light chain - heavy chain) Two or more domains that are joined to each other. In some implementations, the material is long enough to allow complementary pairing between the light and heavy chains. In certain embodiments, the length of the linker is insufficient to allow for complementary pairing between the light and heavy chains. In some embodiments, regardless of the length of the linker, the light bond does not match the heavy chain and does not form a complementary pair. Alternatively, the light and heavy chains can form complementary pairs without being joined by a peptide linker. In certain embodiments, the variable domains that are combined with one another are present in a single compound peptide. In certain embodiments, the variable domains that are combined with each other are present in two (or more) polypeptide strands. Thus, a light chain variable domain may comprise a sequence that binds to an antigen or target, either alone or in combination with a heavy chain antigen binding domain; or both. In addition to the minimal structure of the light chain antigen binding domain, the semi-Ig binding φ $ white second peptide of the present invention may also include other domains. Exemplary examples of the second peptide of the semi-ig-binding protein include, but are not limited to: VD1-(X1)n; VD1-(X1)N-VD2-(X2)N; and VD1-(X1)n- VD2-(X2)n-VD3-(X3)n; In an exemplary embodiment, each VD (or VDL) is independently selected from the group consisting of a light chain variable domain, a double light chain variable domain, and a triple light chain. Variable domain, heavy chain variable domain, double heavy chain variable domain, triple heavy chain variable domain, heavy chain variable domain and light 160877.doc -57-201247704 bond variable domain combination, two heavy chain variable Combination of domain and light chain variable domains, combination of heavy chain variable domain and two light chain variable domains, camelid antibody, domain antibody, cyclin antibody, scFv, receptor and backbone antigen binding protein. For simplicity, a VD herein may be referred to as a variable domain in the context of an exemplary embodiment provided herein but in the context of a peptide comprising a light chain antigen binding domain, it is understood to include a light chain variable as provided herein. Domain, double light key variable domain, 3

, 1 I heavy and light bond variable domain, heavy chain variable domain, double heavy chain variable domain, triple heavy bond variable domain, heavy chain variable domain and light chain variable domain combination, two heavy bonds variable Combination of a domain and a light chain variable domain, a combination of a heavy chain variable domain and two light chain variable domains, a camelid antibody, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. In an exemplary embodiment, the identity of each X depends on its position in the second peptide of the semi-binding protein. The most C-terminal (e.g., XI in the first embodiment, Χ2 in the second embodiment, and χ3 in the third embodiment) may include a light key 悝 localization. When there are other defects (e.g., XI in the second embodiment and X1 and Χ2 in the third embodiment), it includes a linker sequence. Each ν is independently selected from 〇 and 1. In certain embodiments, a half Ig binding protein provided herein includes additional sequences (e.g., a linker sequence, a functional sequence) in a peptide comprising a light bond antigen binding domain. In certain embodiments, the semi-Ig binding protein provided herein includes only the domains set forth above in a peptide comprising a light chain antigen binding domain. In certain embodiments, a peptide containing a light bond antigen binding domain is V. The number of domains is the same as the number of v domains in the peptide containing the heavy chain antigen binding domain. In certain embodiments, the number of V domains in a peptide comprising a light chain antigen binding domain is different from the number of V domains in a peptide comprising a heavy chain antigen binding domain. In certain embodiments 160877.doc-58-201247704, the X in the peptide containing the light chain antigen binding domain does not include the CH1 domain and/or the CH2 domain. In certain embodiments, the binding protein does not include the chi domain and/or the CH2 domain. The X in the peptide containing the light chain antigen binding domain does not include the ch3 domain. Figure 1A provides a schematic representation (top row) of antibodies and immunoglobulin bivalent molecules based on various formats which can be used as a basis for designing a semi-Ig binding protein (bottom row). As used herein, in the case of a semi-Ig binding protein, the monovalent Ig refers to a one-arm Ig' and the bivalent Ig refers to a two-arm Ig, and does not relate to the number of binding sites present. The peptide containing the heavy bond antigen binding domain is paired with a peptide containing a light chain antigen binding domain as shown in various semi-Ig binding protein formats. In the schematic of the semi-Ig binding protein, the semi-DVD binding protein, and the semi-TVD binding protein, the light and heavy chain variable domains are shown as forming complementary pairs of single antigen binding domains. In a semi-RAb-Ig binding protein, the variable domain adjacent to the constant region is shown as a complementary pair and the receptors in the peptide containing the light chain antigen binding domain and the peptide containing the heavy chain antigen binding domain do not interact and An independent binding site is formed. When a semi-Ig binding protein comprises two complementary sequences to form a single binding site comprising two peptides and a sequence that forms an independent binding site on each peptide, preferably the complementary sequence is adjacent to the constant domain and the independent binding site is kept constant. area. Figure IB shows the production of an exemplary half-ig-binding protein from a parental IgG antibody. The naturally occurring IgG dimerizes via a specific interaction domain via interaction of the CH3 domain. The antibody chain is also held together via disulfide bonds present in the hinge region of the antibody. The semi-Ig binding proteins of the present invention are typically produced using known recombinant DNa techniques and antibodies having known nucleotide and/or amino acid sequences, however, the particular method of producing the semi-Ig binding proteins of the present invention does not limit the invention. Mutagenesis (usually site-directed mutagenesis, but also random induction 160877.doc • 59-201247704 variant) is used to alter one, two or three cysteines that form disulfide bonds in the hinge region. Other amino acids to prevent disulfide bond formation, and/or to disrupt sequences important for interactions between CH3 domain sequences (by making 1, 2, 3, 4, 5 in the ch3 domain) , 6, 7, 8, or more residues are mutated). In some embodiments, CH3 domain dimerization can be achieved by truncating the CH3 domain. As used herein, in certain embodiments, when a wild type constant domain of a species (eg, a human or mouse IgG sequence or variant thereof) binds to protein A, in the case of a half ig binding protein, at least a portion of the CH3 domain It is understood that the CH3 domain is sufficient to allow the half Ig binding protein to bind to protein A. As used herein, in certain embodiments, it is understood that portions of the CH3 domain are at least about 70% identical to the full length CH3 domain from the same species, at least about 75% - resulting in at least about 80% - at least about 85% Consistent, at least about 9〇% consistent, at least about 95°/. Consistent, at least about 98% consistent or at least about 99% consistent. In certain embodiments, at least a portion of the CH3 domain is understood to be a polypeptide comprising at least 20 amino acid residues of the wild type CH3 domain. In certain embodiments, at least a portion of the CH3 domain is understood to be a polypeptide comprising at least 20 contiguous amino acid residues (possibly adjacent or possibly not adjacent) of the wild-type sputum domain. As used herein, in certain embodiments, at least a portion of the CH3 domain is understood to interact with the & acceptor or RnFc receptor (: 113 domain. In some embodiments, at least a portion of the CH3 domain includes the listed features Various combinations. As used herein, "CH3-CH3 dimerization" is understood to mean the specific interaction of two CH3 domains with each other. The beta-specific interaction can be driven by the amino acids in both the CH3 domain and the CH3 domain. The interaction may be covalently promoted by the two CH3 domain portions of the inconsistent chain of the bacterioglobulin constant chain (for example, a disulfide bond in the hinge region that closely contacts the CH3 domain) or non-covalent. The interaction is driven. The CH3-CH3 one-polymerized contact zone is a contact zone defined by Dall'Acqua (Biochem. 37: 9266-9273, 1998, which is incorporated herein by reference) and includes the CH3 domain according to Kabat Numbered amino acid positions below: Q347, Y349, T350, L351, T366, L368, K370, K392, T394, P395, V397, L398, D399, F405, Y407 and K409. The term "polypeptide" as used herein. Means any polymeric bond of an amino acid. The terms "peptide" and "protein" are used interchangeably with the term polypeptide and also refer to the polymeric chain of an amino acid. The term "polypeptide" encompasses natural or artificial proteins, protein fragments and protein sequences. Polypeptide analogs. Polypeptides may be monomers or polymers. Unless otherwise indicated, the use of "polypeptides" herein is intended to encompass polypeptides, fragments and variants thereof (including fragments of variants). The fragment optionally contains at least one adjacent or non-linear epitope of the polypeptide. The exact boundaries of the at least one epitope can be determined using the general techniques of the art. The fragment comprises at least about 5 adjacent amino acids. , such as at least about 10 adjacent amino acids, at least about 15 adjacent amino acids, or at least about 20 adjacent amino acids. The variant system of the polypeptide is as described herein. The term "isolated protein" or " An isolated polypeptide" is a protein or a kiln peptide that is not associated with a natural association component associated with its native state, depending on the origin or source of the derivative; Contains no other eggs from the same species. 160877.doc •61 · 201247704 White matter; expressed by cells of different species; or not found in nature. Therefore, chemically synthesized or synthesized in cellular systems other than cells of natural origin The eve will be "isolated" from its natural association component. Protein purification techniques well known in the art can also be used to isolate the protein substantially free of natural association components. For example, The protein determined by conventional methods in the art may be 90% pure, 95% pure, 97% pure, 98% pure, 99% pure or 99% pure, ie, free of other naturally occurring proteins or nucleic acids. Component. The term "recovery" as used herein refers to the process of separating a chemical substance, such as a polypeptide, substantially free of natural association components, for example, using protein purification techniques well known in the art. As used herein, "biological activity" refers to any one or more of the intrinsic biological properties of a molecule (a molecule found in vivo or a molecule provided or realized by recombinant means). Biological properties include, but are not limited to, binding to a receptor, inducing cell proliferation, inhibiting cell growth, inducing other cytokines, inducing apoptosis, and enzymatic activity. Biological activity also includes the activity of Ig molecules. The term "specifically binds" as used herein in reference to the interaction of a binding protein, antibody, protein or peptide with a second chemical means that the interaction depends on the particular structure on the chemical (eg, an antigenic determinant or epitope) The presence of; for example, antibodies generally recognize and bind to specific protein structures rather than proteins. If the antibody is specific for the epitope "A", the presence of a molecule containing the epitope (or free, unlabeled A) in the reaction containing the labeled "A" will reduce the labeled A binding to the antibody. The amount. 160877.doc • 62- 201247704 As used herein, the term "antibody" broadly refers to any immunoglobulin (Ig) molecule comprising four polypeptide chains (two heavy (H) chains and two light (L) chains), or It retains any functional fragment, mutant, variant or derivative of the essential epitope binding characteristics of the Ig molecule. Such mutant, variant or derived antibody formats are known in the art, and non-limiting examples thereof are discussed below. In full length antibodies, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region contains three domains, CHI, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region contains a domain CL. The VH and VL regions can be further subdivided into highly denatured regions (referred to as complementarity determining regions (CDRs), which alternate with more conserved regions (referred to as framework regions (FR)). Each VH and VL consists of three CDRs and four FRs arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The immunoglobulin molecule can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), classes (e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2) or subclasses. The term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain which can be produced by papain digestion of intact antibodies. The Fc region may be a native sequence Fc region or a variant Fc region. The Fc region of an immunoglobulin generally comprises two constant domains (CH2 domain and CH3 domain), and optionally comprises a CH4 domain" substitution amino acid residue in the Fc portion. The function of changing the antibody effect is known in the art (U.S. Patent Nos. 5,648,260 and 5,624,821). The Fc portion of the antibody mediates several important effector functions, such as cytokine induction, antibody dependence • 63·160877.doc 201247704 Sex cell-mediated cytotoxicity (adcc), phagocytosis, complement dependent cytotoxicity (CDC), and Half-life/clearance rate of antibodies and antigen-antibody complexes. Depending on the therapeutic goal, 'in some cases' these effector functions are required for therapeutic antibodies, but in other cases, such effector functions may be unnecessary or even deleterious. Certain human IgG isotypes (especially igG1 and IgG3) mediate ADCC and CDC via binding to FcyR and complement Clq, respectively. The neonatal Fc receptor (FcRn) is a key component in determining the half-life of the antibody cycle. In another embodiment, at least one amino acid residue in the constant region of the antibody (e.g., the Fc region of the antibody) is displaced such that the effector function of the antibody is altered. Dimerization of two identical heavy bonds of immunoglobulins is mediated by dimerization of the CH3 domain and is stabilized by disulfide bonds in the hinge region (Huber et al. (1976)

Nature 264: 415-20; Thies et al. (1999) J. Mol. Biol. 293: 67-79). Mutation of the cysteine residues in the hinge region to prevent the formation of heavy chain-heavy chain disulfide bonds will depolymerize the CH3 domain to destabilize. Residues responsible for CH3 dimerization have been identified (Dall'Acqua (1998) Biochem. 37: 9266-73). Therefore, it is possible to produce a monovalent half-ig binding protein. Interestingly, these monovalent half-ig-binding proteins of both IgG and IgA subclasses have been found in nature (Seligman (1978) Ann. Immunol. 129: 855-70; Biewenga et al., (1983) Clin. Exp. Immunol. 51: 395-400). The FcRn:Ig Fc region has been determined to have a stoichiometry of 2:1 (West et al. (2000) Biochem. 39: 9698-708) and half of the Fc is sufficient to mediate FcRn binding (Kim et al. (1994) Eur. J Immunol. 24: 542-548, which is incorporated herein by reference. Mutations that disrupt the dimerization of the CH3 domain may not have a major adverse effect on their FcRn binding, since residues important for CH3 dimerization are 160877.doc -64- 201247704 located within the CPI3 beta sheet structure & p gate P The region responsible for FcRn binding on the interface is located outside the CH2-CH3 domain and on the interface of the external interface P. &,, the half 1 § binding protein may have certain advantages in tissue penetration due to its small size compared to conventional antibodies. In the examples, the substitution region of the binding protein of the present invention (for example, & region) At least one amino acid residue to disrupt the dimerization of the heavy bond, thereby producing a semi-Ig binding protein.

As used herein, the term "antigen-binding portion" of an antibody (or simply an antibody file) refers to a plurality of fragments of a binding protein that retain the ability to specifically bind to an antigen. For example, antigen binding of antibodies has been shown to be performed by fragments of full length antibodies. Such binding protein embodiments can also be bispecific, dual specific or multispecific formats; they specifically bind to two or more different antigens. Examples of the binding fragments encompassed by the term "antigen-binding portion" of an antibody include: (i) a Fab fragment, that is, a monovalent fragment consisting of VL, VH, CL, and CH1 domains; (u) a F(ab,)2 fragment. , that is, a bivalent fragment comprising two Fab fragments joined by a disulfide bridge in the hinge region; indicating that the disulfide bond is sufficient to mediate dimerization; (iii) an Fd fragment consisting of the VH and CH1 domains; Fv fragment's vl and VH domain group from a single arm of the antibody

a (v)dAb fragment (Ward (1989) Nature 341:544-546; and PCT Publication No. WO 90/05144 A1), which comprises a single variable domain; and (vi) an isolated complementarity determining region (CDR) In addition, although the two domains of the Fv fragment (VL and VH) are encoded by independent genes, they can be joined by synthetic linkers using recombinant methods, enabling the preparation of a single protein chain in which the VL region and the VH region are paired to form a monovalent molecule. (referred to as single-chain Fv (scFv); see, for example, Bird et al, (1988) Science 242: 423-426; and Huston et al, (1988) 160877. doc • 65-201247704

Proc. Natl. Acad. Sci. USA 85:5 879-5 883). Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. "Other forms of single-chain antibodies, such as bifunctional antibodies, are also contemplated. A bifunctional antibody is a bivalent, bispecific antibody in which the VH domain and the VL domain are expressed on a single polypeptide chain, but are used in a linkage that is too short to allow pairing between the two domains on the same bond, thereby forcing such The domain is paired with a complementary domain of another strand and forms two antigen binding sites (see, eg, Holliger, P. et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ Et al. (1994) Structure 2: 1121-1123). Such antibody binding moieties are known in the art (Kontermann and Dubel ed., Antibody Engineering (2001) Springer-Verlag. New York. page 790 (ISBN 3-540-41354-5)). In addition, a 'single bond antibody also includes a "line antibody" comprising a pair of conjugated Fv segments (VH-CHl-VH-CHl) that form a pair of antigen binding regions together with a complementary light chain polypeptide (Zapata et al., 1995). ) Protein Eng. 8(10): 1057-1062; and U.S. Patent No. 5,641, 87). As used herein, a heavy chain antigen binding domain (referred to herein as VD or VDH) is intended to include a heavy chain variable domain, a double heavy chain variable domain, a triple heavy chain variable domain, a light chain variable domain, a double light chain Variable domain, triple light chain variable domain, combination of heavy bond variable domain and light chain variable domain, combination of two heavy chain variable domains and light bond variable domains, heavy chain variable domain and two light chains Combinations of variable domains, domain antibodies, camelid antibodies, scFv, receptor and backbone antigen binding proteins. It will be appreciated that the heavy chain antigen binding domain may or may not be independent of the paired light chain, double light chain or triple light 160877.doc • 66 - 201247704 bond antigen present on the second polypeptide of the binding protein of the invention as appropriate . For example, it is contemplated that the domain antibody, scFv or receptor will be independent; any amino acid sequence on the haptic bond will bind to the target. When the T protein of the present invention forms a functional antigen binding site, if the heavy chain antigen binding domain does not specifically bind to the target antigen independently (ie, does not provide a functional antibody binding site separately), then there should be a second The polypeptide provides a complementary light bond variable domain to provide a functional antibody binding site. As used herein, a light chain antigen binding domain (referred to herein as vd or 乂〇[) is intended to include a light chain variable domain, a dual light chain variable domain, a triple light chain variable domain, a ^^ bond variable domain, a double a heavy chain variable domain, a triple heavy chain variable domain, a combination of a heavy bond variable domain and a light chain variable domain, a combination of two heavy chain variable domains and a light chain variable domain, a heavy chain variable domain and two The light chain variable domain group, the Lucifer antibody, the domain antibody, the roller antibody, the scFv, the receptor and the backbone antigen binding protein should understand that the 'light chain antigen binding domain may or may not be present independently of this a pair of heavy chains on the other polypeptide of the binding protein, double

The heavy or triple heavy chain binds to the antigen. For example, it is contemplated that a domain antibody, MV or body will bind to any amino acid sequence independent of the second polypeptide chain. ,. . As used herein, "VD" alone is understood to mean a heavy chain antigen binding domain or a light chain antigen binding domain, unless the context clearly dictates otherwise. The term "multispecific binding protein" refers to a binding protein that binds two or more related or unrelated targets. As used herein, bispecific and multispecific are also understood to have two or more binding sites for the same antigen or epitope. The semi-DVD 1 § binding protein may be monospecific, ie bind to one antigen; or be multispecific, ie bind to two 160877.doc •67-201247704 or more than two antigens (see eg Figure ΙΑ) ^ For example, a semi-Ig binding protein derived from a naturally occurring bivalent IgG will be monospecific. A half Ig binding protein derived from a DVD binding protein can be monospecific or bispecific. Each half of the DVD-Ig binding protein comprises a heavy binding protein polymorphism and a light chain DVD binding protein polypeptide' and two antigen binding sites. Each of the binding sites includes a heavy chain variable domain and a light chain variable domain, wherein each of the antigen binding sites has a total of six CDRs involved in antigen binding. However, a half ig binding protein can also be bispecific by including two heavy bond antigen binding domains, wherein each heavy chain antigen binding domain is independent of another heavy chain antigen binding domain or a complementary light chain binding target antigen. In another embodiment, a semi-Ig binding protein can be bispecific by including a heavy bond antigen binding domain and a light bond antigen binding domain, wherein each antigen binding domain binds to the antigen of interest independently of the other antigen binding domain. In this particular embodiment, each antigen binding domain can be, for example, s c F v or a receptor. As illustrated in Figure 1A, the trispecific half ig binding protein can also be derived from tvd binding protein and RAb. In certain embodiments, a variable domain can bind its antigen at the same time. In certain embodiments, the antigen competes for binding to a variable domain. Other bispecific, trispecific, tetraspecific, etc. halves can be generated by combining different heavy chain antigen binding domains and light chain antigen binding domains using conventional molecular biotechnology, such as the techniques provided herein. Methods for detecting the binding of a semi-Ig binding protein simultaneously or independently to one or more antigens of interest can also be carried out using conventional methods, such as those provided herein. As used herein, "double variable domain immunoglobulin" or DVD-Ig" and its analogs are included in the drawings and are shown in U.S. Patent Publication Nos. 20100260668 and 20090304693, all of which are incorporated by reference. The manner of 68-201247704 is incorporated herein, including the binding proteins of the structures provided in the Sequence Listing). DVD-IgTM comprises a pair of heavy chain DVD polypeptides and a light chain DVD polypeptide, wherein each pair of heavy and light chains provides two antigen binding sites. Each binding site includes a total of six CDRs involved in antigen binding per antigen binding site. DVD-Ig typically has two arms that are combined with each other and at least partially dimerized by the CH3 domain, wherein each arm of the DVD is bispecific, thereby providing an immunoglobulin having four binding sites. As used herein, it is to be understood that "triple variable domain immunoglobulin" or "TVD-Ig" and its analogs include U.S. Provisional Patent Application No. 61, which is incorporated herein by reference in its entirety. / 426 133 ; and the binding protein of the structure provided by the US Patent Application Serial No. Both applications are in the name of the same assignee. All of the above-mentioned applications (including the sequence listing) are incorporated herein by reference. β τν] χ-protein contains a pair of heavy chain-binding protein polypeptides And the _TVD binding protein is ruined, wherein each pair of heavy and light bonds provides an antigen binding site. Each binding site comprises a total of "ten six antigen-binding CDRs per antigen binding site φ. The TVD binding protein may have two arms that are dimerized with each other, at least in part, from the CH3 domain, wherein the protein. Each arm is trispecific to provide a binding protein with a stone binding site. "As used herein, it is understood that "receptor-antibody immunoglobulin" or two Ab'Ig" and its analogs include Figure 1A. The proteins shown in the structure are also shown (see also XJS 2002/0127231, the entire contents of which are incorporated herein by reference). RAbIg comprises heavy chain polypeptides 160877.doc-69 - 201247704 and light chain RAb polypeptides which together form a total of three antigen binding sites. By pairing the heavy and light chain antibody variable domains present in each of the heavy chain RAb polypeptide and the light chain RAb polypeptide to form a single binding site with a total of 6 CDRi to provide a first antigen binding site, forming an antigen binding site point. The double bond RAb polypeptide and the light chain RAb polypeptide each comprise a receptor sequence that independently binds to the ligand, thereby providing a second and third "antigen" binding site. RAb-Ig typically has two arms that bind to each other and at least partially dimerize from the CH3 domain, wherein each arm of RAb-Ig is trispecific, thereby providing an immunoglobulin having six binding sites. The term "bispecific antibody" as used herein and different from "bispecific semi-Ig binding protein" or "bispecific (semi-Ig) binding protein" refers to a full-length antibody produced by the following method: a four-source fusion tumor (quadroma) technique (see Milstein, C. and Cuello, AC (1983) Nature 305 (5934): pp. 537-540); chemical binding of two different monoclonal antibodies (see Staerz, UD et al., (1985) Nature 3 14 (6012): 628-631); or introduce a knob-into-hole or similar method in the Fc region that does not inhibit the mutation of CH3-CH3 dimerization (see Holliger, P. et al. (1993) Proc. Natl. Acad. Sci USA 90(14): 6444-6448), which produces a variety of different immunoglobulin species', only one of which is a functional bispecific antibody. According to molecular function, a bispecific antibody binds an antigen (or epitope) on one of its two binding arms (a pair of HC/LC) and on its second arm (a pair of different HC/LC) Binding to different antigens (or epitopes). According to this definition, a bispecific antibody has two different antigen binding arms (both in terms of specificity and CDR sequences) and is monovalent for each antigen to which it binds. 160877.doc •70· 201247704 The term “bispecific antibody” as used herein and different from the bispecific semi-Ig binding protein or bispecific binding protein means that each of the two binding arms (one pair of HC/LC) can be used. A full length antibody that binds two different antigens (or epitopes) (see pct publication No. 02/02773). Thus, a bispecific binding protein has two identical antigen binding arms of the same specificity and the same CDR sequence and is bivalent for each antigen to which it binds. A "functional antigen binding site" of a binding protein is a binding protein that binds to a target, an antigen or a ligand. The antigen binding affinity of the antigen binding site is not necessarily as strong as the parent binding protein from which the antigen binding site is derived, but the ability to bind the antigen must be able to use any of a variety of methods known for assessing binding of the binding protein to the antigen. One is measuring. Furthermore, the antigen binding affinities of the various antigen binding sites of the multispecific binding proteins herein need not be the same in amount. The term "cytokine" is a generic term for a protein that is released by one cell population and acts as an intercellular mediator on another cell population. Examples of such cytokines are lymphoid hormones, mononuclear hormones and traditional polypeptide hormones. Cytokines include growth hormones such as human growth hormone, N_曱 thiamine thioglycan human growth hormone and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; relaxin; glycoprotein hormones, such as Follicle stimulating hormone (FSH), gonadotropin (TSH) and luteinizing hormone (LH) 'liver growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor alpha and tumor necrosis Factor_β;mullerian-inhibiting substance; mouse gonadotropin-related peptide; inhibin; activin; vascular endothelial growth factor; integrin; 160877.doc -71 · 201247704 thrombopoietin (ΤΡΟ Nerve growth factor, such as NGF-α; platelet growth factor; placental growth factor; transforming growth factor (TGF), such as TGF-ct and TGF-β; insulin-like growth factor-1 and insulin-like growth factor-11; red blood cells Creatine (ΕΡΟ); osteoinductive factor; interferon, such as interferon-α, interferon-β and interferon-γ; community stimulating factor (CSF), such as Phagocyte-CSF (M-CSF), granulocyte macrophage-CSF (GM-CSF) and granulocyte-CSF (G-CSF); interleukin (IL), such as IL-1, IL-2, IL -3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-15, IL-18 , IL-21, IL-22, IL-23 and IL-33; tumor necrosis factors such as TNF-α or TNF-β; and other polypeptide factors, including LIF and kit ligands (KL) » as used herein, The term cytokine includes proteins from natural sources or recombinant cell cultures, as well as biologically active equivalents of native sequence cytokines.

The term "linker" is used to mean a polypeptide comprising two or more peptide-bonded amino acid residues and for ligation of one or more antigen-binding portions. Such linker polypeptides are well known in the art (see, for example, Holliger, P. et al., (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448; Poljak, RJ et al., (1994) Structure 2:1 121-1123). Exemplary linkers include, but are not limited to, ASTKGPSVFPLAP (SEQ ID NO: 46); ASTKGP (SEQ ID NO: 48); TVAAPSV FIFPP (SEQ ID NO: 50); TVAAP (SEQ ID NO: 52); AKTTPKLEEGEFSEAR (SEQ ID NO: 94); AKTTPKLEEGE FSEARV (SEQ ID NO: 95); AKTTPKLGG (SEQ ID NO: 96); SAKTTPKLGG (SEQ ID NO: 97); SAKTTP (SEQ ID 160877.doc • 72· 201247704

NO: 98); RADAAP (SEQ ID NO: 99); RADAAPTVS (SEQ ID NO: 100); RADAAAAGGPGS (SEQ ID NO: 101); RADAAAA(G4S)4 (SEQ ID NO: 102); SAKTTPKLEEGEF SEARV (SEQ ID NO: 103); ADAAP (SEQ ID NO: 104); ADAAPTVSIFPP (SEQ ID NO: 105); QPKAAP (SEQ ID NO: 106); QPKAAPSVTLFPP (SEQ ID NO: 107); AKTTPP (SEQ ID NO: 108); AKTTPPSVTPLAP (SEQ ID NO: 109); AKTTAP (SEQ ID NO: 110); AKTTAPSVYPLAP (SEQ ID NO: 111); GGGGSGGGGSGGGGS (SEQ ID NO: 112); GENKVEYAPALMALS (SEQ ID NO: 113); GPAKE LTPLKEAKVS (SEQ ID NO: 114); GHEAAAVMQVQYPAS (SEQ ID NO: 115); TVAAPSVFIFPPTVAAPSVFIFPP (SEQ ID NO: 116); and ASTKGPSVFPLAPASTKGPSVFPLAP (SEQ ID NO: 117). An immunoglobulin constant domain refers to a heavy or light chain constant domain. Human IgG heavy and light chain constant domain amino acid sequences are known in the art. As used herein, the term 'monoclonal antibody' or 'mAb' refers to an antibody obtained from a population of substantially homogeneous antibodies, ie, the individual antibodies that make up the population are identical except for the naturally occurring mutations that may be present in minor amounts. . Individual antibodies are highly specific for a single antigen. Furthermore, unlike the multi-drug antibody preparations which typically include different antibodies against different determinants (antigenic determinants), each mAb is directed against a single determinant on the antigen. The modifier "single plant" should not be construed as requiring the production of antibodies by any particular method. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from the germline immunoglobulin sequences of human 160877.doc • 73-201247704. Human antibodies of the invention may, for example, include amino acid residues not encoded by human germline immunoglobulin sequences in CDRs and, in particular, CDR3 (eg, by in vitro random or site-specific mutagenesis or by in vivo Mutations introduced by somatic mutations). However, as used herein, the term "human antibody" is not intended to include antibodies that have been ligated into the human framework sequence from the germline of another mammalian species, such as a mouse. The term 'recombinant human antibody' as used herein is intended to include all human antibodies prepared, expressed, produced or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into a host cell (see Section π C below). Further described in the above), antibodies isolated from recombinant human antibody libraries (Hoogenboom, HR (1997) TIB Tech 15: 62-70; Azzazy, Η· and Highsmith, WE (2002) Clin. Biochem. 35: 425-445 ; Gavilondo, JV and Larrick, JW (2002)

BioTechniques 29: 128-145; Hoogenboom, H. and Chames, P. (2000) Immun〇i. T〇cjay 21: 371-3 78, which is incorporated herein by reference), from human immunoglobulin genes An antibody isolated from a transgenic animal (eg, a mouse) (see Taylor, LD et al., (1992) Nucl. Acids

Res. 20: 6287-6295; Kellermann, SA· and Green, LL (2002) Cur. 〇pin. in Biotechnol. 13: 593-597 ; Little, M. et al., (2000) Immun〇l. Today 21: 364-370), or an antibody produced, expressed, produced or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. The recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. 160877.doc • 74· 201247704 However, in certain embodiments, in vitro mutagenesis is performed on such recombinant human antibodies (or in vivo somatic mutagenesis when using a human ig sequence of a transgenic animal) And, therefore, the amino acid sequence of the VH and VL regions of the recombinant antibody is derived from the VH and VL sequences of the human germline and is related to the VH and VL sequences of the human germline, but may not be naturally present in the human antibody. Sequence within the pedigree. "Affinity maturation" antibodies are antibodies that have one or more changes in one or more CDRs that result in an improvement in the affinity of the antibody for the antigen compared to a parent antibody that does not have a change in the other. The exemplary affinity matured antibody will have a sensitivity to the target antigen or to the picomolar concentration. Affinity matured anti-systems are prepared by procedures known in the art. Marks et al. (1992) Bio/Technology 1 : 779-783 describe affinity maturation by VH and VL domain shuffling. The following documents describe random mutagenesis of CDR and/or framework residues: Barbas et al, (1994) Proc Nat. Acad. Sci. USA 91: 3809-3813; Schier et al, (1995) Gene 169: 147- 155; Yelton et al., (1995) J. Immunol. 155: 1994-2004; Jackson et al., (1995) J. Immunol. 154(7): 3310-9; and Hawkins et al., (1992) J. Mol .

Biol. 226: 889-896; and selective mutation of an activity-enhancing amino acid residue at a selective mutagenesis position, contact or hypermutation position is described in U.S. Patent No. 6,914,128. The term "reporting antibody" refers to an antibody comprising a heavy chain and a light bond variable region sequence from one species and a constant region sequence from another species, such as having a murine heavy chain and a light chain variable region linked to human constant Antibody to the area. 160877.doc •75· 201247704 The term "CDR-grafted antibody" refers to a sequence comprising heavy and light chain variable regions from one species but wherein the sequence of one or more of the CDR regions of VH and/or VL is passed through another species An antibody that is substituted with a CDR sequence, such as an antibody having one or more murine CDRs (e.g., CDR3) that have been replaced by human CDR sequences in the murine heavy and light chain variable regions. The term "humanized antibody" refers to a sequence comprising a heavy bond and a light bond variable region from a non-human species (eg, a mouse), but wherein at least a portion of the VH and/or VL sequence has been altered to be more "humanoid", An antibody that is more similar to the variable sequence of the human germline. One type of humanized antibody is a CDR-implanted antibody in which human CDR sequences are introduced into non-human VH and VL sequences to replace corresponding non-human CDR sequences. A "humanized antibody" is also an antibody or a variant or derivative thereof that immunospecifically binds to a related antigen and comprises an FR region substantially having a human antibody amino acid sequence and a CDR region substantially having a non-human antibody amino acid sequence. , analog or fragment. As used herein, the term "substantially" in the context of a CDR refers to having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence of a CDR of a non-human antibody. - the CDRs of the amino acid sequence. A humanized antibody comprises substantially all of at least one and usually two variable domains (Fab, Fab', F(ab,)2, FabC, Fv), wherein all or substantially all of the CDR regions correspond to non-human immunoglobulins The CDR regions of the (ie, donor antibody) and all or substantially all of the FR regions are the Fr regions of the human immunoglobulin consensus sequence. In one embodiment, the humanized antibody also comprises at least a portion of an immunoglobulin Fc region (typically the Fc region of a human immunoglobulin). In some embodiments, a humanized antibody comprises a light chain and at least a variable domain of a heavy chain. The antibody may also include the CHI, hinge region, CH2, CH3 and CH4 regions of the heavy chain, 160877.doc -76· 201247704. In some embodiments, the humanized antibody contains only humanized light bonds. In some embodiments, the humanized antibody contains only a humanized heavy chain. In a particular embodiment, the humanized antibody only contains a humanized variable domain of a light chain and/or a humanized heavy chain. The terms "Kabat number", "Kabat definition" and "Kabat mark" are used interchangeably herein. The terms recognized in the art refer to amino acids which are more variable (i.e., hypervariable) compared to other amino acid residues in the heavy and light bond variable regions of the antibody or antigen binding portion thereof. Numbering system for residues (Kabat et al., (1971) Ann. NY Acad, Sci. 190:382-391; and Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, 5th edition, US Health and Safety US Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable region ranges from amino acid positions 31 to 35 of CDR1, amino acid positions 50 to 65 of CDR2, and amino acid positions 95 to 102 of CDR3. For the light chain variable region, the hypervariable region ranges from amino acid positions 24 to 34 of CDR1, amino acid positions 50 to 56 of CDR2, and amino acid positions 89 to 97 of CDR3. As used herein, the term "CDR" refers to a complementarity determining region within a variable sequence of an antibody. There are three CDRs in each of the variable regions of the heavy and light chains, and the three CDRs of each variable region are designated CDR1, CDR2 and CDR3. As used herein, the term "CDR set" refers to three CDRs that are present in a single variable region and can bind to one of a population of antigens. The exact boundaries of these CDRs have been defined differently for different systems. The system described by Kabat (Kabat et al., (1987; 1991) 160877.doc •77-201247704

Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md.) not only provides a clear residue numbering system for any variable region of an antibody, but also provides the exact definition of the three CDRs. Residue boundary. These CDRs can be referred to as Kabat CDRs. Chothia and colleagues (Chothia and Lesk (1987) J. Mol. Biol. 196:901 ·917 ; and Chothia et al., (1 989) Nature 342:877-883) found that some sub-parts within the Kabat CDR are Amino acid sequence levels are extremely diverse, and almost identical peptide backbone configurations are used. These subsections are designated as LI, L2 and L3 or HI, H2 and H3' where "L" and "H" represent the light key zone and the heavy key zone, respectively. These regions may be referred to as Chothia CDRs, which have boundaries that overlap with the Kabat CDRs. Other boundaries defining CDRs that overlap with Kabat CDRs have been described by Padlan (1995) FASEB J. 9: 133-139 and MacCallum (1996) J. Mol. Biol. 262(5): 732-45. Other CDR boundary definitions may not strictly follow one of the systems in this article, but will still overlap with the Kabat CDRs, although depending on the particular residue or group of residues or even the entire CDR does not significantly affect the prediction of antigen binding or experimental results, it may Shorten or lengthen. The methods used herein may utilize CDRs as defined by any of these systems, but certain embodiments use CDRs as defined by Kabat or Chothia. As used herein, the term "framework" or "framework sequence" refers to the variable sequence minus the remaining sequence of the CDRs. Since the exact definition of CDR sequences can be determined by different systems, the meaning of the framework sequences follows a correspondingly different interpretation. The six CDRs (CDR-L1, CDR-L2 and CDR-L3 of the light chain, and the CDR-H1, CDR-H2 and CDR-H3 of the heavy chain) also have a framework region on the light and heavy chains 160877.doc - 78-201247704 is divided into four sub-regions (FR1, FR2, FR3, and FR4) in each chain, wherein CDR1 is located between FR1 and FR2, CDR2 is located between FR2 and FR3, and CDR3 is located between FR3 and FR4. In the case where a specific sub-region is not designated as FR1, FR2, FR3 or FR4, the framework region refers to the combination FR in the variable region of the single immunoglobulin chain naturally present in the epoch. As used herein, FR represents one of four sub-regions, and FRs represents two or more sub-regions of the four sub-regions that make up the framing region. As used herein, the term "growth antibody gene" or "gene fragment" refers to a non-lymphocyte-encoded immunoglobulin sequence that has not undergone a genetic rearrangement and mutation to express the maturation process of a particular immunoglobulin. (See, for example, Shapiro et al., (2002) Crit. Rev. Immunol. 22(3): 183-200; Marchalonis et al., (2001) Adv. Exp. Med. Biol. 484: 13-30). One of the advantages provided by various embodiments of the present invention is based on the recognition that the germline antibody gene is more likely than the mature antibody gene to retain the essential amino acid sequence structure characteristic of the individual in the species, and thus when used to treat the species, It is too likely to be identified as coming from a foreign source. As used herein, the term "neutralization" refers to counteracting the biological activity of an antigen when the binding protein specifically binds to the antigen. In one embodiment, the neutralizing binding protein binds to an antigen/target (eg, a cytokine, kinase, growth factor, cell surface protein, soluble protein, phosphatase, or receptor ligand) and reduces its biological activity by at least about 20 %, 40%, 60%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more. The term "activity" includes activities such as the binding specificity and affinity of a half Ig for one or more antigens, targets or ligands. 160877.doc • 19- 201247704 The term "antigenic determinant" includes any polypeptide determinant that specifically binds to a binding protein immunoglobulin or a tau cell receptor. In certain embodiments, the antigenic sulfhydryl determinant comprises a chemically active surface group of a molecule (such as an amino acid, a sugar side chain, a fluorenyl group or an anthracenyl group), and in certain embodiments, It has specific three-dimensional structural features and/or specific charge characteristics. The antigenic determinant is the region of the antigen that is bound by the binding protein. An epitope is thus composed of an amino acid residue of a region known to bind to an antigen (or a fragment thereof) at a complementary site on a particular binding partner. An antigenic fragment may contain more than one antigenic thiol group. In certain embodiments, when an antibody recognizes its antigen of interest in a complex mixture of proteins and/or macromolecules, the antibody specifically binds to the antigen. For example, an antibody "binds to the same epitope" if it cross-competes (an antibody prevents the binding or regulation of another antibody). In addition, the structural definition of the epitope (overlapping, similar, identical) provides a lot of information, but because the functional definition covers structural (combination) and functional (adjustment, competition) parameters, it is usually more relevant. As used herein, the term 'surface plasmon resonance' refers to the detection of biosensor matrices by, for example, the use of the BIAcore® system (BIAcore International AB, a GE Healthcare company, Uppsala, Sweden and Piscataway, NJ). Changes in protein concentration to analyze optical phenomena of immediate biospecific interactions. For more description, see J5nsson, υ· et al., (1993) Ann. Biol. Clin. 51: 19-26; Jdnsson, U. et al., (1991) Biotechniques 1 1: 620-627; Johnsson, B. (1995) J. Mol. Recognit. 8: 125-131; and Johnnson, Β· et al., (1991) Anal. Biochem. 198: 268-277. 160877.doc • 80 - 201247704 As used herein, the term "κ〇η" is intended to mean that a binding protein (e.g., an antibody) associates with an antigen to form an association rate constant, e.g., an antibody/antigen complex, as known in the art. "Km" is also known as the term "association rate constant" or "ka" and is used interchangeably herein. This value indicating the rate of binding of the antibody to its target antigen or the rate of complex formation between the antibody and the antigen is also expressed by the following equation: antibody ("Ab") + antigen ("Ag") 4Ab_Ag. As used herein, the term "Koff" is intended to mean the dissociation rate constant of a binding protein (e.g., an antibody) that is dissociated from, for example, an antibody/antigen complex, as is known in the art. "K〇ff" is also known as the term "dissociation rate constant" or "kd", which are used interchangeably herein. This value indicates the rate at which the antibody dissociates from its target antigen or the Ab-Ag complex separates into free antibody and antigen over time, as shown by the following equation: Ab+Ag<-Ab-Ag. The term "equilibrium dissociation constant" or r KD as used interchangeably herein refers to the value obtained at equilibrium in titration or by dividing the dissociation rate constant (koff) by the association rate constant (value obtained by U). The association rate constant, the dissociation rate constant, and the equilibrium dissociation constant are used to indicate the binding affinity of a binding protein (e.g., an antibody) to an antigen. Methods for determining association and dissociation rate constants are well known in the art. The technology provides the high sensitivity and ability to check samples in a balanced physiological buffer. Other experimental methods and instruments such as BIAcore® (biomolecular interactions) can be used.

Analytical) analytical methods (eg available from BIAcore International AB, GE

Healthcare company, Uppsala, Sweden purchased instruments). Another 160877.doc • 81 · 201247704 can also be analyzed using KinExA® (Kinetic Exclusion Assay) available from Sapidyne Instruments (Boise, Idaho). "Marker" and "detectable label" mean linked to a specific binding partner (such as an antibody or analyte) to, for example, enable a reaction between a member of a specific binding pair (such as an antibody and an analyte) to be detected. The portion, and the specific binding partners (such as antibodies and analytes) so labeled, are referred to as "detectable labels." Thus, as used herein, the term "labeled binding protein" refers to a protein that has been labeled to provide identification of the binding protein. In one embodiment, the detectable label is capable of producing a signal detectable by visual or instrumental means, for example, incorporating a radiolabeled amino acid or linked to a labelable avidin (eg, containing The polypeptide of the biotin-based moiety detected by optical or colorimetric detection of fluorescently labeled or enzymatically active streptavidin. Examples of labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (eg, 3H, i4c, 35s, 90Y, "Tc, 丨丨丨In, 丨25!, 丨77Lu, 16611〇) 153Sm); chromogen, fluorescein 4 shows δ (such as FITC, rhodamine and steel metal phosphors); enzyme labeling (such as horseradish peroxidase, luciferase and alkaline phosphatase) a chemiluminescent label; a biotinyl group; a predetermined polypeptide epitope recognized by a secondary reporter (eg, a leucine zipper pair sequence, a secondary antibody binding site, a metal binding domain, and an epitope tag); A magnetizer for ruthenium chelate. Representative examples of labels commonly used in immunoassays include portions that produce light (e.g., acridinjum compounds) and fluorescent buttons (e.g., luciferin). Other markings are described herein. In the case of 160877.doc •82· 201247704, the part itself may not be detectable but may become detectable after reacting with another part. The use of "detectable mark" is intended to cover the latter type of detectable mark.

The term "conjugate" refers to a binding protein (such as an antibody) chemically linked to a second chemical moiety, such as a therapeutic or cytotoxic agent. The term "agent" is used herein to mean a compound, a mixture of compounds, a biomacromolecule or an extract prepared from a biological material. In one embodiment, the therapeutic or cytotoxic agent includes, but is not limited to, pertussis toxin, taxol, cytochalasin B, gramicidin D, bromination Ethidium, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colostine Colchicin), doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D (actinomycin D), 1-dehydrocinosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol and puromycin ( Puromycin) and its analogs or homologs. When used in the context of immunoassays, the conjugate antibody is a detectably labeled antibody that is used as a detection antibody. As used herein, the terms "crystal" and "crystal" refer to a binding protein (e.g., an antibody) or antigen-binding portion thereof that exists in the form of a crystal. A crystal is a solid form of a substance that is different from other forms such as an amorphous solid state or a liquid crystal state. A crystal system consists of a regular three-dimensional array of atoms, ions, molecules (e.g., proteins, such as antibodies) or molecular assemblies (e.g., antigen/antibody complexes). These three-dimensional arrays are arranged according to the specific mathematical relationships well understood in the art. The basic unit or building block in the crystal is called an asymmetric unit. The asymmetric unit repeatedly provides the "unit cell" of the crystal in an arrangement consistent with the well-defined crystallographic symmetry. Crystals are provided by regular translational repeat unit cells in all three dimensions. See

Giege, R. and Ducmix, A_ Barrett, Crystallization of Nucleic

Acids and Proteins, a Practical Approach, 2nd edition, page SOl-iS, Oxford University Press, New Y〇rk, New York, (1999). A "polynucleotide" means a polymeric form of two or more nucleotides (ribonucleotides or deoxynucleotides or modified forms of either type of nucleotide). The term includes both single and double strand forms of DNA. The term "isolated polynucleotide" shall mean a polynucleotide (eg, a genomic, cDNA or synthetic source, or some combination thereof), in view of its source 'the isolated polynucleotide" is not It is found in nature that all or a portion of the polynucleotides together with the "isolated polynucleotide" are associated; operably linked to a polynucleotide that is not ligated in nature; or not as large in nature One part of the sequence exists. The term "vector" is intended to mean a nucleic acid molecule capable of transporting another nucleic acid to which it is linked. One type of vector is "plastid", which refers to a circular double stranded DNA loop that can be ligated internally to other DNA segments. Another type of vector is a viral vector in which additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., a bacterial vector having a bacterial origin of replication of 160877.doc-84 - 201247704 and a free mammalian vector). Other vectors (e. g., non-episomal mammalian vectors) can be integrated into the genome of the host cell after introduction into the host cell and thereby replicated with the host genome. In addition, certain vectors are capable of directing the performance of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors suitable for use in recombinant DNA techniques are typically in plastid form. Since the plastid is the most common form of the carrier, in this specification, "plastid" and "carrier" are used interchangeably. However, the invention is intended to include such other forms of expression vectors, such as viral vectors that are equivalent (e.g., replication defective retroviruses, adenoviruses, and adeno-associated viruses). The term "operably linked" means that the components are joined together to function in their intended manner. The control sequences are "operably linked" to the coding sequence such that they can be joined in a manner that achieves the performance of the coding sequence under conditions compatible with the control sequences. The sequence of "operably linked" includes expression control sequences adjacent to the relevant gene, and expression control sequences for trans-acting or remote-action controlling related genes. As used herein, the term "expression control sequence" refers to a polynucleotide sequence which is necessary for the performance and processing of the coding sequences to which it is ligated. Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that increase translation efficiency (ie, Kozak common sequences) a sequence that enhances protein stability; and, if necessary, a sequence that enhances protein secretion. The nature of these control sequences differs depending on the host organism I60S77.doc • 85 - 201247704: in prokaryotes, 'these control sequences are generally: ribosome binding sites and transcription termination sequences; in eukaryotic orders,: etc. Control sequences generally include a promoter and a transcription termination sequence. The term "control: sequence" is intended to include components that are necessary for expression and processing, and may also include other components that are suitably present, such as leader sequences and fusion partner sequences. "Transformation" refers to any process that allows foreign DNA to enter a host cell. Transformation can be carried out under natural or "conditions" using a variety of methods well known in the art. (d) can be carried out by any known method for inserting foreign nucleic acid sequences into prokaryotic or eukaryotic host cells. Selection based on the host cell being transformed, and may include, but is not limited to, viral infection, electroporation, lipofection, and particle bombardment. The "transformed" cells, including the inserted DNA, are capable of autonomously replicating plastids. Or a stable transformed cell that replicates in the form of a part of the host chromosome. It also includes cells that transiently express the inserted DNA or RNA for a limited period of time. The term "recombinant host cell" (or simply "host cell") is intended to mean a cell into which foreign DNA has been introduced. It is to be understood that the terms are intended to refer not only to the particular individual cell' but also to the progeny of that cell. Because certain modifications may occur in the progeny due to either mutation or environmental influences, the progeny may not actually be identical to the parent cell' but are still included within the scope of the term "host cell" as used herein. In one embodiment, the host cell comprises prokaryotic cells and eukaryotic cells selected from any of the biological worlds. In another embodiment, the eukaryotic cells include protist, fungal, plant, and animal cells. In another embodiment, the host cell includes, but is not limited to, a prokaryotic cell strain 160877.doc-86-201247704 Enterobacter coi; mammalian cell lines CHO, HEK 293, COS, NSO, SP2, and PER.C6 Insect cell line Sf9; and fungal cell wine yeast (Sacc/zarowyce·? cereWiz'ae). Recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation can be performed using standard techniques (e.g., electroporation, lipofection). Enzymatic reactions and purification techniques can be performed according to the manufacturer's instructions or as commonly done in such techniques or as described herein. The above-described techniques and procedures are generally performed as described in the well-known methods well known in the art and in the various comprehensive references and the more specific references which are cited and discussed throughout the specification. See, for example, Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual (2nd Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY). "Transgenic organisms" as known in the art means An organism comprising a cell of a transgenic gene, wherein the transgenic gene introduced into the organism (or the ancestor of the organism) exhibits a polypeptide that is not naturally expressed in the organism. A "transgenic gene" is stably and operably integrated into the genome of a cell that will develop into a transgenic organism, thereby directing the encoded gene product to be expressed in one or more cell types or tissues of the transgenic organism. DNA constructs. The terms "modulate" and "modulate" are used interchangeably and, as used herein, refers to a change or change in the activity of a related molecule, such as the biological activity of a cytokine. Modulation can be an increase or decrease in the magnitude of a certain activity or function of the relevant molecule. Exemplary activities and functions of the molecule include, but are not limited to, binding characteristics, enzymatic activity, cellular receptor activation, and signal transduction. 160877.doc • 87 - 201247704 Accordingly, the term "modulator" is a compound that is capable of altering or altering the activity or function of a related molecule, such as the biological activity of a cytokine. For example, a modulator can cause an increase in or decrease in the magnitude of a certain activity or function of a molecule compared to the amount of activity or function observed in the absence of such a modulator. In certain embodiments, the modulator is an inhibitor that reduces the magnitude of at least one activity or work of the molecule. Exemplary inhibitors include, but are not limited to, proteins, peptides, antibodies, peptibody, carbohydrates or small organic molecules. Peptibodies are described, for example, in PCT Publication No. WO No. 1/83525. The term "agonist" refers to a modulator that, when contacted with a related molecule, causes an increase in the magnitude of a certain activity or function of the molecule compared to the amount of activity or function observed in the absence of an agonist. Related specific agonists can include, but are not limited to, polypeptides, nucleic acids, carbohydrates, and any other molecule that binds to an antigen. The terms "inhibitor" and "inhibitor" refer to the amount of activity or function of a knife that is caused by contact with a molecule in question as compared to the activity or function observed in the presence of a non-reagent. Reduced regulator. Related specific antagonists include antagonists that block or modulate the biological or immunological activity of the antigen. Antagonists and inhibitors of the antigen can include, but are not limited to, proteins, nucleic acids, carbohydrates, and any other molecule that binds to the antigen. The term "effective amount" as used herein means sufficient to reduce or ameliorate the severity and/or duration of a condition or one or more symptoms thereof; to inhibit or prevent the progression of the condition; to cause the condition to subside; to inhibit or prevent the condition from being associated with - or Recurrence, progression, onset, or progression of multiple symptoms; detection of a condition, or enhancement of the prophylactic or therapeutic effect of a refractory or therapeutic agent: setting. An effective amount may require more than - times. "者" and "individual" are used interchangeably herein to refer to animals, = extracts, including primates (such as humans, non-human primates such as monkeys and chimpanzees), non-primates (such as mothers). Cows, rats, tracing, dogs,:: two rabbits: sheep, hamsters, guinea pigs and recorded), birds (such as ducks or geese) and sharks patients or individuals are humans such as human beings who are treating or evaluating diseases or diseases A human being at risk of a disease, disorder or condition, a human suffering from a disease, disorder or condition, and/or a human being being treated for a disease, disorder or condition. As used herein, the term "sample" is used in its broadest sense. As used herein, 'a biological sample' includes, but is not limited to, any amount of material from a living "g" or previously a living being. These living things include (but are not) human, mouse, large Rats, monkeys, dogs, rabbits, and other animals. These include, but are not limited to, blood (eg, whole blood), plasma, serum, urine, T Χ 'month ^, endothelial cells, white blood cells, monocytes, others Cells, organs, tissues, bone marrow, lymph nodes, and spleen. Knives and at least one component" generally refer to a method that can be included in a kit to analyze a pattern as described herein and in other methods known in the art. Capture binding proteins (such as patient |, serum or plasma samples) (example: detection or conjugate binding protein (eg antibody), control, cup-to-cup, a series of control standards, sensitivity group (sensitivity) ) gluten, buffer, diluent, salt, enzyme, enzyme cofactor, 160877.doc -89· 201247704 detection reagent, pretreatment reagent μ ^ ^ _ quality (for example in solution), stop cold liquid and Its analogue. So this is in this In the context of the invention, at least one component and "New" may encompass a polypeptide as described above or other composition that analyzes an L=3 analyte, such as a polypeptide, as appropriate: Immobilized on an anti-analyte (eg, anti-polypeptide) antibody on a solid support. Some components may be in a form, in a liquid/liquid form, or lyophilized for use in the assay after recovery. "A composition that is known to be free of analyte ("negative control") or contains an analyte ("positive control"). The positive control may contain analytes of known degree. "Control", " "Positive control" and "control standard" are used interchangeably herein to refer to a composition comprising a known concentration of the analyte. "Positive control" can be used to determine analytical performance characteristics and to be an agent (eg, an analyte) The applicable indicator of the integrity. The predetermined cutoff value and the "predetermined degree" generally refer to the analytical cutoff value used to evaluate the diagnostic/prognostic/therapeutic efficacy results by comparing the analytical results with a predetermined cutoff value/degree. Scheduled cut The value/degree has been linked or associated with various clinical parameters (eg, severity of disease, progression/no progression/improvement, etc.) Although the invention may provide an exemplary predetermined degree, it is well known that cutoff values may be of a nature of immunoassay ( For example, the antibodies used, etc. vary. In addition, the invention is adapted to other immunoassays to obtain specific cutoff values for immunoassays for their other immunoassays based on the present invention, which are well known to those skilled in the art. Within the skill range. Although the exact value of the cut-off value/degree can vary between analyses, the relevance (if any) as described herein should be generally applicable. 160877.doc •90· 201247704 eg::文所The "pretreatment reagent" (for example, Lotion, Shen Dian, and/or solubilizing reagent) used in the diagnostic assay is used to dissolve any cells and/or to be present in the sample of the test sample, 6 Α ^ m , Any analyte solubilization in 7: step description, not all samples must pre-exist in the presence of η-related peptides. Solubilization may allow the analyte to bind from any endogenous White released. The pretreatment reagent can be (no separation step required) or heterogeneous (requires separation step). in

When the reagent is pretreated, the self-test sample W is removed before the next step of the analysis to remove any precipitated analyte binding protein. "Quality Control Reagents" are described in the Q ELISA method and kits = including but not limited to the control standards, controls, and sensitivity groups. A calibration (standard) curve for the concentration of a control standard or a "standard" (eg, - or a plurality of 'incorporated for interpolating an analyte (such as a binding protein (eg, anti-) or knife extract). Use a single control standard that is close to 2 positive/negative cut-off values. A variety of control standards (ie, more than one control standard or different amounts of standard) can be used in combination to form a "sensitivity group." Means the monthly or probability of occurrence of a particular event at a time or at a future point in time. "Risk stratification" means allowing a physician to classify a patient as having a low degree of developing a particular disease, disorder or condition. A series of known clinical risk factors with high or highest risk. I specific I. is between members of a specific binding pair (eg, an antigen (or a fragment thereof) protein (eg, an antibody) (or an antigen-reactive fragment thereof)) In the case of interaction, it refers to the selective reactivity of the interaction. The phrase "specificity 160877.doc 201247704, ''n combination" and similar phrases refers to binding proteins (eg antibodies) (or antigens thereof) Reactive green tablets & ) specifically bind to an analyte (or a fragment thereof) and do not specifically bind to other entities. Specific binding is understood to mean binding to a particular antigen, antigenic thiol ligand or binding partner. The preference is at least 103, 104, 105, 106, 1〇7, 1〇8, ^ times higher than the preference for the control non-specific antigen, epitope, receptor ligand or binding partner. In certain embodiments In combination, binding by BiaC〇re8 and specific binding is understood to be combined with a KD value of less than 1×1 〇_6 。. In some embodiments, it can be understood to be less than 1×10-4 Μ, 1χ10-5. Μ, 1χ1〇·7 Μ, 1χ1().8 Μ, 1χΐ().9 Μ, 1Xl〇w Μ, ixio-u Μ or 1χ1〇.12 Mi, value combination. Method of selecting appropriate non-specific control Within the capabilities of those skilled in the art, a specific binding partner is a member of a specific binding pair. A specific binding pair comprises two different molecules that specifically bind to each other chemically or physically. Antigens and binding proteins of analytical methods (eg immunoassays) In addition to specific binding pairs, for example, antibodies, other specific binding pairs may also include biotin and avidin (or streptavidin); carbohydrates and lectins; complementary nucleotide sequences; effector molecules and receptors Molecular 'cofactors and enzymes; enzyme inhibitors and enzymes; and analogs thereof. Further 'specific binding pairs' may include members of analogs that are initially specific binding members, such as analytes - analogs. Immunoreactivity specificity Binding members include isolated or recombinantly produced antigens, antigenic fragments and antibodies (including single and multiple antibodies), as well as complexes, fragments and variants thereof (including variant fragments). 160877.doc -92- 201247704 As used herein, 'variant' means that the amino acid sequence differs from the established polymorphism by the addition (eg, insertion), deletion or conservative substitution of an amino acid (eg, c-Met, CD-28, CD_3, CD_19). , IL_18, BNP, NGAL, Tnl or HIV polypeptides, or anti-polypeptide antibodies), but retain the biological activity of a given polypeptide (eg variant IL-18 can compete with anti-IL_18 antibodies for binding Polypeptide of IL-18). Conservative substitutions of amino acids are believed in the art (i.e., different amino acid substitutions of amino acids with similar properties (e.g., hydrophilicity and extent and distribution of charged sites) generally involve minor changes. As understood by φ in the art, these minor changes can be identified, in part, by considering the hydropathic index of the amino acid (see, for example, Kyte et al. (1982) J. Mol. Biol. 157: 105-132). The hydrophilicity index of an amino acid is based on considerations of its hydrophobicity and charge. Amino acids having a similar hydropathic index are known in the art to be substituted and still retain protein function. In one aspect, the amino acid having a hydropathic index of ±2 is substituted. The hydrophilicity of the amino acid can also be used to reveal substitutions that will result in proteins that retain biological function. Consideration of the hydrophilicity of the amino acid in the case of peptides allows calculation of the maximum local average hydrophilicity of the peptide, which is a suitable indicator that has been reported to be closely related to antigenicity and immunogenicity (see, e.g., U.S. Patent No. 4,554, No. 101). As understood in the art, 'substitution with an amino acid having a similar hydrophilicity value results in a peptide that retains biological activity (e.g., immunogenicity). In one aspect, amines having a hydrophilicity value within ±2 are used. The base acid is substituted. The hydrophobicity index and the hydrophilicity value of the amino acid are all affected by the specific side chain of the amino acid. Consistent with the observations of the 'biological acid-compatible amino acid substitutions are understood to depend on the relative similarity of the amino acids, and especially the side chains of their amino acids (eg by hydrophobicity, pro 160877.doc •93_ 201247704 Water ± ± charge, size and other properties revealed). "Variant" can also be used to describe a polypeptide that has been differentially processed (such as by proteolysis, 4 acidification, or other post-translation), but still retains biological activity or antigenic reactivity (eg, the ability to bind IL-18). Or a fragment thereof. Unless the context dictates otherwise, the use of the "variant" herein is intended to cover a segment of the variant. I. Generation of a semi-Ig binding protein The present invention relates to a semi-Ig binding protein which can bind to one or more targets and a process for the preparation thereof. In one embodiment, the half 4 binding protein of the invention comprises a minimally structured immunoglobulin-based binding protein having a C-terminally joined to the N-terminal heavy chain antigen binding domain of at least a portion of the immunoglobulin cH3 domain, wherein the CH3 domain is Preferably, one or more mutations are included in the CH3/CH3 contact region to inhibit CH3-CH3 dimerization. Half ig may be referred to herein as a "binding protein." The semi-Ig comprises a functional antigen binding site which can be provided by a single heavy bond antigen binding bond or form a functional antigen binding site by complementary pairing of a peptide comprising a heavy bond antigen binding domain with a light chain antigen binding domain. Click to provide. In addition to the minimal structure of the heavy chain antigen binding domain joined to at least a portion of the CH3 domain, the semi-Ig binding protein of the invention may also include other domains. For the sake of simplicity, the antigen binding domain present in the peptide comprising the CH3 domain is referred to as the heavy chain antigen binding domain, although the antigen binding domain need not be derived from the antibody heavy chain. In certain embodiments, the peptide comprising the heavy chain antigen binding domain does not comprise a CH1 domain and/or a CH2 domain. Exemplary embodiments of the semi-Ig include, but are not limited to: VD1-(X1)n-X2; VD1-(X1)N-VD2-(X2)N-X3; and 160877.doc-94-201247704 VD1-( X1) n-VD2-(X2)n-VD3-(X3)n-X4; In an exemplary embodiment, each VD (or VDH) line is independently selected from a heavy chain variable domain, a double heavy chain variable domain, Triple heavy chain variable domain, light chain variable domain, double light chain variable domain, triple light chain variable domain, heavy chain variable domain and light chain variable domain combination, two heavy chain variable domains and light Combination of chain variable domains, combination of heavy chain variable domains and two light chain variable domains, domain antibodies, camelid antibodies, scFv, receptor and backbone antigen binding proteins. For simplicity, a VD herein may be referred to as a variable domain, but in the context of a peptide comprising a heavy chain antigen binding domain, it is to be understood to include a heavy chain variable domain, a double heavy chain variable domain, as provided herein, Triple heavy chain variable domains, domain antibodies, scFv, receptor and backbone antigen binding proteins. And X. . In an exemplary embodiment, the consistency of each X depends on its position in the half Ig. The most C-terminal X (for example, X2 in the first embodiment, X3 in the second embodiment, and X4 in the third embodiment) may include a polypeptide having at least a portion of the CH3 domain in the CH3/CH3 contact region. The residue within the residue has at least one mutation that inhibits the dimerization of CH3-CH3. The second-end C-terminus X (for example, XI in the first embodiment, X2 in the second embodiment, and X3 in the third embodiment) may include a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain, or a linker. . In certain embodiments, the second and last C-terminus X further comprise a sequence of hinge regions that can be a linker. In some embodiments, the hinge region is preferably between the CH1 and CH2 domains when both the CH1 and CH2 domains are present. When there is CH1 and 160877.doc -95- 201247704 CH3 domain and no CH2 domain exists, the 'hinge zone is better between (:111 and (:yang). When there is CH2 and CH3 domain and no CH1 domain exists, hinge The region is preferably located at the N-terminus of the CH2 domain. When there are other defects (for example, χι in the second embodiment and χι&χ2 in the third embodiment), it includes a linker sequence. Each X is independently 〇 or 1 In certain embodiments, a half Ig binding protein provided herein includes additional sequences (eg, linker sequences, functional sequences) in a peptide comprising a heavy chain antigen binding domain. In certain embodiments, the half provided herein The binding protein only includes the domain proposed above in the peptide containing the heavy bond antigen binding domain. In certain embodiments, the X in the peptide containing the heavy chain antigen binding domain does not include the CH1 domain and/or the 〇^2 domain. In certain embodiments, the peptide binding protein comprising a heavy chain antigen binding domain does not comprise a CH1 domain and/or a CH2 domain. Furthermore, the semi-Ig binding protein may comprise a first peptide chain comprising at least a light chain antigen binding domain. The bond antigen binding domain should be understood to include the light bond variable domain, double a light chain variable domain, a triple light chain variable domain, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. Thus, a light chain variable domain may include an antigen that can bind to the antigen alone or in combination with a peptide comprising a heavy chain antigen binding domain. Or a sequence of the target; or both. In addition to the minimal structure of the light chain antigen binding domain, the second peptide of the semi-Ig binding protein of the present invention may include other domains. An illustration of the second peptide of the half 1 § binding protein Examples include, but are not limited to: VD1-(X1)n; VD1-(X1)N-VD2-(X2)N; and VD1-(X1)n-VD2-(X2)n-VD3-(X3 In an exemplary embodiment, each VD (or VDL) is independently selected from the group consisting of a light bond 160877.doc-96·201247704, a domain only, a dual light chain variable domain, a triple light chain variable domain, a domain antibody , scFv, sacred body and skeletal antigen binding protein. For the sake of simplicity, herein may be referred to as a variable domain 'but in the case of a peptide comprising a light chain antigen binding domain, the solution is to include a light bond variable as provided herein Domain, dual light bond variable domain heavy light chain variable domain, domain antibody, scFv, receptor and backbone antigen binding protein beta in an exemplary embodiment The identity of each X depends on its position in the second peptide of the half-ig binding protein. The x at the most c-terminus (for example, X1 in the first embodiment, X2 in the second embodiment, and the third embodiment) X3) may include a light chain '艮定定°° when there are other Xs (for example, XI in the second embodiment and X1 and X2 in the third embodiment) 'which includes a linker sequence. Each N is independently selected from 0 And 1 ° In certain embodiments, the half of the binding protein provided herein includes other sequences (eg, linker sequences, functional sequences) in the peptide containing the light bond antigen binding domain, and peptides in the light chain antigen-binding domain. In certain embodiments, 'X does not include a CH1 domain and/or a CH2 domain and/or a variable light chain. In certain embodiments of the peptide comprising a light chain antigen peptide binding domain, the binding protein does not comprise a CH1 domain and/or a CH2 domain. In certain embodiments, the half ig binding proteins provided herein include only the domains raised above in peptides containing a light chain antigen binding domain. In certain embodiments, the number of V domains in a peptide comprising a light chain antigen binding domain is the same as the number of V domains in a peptide comprising a heavy chain antigen binding domain. In certain embodiments, the number of V domains in a peptide comprising a light chain antigen binding domain is different from the number of V domains in a peptide comprising a heavy chain antigen binding domain. Figure 1A provides a schematic representation of the bivalent molecules of antibodies and immunoglobulins based on various formats (top row), which can be used as a basis for designing a semi-Ig binding protein (bottom row) 160877.doc • 97· 201247704. As shown in the various semi-Ig formats, the form containing the heavy bond antigen binding domain is paired with a peptide containing a light chain antigen binding domain. In the schematic of the half 1 § binding protein, the semi-dvd binding protein, and the semi-TVD binding protein, the light chain and heavy chain variable domains are not complementary pairs that form a single antigen binding domain. In a semi-RAb_ig binding protein, the variable domain adjacent to the constant region is shown as a complementary pair, and the receptors in the peptide containing the light chain antigen binding domain and the peptide containing the heavy chain antigen binding domain do not interact and form an independent Binding site. When a semi-Ig binding protein comprises two complementary sequences to form a single binding site comprising two peptides, and a sequence that forms an independent binding site on each peptide, preferably the complementary sequence is adjacent to the constant domain and the independent binding site is kept away from the constant area. Figure 1B shows an exemplary half of the igG antibody produced by the parent. Binding protein. The naturally occurring IgG dimerizes via a specific interaction domain via interaction of the CH3 domain. The antibody chain is also maintained via the disulfide bond present in the antibody hinge region. The semi-Ig binding proteins of the present invention are typically produced using known recombinant DNA techniques and antibodies having known nucleotide and/or amino acid sequences, however, the particular method of producing the semi-Ig binding proteins of the present invention does not limit the invention. Mutagenesis (usually site-directed mutagenesis, but also random mutagenesis as appropriate) is used to change one, two or three semi-proline to form a disulfide bond in the hinge region to another amino acid To prevent the formation of disulfide bonds and/or to disrupt sequences important for interactions between CH3 domain sequences (by making one, two, three, four, five, six, seven in the ch3 domain) , 8 or more residues are mutated). In some embodiments, CH3 domain dimerization can be achieved by truncating the CH3 domain. As used herein, in certain embodiments, when the wild-type value of a species localizes I60877.doc-98-201247704 (eg, a human or mouse IgG sequence or variant thereof) binds to protein A, a half Ig binding protein In the case, at least a portion of the CH3 domain is understood to be sufficient to allow a half 1 g of binding protein to bind to the CH3 domain portion of protein A. As used herein, in certain embodiments, it is understood that a portion of the CH3 domain is at least about 70% identical to the full length CH3 domain from the same species, at least about 75%, and at least about 80%, at least about 85. /. Consistently, at least about consistent, at least about 95%, at least about 98% identical or at least about 99% identical. As used herein, in certain embodiments, at least a portion of the CH3 domain is understood to be the CH3 domain that interacts with the RnFc receptor. As used herein, "CH3-CH3 dimerization" is understood to mean the specific interaction of two CH3 domains with each other. The specific interaction can be driven by a covalent interaction (e.g., formation of a disulfide bond in the hinge region that closely contacts the CH3 domain) or non-covalent interactions that promote the specific binding of the two CH3 domain portions of the immunoglobulin invariant chain to each other. The semi-Ig binding proteins of the invention can be produced using a variety of techniques. The present invention provides expression vectors, host cells and methods for producing binding proteins. A. Producing a parental molecule for selecting a sequence to be incorporated into a semi-Igg protein. The variable domain of a single-binding protein half-Ig binding protein can be derived from a parental binding protein (such as an antibody, including a binding antigen). Strain and mAb) were obtained. These antibodies may be present naturally or may be produced by recombinant techniques. MAbs can be prepared using a variety of techniques known in the art, including the use of fusion sigma tumor recombination and sputum cell presentation techniques or combinations thereof. For example, mAbs can be produced using fusion tumor technology, including those known in the art and taught, for example, in the teachings of 160877.doc-99-201247704, Harl〇w et al., (ι988)

Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed.); Hammerling et al., (1981)

Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, Ν·Υ·). As used herein, the term "monoclonal antibody" is not limited to antibodies produced by fusion tumor technology. The term "monoclonal antibody" refers to an antibody derived from a single pure line, including any eukaryotic, prokaryotic or bacteriophage-derived line, rather than the method by which it is produced. As discussed in Example 1 below, fusion tumors were selected, colonized and further screened for the desired characteristics, including vigorous fusion tumor growth, high antibody production, and desired antibody characteristics. The fusion tumor can be cultured and expanded in vivo in an isogenic animal, an animal lacking an immune system (e.g., a nude mouse) or cultured and expanded in vitro in a cell culture. Methods for selecting, breeding, and expanding fusion tumors are well known to those of ordinary skill. In a particular embodiment, the fusion tumor is a mouse fusion tumor. In another embodiment, the fusion tumor is produced in a non-human, non-mouse species, such as a rat, sheep, pig, mountain sheep, cow or horse. In another embodiment, the fusion tumor is a human fusion tumor in which human non-secretory myeloma is fused to a human cell that exhibits an antibody that binds to a particular antigen.

For example, U.S. Patent No. 5,627,052, PCT Publication No. WO 92/02551 and Babcock, J.S. et al., (1996) Proc. Natl. Acad. Sci. USA

Recombinant mAbs were also produced from a single isolated lymphocyte using the procedure described in the art as the Selected Lymphocyte Antibody Method (SLAM), 93:7843-7848. In this method, a single cell secreting the relevant antibody (for example, a lymphocyte derived from an immunized animal) is identified, and the heavy chain and light chain variable region are self-retained by reverse transcriptase PCR 160877.doc -100· 201247704 cDNA. In the case of a suitable immunoglobulin constant region (e. g., a human constant region), such variable regions can then be expressed in mammalian host cells such as COS or CH0 cells. The transfected cells can then be in vitro, for example, by panning a host cell transfected with an amplified immunoglobulin sequence derived from a lymphocyte selected from a living body to isolate a cell expressing an antibody against the relevant antigen. Further analysis and selection. The amplified immunoglobulin sequences can be further manipulated in vitro, such as by in vitro affinity maturation methods, such as those described in PCT Publication Nos. WO 97/29131 and WO 00/56772. Individual antibodies are also produced by immunizing non-human animals containing some or all of the human immunoglobulin loci with related antigens. In one embodiment, the non-human animal is a XEN0M0USE® transgenic mouse, which is an engineered mouse strain comprising a large fragment of a human immunoglobulin locus and lacking a mouse antibody. See, for example, Green et al., (1994) Nature Genet. 7: 13-21; and U.S. Patent Nos. 5,916,771; 5,939,598; 5,985,615; 5,998,209; 6,075,181; 6,091,001; 6,114,598; and 6,130,364. See also PCT Publication No. WO 91/10741; WO 94/02602; WO 96/34096; WO 96/33735; WO 98/16654; WO 98/24893; /50433; WO 99/45031; WO 99/53049; WO 00/09560; and WO 00/037504. XENOMOUSE® transgenic mice produce adult human-like lineages of fully human antibodies and produce antigen-specific human monoclonal antibodies. XENOMOUSE® transgenic mice contain approximately 80% of human antibodies via the introduction of the human heavy chain gene 160877.doc -101 - 201247704 and the megabase-scale germline configuration of the YAC fragment of the χ light-key locus. pedigree. See Mendez et al., (1997) Nature Genet. 15: 146-156; Green and Jakobovits (1998) J. Exp. Med. 188: 483-495 ° Parental antibodies can also be prepared using in vitro methods, in which antibodies are screened. The library is used to identify antibodies with the desired binding specificity. The method of screening recombinant antibody libraries is well known in the art and includes, for example, the methods described in Ladner et al., U.S. Patent No. 5,223,409; PCT Publication No. WO 92/18619; WO 91/17271 No. WO 92/20791; No. WO 92/15679; No. WO 93/01288; No. WO 92/01047; WO 92/09690; and WO 97/29131; Fuchs et al. 1991) Bio/Technology 9: 1370-1372; Hay et al., (1992) Hum. Antibod·Hybridomas 3: 81-85; Huse et al., (1989) Science 246: 1275-1281; McCafferty et al., (1990) Nature 348: 552-554; Griffiths et al., (1993) EMBO J. 12: 725-734; Hawkins et al., (1992) J. Mol. Biol. 226: 889-896; Clackson et al., (1991) Nature 352: 624-628; Gram et al., (1992) Proc. Natl. Acad. Sci. USA 89: 3576-3580; Garrad et al., (1991) Bio/Technology 9: 1373-1377;

Hoogenboom et al., (1991) Nucl. Acid Res. 19: 4133-4137; and Barbas et al., (1991) Proc. Natl. Acad. Sci. USA 88: 7978-7982; and U.S. Patent Publication No. 2003/0186374 number. A2. scFv and in vitro produced binding proteins Parental binding proteins (such as antibodies) of the present invention can be produced using various phage display methods known in the art. In phage display, a functional antibody domain is presented on the surface of a phage particle carrying a polynucleotide sequence encoding the same. In particular, the phage can be utilized to present an antigen binding domain that is expressed by a lineage or a combinatorial antibody library (e.g., human or murine). The antigen can be used, for example, using a labeled antigen or an antigen bound or captured on a solid surface or bead to select or identify a phage that exhibits an antigen binding domain that binds to the associated antigen. The phage used in these methods are usually filamentous phage including fd and M13 binding domains expressed by phage which are recombinantly fused from Fab, Fv or disulfide stabilized Fv antibody domain to phage gene III or gene VIII protein. Examples of phage display methods that can be used to prepare antibodies of the invention include those disclosed in Brinkman et al, (1995) J. Immunol. Methods 182: 41-50; Ames et al, (1995) J. Immunol. 184: 177-186; Kettleborough et al., (1994) Eur. J. Immunol. 24: 952-958; Persic et al., (1997) Gene 187: 9-18; Burton et al., (1994) Advances in

Immunol. 57: 191-280; PCT Application No. PCT/GB91/01134; PCT Publication No. WO 90/02809; WO 91/10737; WO 92/01047; WO 92/18619; WO 93/11236; WO 95/15982; and WO 95/20401; and U.S. Patent No. 5,698,426; 5,223,409; 5,403,484; 5,580,717; 5,427,908; 5,750,753; Nos. 5, 821, 698; 5, 427, 908; 5, 516, 637; 5, 780, 225; 5, 658, 727; 5, 733, 743; and 5, 969, 108. As described in the references provided herein, after phage selection, the antibody coding region can be isolated from 160877.doc •103·201247704 sputum and used to generate intact antibodies including human antibodies or any other desired antigen-binding fragment And, for example, described in detail below, 'behaves in any desired host including mammalian cells, insect cells, plant cells, yeast, and bacteria. For example, techniques for recombinant production of Fab, Fab' and F(ab')2 fragments can also be used using methods known in the art, such as PCT Publication No. WO 92/22324; Mullinax et al., (1992) BioTechniques 12(6): 864-869; Sawai et al., (1995) AJRI 34: 26-34; and Better et al., (1988) Science 240: 1041-1043. Examples of techniques that can be used to generate single-chain Fvs and antibodies include U.S. Patent Nos. 4,946,778 and 5,258,498; Huston et al., (1991), Methods Enzymol. 203:46-88; Shu et al., (1993) Proc. Natl Acad. Sci. USA 90: 7995-7999; and the technique described in Skerra et al., (1988) Science 240: 1038-1040. As an alternative to screening recombinant antibody libraries by phage display, other methods known in the art for screening large combinatorial libraries can be used to identify parent antibodies. One type of alternative expression system is the expression system for expressing recombinant antibody libraries as RNA-protein fusions as described in the following literature: PCT Publication No. WO 98/3 1700' and Roberts, RW and Szostak, JW (1997) Proc. Natl. Acad. Sci. USA 94: 12297-12302. In this system, a covalent fusion is formed between the mRNA and its encoded peptide or protein by in vitro translation of synthetic mRNA with puromycin (a peptide receptor antibiotic) at the 3' end. Thus 'can be based on the nature of the encoded peptide or protein (eg antibody or portion thereof) (such as binding of an antibody or portion thereof to a dual 160877.doc 201247704 specific antigen)' from a complex mixture of mRNAs (eg combinatorial libraries) Set specific mRNAs. The nucleic acid sequence encoding the antibody or portion thereof recovered from screening such libraries can be expressed by recombinant means as described herein (e.g., in a mammalian host cell) and, in addition, can be initially selected by further rounds Screening of mutated mRNA-peptide fusions in the sequence or further affinity maturation of the nucleic acid sequences by other methods of affinity maturation of the recombinant antibodies in vitro as described herein. In another method, a parental antibody can also be produced using the yeast presentation method φ known in the art. In yeast presentation, the antibody domain is tied to the yeast cell wall using genetic methods and presented on the yeast surface. In particular, the yeast can be utilized to present an antigen binding domain that is expressed by a lineage or combinatorial antibody library (e. g., human or murine). Examples of yeast delivery methods that can be used to prepare parental antibodies include those disclosed in U.S. Patent No. 6,699,658. A3·Humanized and engineered binding proteins

The binding proteins (e.g., antibodies) described herein can be further modified to produce CDR-grafted antibodies and humanized parent antibodies. The CDR-grafted parent antibody comprises a heavy chain and light chain variable region sequence from a human antibody, wherein one or more CDR regions of Vh& / or V1 are replaced by a murine antibody that binds to the relevant antigen. The framework sequences from any human antibody can be used as a template for cDR transplantation. The framework regions can be selected in unit form, i.e., a combination of naturally occurring FR1, FR2 &FR3; or independently selected, e.g., based on homology to individual FRs of the parent antibody. However, linear cross-linking on this framework typically results in a loss of binding affinity for the antigen to some extent. The higher the homology of the human antibody to the original murine antibody, the less likely it is to introduce the murine CDRs and the human framework in CDR 160877.doc-105·201247704 to introduce distortions that may reduce affinity. Thus, in one embodiment, a human variable framework selected to replace a murine variable framework other than a CDR has at least 65% sequence identity to a murine antibody variable region framework. In one embodiment, the human and murine variable regions have at least 70% sequence identity in addition to the CDRs. In a particular embodiment, the human and murine variable regions have at least 75% sequence identity in addition to the CDRs. In another embodiment, the human and murine variable regions have at least 80% sequence identity in addition to the CDRs. Methods for producing such antibodies are known in the art (see EP 239,400; PCT Publication No. WO 91/09967; and U.S. Patent Nos. 5,225,539; 5,530,101; and 5,585,089); Or surface resurfacing (EP 592,106; EP 519,596; Padlan (1991) Mol. Immunol. 28(4/5): 489-498; Studnicka et al., (1994) Prot. Engineer. 7(6): 805-814; and Roguska et al, (1994) Proc. Acad. Sci. USA 91: 969-973); strand shuffling (U.S. Patent No. 5,565,352); and anti-idiotypic antibodies. A humanized antibody is an antibody molecule from a non-human species that binds to a desired antigen and has one or more CDRs from a non-human species and a framework region from a human immunoglobulin molecule. Human Ig sequences are known to be disclosed, for example, at www.ncbi.nlm.nih.gov/entrez-/query.fcgi; www.atcc.org/phage/hdb.html; www.sciquest.com; www.abcam.com; www .antibodyresource.com/onlinecomp.html ; www.public, iastate.edu/.about.pedro/research tools.html ; www.mgen.uni-heidelberg.de/SD/IT/IT.html ; www.whfreeman.com /immunology/ CH-05/kuby05.htm ; www.library.thinkquest.org/12429/Immune/ 160877.doc -106- 201247704

Antibody.html ; www.hhmi.org/grants/lectures/1996/vlab » www.path.cam.ac.uk7.about.mrc7/m-ikeimages.html ; www. antibodyresource.com ; mcb.harvard.edu/ BioLinks/Immuno-logy.html. ; www.immunologylink.com ; pathbox.wustl. edu/

.about.hcenter/index.-html ; www.biotech.ufl.edu/.about.hcl ; www.pebio.com/pa/340913/340913.html- ; www.nal.usda.gov/ awic/pubs/ Antibody ; www.m.ehime-u.acjp/.about.yasuhito-/ Elisa.html ; www.biodesign.com/table.asp ; www.icnet.uk/ axp/facs/davies/lin-ks.html ; Www. biotech, ufl.edu/. about, fccl/protocol.html ; www.isac-net.org/sites_geo.html ; aximtl. imt.uni-marburg.de/.about.rek/AEP-Start.html ; Baserv.uci. kun.nl/.about.jraats/linksl.html ; www.recab.uni-hd.de/immuno. bme.nwu.edu/ ; www.mrc-cpe.cam.ac.uk/imt- Doc/pu-blic/INTRO. html ; www.ibt.unam.mx/vir/V_mice.html ; imgt.cnusc.fr:8104/ ; www.biochem.ucl.ac.uk/.about.martin/abs/ Index.html ; antibody. bath.ac.uk/ ; abgen.cvm.tamu.edu/lab/wwwabgen.html ; www. unizh.ch/.about.honegger/AHOsem-inar/SlideO 1 .html ; www. cryst .bbk.ac.uk/.about.ubcg07s/ ; www.nimr.mrc.ac.uk/CC/ccaewg/ ccaewg.htm ; www.path.cam.ac.uk/.about.mrc7/h-umanisation/ TAHHP.html ; www.ibt.unam.mx/vir/structure/stat_aim.html ; www.biosci.missouri .edu/smithgp/index.html ; www.cryst. bioc.cam.ac.uk/.abo-ut.fmolina/Web-pages/Pept/spottech.html ; www.jerini.de/fr roducts.htm ; www .patents.ibm.com/ibm.html ; and Kabat et al., Sequences of Proteins of Immunological 160877.doc -107- 201247704

Interest, U.S. Dept. Hea] th (1983). Such input sequences can be used to reduce immunogenicity or to reduce, enhance or improve binding, affinity, association rate, dissociation rate, affinity, specificity, half-life, or any other suitable feature known in the art. "X replaces framework residues in the human framework regions with corresponding residues from CDR donor antibodies to alter (e. g., improve) antigen binding. Such framework substitutions are identified by methods well known in the art, such as by establishing a model for interaction of cDR with framework residues to identify framework residues important for antigen binding and for sequence comparison to identify unusual locations. Framework residues. (See, e.g., U.S. Patent No. 5,5,85,089; Riechmann et al., (1988) Nature 332:323). Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs can be utilized that illustrate and display possible three-dimensional conformational structures of selected candidate immunoglobulin sequences. The tests shown therein permit the analysis of the possible role of the residues in the functioning of the candidate immunoglobulin sequences', i.e., the analysis of the residues affecting the ability of the candidate immunoglobulin to bind its antigen. In this manner, FR residues can be selected from the common and input sequences and combined to achieve the desired antibody characteristics, such as increased affinity for the antigen of interest. In general, 'CDR residues are directly and to the greatest extent involved in affecting antigen binding. Antibodies can be humanized using a variety of techniques known in the art, such as, but not limited to, Jones et al, (1986) Nature 321 : 522; Verhoeyen et al, (1988) Science 239: 1534; Sims et al, ( 1993) J. Immunol. 151: 2296; Chothia and Lesk (1987) J. Mol. Biol. 196: 901; Carter et al., (1992) Proc. Natl. Acad. Sci. USA 89: 4285; Presta et al. (1993) J. Immunol. 151: 160877.doc -108- 201247704 2623; Padlan (1991) Mol. Immunol. 28(4/5): 489-498; Studnicka et al., (1994) Prot. Engineer. 7( 6): 805-814;

Roguska et al., (1994) Proc. Natl. Acad. Sci. USA 91: 969-973; PCT Publication No. WO 91/09967; US 98/16280; US 96/18978; US 91/ No. 09630; US 91/05939; US 94/01234; GB 89/01334; GB 91/01134; GB 92/01755; WO 90/14443; WO 90/14424 And WO 90/14430; European Patent Publication No. EP 229,246; EP 592,106; EP 519,596; and EP 239,400; and U.S. Patent No. 5,565,332; 5,723,323; 5,976,862 No. 5,824,514; 5,817,483; 5,814,476; 5,763,192; 5,723,323; 5,766,886; 5,714,352; 6,204,023; 6,180,370; 5,693,762; 5,530,101 No. 5,585,089; 5,225,539; and the technique described in 4,816,567 A4. Exemplary Single Variable Domains Exemplary single variable domains for use in the semi-Ig binding proteins of the invention include the following variable domain sequences . The single variable domain set forth below is included in the DVD provided in U.S. Patent No. 6,612,181, the disclosure of which is incorporated herein in its entirety. 160877.doc •109· 201247704 Table 1: List of amino acid sequences for the VH and VL regions of antibodies used to generate semi-Ig binding proteins

SEQ ID NO. The variable region protein sequences of the domain name 1234567890123456789012345678901234567890 118 AB081VH VH HIV (sequence 1) QVQLQQSGAELMKPGASVKISCKASGYTETSYWIEWIKQR PGHGLEWIGEILPGTGSLNNNEKFRDKATFTADTSSNTAY MQLSSLTSEDSAVYYCARGYRYDGWFAYWGQGTLVTVSA 119 AB081VL VL HIV (sequence 1) DIQMTQSPASLSASVGETVTITCRTSENIYSYLAWYQQKP GKSPHLLVYNTKTLAEGVPSRFSGSGSGTQFSLKINSLQP EDFGSYYCQHHYDSPLTFGSGTKLELKR 120 AB082VH VH NGAL (sequence 1) EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISRDTAR NTLYLQMTSLKSEDTAMYYCARHFGDYSYFDYWGQGTTLT VSS 121 AB082VL VL NGAL (SEQ ID 1 > DIQMTQSPASLSASVGETVTITCRASENFYSYLAWYQQKQ GKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFSLKINSLQP EDFGTYYCQHHYDIPLTFGAGTKLELKR 122 AB083VH VH NGAL (sequence 2 > KIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNWVKQA PGKGLKWMGWININTGEPTYAEEFKGRFAFSLETSATTAF LQINNLKNEDTATYLCARDSYSGGFDYWGQGTIVTVSS 123 AB083VL VL NGAL (sequence 2) DIVMTQSPSSLSVSAGEKVTLSCKSSQSLLISGDQKNYLA WYQQKPGQPPKLLIYGASTRDSGVPDRFTGSGSGADFTLT ISSVQAEDLAVYYCQNDHSFPPTFGAGTKLELKR 124 AB084 VH VH HIV (sequence 2) QIQLVQSGPELKKPGETVKISCKASGYTFTDYSMHWVKQA PGKGLKWMGWIHTETGEPRYVDDFKGRFAFSLETSASTAY LQINNLKNEDTATYFCARDSYYFGSSYYFDYWGQGTTLTV SS 125 AB084VL VL HIV (sequence 2) DTVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWYQQKP GQSPKLLIYSASYRYTGVPDRFTGSGSGMDFTFTISSVQA EDLAVYYCQQHYSTPLTFGAGTKLELER 126 AB085VH ▽ H HIV (sequence 3) EVQLQQSGPELVKPGASMKISCKASDYSFTAYTIHWMKQS HGKNLEWIGLINPYNGGTSYNQKFQGRATLTVDKSSSIAY MELLSLTSEDSAVYYCARRGYDREGHYYAMDYWGQGTSVT VSS 127 AB085VL VL HIV (sequence 3) DIQMTQSPASLAASVGETVTITCRASENIYTFLAWYQQKQ GKSPQLLVYTTKTLAEGVPSRFSGSGSGTQFSLKIKSLQP EDFGSYYCQHHYGLPLTFGAGTKLELKR 128 AB086VH VH HIV (SEQ ID 4) EVQLQQSGPELVQPGASMKISCKASGYSFTDYTMNWVKQS HGKNLEWIGLINPYNGGSRYNQKFMAKATLTVDKSSNTAY MELLSVTSEDSAVYYCARDAGYFGSGFYFDYWGQGTTLTV SS 129 AB086VL VL HIV (sequence 4) DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWYQQKP GQSPKLLIYSASYRSTGVPDRFTGSGSGTDFTFTISSVQA EDLAVYYCQQHYSTPTFGAGTKLELKR 130 AB088VH VH IL-18 QVQLQQPGSELVRPGASVKLSCKASGYTFTSYWMHWVKQR PGQGLEWIGNIYPGTVNTNYDEKFKNKATLTVDTSSSTAY -110 · 160877.doc 201247704

SEQ ID NO · variable region protein sequences of the domain name 1234567890123456789012345678901234567890 MLLSSLTSEDSAVYYCTRDYYGGGLNYWGQGTTLTVSS 131 AB088VL VL IL-18 SIVMTQTPKFLLVSAGDRVTITCKASQSVSNDVAWFQQKP GQSPKLLIYYASNRYAGVPDRFTGSGFGTDFTFTISTVQA EDLAVYFCHQDYSSPRTFGGGTKLEIKR 132 AB089VH VH BNP (sequence 1) QIQLVQSGPELRKPGETVKISCKGSGYTFTHYGINWVKQT PRKDLKWMGWINTHTGEAYYADDFKGRFAFSLETSANTAY LQINNLNNGDMGTYFCTRSHRFGLDYWGQGTSVTVSS 133 AB089VL VL BNP (sequence 1) DNVLTQSPPSLAVSLGQRATISCKANWPVDYNGDSYLNWY QQKPGQPPKFLIYAASNLESGIPARFSGSGSGTDFNLNIH PVEEEDAATYYCQQSNEDPFTFGSGTKLEIKR 134 AB090VH VH BNP (sequence 2) QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNWVKQR PEQGLEWIGRIDPYDSETHYNQKFKDKAILTVDKSSSTAF VQLTSLTSEDSAVYYCVSDGYWGAGTTVTVSS 135 AB090VL VL BMP (sequence 2) DVVMTQTPLTLSVTTGQPASISCKSSQSLLDSDGKTYLNW LFQRPGESPKLLIYWSKLESGVPDRFTGSGSGTDFTLKI SRVEAEDLGVYYCLQATHFPWTFGGGTKLEIKR 136 AB092VH VH BNP (sequence 4) QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNWVKQR PEQGLEWIGRIDPYDSETHYNQKFKDKAILTVDKSSSTAF VQLTSLTSEDSAVYYCVSDGYWGAGTTVTVSS 137 AB092VL VL BNP (sequence 4) DWMTQTPLTLSVTTGQPASISCKSSQSLLDSDGKTYLNW LFQRPGESPKLLIYVTDILESGVPDRFTGSGSGTDFTLKI SRVEAEDLGVYYCLQATHFPWTFGGGTKLEIKR 138 AB093VH VH Tnl EVQLQQSGPDLVKPGASVRISCKASGYTFTDYNLHWVKQS HGKSLEW station GYIYPYNGITGYNQKFKSKATLTVDSSSNTAY MDLRSLTSEDSAVYFCARDAYDYDYLTDWGQGTLVTVSA 139 AB093VL VL Tnl DILLTQSPVILSVSPGERVSFSCRTSKNVGTNIHWYQQRT NGSPRLLIKYASERLPGIPSRFSGSGSGTDFTLSINSVES EDIADYYCQQSNNWPYTFGGGTKLEIKR

Other single variable domain sequences are provided in the examples below. A5. Dual variable domain binding protein The semi-DVD-Ig binding protein of the present invention can be produced by selecting a variable domain from the single antibody identified above or a single variable domain provided herein; and using the above method. Alternatively, the semi-DVD-Ig binding protein can be produced using the sequences provided in U.S. Patent Publication Nos. 20,100,260,668 and 2009030 4,469. The sequences may also be selected from the table below or other sequences provided below. It will be appreciated that the single variable domain may be selected from the dual variable domains used in other semi-Ig binding proteins of the invention. Alternative linker sequences from the sequences shown in bold can be used to join variable domains. • 111-160877.doc 201247704 Exemplary dual variable domains for use in the half-ig binding proteins of the invention include the following dual variable domain sequences for binding to a specified protein. The linker sequence is shown in bold. Table 2: Dual variable domain sequences with both first and second variable domains for binding to HIV

SEQ ID NO DVD variable domain, said variable domain, said variable domain outside the said linker sequence 123456789012345678901234567890123456 140 DVD715H AB081VH HG- short AB081VH QVQLQQSGAELMKPGASVKISCKASGYTFTSYWIEW IKQRPGHGLEWIGEILPGTGSLNNNEKFRDKATFTA DTSSNTAYMQLSSLTSEDSAVYYCARGYRYDGWFAY WGQGTLVTVSAASTKGPQVQLQQSGAELMKPGASVK ISCKASGYTFTSYWIEWIKQRPGHGLEWIGEILPGT GSLNNNEKFRDKATFTADTSSNTAYMQLSSLTSEDS AVYYCARGYRYDGWFAYWGQGTLVTVSA 141 DVD715L AB081VL LK- short AB081VL DIQMTQSPASLSASVGETVTITCRTSENIYSYLAWY QQKPGKSPHLLVYNTKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGSYYCQHHYDSPLTFGSGTKLELKR TVAAPDIQMTQSPASLSASVGETVTITCRTSENIYS YLAWYQQKPGKSPHLLVYNTKTLAEGVPSRFSGSGS GTQFSLKINSLQPEDFGSYYCQHHYDSPLTFGSGTK LELKR 142 DVD716H AB081VH HG- long AB081VH QVQLQQSGAELMKPGASVKISCKASGYTFTSYWIEW IKQRPGHGLEWIGEILPGTGSLNNNEKFRDKATFTA DTSSNTAYMQLSSLTSEDSAVYYCARGYRYDGWFAY WGQGTLVTVSAASTKGPSVFPLAPQVQLQQSGAELM KPGASVKISCKASGYTFTSYWIEWIKQRPGHGLEWI GEILPGTGSLNNNEKFRDKATFTADTSSNTAYMQLS SLTSEDSAVYYCARGYRYDGWFAYWGQGTLVTVSA 143 DVD716L AB081VL LK- AB081VL DIQMTQSPASLSASVGETVTITCRTSENIYSYLAWY QQKPGKSPHLLVYNTKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGSYYCQHHYDSPLTFGSGTKLELKR TVAAPSVFIFPPDIQMTQSPASLSASVGETVTITCR TSENIYSYLAWYQQKPGKSPHLLVYNTKTLAEGVPS RFSGSGSGTQFSLKINSLQPEDFGSYYCQHHYDSPL TFGSGTKLELKR 144 DVD717H AB081VH HG- length X2 AB081VH QVQLQQSGAELMKPGASVKISCKASGYTFTSYWIEW IKQRPGHGLEWIGEILPGTGSLNNNEKFRDKATFTA DTSSNTAYMQLSSLTSEDSAVYYCARGYRYDGWFAY WGQGTLVTVSAASTKGPSVFPLAPASTKGPSVFPLA PQVQLQQSGAELMKPGASVKISCKASGYTFTSYWIE WIKQRPGHGLEWIGEILPGTGSLNNNEKFRDKATFT ADTSSNTAYMQLSSLTSEDSAVYYCARGYRYDGWFA YWGQGTLVTVSA 145 DVD717L AB081VL LK- length X2 AB081VL DIQMTQSPASLSASVGETVTITCRTSENIYSYLAWY QQKPGKSPHLLVYNTKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGSYYCQHHYDSPLTFGSGTKLELKR

•112· 160877.doc 201247704

TVAAPSVFIFPPTVAAPSVFIFPPDIQMTQSPASLS ASVGETVTITCRTSEMIYSYLAWYQQKPGKSPHLLV YNTKTLAEGVPSRFSGSGSGTQFSLKINSLQPEDFG SYYCQHHYDSPLTFGSGTKLELKR 146 DVD746H AB081VH HG- long AB085VH QVQLQQSGAELMKPGASVKISCKASGYTFTSYWIEW IKQRPGHGLEWIGEILPGTGSLNNNEKFRDKATFTA DTSSNTAYMQLSSLTSEDSAVYYCARGYRYDGWFAY WGQGTLVTVSAASTKGPSVFPLAPEVQLQQSGPELV KPGASMKISCKASDYSFTAYTIHWMKQSHGKNLEWI GLINPYNGGTSYNQKFQGRATLTVDKSSSIAYMELL SLTSEDSAVYYCARRGYDREGHYYAMDYWGQGTSVT VSS 147 DVD746L AB081VL LK- long AB085VL DIQMTQSPASLSASVGETVTITCRTSENIYSYLAWY QQKPGKSPHLLVYNTKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGSYYCQHHYDSPLTFGSGTKLELKR TVAAPSVFIFPPDIQMTQSPASLAASVGETVTITCR ASENIYTFLAWYQQKQGKSPQLLVYTTKTLAEGVPS RFSGSGSGTQFSLKIKSLQPEDFGSYYCQHHYGLPL TFGAGTKLELKR 148 DVD747H AB081VH HG- long X2 AB085VH QVQLQQSGAELMKPGASVKISCKASGYTFTSYWIEW IKQRPGHGLEWIGEILPGTGSLNNNEKFRDKATFTA DTSSNTAYMQLSSLTSEDSAVYYCARGYRYDGWFAY WGQGTLVTVSAASTKGPSVFPLAPASTKGPSVFPLA PEVQLQQSGPELVKPGASMKISCKASDYSFTAYTIH WMKQSHGKNLEWIGLINPYNGGTSYNQKFQGRATLT VDKSSSIAYMELLSLTSEDSAVYYCARRGYDR EGHY YAMDYWGQGTSVTVSS 149 DVD747L AB081VL LK- length X2 AB085VL DIQMTQSPASLSASVGETVTITCRTSENIYSYLAWY QQKPGKSPHLLVYNTKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGSYYCQHHYDSPLTFGSGTKLELKR TVAAPSVFIFPPTVAAPSVFIFPPDIQMTQSPASLA ASVGETVTITCRASENIYTFLAWYQQKQGKSPQLLV YTTKTLAEGVPSRFSGSGSGTQFSLKIKSLQPEDFG SYYCQHHYGLPLTFGAGTKLELKR 150 DVD748H AB085VH HG- long AB081VH EVQLQQS G PELVKPGASMKISCKAS DYS FTAYTIHW MKQSHGKNLEWIGLINPYNGGTSYNQKFQGRATLTV DKSSSIAYMELLSLTSEDSAVYYCARRGYDREGHYY AMDYWGQGTSVTVSSASTKGPSVFPLAPQVQLQQSG AELMKPGASVKISCKASGYTFTSYWIEWIKQRPGHG LEWIGEILPGTGSLNNNEKFRDKATFTADTSSNTAY MQLSSLTSEDSAVYYCARGYRYDGWFAYWGQGTLVT VSA 151 DVD748L AB085VL LK- long AB081VL DIQMTQSPASLAASVGETVTITCRASENIYTFLAWY QQKQGKSPQLLVYTTKTLAEGVPSRFSGSGSGTQFS LKIKSLQPEDFGSYYCQHHYGLPLTFGAGTKLELKR TVAAPSVFIFPPDIQMTQSPASLSASVGETVTITCR TSENIYSYLAWYQQKPGKSPHLLVYNTKTLAEGVPS RFSGSGSGTQFSLKINSLQPEDFGSYYCQHHYDSPL TFGSGTKLELKR 152 DVD749H AB085VH HG-Long X2 AB081VH EVQLQQSGPELVKPGASMKISCKAS DYS FTAYTIHW MKQSHGKNLEWIGLINPYNGGTSYNQKFQGRATLTV DKS SSIAYMELLSLTSEDSAVYYCARRGYDREGHYY AMDYWGQGTS VTVS SASTKGPSVFPLAPASTKGPSV FPLAPQVQLQQSGAELMKPGASVKISCKASGYTFTS 160877.doc •113· 201247704

YWIEWIKQRPGHGLEWIGEILPGTGSLNNNEKFRDK ATFTADTSSNTAYMQLSSLTSEDSAVYYCARGYRYD GWFAYWGQGTLVTVSA 153 DVD749L AB085VL LK- length X2 AB081VL DIQMTQSPASLAASVGETVTITCRASENIYTFLAWY QQKQGKSPQLLVYTTKTLAEGVPSRFSGSGSGTQFS LKIKSLQPEDFGSYYCQHHYGLPLTFGAGTKLELKR TVAAPSVFIFPPTVAAPSVFIFPPDIQMTQSPASLS ASVGETVTITCRTSENIYSYLAWYQQKPGKSPHLLV YNTKTLAEGVPSRFSGSGSGTQFSLKINSLQPEDFG SYYCQHHYDSPLTFGSGTKLELKR 154 DVD744H AB084VH HG- long AB084VH QIQLVQSGPELKKPGETVKISCKASGYTFTDYSMHW VKQAPGKGLKWMGWIHTETGEPRYVDDFKGRFAFSL ETSASTAYLQINNLKNEDTATYFCARDSYYFGSSYY FDYWGQGTTLTVSSASTKGPSVFPLAPQIQLVQSGP ELKKPGETVKISCKASGYTFTDYSMHWVKQAPGKGL KWMGWIHTETGEPRYVDDFKGRFAFSLETSASTAYL QINNLKNEDTATYFCARDSYYFGSSYYFDYWGQGTT LTVSS 155 DVD744L AB084VL LK- long AB084VL DTVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWY QQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGMDFT FTISSVQAEDLAVYYCQQHYSTPLTFGAGTKLELER TVAAPSVFIFPPDTVMTQSHKFMSTSVGDRVSITCK ASQDVSSAVAWYQQKPGQSPKLLIYSASYRYTGVPD RFTGSGSGMDFTFTISSVQAEDLAVYYCQQHYSTPL TFGAGTKLELER 156 DVD745H AB084VH HG- short AB084VH QIQLVQSGPELKK PGETVKISCKASGYTFTDYSMHW VKQAPGKGLKWMGWIHTETGEPRYVDDFKGRFAFSL ETSASTAYLQINNLKNEDTATYFCARDSYYFGSSYY FDYWGQGTTLTVSSASTKGPQIQLVQSGPELKKPGE TVKISCKASGYTFTDYSMHWVKQAPGKGLKWMGWIH TETGEPRYVDDFKGRFAFSLETSASTAYLQINNLKN EDTATYFCARDSYYFGSSYYFDYWGQGTTLTVSS 157 DVD745L AB084VL LK * short AB084VL DTVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWY QQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGMDFT FTISSVQAEDLAVYYCQQHYSTPLTFGAGTKLELER TVAAPDTVMTQSHKFMSTSVGDRVSITCKASQDVSS AVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGS GMDFTFTISSVQAEDLAVYYCQQHYSTPLTFGAGTK LELER 158 DVD750H AB086VH HG- long AB086VH EVQLQQSGPELVQPGASMKISCKASGYSFTDYTMNW VKQSHGKNLEWIGLINPYNGGSRYNQKFMAKATLTV DKSSNTAYMELLSVTSEDSAVYYCARDAGYFGSGFY FDYWGQGTTLTVSSASTKGPSVFPLAPEVQLQQSGP ELVQPGASMKISCKASGYSFTDYTMNWVKQSHGKNL EWIGLINPYNGGSRYNQKFMAKATLTVDKSSNTAYM ELLSVTSEDSAVYYCARDAGYFGSGFYFDYWGQGTT LTVSS 159 DVD750L AB086VL LH- long AB086VL DIVMTQSHKFMSTSVGDRVSITCKASQDVSTAVAWY QQKPGQSPKLLIYSASYRSTGVPDRFTGSGSGTDFT FTISSVQAEDLAVYYCQQHYSTPTFGAGTKLELKRT VAAPSVFIFPPDIVMTQSHKFMSTSVGDRVSITCKA SQDV STAVAWYQQKPGQSPKLLIYSASYRSTGVPDR FTGSGSGTDFTFTISSVQAEDLAVYYCQQHYSTPTF GAGTKLELKR

160877.doc •114- 201247704 Table 3: Dual variable domain sequences with both first and second variable domains for binding to NGAL

SEQ ID NO DVD variable domain, said variable domain, said variable domain outside the said linker sequence 123456789012345678901234567890123456 160 DVD719H AB082VH HG- long AB082VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPSVFPLAPEVQLVESGGGLV QPGGSLKLSCAASGFTFNNYYMSWVRQTPERRLEWV AYISSSGGSTYYSDSVRGRFTISRDTARNTLYLQMT SLKSEDTAMYYCARHFGDYSYFDYWGQGTTLTVSS 161 DVD719L AB082VL LK- long AB082VL DIQMTQSPASLSASVGETVTITCRASENFYSYIAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPSVFIFPPDIQMTQSPASLSASVGETVTITCR ASENFYSYLAWYQQKQGKSPQLLVYNAKTLAEGVPS RFSGSGSGTQFSLKINSLQPEDFGTYYCQHHYDIPL TFGAGTKLELKR 162 DVD720H AB082VH HG- short AB082VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPEVQLVESGGGLVQPGGSLK LSCAASGFTFNNYYMSWVRQTPERRLEWVAYISSSG GSTYYSDSVRGRFTISRDTARNTLYLQMTSLKSEDT AMYYCARHFGDYSYFDYWGQGTTLTVSS 163 DVD720L AB082VL LK-short AB082VL DIQMTQSPASLSASVGETVTITCRASENFYSYLAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPDIQMTQSPASLSASVGETVTITCRASENFYS YLAWYQQKQGKSPQLLVYNAKTLAEGVPSRFSGSGS GTQFSLKINSLQPEDFGTYYCQHHYDIPLTFGAGTK LELKR 164 DVD721H AB083VH HG- long AB083VH KIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNW VKQAPGKGLKWMGWININTGEPTYAEEFKGRFAFSL ETSATTAFLQINNLKNEDTATYLCARDSYSGGFDYW GQGTIVTVSSASTKGPSVFPIAPKIQLVQSGPELKK PGETVKISCKASGYTFTNYGMNWVKQAPGKGLKWMG WININTGEPTYAEEFKGRFAFSLETSATTAFLQINN LKNEDTATYLCARDSYSGGFDYWGQGTIVTVSS 165 DVD721L AB083VL LK- long AB083VL DIVMTQSPSSLSVSAGEKVTLSCKSSQSLLISGDQK NYLAWYQQKPGQPPKLLIYGASTRDSGVPDRFTGSG SGADFTLTISSVQAEDLAVYYCQNDHSFPPTFGAGT KLELKRTVAAPSVFIFPPDIVMTQSPSSLSVSAGEK VTLSCKSSQSLLISGDQKNYLAWYQQKPGQPPKLLI YGASTRDSGVPDRFTGSGSGADFTLTISSVQAEDLA VYYCQNDHSFPPTFGAGTKLELKR 166 DVD722H AB083VH HG- short AB083VH KIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNW VKQAPGKGLKWMGWININTGEPTYAEEFKGRFAFSL ETSATTAFLQINNLKNEDTATYLCARDSYSGGFDYW GQGTIVTVSSASTKGPKIQLVQSGPELKKP GETVKI -115- 160877.doc 201247704

SCKASGYTFTNYGMNWVKQAPGKGLKWMGWININTG EPTYAEEFKGRFAFSLETSATTAFLQINNLKNEDTA TYLCARDSYSGGFDYWGQGTIVTVSS 167 DVD722L AB083VL LK- short AB083VL DIVMTQSPSSLSVSAGEKVTLSCKSSQSLLISGDQK NYLAWYQQKPGQPPKLLIYGASTRDSGVPDRFTGSG SGADFTLTISSVQAEDLAVYYCQNDHSFPPTFGAGT KLELKRTVAAPDIVMTQSPSSLSVSAGEKVTLSCKS SQSLLISGDQKNYLAWYQQKPGQPPKLLIYGASTRD SGVPDRFTGSGSGADFTLTISSVQAEDLAVYYCQND HSFPPTFGAGTKLELKR 168 DVD723H AB082VH HG- long AB083VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPSVFPLAPKIQLVQSGPELK KPGETVKISCKASGYTFTNYGMNWVKQAPGKGLKWM GWININTGEPTYAEEFKGRFAFSLETSATTAFLQIN NLKNEDTATYLCARDSYSGGFDYWGQGTIVTVSS 169 DVD723L AB082VL LK- long AB083VL DIQMTQSPASLSASVGETVTITCRASENFYSYLAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPSVFIFPPDIVMTQSPSSLSVSAGEKVTLSCK SSQSLLISGDQKNYLAWYQQKPGQPPKLLIYGASTR DSGVPDRFTGSGSGADFTLTISSVQAEDLAVYYCQN DHSFPPTFGAGTKLELKR 170 DVD724H AB082VH HG- Short AB083VH EVQLVESGGGLVQ PGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPKIQLVQSGPELKKPGETVK ISCKASGYTFTNYGMNWVKQAPGKGLICWMGWININT GEPTYAEEFKGRFAFSLETSATTAFLQINNLKNEDT ATYLCARDSYSGGFDYWGQGTIVTVSS 171 DVD724L AB082VL LK- short AB083VL 'DIQMTQSPASLSASVGETVTITCRASENFYSYLAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPDIVMTQSPSSLSVSAGEKVTLSCKSSQSLLI SGDQKNYLAWYQQKPGQPPKLLIYGASTRDSGVPDR FTGSGSGADFTLTISSVQAEDLAVYYCQNDHSFPPT FGAGTKLELKR 172 DVD725H AB083VH HG- long AB082VH KIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNW VKQAPGKGLKWMGWININTGEPTYAEEFKGRFAFSL ETSATTAFLQINNLKNEDTATYLCARDSYSGGFDYW GQGTIVTVSSASTKGPSVFPLAPEVQLVESGGGLVQ PGGSLKLSCAASGFTFNNYYMSWVRQTPERRLEWVA YISSSGGSTYYSDSVRGRFTISRDTARNTLYLQMTS LKSEDTAMYYCARHFGDYSYFDYWGQGTTLTVSS 173 DVD725L AB083VL LK- long AB082VL DIVMTQSPSSLSVSAGEKVTLSCKSSQSLLISGDQK NYLAWYQQKPGQPPKLLIYGASTRDSGVPDRFTGSG SGADFTLTISSVQAEDLAVYYCQNDHSFPPTFGAGT KLELKRTVAAPSVFIFPPDIQMTQSPASLSASVGET VTITCRASEN FYSYLAWYQQKQGKSPQLLVYNAKTL AEGVPSRFSGSGSGTQFSLKINSLQPEDFGTYYCQH HYDIPLTFGAGTKLELKR 174 DVD726H AB083VH HG- Short AB082VH KIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNW VKQAPGKGLKWMGWININTGEPTYAEEFKGRFAFSL

160877.doc •116· 201247704

ETSATTAFLQINNLKNEDTATYLCARDSYSGGFDYV GQGTIVTVSSASTKGPEVQLVESGGGLVQPGGSLKL SCAASGFTFNNYYMSWVRQTPERRLEWVAYISSSGG STYYSDSVRGRFTISRDTARNTLYLQMTSLKSEDTA MYYCARHFGDYSYFDYWGQGTTLTVSS 175 DVD726L AB083VL LK- short AB082VL DIVMTQSPSSLSVSAGEKVTLSCKSSQSLLISGDQK NYLAWYQQKPGQPPKLLIYGASTRDSGVPDRFTGSG SGADFTLTISSVQAEDLAVYYCQNDHSFPPTFGAGT KLELKRTVAAPDIQMTQSPASLSASVGETVTITCRA SENFYSYLAWYQQKQGKSPQLLVYNAKTLAEGVPSR FSGSGSGTQFSLKINSLQPEDFGTYYCQHHYDIPLT FGAGTKLELKR Table 4:? Used in conjunction with dual variable domain sequences of NGAL and IL-18 • • SEQ ID NO DVD external variable variable domain names domain Name variable domain sequences within the linker, said 123456789012345678901234567890123456 176 DVD727H AB082VH HG- short AB088VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPQVQLQQPGSELVRPGASVK LSCKASGYTFTSYWMHWVKQRPGQGLEWIGNIYPGT VNTNYDEKFKNKATLTVDTSSSTAYMLLSSLTSEDS AVYYCTRDYYGGGLNYWGQGTTLTVSS 177 DVD727L AB082VL LK- short AB088VL DIQMTQSPASLSASVGETVTITCRASENFYSYLAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPSIVMTQTPKFLLVSAGDRVTITCKASQSVSN DVAWFQQKPGQSPKLLIYYASNRYAGVPDRFTGSGF GTDFTFTISTVQAEDLAVYFCHQDYSSPRTFGGGTK LEIKR 178 DVD728H AB082VH HG- long AB088VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPSVFPLAPQVQLQQPGSELV RPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWI GNIYPGTVNTNYDEKFKNKATLTVDTSSSTAYMLLS SLTSEDSAVYYCTRDYYGGGLNYWGQGTTLTVSS 179 DVD728L AB082VL LK- long AB088VL DIQMTQSPASLSASVGETVTITCRASENFYSYLAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPSVFIFPPSIVMTQTPKFLLVSAGDRVTITCK ASQSVSNDVAWFQQKPGQSPKLLIYYASNRYAGVPD RFTGSGFGTDFTFTISTVQAEDLAVYFCHQDYSSPR TFGGGTKLEIKR 180 DVD729H AB082VH HG- length X2 AB088VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSSASTKGPSVFPLAPASTKGPSVFPLA PQVQLQQPGSELVRPGASVKLSCKASGYTFTSYWMH WVKQRPGQGLEWIGNIYPGT VNTNYDEKFKNKATLT •117· 160877.doc 201247704

VDTSSSTAYMLLSSLTSEDSAVYYCTRDYYGGGLNY WGQGTTLTVSS 181 DVD729L AB082VL LK- long X2 AB088VL DIQMTQSPASLSASVGETVTITCRASENFYSYLAWY QQKQGKSPQLLVYNAKTLAEGVPSRFSGSGSGTQFS LKINSLQPEDFGTYYCQHHYDIPLTFGAGTKLELKR TVAAPSVFIFPPTVAAPSVFIFPPSIVMTQTPKFLL VSAGDRVTITCKASQSVSNDVAWFQQKPGQSPKLLI YYASNRYAGVPDRFTGSGFGTDFTFTISTVQAEDLA VYFCHQDYSSPRTFGGGTKLEIKR 182 DVD730H AB088VH HG- short AB082VH QVQLQQPGSELVRPGASVKLSCKASGYTFTSYWMHW VKQRPGQGLEWIGNIYPGTVNTNYDEKFKNKATLTV DTSSSTAYMLLSSLTSEDSAVYYCTRDYYGGGLNYW GQGTTLTVSSASTKGPEVQLVESGGGLVQPGGSLKL SCAASGFTFNNYYMSWVRQTPERRLEWVAYISSSGG STYYSDSVRGRFTISRDTARNTLYLQMTSLKSEDTA MYYCARHFGDYSYFDYWGQGTTLTVSS 183 DVD730L AB088VL LK- short AB082VL SIVMTQTPKFLLVSAGDRVTITCKASQSVSNDVAWF QQKPGQSPKLLIYYASNRYAGVPDRFTGSGFGTDFT FTISTVQAEDLAVYFCHQDYSSPRTFGGGTKLEIKR TVAAPDIQMTQSPASLSASVGETVTITCRASENFYS YLAWYQQKQGKSPQLLVYNAKTLAEGVPSRFSGSGS GTQFSLKINSLQPEDFGTYYCQHHYDIPLTFGAGTK LELKR 184 DVD731H AB088VH HG- AB024VH QVQLQQPGSELVRPGASVKLSCKASGYTFTSYWMHW VKQRPGQGLEWIGNIYPGTVNTNYDEKFKNKATLTV DTS SSTAYMLLSSLTSEDSAVYYCTRDYYGGGLNYW GQGTTLTVSSASTKGPSVFPIAPEVQLVESGGGLVQ PGGSLKLSCAASGFTFNNYYMSWVRQTPERRLEWVA YISSSGGSTYYSDSVRGRFTISRDTARNTLYLQMTS LKSEDTAMYYCARHFGDYSYFDYWGQGTTLTVSS 185 DVD731L AB088VL LK- long AB082VL SIVMTQTPKFLLVSAGDRVTITCKASQSVSNDVAWF QQKPGQSPKLLIYYASNRYAGVPDRE'TGSGF'GTDFT FTISTVQAEDLAVYFCHQDYSSPRTFGGGTKLEIKR TVAAPSVFIFPPDIQMTQSPASLSASVGETVTITCR ASENFYSYLAWYQQKQGKSPQLLVYNAKTLAEGVPS RFSGSGSGTQFSLKINSLQPEDFGTYYCQHHYDIPL TFGAGTKLELKR 186 DVD732H AB088VH HG- length X2 AB082VH QVQLQQPGSELVRPGASVKLSCKASGYTFTSYWMHW VKQRPGQGLEWIGNIYPGTVNTNYDEKFKNKATLTV DTSSSTAYMLLSSLTSEDSAVYYCTRDYYGGGLNYW GQGTTLTVSSASTKGPSVFPLAPASTKGPSVFPLAP EVQLVESGGGLVQPGGSLKLSCAASGFTFNNYYMSW VRQTPERRLEWVAYISSSGGSTYYSDSVRGRFTISR DTARNTLYLQMTSLKSEDTAMYYCARHFGDYSYFDY WGQGTTLTVSS 187 DVD732L AB088VL LK- length X2 AB082VL SIVMTQT PKFLLVSAGDRVTITCKASQSVSNDVAWF QQKPGQSPKLLIYYASNRYAGVPDRFTGSGFGTDFT FTISTVQAEDLAVYFCHQDYSSPRTFGGGTKLEIKR TVAAPSVFIFPPTVAAPSVFIFPPDIQMTQSPASLS ASVGETVTITCRASENFYSYLAWYQQKQGKSPQLLV YNAKTLAEGVPSR FSGSGSGTQFSLKINSLQPEDFG TYYCQHHYDIPLTFGAGTKLELKR

160877.doc -118- 201247704 Table 5: Dual variable domain sequences with both first and second variable domains for binding to BNP

SEQ ID NO DVD variable domain, said variable domain name outside the variable domain name linker sequence 123456789012345678901234567890123456 188 DVD733H AB089VH HG- long AB089VH QIQLVQSGPELRKPGETVKISCKGSGYTFTHYGINW VKQTPRKDLKWMGWINTHTGEAYYADDFKGRFAFSL ETSANTAYLQINNLNNGDMGTYFCTRSHRFGLDYWG QGTSVTVSSASTKGPSVFPLAPQIQLVQSGPELRKP GETVKISCKGSGYTFTHYGINWVKQTPRKDLKWMGW INTHTGEAYYADDFKGRFAFSLETSANTAYLQINNL NNGDMGTYFCTRSHRFGLDYWGQGTSVTVSS 189 DVD733L AB089VL LK- long AB089VL DNVLTQSPPSLAVSLGQRATISCKANWPVDYNGDSY LNWYQQKPGQPPKFLIYAASNLESGIPARFSGSGSG TDFNLNIHPVEEEDAATYYCQQSNEDPFTFGSGTKL EIKRTVAAPSVFIFPPDNVLTQSPPSLAVSLGQRAT ISCKANWPVDYNGDSYLNWYQQKPGQPPKFLIYAAS NLESGIPARFSGSGSGTDFNLNIHPVEEEDAATYYC QQSNEDPFTFGSGTKLEIKR 190 DVD734H AB089VH HG- short AB089VH QIQLVQSGPELRKPGETVKISCKGSGYTFTHYGINW VKQTPRKDLKWMGWINTHTGEAYYADDFKGRFAFSL ETSANTAYLQINNLNNGDMGTYFCTRSHRFGLDYWG QGTSVTVSSASTKGPQIQLVQSGPELRKPGETVKIS CKGSGYTFTHYGINWVKQTPRKDLKWMGWINTHTGE AYYADDFKGRFAFSLETSANTAYLQINNLNNGDMGT YFCTRSHRFGLDYWGQGTSVTVSS 191 DVD734L AB089VL LK~ short AB089VL DNVLTQSPPSLAVSLGQRATISCKANWPVDYNGDSY LNWYQQKPGQPPKFLIYAASNLESGIPARFSGSGSG TDFNLNIHPVEEEDAATYYCQQSNEDPFTFGSGTKL EIKRTVAAPDNVLTQSPPSLAVSLGQRATISCKANW PVDYNGDSYLNWYQQKPGQPPKFLIYAASNLESGIP ARFSGSGSGTDFNLNIHPVEEEDAATYYCQQSNEDP FTFGSGTKLEIKR 192 DVD735H AB090VH HG- long AB090VH QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNW VKQRPEQGLEWIGRIDPYDSETHYNQKFKDKAILTV DKSSSTAFVQLTSLTSEDSAVYYCVSDGYWGAGTTV TVSSASTKGPSVFPLAPQVQLQQPGAELVRPGASVK LSCKASGYTFTSYWMNWVKQRPEQGLEWIGRIDPYD SETHYNQKFKDKAILTVDKSSSTAFVQLTSLTSEDS AVYYCVSDGYWGAGTTVTVSS 193 DVD735L AB090VL LK- long AB090VL DVVMTQTPLTLSVTTGQPASISCKSSQSLLDSDGKT YLNWLFQRPGESPKLLIYWSKLESGVPDRFTGSGS GTDFTLKISRVEAEDLGVYYCLQATHFPWTFGGGTK LEIKRTVAAPSVFIFPPDVVMTQTPLTLSVTTGQPA SISCKSSQSLLDSDGKTYLNWLFQRPGESPKLLIYV VSKLESGVPDRFTGSGSGTDFTLKISRVEAEDLGVY YCLQATHFPWTFGGGTKLEIKR 194 DVD736H AB090VH HG- long AB089VH QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNW VKQRPEQGLEWIGRIDPYDSETHYNQKFKDKAILTV -119 · 160877.doc 201247704

DKSSSTAFVQLTSLTSEDSAVYYCVSDGYWGAGTTV TVSSASTKGPSVFPLAPQIQLVQSGPELRKPGETVK ISCKGSGYTFTHYGINWVKQTPRKDLKWMGWINTHT GEAYYADDFKGRFAFSLETSANTAYLQINNLNNGDM GTYFCTRSHRFGLDYWGQGTSVTVSS 195 DVD736L AB090VL LK- long AB089VL DVVMTQTPLTLSVTTGQPASISCKSSQSLLDSDGKT YLNWLFQRPGESPKLLIYWSKLESGVPDRFTGSGS GTDFTLKISRVEAEDLGVYYCLQATHFPWTFGGGTK LEIKRTVAAPSVFIFPPDNVLTQSPPSLAVSLGQRA TISCKANWPVDYNGDSYLNWYQQKPGQPPKFLIYAA SNLESGIPARFSGSGSGTDFNLNIHPVEEEDAATYY CQQSNEDPFTFGSGTKLEIKR 196 DVD737H AB090VH HG- long X2 AB089VH QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNW VKQRPEQGLEWIGRIDPYDSETHYNQKFKDKAILTV DKSSSTAFVQLTSLTSEDSAVYYCVSDGYWGAGTTV TVSSASTKGPSVFPLAPASTKGPSVFPLAPQIQLVQ SGPELRKPGETVKISCKGSGYTFTHYGINWVKQTPR KDLKWMGWINTHTGEAYYADDFKGRFAFSLETSANT AYLQINNLNNGDMGTYFCTRSHRFGLDYWGQGTSVT VSS 197 DVD737L AB090VL LK- long X2 AB089VL DWMTQTPLTLSVTTGQPASISCKSSQSLLDSDGKT YLNWLFQRPGESPKLLIYWSKLESGVPDRFTGSGS GTDFTLKISRVEAEDLGVYYCLQATHFPWTFGGGTK LEIKRTVAAPSVFIFPPTVAAPSVFIFPPDNVLTQS PPSLAVSLGQRATISCKANWPVDYNGDSYLNWYQQK PGQPPKFLIYAASNLESGIPARF SGSGSGTDFNLNI HPVEEEDAATYYCQQSNEDPFTFGSGTKLEIKR 198 DVD738H AB089VH HG- long AB090VH QIQLVQSGPELRKPGETVKISCKGSGYTFTHYGINW VKQTPRKDLKWMGWINTHTGEAYYADDFKGRFAFSL ETSANTAYLQINNLNNGDMGTYFCTRSHRFGLDYWG QGTSVTVSSASTKGPSVFPLAPQVQLQQPGAELVRP GASVKLSCKASGYTFTSYWMNWVKQRPEQGLEWIGR IDPYDSETHYNQKFKDKAILTVDKSSSTAFVQLTSL TSEDSAVYYCVSDGYWGAGTTVTVSS 199 DVD738L AB089VL LK- long AB090VL DNVLTQSPPSLAVSLGQRATISCKANWPVDYNGDSY LNWYQQKPGQPPKFLIYAASNLESGIPARFSGSGSG TDFNLNIHPVEEEDAATYYCQQSNEDPFTFGSGTKL EIKRTVAAPSVFIFPPDVVMTQTPLTLSVTTGQPAS ISCKSSQSLLDSDGKTYLNWLFQRPGESPKLLIYVV SKLESGVPDRFTGSGSGTDFTLKISRVEAEDLGVYY CLQATHFPWTFGGGTKLEIKR 200 DVD739H AB089VH HG- length X2 AB090VH QIQLVQSGPELRKPGETVKISCKGSGYTFTHYGINW VKQTPRKDLKWMGWINTHTGEAYYADDFKGRFAFSL ETSANTAYLQINNLNNGDMGTYFCTRSHRFGLDYWG QGTSVTVSSASTKGPSVFPLAPASTKGPSVFPLAPQ VQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNWV KQRPEQGLEWIGRIDPYDSETHYNQKFKDKAILTVD KSSSTAFVQLTSLTSEDSAVYYCVSDGYWGAGTTVT VSS 201 DVD739L AB089VL LK - Long X2 AB090VL DNVLTQSPPSLAVSLGQRATISCKANWPVDYNGDSY LNWYQQK PGQPPKFLIYAASNLESGIPARFSGSGSG TDFNLNIHPVEEEDAATYYCQQSNEDPFTFGSGTKL

160877.doc •120· 201247704

EIKRTVAAPSVFIFPPTVAAPSVFIFPPDVVMTQTP LTLSVTTGQPASISCKSSQSLLDSDGKTYLNWLFQR PGESPKLLIYVVSKLESGVPDRFTGSGSGTDFTLKI SRVEAEDLGVYYCLQATHFPWTFGGGTKLEIKR 202 DVD742H AB092VH HG- long AB092VH QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWMNW VKQRPEQGLEWIGRIDPYDSETHYNQKFKDKAILTV DKSSSTAFVQLTSLTSEDSAVYYCVSDGYWGAGTTV TVSSASTKGPSVFPLAPQVQLQQPGAELVRPGASVK LSCKASGYTFTSYWMNWVKQRPEQGLEWIGRIDPYD SETHYNQKFKDKAILTVDKSSSTAFVQLTSLTSEDS AVYYCVSDGYWGAGTTVTVSS 203 DVD742L AB092VL LK- long AB092VL DVVMTQTPLTLSVTTGQPASISCKSSQSLLDSDGKT YLNWLFQRPGESPKLLIYVTDILESGVPDRFTGSGS GTDFTLKISRVEAEDLGVYYCLQATHFPWTFGGGTK LEIKRTVAAPSVFIFPPDWMTQTPLTLSVTTGQPA SISCKSSQSLLDSDGKTYLNWLFQRPGESPKLLIYV TDILESGVPDRFTGSGSGTDFTLKISRVEAEDLGVY YCLQATHFPWTFGGGTKLEIKR

Table 6: Dual variable domain sequences with both first and second variable domains for binding to Tnl

SEQ ID NO DVD variable domain, said variable domain, said variable outer linker domain name within the sequence 123456789012345678901234567890123456 204 DVD743H AB093VH HG- long AB093VH EVQLQQSGPDLVKPGASVRISCKASGYTFTDY NLHWVKQSHGKSLEWIGYIYPYNGITGYNQKF KSKATLTVDSSSNTAYMDLRSLTSEDSAVYFC ARDAYDYDYLTDWGQGTLVTVSAASTKGPSVF PLAPEVQLQQSGPDLVKPGASVRISCKASGYT FTDYNLHWVKQSHGKSLEWIGYIYPYNGITGY NQKFKSKATLTVDSSSNTAYMDLRSLTSEDSA VYFCARDAYDYDYLTDWGQGTLVTVSA 205 DVD743L AB093VL LK- long AB093VL DILLTQSPVILSVSPGERVSFSCRTSKNVGTN IHWYQQRTNGSPRLLIKYASERLPGIPSRFSG SGSGTDFTLSINSVESEDIADYYCQQSNNWPY T FGGGTKLEIKRTVAAPSVFIFPPDILLTQS P VILSVSPGERVSFSCRTSKNVGTNIHWYQQRT NGSPRLLIKYASERLPGIPSRFSGSGSGTDFT LSINSVESEDIADYYCQQSNNWPYTFGGGTKL EIKR Other dual variable domain sequences are provided in the following examples. A5. Triple variable domain binding protein The invention can be produced by selecting a single variable domain and a dual variable domain from the monoclonal antibodies identified above, and/or the single variable domain and the dual variable domain provided herein. - 160877.doc 201247704 Half of the TVD-Ig binding protein. Alternatively, the semi-TVD_Ig binding protein can be generated using the sequences provided in U.S. Patent Publication Nos. 20,100,260,668 and 2009030,469. The sequence may also be selected from the sequences provided herein. It will be understood that the single variable domain may be selected from the dual variable domains used in other semi-Ig binding proteins of the invention. An exemplary three-variable or triple-variable domain for use in a semi-Ig binding protein of the invention is provided in US Provisional Patent Application No. 61/426,133, filed on Dec. 22, 2010; Patent application - in the middle. Both applications are filed in the name of the same assignee. The entire content of each of the aforementioned applications, including the sequence listing, is incorporated herein by reference. A6. Domain antibody The half Ig binding protein of the present invention also includes a heavy chain antigen binding domain and a light chain antigen, 5 domain, wherein the antigen binding domain is a domain antibody. Domain antibodies are methods known in the art and which are useful for screening domain antibodies that bind to a particular epitope, such as, for example, 7,829,096, which is incorporated herein by reference. A number of domain antibody sequences are publicly available, for example, in U.S. Patent Nos. 7,696,32, and 7,829,096, and U.S. Patent Publications 2, i 002, 666, s, s, s, s, s, s, s, s, s, s, s, s. A7. Receptor immunoglobulin The semi-Ig binding protein of the present invention may further comprise a heavy chain antigen binding domain and a light chain antigen binding domain, wherein the antigen binding domain is a receptor sequence. A number of receptor sequences are known in the art and can be identified using any of BLAST or a library of materials publicly available from 160877.doc-122.201247704. Acceptor sequences include, for example: 1. CTLA-4: AMHVAQPAVVLASSRGIASFVCEYASPGKATEVRVTVLRQADSQVTEVCAATYMMGNE LTFLDDS1CTGTSSGNQVNLTIQGLRAMDTGLYICKVELMYPPPYYLGIGNGTQIYVIDPE PCPDSD (SEQ ID NO: 206) 2. TNFRSF1B (synonym: CD120b, p75, TNFR2) AQVAFTPYAPEPGSTCRLREYYDQTAQMCCSKCSPGQHAKVFCTKTSDTyCDSCEDSTY TQLWNWVPECLSCGSRCSSDQVETQACTREQNRICTCRPGWYCALSKQEGCRLCAPLRK CRPGFGVARPGTETSDVVCKPCAPGTFSNTTSSTDICRPHQIC (SEQ ID NO: 207) Φ may be used to produce semi- The same molecular biology techniques used for Ig binding proteins (including other variable domain sequences) incorporate the acceptor sequences into the semi-Ig binding proteins of the invention. A8. Skeletal antigen binding protein The semi-Ig binding protein of the present invention comprises a heavy chain antigen binding domain and a light chain antigen binding domain, wherein the antigen binding domain is a backbone antigen binding protein. Skeletal antigen binding proteins are known in the art, for example, fibronectin and designed ankyrin-repeat φ protein (DARPin) have been used as surrogate backbones for antigen binding domains, see, for example, Gebauer and Skerra. , Engineered protein scaffolds as next-generation antibody therapeutics. Curr Op in Chem Biol 13:245-255 (2009) and Stumpp et al, Darpins: A new generation of protein therapeutics. Drug Discov Today 13: 695-701 (2008), Both of these documents are incorporated herein by reference in their entirety. B. Guidelines for selection of parental variable domains and receptors for semi-Ig binding proteins 160877.doc -123- 201247704 An embodiment of the invention relates to the selection of a parental binding protein (eg, an antibody) having a half-jg binding protein A variable domain and/or receptor of one or more properties is desired. The desired properties are selected from one or more binding protein parameters such as antigen specificity, affinity for antigen, potency, biological function, epitope recognition stability, solubility 'production efficiency, immunogenicity, pharmacokinetics, bioavailability , tissue cross-reactivity and orthologous antigen binding. B1. Affinity to Antigen The desired affinity of a therapeutic binding protein, such as a mAb, may depend on the nature of the antigen and the endpoint to be treated. For example, in one embodiment, when a monoclonal antibody blocks a cytokine-cytokine receptor interaction, it has a more affinity (Kd = 0.01-0.50 PM) because these interactions are usually of high affinity. Interaction (eg <pM_<nM range). In such cases, the affinity of the mAb for its target should be equal to or higher than the affinity of the cytokine (ligand) for its receptor. On the other hand, a mAb having a small affinity (>nM range) can be therapeutically effective for, for example, scavenging latent pathogenic proteins in the circulation, such as a single plant that binds, sequesters, and scavenges circulating substances of Αβ-type amyloid. antibody. In other situations,! |Reducing the affinity of existing high-affinity mAbs by site-directed decoy or using a lower affinity for the target can be used to avoid potential side effects, such as high-affinity mAbs may sequester/neutralize all of their intended targets, thereby completely consuming / Eliminate the function of the protein being targeted. In this case, the low affinity mAb can sequester/neutralize a portion of the target that may cause disease symptoms (pathological or overproduced levels), allowing a portion of the target to continue performing its normal physiological function. Therefore, it is possible to reduce the dose by 160877.doc •124·201247704 to adjust the dose and/or reduce side effects. The affinity of the parent binding protein may affect the proper targeting of cell surface molecules to achieve the desired therapeutic result. For example, if a target is expressed on cancer cells at a high density and is expressed on normal cells at a low density, then a lower affinity binding protein will bind a greater number of targets on tumor cells than on normal cells. ADCC or CDC eliminates tumor cells and thus may produce the desired therapeutic effect. Thus, selecting a binding protein (such as a mAb) with the desired affinity may be related to both the soluble target and the surface target. Signaling through interaction of the receptor with its ligand may depend on the affinity of the acceptor-ligand interaction. Similarly, it is believed that the affinity of the binding protein to the surface receptor can be used to determine the nature of intracellular signaling and to bind the protein line to deliver an agonist signal or to deliver an antagonist signal. The affinity-based nature of protein-mediated signaling can affect its side-effect profile. Therefore, it is necessary to carefully determine the desired affinity and desired function of the therapeutic binding protein by in vitro and in vivo experiments. The desired Kd of the binding protein (e.g., antibody) can be determined experimentally depending on the desired therapeutic result. In one embodiment, a parental binding protein (e.g., an antibody) having an affinity for a particular antigen (Kd) equal to or greater than the desired affinity of the half Ig binding protein for the same antigen is selected. When the semi-Ig binding protein comprises two or more functional antigen binding sites, the parent binding proteins for a given half 1 § binding protein may be the same or different. In one embodiment, it is different. Antigen binding affinity and kinetics were assessed by BIACOreTM or another similar technique. In one embodiment, the dissociation constant (Kd) of the parent binding protein to its antigen is selected from the group consisting of: up to about 1〇·7 m·, up to 160877.doc •125·201247704 约·8 Μ ; at most Joΐ〇·9 Μ; at most about ίο·]0 Μ; at most about ίο·"Μ; at most about 1〇-丨2 Μ; and at most about 1〇·η Μ. The first parent binding protein for obtaining VD1 and the second parent binding protein for obtaining VD2 may have similar or different affinities (KD) for the respective antigens. As measured by surface plasmon resonance, the association rate constant of the parent binding protein to the antigen (K(jn) is selected from the group consisting of at least about 102; at least about 1〇3; at least about ΙΟ4 Μ·, -1 ; at least about 5 Μ, -1 ; and at least about 1 〇 6 Μ · ν. When the semi-Ig binding protein includes two or more functional antigen binding sites, 'the first pro for obtaining VD1 The binding protein and the second parent binding protein for obtaining ν〇2 may have similar or different association rate constants (Κ〇η) for the respective antigens. In one embodiment, such as surface plasmon resonance The dissociation rate constant (K〇ff) of each parental binding protein to the antigen is selected from the group consisting of up to about 1〇-3 S·; up to about 1〇-4 S-1; up to about 1〇_ 5 s_,; and up to about 1〇-6 S-1. The first parental binding protein for obtaining VD1 and the second parental binding protein for obtaining VD2 may have similar or different dissociation rates for their respective antigens. Constant (Kt) ff). B2. Efficacy The affinity/efficacy of the parental binding protein will depend on the outcome of the treatment. For example, for receptor-ligand (R-L) interactions, the affinity (kd) is equal to or greater than R_L kd (pM range). To simply remove pathological circulating proteins, such as clearing various circulating A (3 peptide substances, "can be in the low nM range. In addition, 'kd will also depend on whether the target exhibits multiple copies of the same epitope, such as mAb targeting A conformational epitope in a Αβ oligomer. I60877.doc 201247704 In one embodiment, a parental binding protein that has a neutralizing potency to a particular antigen equal to or greater than a half Ig binding protein to the same antigen is selected. Neutralizing potency can be assessed by target-dependent bioanalysis, in which a suitable type of cell produces a measurable signal (ie, proliferation or cytokine production) in response to a target stimulus, and the target neutralized by the binding protein can The signal is attenuated in a dose-dependent manner.B3. Biologically functional binding proteins (such as monoclonal antibodies) can potentially perform several functions. Some of these functions are listed in Table 7. These functions can be performed by in vitro assays (eg, cell-based) Analytical and biochemical assays and in vivo animal models for evaluation. Table 7: Some potential applications for therapeutic binding proteins Target (category) mechanism of action (target) soluble (cytokine, other) neutralizing activity (eg cytokine) enhanced clearance (eg Αβ oligomer) increased half-life (eg GLP 1) cell surface (receptor, other) efficacious Agents (eg GLP1 R; EPO R, etc.) Antagonists (eg integrin, etc.) Cytotoxicity (CD 20, etc.) Protein deposits enhance clearance/degradation (eg Αβ plaques, phylogenetic protein deposits) Different functional binding proteins as described in Table 7 to achieve the desired therapeutic result. Two or more selected parental binding proteins can then be used in a semi-Ig binding protein format to achieve in a single half of the Ig binding protein. Two or more different functions. For example, a parental binding protein that selectively neutralizes the function of a particular cytokine and a parental binding that enhances clearance of pathological proteins by 160877.doc -127-201247704 Protein to produce a half 1 § binding protein. Similarly, two parental binding proteins (eg, monoclonal antibodies) that recognize two different cell surface receptors can be selected, such as an m Ab has an agonist function for one receptor and another has an antagonist function for the other receptor. These two different binding proteins with different functions can be used to construct a selected binding protein in a single molecule. A single-half Ig binding protein of two different functions (agonists and antagonists). Similarly, two antagonists that block the binding of individual receptor ligands (eg, EGF and IGF) to cell surface receptors The binding protein (eg, monoclonal antibody) can be used in a semi-Ig binding protein format. Instead, an antagonist anti-receptor mAb (eg, anti-EGFR) and a neutralizing anti-soluble mediator (eg, anti-IGF l/2) mAb can be selected. A half ig binding protein was prepared. B4. epitope recognition Different regions of the protein can perform different functions. For example, a particular region of a cytokine interacts with a cytokine receptor to cause receptor activation, while other regions of the protein may be required to stabilize the cytokine. In this case, a binding protein that specifically binds to the receptor interaction region on the cytokine can be selected and thereby block the cytokine-receptor interaction. In some cases, such as certain chemokine receptors that bind multiple ligands, it is possible to bind to an antigenic determinant (region on the chemokine receptor) that interacts with only one ligand. Protein (eg mAb). In other cases, although the binding protein binds to an antigenic thiol group on the target that is not directly responsible for the physiological function of the protein, binding of the binding protein to these regions interferes with the physiological function of the protein (steric hindrance) or alters the conformation of the protein. Not working (eg, 160877.doc •128-201247704 mAb binding to receptors with multiple ligands alters the receptor conformation so that all ligands fail to bind). An anti-cytokine binding protein (e.g., a monoclonal antibody) that blocks the binding of a cytokine to its receptor but blocks signal transduction (e.g., 125-2H, an anti-IL-18 mAb) has also been identified. Examples of epitopes and binding protein functions include, but are not limited to, blocking receptor-ligand (RL) interactions (eg, neutralizing mAbs that bind to R-interaction sites); for example, causing a decrease in R-binding or No steric hindrance of R-bonding. Although the binding protein binds to the target at a site other than the receptor binding site, it still induces conformational changes and eliminates function (eg, Xolair®), eg, binds to R but blocks signaling (125-2H). Interfering with the receptor binding and function of the target. In one embodiment, the parent binding protein requires targeting an appropriate epitope to achieve maximum efficacy. This epitope should be maintained in the semi-Ig binding protein. The binding epitope of a binding protein (eg, a mAb) can be determined by several methods, including co-crystallization, mAb-antigen complex restriction mass spectrometry coupled with mass spectrometry peptide mapping (Legros, V et al. , (2000) Protein Sci. 9:1002-10), phage-present peptide libraries (O'Connor, KH et al., (2005) J. Immunol. Methods. 299:21-35) and mutagenesis (Wu C. Et al., (2003) J. Immunol. 170:5571-7, herein incorporated by reference. Β 5. Physical Chemistry and Pharmacological Properties Therapeutic treatment with binding proteins (eg, antibodies) usually requires high doses, usually several milligrams per kilogram (due to the molecular weight usually larger) 160877.doc -129- 201247704

And the performance by quality is lower). In order to modulate patient compliance and to properly treat chronic disease therapy and outpatient treatment, it is desirable to administer therapeutic binding proteins either subcutaneously (S.C.) or intramuscularly (i.m.). For example, the maximum desirable volume for subcutaneous administration is about 约, and therefore, a concentration of mg/mL is required to limit the number of injections per dose. In one embodiment, the therapeutic binding protein is administered in a single dose. However, the development of such formulations is limited by protein protein interactions (e. g., potentially increasing the aggregation of immunogenic risks) and limiting factors (e.g., viscosity) during processing and delivery. Therefore, the large number of clinically required and related developmental constraints may limit the full utilization of antibody formulation stagnation and subcutaneous administration in high dose regimens. It is clear that the physical and chemical properties of the protein and protein solutions (such as stability, solubility and viscosity characteristics) are extremely important. Β5·1· Stability A "stable" binding protein formulation is one in which the binding protein retains its physical stability and/or chemical stability and/or biological activity upon storage. The stability of the selected time period can be measured at the selected temperature. In embodiments, the binding protein in the formulation is stable for at least 1 month at room temperature (about 3 〇 c > c) or at 4 ° C for at least 1 month and/or at about 2 to 8 t: for at least i years, Such as to J 2 years. Further, in one embodiment, the formulation is stable after freezing the formulation (to, for example, -70. 〇 and thawing (hereinafter referred to as "freeze/thaw cycle"). In another example, "stable" blending The formulation may be less than about 10% and less than about 5% of the protein in the form of aggregates. The formulation 0 is stable in vitro for a long time at various temperatures. The binding protein is 160877.doc-130· 201247704 Required. This can be achieved by rapid screening of parental binding proteins that are stable in vitro for 2-4 weeks at elevated temperatures (eg, under 4>c), and then assessed for stability. During storage at -8 ° C, the protein exhibits stability for at least 12 months, for example at least 24 months. Stability (percent of intact monomer molecules) can be used in a variety of techniques (such as cation exchange chromatography, size exclusion chromatography) Evaluation, SDS-PAGE, and Bioactivity Testing. For a more comprehensive list of analytical techniques that can be used to analyze covalent and conformational modifications, see, for example, Jones, AJS (1993) Analytical methods f〇r such as assessment of protein Formulation and delivery systems ; Formulation and delivery of peptides and proteins, Cleland and Langer, eds., 1st edition, ACS, Washington, pp. 22-45, and Pearlman, R. and Nguyen, Τ· Η. (1990) Analysis of Protein drugs, Peptide and protein drug delivery, edited by Lee, i-th edition

Marcel Dekker, Inc., New York, pp. 247-page 301. Heterogeneity and 5^ collective formation. The stability of the binding protein may allow the formulation to exhibit less than about 10%, such as less than about 5%, such as less than about 2%, or in the range of 0.5% to 1.5% or Fewer GMP antibody species are present in aggregate form. Size Exclusion Chromatography is a sensitive, reproducible and extremely robust method for detecting protein aggregates. In addition to low aggregate content, in one embodiment, the binding protein must be chemically stable. Chemical stability can be determined by ion exchange chromatography (e.g., cation or anion exchange chromatography), hydrophobic interaction chromatography, or other methods such as isoelectric focusing or capillary electrophoresis. For example, the chemical stability of the binding protein can be such that after storage at 2-8 ° C for at least 12 months, compared to the storage of the binding protein solution before storage test 160877.doc -131 - 201247704, in cation exchange chromatography The peak representing the unmodified binding protein may increase by no more than 20%, such as by no more than 10%, or by no more than 5%. In one embodiment, the parent binding protein exhibits structural integrity, proper disulfide bond formation, and proper folding. Chemical instability due to secondary or tertiary structural changes in the binding protein can affect binding protein activity. For example, as indicated by the activity of the binding protein, the antibody activity can be reduced by no more than 50% after storage at 2-8 ° C for at least 12 months compared to the binding protein solution prior to storage testing. Such as reducing no more than 30%, or reducing no more than 10%, or reducing no more than 5% or 1 ° /. . The binding protein activity can be determined using a suitable antigen binding assay. B5.2. Solubility The "solubility" of binding proteins (eg, mAbs) is associated with the production of correctly folded monomeric IgG. Therefore, the solubility of IgG can be evaluated by HPLC. For example, soluble (monomer) IgG will produce a single peak on the HPLC chromatogram, while insoluble (e.g., multimeric and aggregate) IgG will produce a plurality of peaks. Therefore, those skilled in the art will be able to detect an increase or decrease in IgG solubility using conventional HPLC techniques. For a more comprehensive list of analytical techniques that can be used to analyze solubility, see Jones, A. G. Dep. Chem. Biochem. Eng·, Univ. Coll. London, London, UK. Shamlou, P. Ayazi.

Process. Solid-Liq. Suspensions (1993), 93-117. Publisher: Butterworth-Heinemann, Oxford, UK; and Pearlman and

Nguyen (1990) Advances in Parenteral Sciences, 4 (Pept. Protein Drug Delivery), 247-30 1). Therapeutic binding proteins or immunizations 160877.doc -132- 201247704 The solubility of globulin molecules is critical for the high concentration normally required for formulation into a sufficient amount. As outlined herein, a solubility greater than 1 mg/mL may be required to accommodate effective binding protein administration. For example, antibody solubility may be no less than about 5 mg/mL at the beginning of the study, such as no less than about 25 mg/mL at the end of the process science, such as no less than about 1 mg/mL, or no less than about 150. Mg/mL. It will be apparent to those skilled in the art that the intrinsic properties of protein molecules are important for the physicochemical properties of protein solutions (e.g., stability, solubility, viscosity). However, those skilled in the art will appreciate that there are a variety of excipients that can be used as additives to beneficially affect the characteristics of the final protein formulation. Such excipients may include: (1) a liquid solvent, a cosolvent (eg, an alcohol such as ethanol); (ii) a buffer (eg, phosphate, acetate, citrate, and amino acid buffer); (iH) An alcohol or a sugar alcohol (such as sucrose, trehalose 'fructose, raffinose, mannitol, sorbitol, and dextran); (iv) a surfactant (eg, polysorbate 2, polysorbate 40 , Polysorbate 60 and polysorbate 80, and poloxamer) '(v) isotonicity regulators (for example, salt and sugar alcohols such as NaCi), and (V1) other Shape agents (eg, preservatives, chelating agents, antioxidants, chelating materials (eg, EDTA), biodegradable polymers, and carrier molecules (eg, HAS and PEG)). The viscosity is related to the production of δ protein and the processing of protein binding (such as dialysis/ultrafiltration) filling-refining process (pumping state, filtering state) and transfer mode (can/mainness, perfect device transfer) Important parameters. The low viscosity allows the liquid solution of the binding protein to have a higher concentration. This makes it possible to administer the same dose in a small volume. The small injection volume inherently has a less painful feeling of injection. 160877.doc -133· 201247704 Advantages and the solution does not necessarily have to be isotonic to alleviate the pain of the patient during injection. The viscosity of the antibody solution can be such that at a shear rate of 100 Q/S), the binding protein solution has a viscosity of less than 200 mPas, such as less than 125 mPas, such as less than 70 mPas, such as less than 25 mPas or even less than 10 mPas. B5.3. Production Efficiency In one embodiment, the production of a semi-Ig binding protein that is efficiently expressed in mammalian cells, such as Chinese hamster ovary cells (CHO), will require at least the pro-existing pro-expression in mammalian cells. This plant binds to the protein. The yield from a stable cyanobacterial cell line (i.e., CHO) should be above about 0.5 g/L, such as above about 1 g/L, such as in the range of about 2-5 g/L or more. Kipriyanov, SM and Little M. (1999) Mol. Biotechnol. 12: 173-201; Carroll, S. and Al-Rubeai, Μ (2004) Expert. 〇pin. Biol. Ther. 4: 1821-9). The production of binding proteins and 1 § fusion proteins (such as semi-Ig binding proteins) in mammalian cells is affected by several factors. The transcription of the expression vector by incorporation of a strong promoter, enhancer and selection marker maximizes the transcription of the relevant gene from the integrated vector copy. The identification of a vector integration site that allows for high levels of gene transcription enhances the performance of the protein from the vector (Wurm et al. (2004) Nature Biotechnol. 22(1 1): 1393-1398). In addition, the production is affected by the ratio of antibody heavy chain to light chain and various steps in the process of protein assembly and secretion (jiang et al., (2〇〇6) Bi〇techn〇l pr〇g 22(1): 313- 8). B6. Immunogenicity Administration of a therapeutic semi-Ig binding protein may cause certain immune responses to occur. 160877.doc 201247704 (i.e., to form an endogenous antibody to a therapeutic semi-binding protein). The potential elements that may induce immunogenicity should be analyzed during the selection of the parental binding protein, and steps to reduce this risk may be taken to optimize the parental binding protein prior to the construction of the semi-"binding protein. It has been found to be derived from mouse binding. Proteins, such as antibodies, are highly immunogenic in patients. The production of chimeric antibodies comprising mouse variable regions and human constant regions is a logical step in reducing the immunogenicity of therapeutic antibodies. (1) Nature, 332: 323. 327, for the treatment of sex antibodies, can be transferred to a human antibody framework (remodeling/CDR grafting/humanization) by • transferring the murine CDR sequences To reduce immunogenicity. Another method is called "surface reshaping" or "face decoration", which starts with the rodent variable light chain domain and heavy chain domain, and only changes the surface-structured amino acid to human. The amino acid is frameworkd while the cdr and inline amino acids are still derived from the parental rodent binding protein (R〇guska et al. (1996) Prot. Engineer 9:895-904). In another type of humanization,

One technique only transplants a "specificity determining region" (SDR) rather than a whole CDR, which is defined as a subset of the cDR ruthenium involved in the binding of the antibody to its target (Kashmiri et al., (2005) Methods 36 ( 1): 25-34). This makes it necessary to identify SDR by analyzing the three-dimensional structure of the antibody-target complex or by performing a mutational analysis of the antibody CDR residues to determine the residues that interact with the target. Alternatively, a fully human antibody can have reduced immunogenicity compared to murine, chimeric or humanized antibodies. Another way to reduce the immunogenicity of therapeutic binding proteins is to eliminate certain specific sequences that are predicted to be immunogenic. In one method, a first-generation biologic was tested in humans and found to have unacceptable immunogenicity 160877.doc -135-201247704 after 'B cell epitopes can be localized and then altered to avoid immunodetection . Another method uses a method of predicting and removing potential T cell epitopes. Computational methods have been developed to scan and identify peptide sequences of biotherapeutic agents that are likely to bind MHC proteins (Desmet et al., (2005) Proteins 58: 53-69). Alternatively, methods based on human dendritic cells can be used to identify CD4+ T cell epitopes in potential protein allergens (Stickler et al., (2000) J. lmmunother 23: 654_6〇; s 丄Morrison and j. Schlom (1990) Important Adv· Oncol· 3-18 ; Riechmann et al. (1988) Nature 332: 323-327; Roguska et al., (1996) Protein Engineer. 9: 895-904; Kashmiri et al., (2005) Methods 36(1): 25-34; Desmet et al., (2005) Proteins 58: 53-69; and Stickler et al., (2000) j. Immun〇therapy 23: 654_ 60.). Β7· In vivo efficacy To produce a half-binding protein with the desired in vivo efficacy, it is important to produce and select a binding protein that has similar in vivo efficacy when administered in combination. However, in some cases, semi-匕 binding Proteins may exhibit in vivo efficacy that cannot be achieved by a combination of two independent binding proteins. For example, a semi-Ig binding protein allows two targets to be in close proximity, resulting in an activity that cannot be achieved by a combination of two independent binding proteins. Biological Function The parental binding protein described in Section B3 of this document that q has a desired feature in a semi-Ig binding protein can be based on, for example, pharmacokinetics; tissue distribution; soluble target versus cell surface target; and target concentration (soluble)/ Density (surface) factors are selected. Also 160877.doc * 136 - 201247704 Β8. In vivo tissue distribution is to produce a semi-1 binding protein with the desired tissue distribution in vivo, in one embodiment 'must be selected to have a similar desired living body a parental binding protein of the inner tissue distribution pattern. In this regard, the parental binding protein can be Like a binding protein or a different binding protein. Or, based on a mechanism of a dual specific targeting strategy, in other cases it may not be necessary to select a parent binding protein that has a similar distribution in vivo when administered in combination (eg, in a The binding component directs the semi-Ig binding protein to a specific site, thereby directing the second binding component to half of the same target site. For example, one of the semi-Ig binding proteins binds specifically to the pancreas (islet cells), and another specificity can direct GLP1 to the pancreas to induce insulin. B9. Homotype: In one embodiment, in order to produce a semi-Ig binding protein having desirable properties including, but not limited to, isoforms, effector functions, and circulating half-lives, depending on the therapeutic utility and the desired endpoint of treatment, the selection has the appropriate Fc. A parental binding protein (eg, a mAb) that functions as a function. The parent binding proteins may be the same or different. There are five major heavy chain classes or isotypes, some of which have several subtypes, and such classes or isotypes determine the effector function of the antibody molecule. These effector functions are present in the hinge region, CH2 and CH3 domains of the antibody molecule. However, residues in other parts of the antibody molecule may also have an effect on the function of the effect. Hinge region Fc effector functions include: (1) antibody-dependent cellular cytotoxicity; (ii) complement (Clq) binding, activation and complement-dependent cytotoxicity (CDC); (iii) phagocytosis/clearance of antigen-antibody complexes; Iv) In some cases, cytokines are released. These F c effector functions of the antibody molecule mediate the e-IgG 1 isotype antibody by the interaction of the F c region with a group of specific 160877.doc -137-201247704 heterologous cell surface receptors, while IgG2 and IgG4 have The minimum effect function or no effect function. The effector function of IgG antibodies is mediated via interaction with three structurally homologous cellular Fc receptor types (and subtypes) (FcgRl, FcgRII and FcgRIII). These effector functions of IgGl can be eliminated by mutating the specific amino acid residues (e.g., L234A, L23 5A) required for FcgR and Clq binding in the downstream hinge region. The amino acid residues in the Fc region (especially the CH2_CH3 domain) also determine the circulating half-life of the antibody molecule. This Fc function is mediated via binding of the Fc region to the neonatal Fc receptor (FcRn). This binding is responsible for recycling the antibody molecule from the acidic lysosome back into the systemic circulation. Whether the antibody should have an active or inactive isoform will depend on the antibody to be treated, and it will be awkward. Some examples of the use of isoforms and the results of the desired treatment are listed below: a) If the desired endpoint is a functional neutralization of soluble cytokines, an inactive isoform can be used; b) if the desired result is to remove pathological proteins, then the activity can be used The same type; c) The right result is to clear the protein aggregate, then the activity isotype can be used; d) If the desired result is to antagonize the surface receptor, use the inactive isotype (Tysabri, igG4; OKT3, mutant IgGl e) The result to the right is to eliminate the target cell, then use the active isotype (Herceptin, IgG1 (and has enhanced effector function)); and f) if the desired result is self-circulation to clear the protein without entering In cns, 160877.doc 201247704 can use IgM isotype (for example, to clear circulating Ab peptide substances). The Fc effector function of a parent binding protein (e. g., a mAb) can be determined by various in vitro methods well known in the art. The choice of the same type as discussed, and thus the choice of effector function, will depend on the endpoint of the treatment to be treated. In cases where it is desirable to simply neutralize the circulating target (e. g., block receptor-ligand interactions), effector functions may not be required. In these cases, the homologous effect of the antibody's elimination effector function or the mutation in the Fc region is desirable. In other situations where the elimination of target cells is the end of treatment • (eg, elimination of tumor cells), the same effect of enhanced effect function 1 or? It is desirable to change or de-fucosylation (de_fuc〇sylati〇n) in Presta, GL (2006) Adv. Drug Deliv. Rev. 58:640-656 and Satoh, M. et al. (2006) Expert 〇pin Bi〇1

Ther· 6: 1161-1173). Similarly, depending on the therapeutic utility, the circulating half-life of an antibody molecule can be shortened/prolonged by introducing a specific mutation in the Fc region to regulate antibody-FcRn interaction (DaU, Acqua, WF et al, (2006) J. Biol. Chem. 281: 23514-23524; Petkova, SB • (2006) # Λ, Internal. Immunol. 18:1759-1769 ; Vaccaro, C. et al., (2007) Proc. Natl. Acad. Sci. USA 103 : 18709- 18714). With regard to semi-Ig binding proteins, it may be desirable to identify published information about various residues that affect the different effector functions of normal therapeutic antibodies. In the semi-Ig binding protein format, it is possible that other (different) Fc region residues other than the Fc region residues identified for regulating antibody effector function may be important. In summary, the key decision in the final half of the § binding protein format (iv) should be based on the disease indication, the treatment target, and the desired treatment endpoint and safety considerations. Exemplary exemplified appropriate heavy and light bond constant regions are listed below, including but not limited to: 〇IgG1-heterotype: Glmz ° IgG1 mutant-A234, A235 〇IgG2-type: G2m(n-) 〇K-Km3 〇 λ

Fc Receptor and Clq Studies: Mutations in the Hinge Domain by (eg, L234A, L235A) Can Eliminate Antibody and Any Over-Performed Targets on the Cell Membrane Cause Improper Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) and Complement Dependence The possibility of cytotoxicity (CDC) is expected that the substituted amino acids present in the IgG 1 hinge region of the half-ig-binding protein attenuate the binding of the half-ig-binding protein to the human Fc receptor (but not FcRn) because The FcgR binding line occurs in the overlapping sites on the 1 〇1 hinge region. This feature of the semi-"binding protein allows its safety profile to be improved compared to antibodies containing wild-type IgG. The binding of the semi-Ig binding protein to the human Fc receptor can be performed by flow cytometry experiments using cell lines ( For example, THP-1, K562) and engineered CHO cell strains expressing FcgRIIb (or other FcgR) were assayed. Compared to IgG1, the control antibody showed a decrease in binding to FcgRI and FcgRIIa, and binding to FcgRIIb. Sexuality is unaffected. The typical complement cascade is triggered by antigen/IgG immune complex binding and activation of Clq, and subsequent inflammatory and/or immunomodulatory responses. The Clq binding site on IgG has been mapped to residues in the IgG hinge region. The binding of Clq to increasing concentrations of the semi-Ig binding protein 160877.doc • 140· 201247704 can be assessed by Clq ELISA. If desired, the selection can not bind C 1 q compared to the binding to wild-type control IgG1. Semi-Ig binding protein. In summary, L234A, L235A hinge region mutation disruption antibody and FcgRI, FcgRIIa &

Binding of Clq' does not affect the interaction of the antibody with FcgRIIb. These data indicate that although the half 1 § binding protein with a mutant Fc will normally interact with the inhibitory FcgRIIb in vivo, it may not interact with the activated FcgRI & FcgRIIa receptor or Clq. Human FcRn binding: The neonatal receptor (FcRn) is responsible for transporting IgG across the placenta and controlling the catabolic half-life of IgG molecules. It may be necessary to increase the terminal half-life of the binding protein to improve efficacy, reduce the dose or frequency of administration, or improve the localization of the target. Alternatively, the opposite may be required' to shorten the terminal half-life of the bound protein to reduce systemic exposure or to improve the target to non-target binding ratio. Adjusting the interaction between IgG and its rescue receptor FcRn2 provides a means to increase or decrease the terminal half-life of IgG. The circulating proteins (including IgG) are absorbed in the fluid phase via the microcapsules of certain cells, such as cells of the vascular endothelium. IgG can bind to FeRn in a slightly acidic condition (pH 6.0-6.5) in the endosome and can be recycled to the cell surface, which is released on the cell surface under almost neutral conditions (ρΗ 7. Pairing site binding site) Positioning revealed that two histidine residues (His310 and His435) conserved between species caused a pH dependence of this interaction. Using phage display technology, it was identified that increased binding to FcRn and prolonged half-life of mouse IgG Mouse fc region mutation (see

Victor, G. et al., (1997) Nature Biotechnol. 15(7): 637-640). Fc region mutations have been identified that increase the binding affinity of human IgG to FcRn at pH 6, 而非, but not at pti 7·4 160877.doc -141 - 201247704 (see Dall, Acqua, F. 荨人, ( 2002) J. Immunol. 169(9): 5171-80). In addition, in one case, a similar increase in pH-dependent binding (up to 27-fold) was observed for rhesus FcRn, and this resulted in a two-fold increase in serum half-life of rhesus monkeys compared to parental IgG (see Hinton). , Paul R. et al., (2004) J. Biol. Chem. 279(8): 6213-6216). These findings indicate that it is feasible to extend the blood stagnation of the antibody therapeutic by adjusting the interaction of the Fc region with FcRn. Conversely, a mutation in the pc region that attenuates interaction with FcRn can shorten antibody half-life. B.10 Pharmacokinetics (ρκ) In one embodiment, a parental binding protein having a similar desired pharmacokinetic profile is selected for the production of a semi-Ig binding protein having the desired pharmacokinetic profile. One consideration is the immunogenic response to individual antibodies (i.e., HAHA 'human anti-human antibody response; HACA, human anti-chimeric antibody response) further complicating the pharmacokinetics of such therapeutic agents. In one embodiment, 'the half-binding protein is constructed using a binding protein (eg, a monoclonal antibody) having minimal or no immunogenicity such that the resulting half 1 § binding protein will also have minimal immunogenicity or Not immunogenic. Some factors that determine the PK of a binding protein (e. g., a mAb) include, but are not limited to, the intrinsic properties of the binding protein (VH amino acid sequence), immunogenicity, FcRn binding, and Fc function. The PK pattern of the selected parental binding protein can be readily determined in rodents 'because the PK pattern of rodents is closely related to the ρκ pattern of binding proteins in cynomolgus monkeys and humans (or former Can be refined -142- 160877.doc 201247704 dense prediction of the latter). The PK type is determined as described in the Examples section. After selecting a parental binding protein having the desired characteristics (and other desirable functional properties as described herein), construct a semi-Ig binding protein and also evaluate the half

The PK properties of Ig binding proteins. Therefore, in the measurement of the semi-Ig binding protein 2PK property, a p-kappa analysis method for determining the sputum type based on the function of the antigen-binding domain derived from the parental binding protein or the binding protein can be used. The sputum form of the semi-Ig binding protein can be determined as described in the Examples provided below. May become half

Other factors of the PK pattern of Ig binding proteins include antigen binding domain (CDR) orientation, linker size, and Fc/FcRn interaction. The ρκ characteristics of the parental binding protein can be assessed by evaluating the following parameters: absorption, distribution, metabolism, and excretion. Semi-Ig binding proteins can be modified using the methods provided herein. Methods for analysing the pharmacokinetics of a semi-Ig binding protein are also within the abilities of those skilled in the art and can be accomplished using methods known in the art and provided herein. Absorption. As far as attack is concerned, therapeutic binding proteins (e.g., monoclonal antibodies) are administered by parenteral routes (e.g., intravenous (IV), subcutaneous (sc), or intramuscular (im). After administration of SC or IM, the mAb is absorbed from the intercellular space mainly through the lymphatic pathway to the whole body circulation towel. Saturated systemic pro-colon proteolytic degradation can produce variable absolute bioavailability after extravascular administration. In general, since the proteolytic capacity is saturated at the same dose, it is observed that the absolute bioavailability increases as the individual antibody dose increases. Because the lymph fluid is slowly discharged into the system, the absorption process of the mAb is usually quite slow, and the duration of absorption can last from several hours to several days. The absolute bioavailability of monoclonal antibodies after % administration is generally in the range of 5% to 1%. 160877.doc 201247704 Distribution · After administration of ιν, monoclonal antibodies usually follow a phase of rapid distribution, followed by a slow elimination phase of the two-phase serum (or plasma) concentration and time profile. In general, a double exponential pharmacokinetic model best describes this pharmacokinetic profile. The volume of distribution of the mAb in the central compartment (Vc) is usually equal to or slightly proportional to the plasma volume (2 liters to 3 liters of serum (plasma) concentration of other parenteral routes (such as IM or SC) versus time) The unique two-phase mode of the state may not be obvious, because the distribution of serum (plasma) concentration/time curve is partially obscured by long absorption, including physicochemical properties, site-specific and target-directed receptor-mediated absorption, tissue binding ability. And a variety of factors including the dose of mAb can affect the biodistribution of mAb. Some of these factors can cause non-linearity in the biodistribution of mAb. Metabolism and excretion: intact individual antibodies cannot be excreted into the urine via the kidney due to molecular size. It is not activated by metabolism (e.g., catabolism). Depending on the particular characteristics of the semi-Ig binding protein of the present invention, it may or may not be excreted in the urine via the kidney, and the cutoff value of renal excretion in # is generally considered to be about 50 kDa. For IgG-based therapeutics, the half-life is usually in the range of hours or I-2 days to more than 20 days. The elimination of semi-Ig binding proteins can be affected by many factors. Including, but not limited to, the affinity of the WFcRn receptor, the immunogenicity of the semi-(8) gemstone protein, the degree of glycosylation of the semi-binding protein, the sensitivity of the semi-Ig binding protein to proteolysis, and receptor-mediated elimination. B. Tissue and Reactive Patterns of Human and Toxicological Species The same staining pattern indicates that toxicological species can be used to assess potential human toxicity. Toxicological species are used to study unrelated toxicity of their animals. Choose to meet the following two criteria Individual binding proteins: (1) tissue staining suitable for I60877.doc • 144· 201247704 combined with known binding protein target performance, and (2) similar staining patterns between human tissues from the same organ and toxicological species tissues. : Immunization and/or binding protein selection usually uses recombinant or synthetic antigens (proteins, carbohydrates or other molecules). Binding to natural counterparts and counter-screening against unrelated antigens are usually used for therapeutic binding proteins (eg antibodies) Part of the screening funnel. However, screening for large numbers of antigens is often impractical. Therefore, use from all major organs Tissue-based cross-reactivity studies can be used to avoid inappropriate binding of binding proteins to any unrelated antigens. Criteria 2: Use human and toxicological species to organize (crab-eating macaques, dogs, possibly rodents and other animals, such as human studies) Comparing tissue cross-reactivity studies conducted in the same 36 or 37 tissues in the same way to help validate the selection of toxicological species. In typical tissue cross-reactivity studies on frozen tissue sections, therapeutic binding proteins can be expressed The expected binding of the antigen is known and/or based on a low degree of interaction (non-specific binding, low degree of binding to similar antigens, interaction based on low charge, etc.) and tissue to a lower degree of binding. In any case, The most relevant toxicological animal species are animal species with the highest degree of coincidence with human and animal tissue. Organizational cross-reactivity studies follow appropriate regulatory guidelines, including EC CPMP guidelines for original production and quality control of mAbs and 1997 US FDA/CBER "Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products For Human Use". Human tissue cryosections (5 μm) obtained at necropsy or biopsy were fixed on an objective lens and dried. Permease staining of tissue sections using the 160877.doc -145- 201247704 avidin-biotin system (FDA Directed J. Coxi/er ζ·« Λβ

Testing of Monoclonal Antibody Products f〇r Human Usej ) ° B.11A Tests for Binding Protein Products for Human Use Tissue cross-reactivity studies are usually conducted in two phases, the first phase consisting of making a human donor 32 tissues (usually: adrenal gland, gastrointestinal tract, prostate, bladder, heart, skeletal muscle, blood cells, kidney, skin, bone marrow, liver, spinal cord, breast, lung, spleen, cerebellum, lymph nodes, testicles, cerebral cortex, ovary, Frozen sections of the thymus, colon, spleen, thyroid, endothelium, accessory squamous gland, ureter, eye, pituitary, uterus, fallopian tube, and placenta. In the second phase, use up to 38 tissues from 3 unrelated adults (including adrenal gland, blood, blood vessels, bone marrow, cerebellum, brain, cervix, esophagus, eye, heart, kidney, large intestine, liver, lung, lymph nodes) , breast, ovary, fallopian tube, pancreas, parathyroid, peripheral nerve, pituitary, placenta, prostate, salivary gland, skin, small intestine, ridge, spleen, stomach, striated muscle, sputum, thymus, thyroid, tonsil, ureter, bladder and uterus ) Conduct a complete cross-reactivity study. Studies are usually performed at a minimum of 2 dose concentrations. Therapeutic binding proteins (ie, test articles) and isotype-matched binding proteins can be biotinylated for avidin-biotin complex (ABC) detection. Other detection methods can include fittings (or The test article in other ways is subjected to three antibody detections, or the unlabeled test article is pre-combined with the labeled anti-human IgG. 160877.doc -146· 201247704 Briefly, human tissue cryosections (approximately 5 μm) obtained during necropsy or biopsy were fixed on an objective lens and dried. Tissue sections were subjected to peroxidase staining using the avidin-biotin system. First (in the case of a pre-combination detection system), the test article is incubated with biotin-labeled secondary anti-human IgG and forms an immune complex. An immunocomplex with a final concentration of the test article of 2 pg/mL and 10 pg/mL was added to the tissue section on the objective lens, and then the tissue section was reacted with the avidin-biotin-peroxidase kit. minute. Subsequently, coating DAB (33, _ two

Aminobenzidine (the substrate of the peroxidase reaction), which was subjected to tissue staining for 4 minutes. Antigen agarose beads were used as positive control tissue sections. Any specific staining is judged to be the expected reactivity (e.g., consistent with antigenic performance) or unintended reactivity based on the known performance of the antigen of interest in question. Any judged-specific stains were scored for intensity and frequency. Antigen or serum competition or blocking studies may further aid in determining whether the observed staining is specific or non-specific. If the selected binding protein is found to meet selection criteria (i.e., tissue staining is suitable and stain matching between human and toxicological animal specific tissues), it can be selected for semi-Ig binding protein production. It is necessary to repeat the tissue cross-reactivity study with the final semi-Ig binding protein construct', but when these studies follow the same protocol as outlined herein, the assessment may be more complicated, when the antigen-binding domain is obtained from more than one The parent binds to the protein and needs to compete with the complex antigen to study the shirt. Anything cannot be explained: especially when combined. Obviously, if you choose the following two parental binding proteins, you can greatly simplify the use of multispecific molecules (such as half 1 § binding protein) for tissue cross-reactivity studies: (1) Lack of unintended tissue cross-reactivity studies and (2) Appropriate similarities in the results of tissue cross-reactivity studies between humans and toxicological animal species. B.12. Specificity and selectivity In order to generate a semi-Ig binding protein with the desired specificity and selectivity, it is necessary to generate and select a parental binding protein having a similar desired specificity and selectivity. In this regard, when more than one antigen binding site is desired, the parent binding protein can be the same binding protein or preferably a different binding protein. Binding studies on the specificity and selectivity of semi-Ig binding proteins can be complicated by the presence of the number of locus sites and the number of target antigen binding sites. In short, for multiple target antigens, use ELISA (enzyme-linked immunosorbent assay) with hemexamic protein, 8, conjugated studies with 8 or other interaction studies, or two species or two Binding of two or more antigens to a half 1 § binding protein. Although BIAc〇re® technology can determine the continuous independent binding of multiple antibodies, it is not comparable to traditional methods (including ELISA) or more modern techniques (such as κίηΕχΑ8). Careful characterization of the binding proteins of each parent is critical. The confirmation of the specific retention of individual binding sites in the half-binding protein after characterization of each binding protein will be greatly simplified. If the two parental binding proteins are selected for specificity and then combined into a half 1g binding protein' then the complex operation to determine the specificity of the semi-Ig binding protein will be greatly simplified. The parental binding protein can be the same binding protein or different Binding protein. 160877.doc 201247704 Antigen-binding protein (eg antigen-antibody) interaction studies can take many forms, including multiple Typical protein-protein interaction studies: ELISA, mass spectrometry, chemical cross-linking, SEC using light scattering, equilibrium dialysis, gel permeation, ultrafiltration, gel chromatography, large-area analytical SEC, micropreparation ultracentrifugation ( Settling equilibrium), spectroscopy, titration microcalorimetry, sedimentation equilibrium (in analytical ultracentrifugation), sedimentation velocity (in analytical centrifugation), and surface plasmonic resonance (including BIAcore®) »Related references include Current Protocols in Protein Science," Coligan, JE, et al. (eds.), Vol. 3, Chapters 19 and 20, published by John Wiley & Sons Inc., and "Current Protocols in Immunology," Coligan, JE et al. Published by John Wiley & Sons Inc., and related references included therein. B.13 Cytokine Release in Whole Blood The interaction of semi-Ig binding proteins with human blood cells can be studied by cytokine release assay (Wing, MG. (1995) Therapeut. Immunol. 2(4): 183-190; "Current Protocols in Pharmacology,"

Enna, SJ et al. (eds.) published by John Wiley & Sons Inc; Madhusudan, S. (2004) Clin. Cancer Res. 10(19): 6528-6534; Cox, J. (2006) Methods 38(4) : 274-282 ; Choi, I. (2001) Eur. J. Immunol. 31(1): 94-106). Briefly, half of the Ig binding proteins of various concentrations were incubated with human whole blood for 24 hours. The concentration tested should cover a wide range including the final concentration of the typical blood content in the patient (including but not limited to 1 ng/ml - l 〇〇 pg / ml). After the incubation, the supernatants and cell lysates were analyzed for the presence of 1L-1Ra, TNF-a, IL-160877.doc • 149·201247704 lb, IL-6 and IL-8. The cytokine concentration profile produced by the binding protein was compared to the negative human IgG control and the positive LPS or PHA control produced. The half Ig binding protein from both the cell supernatant and the cell lysate exhibited a cytokine profile comparable to that of the control human IgG and/or semi-Ig binding protein. In one embodiment, the monoclonal antibodies do not interact with human blood cells to spontaneously release inflammatory cytokines. Cytokine release studies of half-h binding proteins can become complicated when multiple binding sites are available for multiple targets. Briefly, cytokine release studies as described herein measure the effect of intact semi-Ig binding proteins on whole blood or other cellular systems, but can determine which part of the molecule causes cytokine release. Once cytokine release is detected, the purity of the semi-ig-binding protein preparation must be determined, as some co-purified cellular components alone can cause cytokine release. If purity is not an issue, it may be necessary to use a semi-"binding protein cleavage (including but not limited to, removal of the Fc portion, isolation of the binding site, etc.), ,, σ 5 site mutagenesis or other methods for any Observed results are deconvolved. Obviously, if you choose two or more parental antibodies that lack cytokine release, and then combine them into a semi-匕 binding protein, this complex operation can be greatly simplified. 14.14 and used Cross-Reactivity of Other Species in Toxicology Studies In one embodiment, individual binding proteins that are sufficiently responsive to appropriate toxicological species (eg, cynomolgus monkeys) are selected. The parental binding protein needs to be bound to the immediate The source species target (ie, the crab-eating macaque) and initiates an appropriate response (regulation, "activation). In one embodiment, the reactivity (affinity/potency) of the orthologous species target should be within 1 fold of the human target. 160877.doc •150· 201247704 Practically, for a variety of species including mouse, rat, dog, monkey (and other non-human primates) and disease model species (ie sheep for asthma models) The parental binding protein is assessed. The acceptable cross-reactivity of the parental binding protein to the toxicological species allows future toxicological studies of the semi-Ig binding protein with the same species. For each reason, the two parental binding proteins have acceptable cross-reactivity to common toxicological species, thereby allowing toxicological studies of semi-Ig binding proteins with the same species. The parent binding protein may be selected from a variety of binding proteins, such as monoclonal antibodies, which bind to a particular target and are well known in the art. The parent binding protein can be the same binding protein or a different binding protein. Such binding proteins include, but are not limited to, anti-TNF antibodies (U.S. Patent No. 6,258,562), anti-IL-12 and/or anti-IL-12p40 antibodies (U.S. Patent No. 6,914,128); anti-IL-18 antibodies (US) Patent Publication No. 2005/0147610), anti-C5, anti-CBL, anti-CD147, anti-gpl20, anti-VLA-4, anti-CDlla, anti-CD18, anti-VEGF, anti-CD40L, anti-CD-40 (see, for example, PCT Publication No. WO2007/124299), anti-Id, anti-ICAM-1, anti-CXCL13, anti-CD2, anti-EGFR, anti-TGF-P2, anti-HGF, anti-cMet, anti-DLL-4, anti-NPR1, anti-PLGF, anti-ErbB3, anti-E - selectin, anti-Fact VII, anti-Her2/neu, anti-F gp, anti-CD11/18, anti-CD14, anti-ICAM-3, anti-RON, anti-SOST, anti-CD-19, anti-CD80 (see, for example, PCT Publication No. WO 2003/039486), anti-CD4, anti-CD3, anti-CD23, anti-β2-integrin, anti-α4β7, anti-CD52, anti-HLA DR, anti-CD22 (see, for example, U.S. Patent No. 5,789,554), anti-CD20, anti-MIF, anti-CD64 (FcR), anti-TCRaP, anti-CD2, anti-Hep B, anti-CA 125, anti-160877.doc • 151 - 201247704

EpCAM, anti-gpl20, anti-CMV, anti-gpllbllla, anti-IgE, anti-CD25, anti-CD33, anti-HLA, anti-IGF1, 2, anti-IGFR, anti-VNR integrin, anti-IL-la, anti-IL-Ιβ, anti-IL-1 Receptors, anti-IL-2 receptors, anti-IL-4, anti-IL-4 receptors, anti-IL5, anti-IL-5 receptors, anti-IL-6, anti-IL-8, anti-IL-9, anti-IL- 13. Anti-IL-13 receptor, anti-IL-17 and anti-IL-23 (see Presta, LG (2005) J. Allergy Clin. Immunol. 116: 731-6 and www.path.cam.ac.uk/~ Mrc7/humanisation/antibodies.html). The parent binding protein may also be selected from a variety of therapeutic antibodies that are approved for use, in clinical trials, or in development for clinical use. Such therapeutic antibodies include, but are not limited to, rituximab (Rituxan®, IDEC/Genentech/Roche) (see, e.g., U.S. Patent No. 5,736,137), an approved for the treatment of non-Hodgkin Chimeric anti-CD20 antibody to lymphoma; HuMax-CD20, an anti-CD20 currently being developed by Genmab; anti-CD20 antibody, AME-133 (Applied Molecular Evolution), hA20 (Immunomedics, Inc.) as described in U.S. Patent No. 5,500,362. ; HumaLYM (Intracel), and PRO70769 (PCT Application No. PCT/US2003/040426); trastuzumab (Herceptin®, 〇61^1^£^) (see, for example, U.S. Patent No. 5,677,171) a humanized anti-Her2/neu antibody approved for the treatment of breast cancer; pertuzumab (rhuMab-2C4, Omnitarg®) currently being developed by Genentech; anti-Her2 antibody (US Patent No. 4,753,894) Cetuximab (Erbitux®, Imclone) (U.S. Patent No. 4,943,533; (: Ding Publication No. 96/40210), a chimeric for clinical use in various cancers; Anti-EGFR antibody; currently being used by 1 60877.doc -152- 201247704

Abgenix-Immunex-Amgen developed ΑΒΧ-EGF (U.S. Patent No. 6,23, 883); 11 times 3 parent-£0?1' currently developed by 〇61111^ (U.S. Patent No. 7,247,301); 425, EMD55900 , EMD 62000 and EMD 72000 (Merck KGaA) (U.S. Patent No. 5,558,864; Murthy et al., (1987), Arch. Biochem. Biophys. 252(2): 549-60; Rodeck et al., (1987), J. Cell. Biochem. 35(4): 315_20; Kettleborough et al., 1991, Protein Eng. 4(7): 773-83); ICR62 (Institute of Cancer Research) (PCT Publication No. WO 95/20045; Modjtahedi et al, (1993) J. Cell. Biophys. 22(1-3): 129-46; Modjtahedi et al., (1993) Br. J. Cancer 67(2): 247-53; Modjtahedi et al., (1996) Br· J. Cancer 73(2): 228-35; Modjtahedi et al., (2003) Int. J. Cancer 105(2): 273-80); TheraCIM hR3 (YM Biosciences, Canada and Centro de Immunologia Molecular, Cuba) U.S. Patent No. 5,891,996; U.S. Patent No. 6,506,883; Mateo et al., (1997) Immunotechnol. 3(1): 71-81); mAb-806 (Ludwig Institue for Cancer Research, Memorial Sl) oan-Kettering) (Jungbluth et al., (2003) Proc. Natl. Acad. Sci. USA. 100(2): 639-44); KSB-102 (KS Biomedix); MR1-1 (IVAX, National Cancer Institute) (PCT Publication No. WO 01/62931 A2); and SC100 (Scancell) (PCT Publication No. WO 01/88138); alemtuzumab (Campath®, Millenium), a currently approved for treatment Humanized mAb of B-cell chronic lymphocytic leukemia; Moromonab-CD3 (Orthoclone OKT3®), a 160877.doc-153-201247704 Anti-CD3 antibody developed by Ortho Biotech/Johnson & Johnson Ibritumomab tiuxetan (Zevalin®), an anti-CD20 antibody developed by IDEC/Schering AG; gemtuzumab ozogamicin (Mylotarg®), an anti-CD33 developed by Celltech/Wyeth P67 protein) antibody; alefacept (Amevive®), an anti-LFA-3 Fc fusion developed by Biogen;

Abciximab (ReoPro®) developed by Centocor/Lilly; basiliximab (Simulect®) developed by Novartis;

Palivizum>ab (Synagis®) developed by Medimmune; Infliximab (Remicade®), an anti-TNFa antibody developed by Centocor; adalimumab (Humira®) , an anti-TNFa antibody developed by Abbott; Humicade®, a

Anti-TNFa antibody developed by Celltech; golimumab (CNTO-148), a fully human TNF antibody developed by Centocor; etanercept (Enbrel®), a development by Immunex/Amgen P75 TNF receptor Fc fusion; lenercept, a p55 TNF receptor Fc fusion previously developed by Roche; ABX-CBL, an anti-CD147 antibody developed by Abgenix; ABX-IL8, a positive An anti-IL8 antibody developed by Abgenix; ABX-MA1, an anti-MUC18 antibody being developed by Abgenix; Pemtumomab (R1549, 90Y-muHMFGl), an anti-MUC1 being developed by Antisoma; Therex (R1550), an anti-MUC1 antibody being developed by Antisoma; AngioMab (AS1405), which is being developed by Antisoma; HuBC-1, which is being developed by Antisoma; is being used by 160877.doc - 154- 201247704

Thioplatin (AS1407) developed by Antisoma; Antegren® (natalizumab), an anti-α-4-β-l (VLA-4) and α-4-β developed by Biogen -7 antibody; VLA-1 mAb, an anti-VLA-1 integrin antibody being developed by Biogen; LTBR mAb, an anti-lymphocyte toxin beta receptor (LTBR) antibody being developed by Biogen; CAT-152, a type Anti-TGF-P2 antibody developed by Cambridge Antibody Technology; ABT 874 (J695), an anti-IL-12 p40 antibody being developed by Abbott; CAT-192, an anti-TGFpi antibody being developed by Cambridge Antibody Technology and Genzyme; CAT- 213, an anti-Eotaxin1 antibody being developed by Cambridge Antibody Technology; LymphoStat-B®, an anti-Blys antibody being developed by Cambridge Antibody Technology and Human Genome Sciences Inc.; TRAIL-RlmAb, a type of Cambridge Antibody Technology and Human Genome Anti-TRAIL-R1 antibody developed by Sciences, Inc.; Avastin® bevacizumab (rhuMAb-VEGF), an anti-VEGF antibody developed by Genentech; developed by Genentech Anti-HER receptor family antibody; anti-tissue factor (ATF), an anti-tissue factor antibody being developed by Genentech; Xolair® (Omalizumab), an anti-IgE antibody being developed by Genentech; Raptiva® (Efalizumab), an anti-CD1 la antibody being developed by Genentech and Xoma; MLN-02 antibody (formerly LDP-02) being developed by Genentech and Millenium Pharmaceuticals; HuMax CD4, a type of positive An anti-CD4 antibody developed by Genmab; HuMax-IL15, an anti-IL15 antibody from Genmab and Amgen 160877.doc-155-201247704; HuMax-Inflam developed by Genmab and Medarex; HuMax-Cancer, a type by Genmab And anti-type I heparin antibodies developed by Medarex and Oxford GcoSciences; HuMax-Lymphoma developed by Genmab and Amgen; HuMax-TAC being developed by Genmab; IDEC-131 and anti-CD40L antibodies being developed by IDEC Pharmaceuticals; IDEC -151 (Clenoliximab), an anti-CD4 antibody being developed by IDEC Pharmaceuticals; IDEC-114, an anti-CD developed by IDEC Pharmaceuticals 80 antibodies;

IDEC-152, an anti-CD23 that is being developed by IDEC Pharmaceuticals; anti-megaphage cell migration being developed by IDEC Pharmaceuticals

Factor (MIF) antibody; BEC2, an anti-individual genotype antibody being developed by Imclone; IMC-1C11, an anti-KDR antibody being developed by Imclone; DC101, an anti-flk-Ι antibody being developed by Imclone; Imclone developed anti-VE弼 mucin antibody; CEA-Cide® (labetuzumab), an anti-carcinoembryonic antigen (CEA) antibody being developed by Immunomedics; LymphoCide® (Epratuzumab (Epratuzumab) )), an anti-CD22 antibody being developed by Immunomedics; AFP-Cide being developed by Immunomedics; MyelomaCide being developed by Immunomedics; LkoCide being developed by Immunomedics; ProstaCide being developed by Immunomedics; MDX-010, a type Anti-CTLA4 antibody developed by Medarex; MDX-060, a kind of positive

Anti-CD30 antibody developed by Medarex; MDX·070 being developed by Medarex; MDX-018 being developed by Medarex; Osidem® (IDM-l) and anti-160877.doc-156- developed by Medarex and Immuno-Designed Molecules 201247704

Her2 antibody; HuMax®-CD4, an anti-CD4 antibody being developed by Medarex and Genmab; HuMax-ILl5, an anti-IL15 antibody being developed by Medarex and Genmab; CNTO 148, one being developed by Medarex and Centocor/J&J Anti-TNFa antibody; CNTO 1275, an anti-cytokine antibody being developed by Centocor/J&J; MOR101 and MOR102, an anti-intercellular adhesion molecule-1 (ICAM-1) (CD54) antibody being developed by MorphoSys; MOR201 , an anti-fibroblast growth factor receptor 3 (FGFR-3) antibody being developed by MorphoSys; Nuvion® (visilizumab), an anti-CD3 antibody being developed by Protein Design Labs; HuZAF® , an anti-gamma interferon antibody being developed by Protein Design Labs; anti-α5β1 integrin being developed by Protein Design Labs; anti-IL-12 being developed by Protein Design Labs; ING-1, an anti-development developed by Xoma Ep-CAM antibody; Xolair® (Omalizumab), a humanized anti-IgE antibody developed by Genentech and Novartis; and MLN01, an anti-β2 integrin antibody being developed by Xoma. In another embodiment, the therapeutic agent comprises KRN330 (Kirin); huA33 antibody (Α33, Ludwig Institute for Cancer Research); CNTO 95 (aV integrin, Centocor); MEDI-522 (aVP3 integrin, Medimmune); Volociximab (aVpi integrin, Biogen/PDL); human mAb 216 (B cell glycosylation epitope, NCI); BiTE MT103 (bispecific CD19xCD3, Medimmune); 4G7xH22 (bispecific B cell) xFcyRl, Medarex/Merck KGa); rM28 (bispecific

CD28xMAPG, European Patent No. EP1444268); MDX447 (EMD 160877.doc • 157. 201247704 82633) (bispecific CD64xEGFR, Medarex); Cattoxomab (removab) (bispecific EpCAMx anti-CD3, Trion/ Fres); Ertumaxomab (bispecific HER2/CD3, Fresenius Biotech); ogovumab (OvaRex) (CA-125, ViRexx); Rencarex® (WX G250) (carbonic anhydride) Enzyme IX, Wilex); CNTO 888 (CCL2, Centocor); TRC105 (CD105 (endoglin), Tracon); BMS-663513 (CD137 agonist, Brystol Myers Squibb); MDX-1342 (CD19,

Medarex); Sicilizumab (MEDI-507) (CD2, Medimmune); Ofatumumab (Humax-CD20) (CD20, Genmab); Rituxan (CD20, Genentech) ); Vetuzumab (hA20) (CD20, Immunomedics); Epalizumab (CD22, Amgen); Lucuximab (lumiliximab, IDEC 152) (CD23, Biogen); Moromona - CD3 (CD3, Ortho); HuM291 (CD3 fc receptor, PDL Biopharma); HeFi-1 (CD30, NCI); MDX-060 (CD30, Medarex); MDX-1401 (CD30, Medarex); SGN_30 (CD30, Seattle) Genentics); SGN-33 (Lintuziimab) (CD33, Seattle Genentics); Zallimumab (HuMax-CD4) (CD4, Genmab); HCD122 (CD40, Novartis); SGN-40 (CD40, Seattle Genentics); Campathlh (allezumab) (CD52, Genzyme); MDX-1411 (CD70 >Medarex); hLLl (EPB-l) (CD74.38, Immunomedics); Galifizumab (Galiximab, IDEC-144) (CD80, Biogen); MT293 (TRC093/D93) (lysis of 160877.doc -158- 201247704

Collagen, Tracon); HuLuc63 (CSl, PDL Pharma); Ilipizumab (ipilimumab, MDX-010) (CTLA4, Brystol Myers Squibb); Chuanlimumab (Tremelimumab, Ticilimumab, CP -675,2) (CTLA4, Pfizer); HGS-ETR1 (Mapatumumab) (DR4 TRAIL-R1 agonist, Human Genome Science/Glaxo Smith Kline); AMG-655 (DR5, Amgen) ; Apomab (DR5, Genentech); CS-1008 (DR5, Daiichi Sankyo); HGS-ETR2 (lexatumumab) (DR5 TRAIL-R2 agonist, HGS); Ertox (Erbitux) (EGFR, Imclone); IMC-11F8 (EGFR, Imclone); Nimotuzumab (EGFR, YM Bio); Panitumumab (Vectabix) (EGFR, Amgen); Zalutumumab (HuMaxEGFr) (EGFR, Genmab); CDX-110 (EGFRvIII, AVANT Immunotherapeutics); Admitumumab (MT201) (Epcam, Merck); Izumuzumab (edrecolomab, Panorex, 17-lA) (Epcam, Glaxo/Centocor); MORAb-003 (folate receptor a, Morphotech); KW-2871 (neuronectin GD3, Kyowa); MORAb-009 (GP) -9,M Orphotech) ·, CDX-1307 (MDX-1307) (hCGb, Celldex); trastuzumab (Herceptin) (HER2, Celldex); pertuzumab (rhuMAb 2C4) (HER2 (DI) ), Genentech); apozumab (HLA-DR beta chain, PDL Pharma); AMG-479 (IGF-1R, Amgen); anti-IGF-1R R1507 (IGF1-R, Roche); CP 751871 (IGF1-R, Pfizer); IMC-A12(IGF1-R, Imclone); BIIB022(IGF-1R, 160877.doc -159- 201247704

Biogen); Mik-β-1 (IL-2Rb (CD 122) » Hoffman LaRoche); CNTO 328 (IL6, Centocor); anti-KIR (1-7F9) (killer cell Ig-like receptor (KIR) 'Novo); Hu3S193 (Lewis (y), Wyeth, Ludwig Institute of Cancer Research); hCBE-ll (LTpR, Biogen); HuHMFGl (MUCl, Antisoma/NCI); RAV12 (N-linked carbohydrate epitope, Raven); CAL ( Parathyroid hormone-related protein (PTH-rP), University of California); CT-011 (PD1,

CureTech); MDX-1106 (ono-4538) (PD 1 >Medarex/Ono); MAb CT-011 (PD1, Curetech); IMC-3G3 (PDGFRa »

Imclone); bavituximab (bavituximab)

Peregrine); huJ591 (PSMA > Cornell Research Foundation); muJ591 (PSMA, Cornell Research Foundation); GC1008 (TGFb (ubi) inhibitor (IgG4), Genzyme); Remicade (TNFa, Centocor); A27.15 (transferrin receptor,

Salk Institute, INSERN WO 2005/11 1082); Ε2·3 (transferrin receptor, Salk Institute); bevacizumab (Avastin) (VEGF >Genentech); HuMV833 (VEGF » Tsukuba Research · Lab, PCT Publication No. WO/2000/034337, University of

Texas) ; IMC-18F1 (VEGFR1 >Imclone); IMC-1121 (VEGFR2,

Imclone) o C. Construction of a semi-Ig binding protein to design a semi-Ig binding protein that encodes at least one or more parental binding proteins (eg, monoclonal antibodies, scFv, domain antibodies, camelid antibodies, receptors or backbone antigen binding proteins) The sequence of the heavy chain antigen binding domain is directly linked in tandem by a DNA technique of the group 160877.doc-160-201247704 or via a peptide sequence (eg, one or more of a CH1 domain joined in a tandem or via a linker, in the CH2 domain) Short link connection. The semi-Ig binding protein may further comprise a sequence encoding a light bond antigen binding domain which may comprise one or more light bond variable domains, a domain antibody based on a light or heavy chain sequence, a scFv, a receptor or a backbone antigen binding protein, Connected to the light chain constant domain as appropriate. In certain embodiments, each heavy and light chain antigen binding domain can comprise more than one antigen binding domain. (Figure 1A). In certain embodiments, the heavy and light linkages in the first and second polypeptides are variable. The domains together are complementary variable domains and form a single functional antigen binding site. In certain embodiments, the heavy and light chain variable domains in a single polypeptide form a complementary pair to bind a single antigen. In certain embodiments, the heavy and light chain variable domains form a complete independent antigen binding site on each polypeptide. For example, when the heavy and light bond antigen binding domains are independently selected from a domain antibody, a pathway antibody, a receptor, and an scFv, a complete independent antigen binding site is present on each peptide. In certain embodiments, such as shown for the half RAb_Ig in Figure 1A, there are two antigen binding domains in each of the light and heavy chains. An antigenic junction of the first polypeptide can be paired with an antigen binding domain on the second polypeptide to form a single functional antigen binding site; and the second antigen binding domain of each of the first and second polypeptides is separate A functional antigen binding site is formed. In such embodiments, the anti-primitive binding domain is preferably adjacent to the linker or gamma domain present in the binding protein, and the independent functional antigen binding site is remote from the linker or CH3 domain present in the binding protein. The variable antigen binding domain can be assayed using parental antibodies (eg, monoclonal antibodies), DVDs, tvds, "π, domain anti-160877.doc • 161 - 201247704 bodies, receptors or backbones known in the art. The antigen binding protein is obtained, or produced by any of the methods described herein. In one embodiment, the antigen binding domain is a murine heavy or light chain variable domain. In another embodiment, the variable domain is a CDR Transplanting or humanizing a heavy chain variable domain or a light chain variable domain. In one embodiment, the variable domain is a human heavy chain variable domain or a light chain variable domain. In one embodiment, recombinant DNA techniques are employed. The first and second antigen binding domains are directly linked to each other. In another embodiment, the antigen binding domain is joined via a linker sequence. In one embodiment, two antigen binding domains are joined. Three or more antigen binding domains It may also be ligated directly or via a linker sequence. The variable domain may bind to the same antigen or may bind to a different antigen" The semi-Ig molecule of the invention may comprise an immunoglobulin variable domain and a non-immunoglobulin variable domain ( Receptor The ligand binding domain or the active domain of the enzyme. The semi-Ig binding protein may also comprise two or more non-Ig domains. The linker sequence may be a single amino acid or polypeptide sequence. In one embodiment, the linker The sequence is selected from the group consisting of: ASTKGPS VFPLAP (SEQ ID NO: 46); ASTKGP (SEQ ID NO: 48); TVAAPSV FIFPP (SEQ ID NO: 50); TVAAP (SEQ ID NO: 52); AKTTPKLEEGEFSEAR (SEQ ID NO: 94); AKTTPKLEEG EFSEARV (SEQ ID NO: 95); AKTTPKLGG (SEQ ID NO: 96); SAKTTPKLGG (SEQ ID NO: 97); SAKTTP (SEQ ID NO: 98); RADAAP (SEQ ID NO: 99) RADAAPTVS (SEQ ID NO: 100); RADAAAAGGPGS (SEQ ID NO: 101); RADAAAA(G4S)4 (SEQ ID NO: 102); SAKTTPKLEEGEFS EARV (SEQ ID NO: 103); ADAAP (SEQ ID NO: 104) 160877.doc -162- 201247704 ADAAPTVSIFPP (SEQ ID NO: 105); QPKAAP (SEQ ID NO: 106); QPKAAPSVTLFPP (SEQ ID NO: 107); AKTTPP (SEQ ID NO: 108); AKTTPPSVTPLAP (SEQ ID NO: 109); AKTTAP (SEQ ID NO: 110); AKTTAPSVYPLAP (SEQ ID NO: 111); GGGGSGGGGSGGGGS (SEQ ID NO: 112); GENKVEYA PALMALS (SEQ ID NO: 113); GPAKELTPLKEAKVS (SEQ ID NO: 114); GHEAAAVMQVQYPAS (SEQ ID NO: 115); TVAAPSVFIFPPTVAAPSVFIFPP (SEQ ID NO: 116); and ASTKGPSVFPLAPASTKGPSVFPLAP (SEQ ID NO: 117). The choice of linker sequences is based on the analysis of the crystal structure of several Fab molecules. In Fab or antibody molecular structures, there is a natural flexible linkage between the variable domain and the CH1/CL constant domain. This natural linkage comprises about 10-12 amino acid residues, provided by 4-6 residues from the C-terminus of the V domain and 4-6 residues from the N-terminus of the CL/CH1 domain. The DVD Ig of the present invention is produced by using N-terminal 5-6 amino acid residues of CL or CH1 or 11-12 amino acid residues, respectively, as a linker in the light chain of the half Ig and the heavy chain. The N-terminal residue of the CL or CH1 domain (especially the first 5-6 amino acid residues) adopts a ring configuration without a stable secondary structure and thus acts as a flexible linker between the two variable domains. The N-terminal residue of the CL or CH1 domain is a naturally extending sequence of the variable domain due to its part of the Ig sequence, and thus to a large extent minimizes any immunogenicity that may be caused by the linker and junction. . Other linker sequences may include any sequence of any length of the CL/CH1 domain (eg, (:[/(: first 5-12 amino acid residues of the 111 domain) instead of (:1^/(:111 domain) All residues; the light chain linker may be from Ck or (: λ; and the heavy chain linker 160877.doc -163 - 201247704 may be derived from any CHI of the same type, including Cyl, Cy2, Cy3, Cy4, Cal, Ca2, Ce and Ομ. The linker sequence may also be derived from other proteins, such as Ig-like proteins (eg, TCR, FcR, KIR); G/S-based sequences (eg, G4S repeats (SEQ ID NO: 208)); Sequences of regions; and other native sequences from other proteins. D. Production of non-dimerized CH3 domains The invention encompasses peptides having a CH3 domain, wherein dimerization is, for example, fully or partially inhibited. For example, including inhibition of CH3- At least one mutation of at least one mutated CH3 dimerization of CH3 at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70°, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96% At least 97%, at least 98%, or at least 99% are not dimerized via the CH3 domain. Provided herein are methods for generating, testing, and identifying CH3 domains in which dimerization is inhibited, such as in Example 1.1.2, which teaches a large amount of A heavy chain construct having a mutation in the Fc domain of both the hinge region and the CH3 domain. Site-directed mutagenesis is a conventional method in the art. Example 3 provides a test for the CH3 domain in the case of the semi-Ig of the present invention. Method of dimerization. Analysis indicates that mutations in cysteine in the hinge region and adjacent regions (according to Kabat Nos. C220, C226 and C229) are suitable, but not necessary, to substantially inhibit the dimerization of semi-Ig binding protein constructs. The mutation of the amino acid T366, L368, P395, F405, Y407 and K409 in the CH3 domain alone or in combination is suitable for inhibiting the dimerization of the CH3 domain. Surprisingly, the single point mutation in the CH3 domain F405R, Y407R And the combination of K409D and the hinge region mutations C226S and C229S can substantially destroy the dimerization of 160877.doc -164- 201247704 CH3-CH3, resulting in a high percentage of half ig, while C226S and C229S and T366F, L368F, P395A, F405R, Y407R And the group of K409D This results in a higher degree of CH3 dimerization. As demonstrated herein, fewer mutations may be more effective in inhibiting CH3 dimerization. E. Making a semi-Ig binding protein may be by any of a variety of techniques known in the art. The binding proteins of the invention are made. For example, from host cell expression, the expression vector encoding the heavy chain of the semi-Ig binding protein and optionally the light chain is transfected into the host cell by standard techniques. The various forms of the term "transfection" are intended to encompass a variety of techniques commonly used to introduce exogenous DNA into prokaryotic or eukaryotic host cells, such as electroporation, calcium phosphate precipitation, DEAE-dextran transfection, and the like. Although it is possible to express the semi-Ig binding protein of the present invention in a prokaryotic or eukaryotic host cell, the semi-Ig binding protein should be expressed in eukaryotic cells (e.g., mammalian host cells) because of such eukaryotic cells (and especially Mammalian cells are more likely than prokaryotic cells to assemble and secrete appropriately folded and immunologically active half Ig binding proteins. • Exemplary mammalian host cells for use in representing recombinant antibodies of the invention include Chinese hamster ovary (CHO) cells (including dhfr-CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77 : 4216-4220, for example, as described in Kaufman, RJ and Sharp, PA (1982) Mol. Biol. 159: 601-621, for use with DHFR selectable markers), NSO myeloma cells, COS cells, SP2 And PER.C6 cells. When a recombinant expression vector encoding a semi-Ig binding protein is introduced into a mammalian host cell, the host cell is cultured for a period of time sufficient to allow expression of the semi-Ig binding protein 160877.doc-165-201247704 in the host cell or secretion of the semi-Ig binding protein to A period of time in the medium in which the host cells are grown produces a half-ig binding protein. The semi-Ig binding protein can be recovered from the culture medium using standard protein purification methods. In an exemplary system for recombinant expression of a half-binding protein of the invention, a recombinant expression vector encoding a heavy chain of a hemi-Ig binding protein and optionally a light chain of a semi-Ig binding protein by calcium phosphate-mediated transfection Introduced into dhfr_CH〇 cells. In recombinant expression vectors, the half 1 § binding protein heavy and light chain genes are each operably linked to a CMV enhancer/AdMLP promoter regulatory element to drive high level transcription of the gene. The recombinant expression vector also carries the DHFR gene, which allows for the selection of CHO cells that have been transfected with the vector using methotrexate selection/amplification. The selected transitional host cells are cultured to allow expression of the semi-Ig binding protein heavy and light chains and the recovery of intact half-binding proteins from the culture medium. Standard molecular biology techniques are used to prepare recombinant expression vectors, to transfect host cells, to select for transformation, to culture host cells, and to recover semi-Ig binding proteins from the culture medium. The present invention further provides a method of synthesizing a half-binding protein of the present invention by culturing a host cell of the present invention in a suitable medium until synthesis of a half-binding protein of the present invention. The method can further comprise isolating the semi-Ig binding protein from the culture medium. An important feature of the semi-Ig scorpion protein of the present invention is that its τ is produced and purified in the same manner as the conventional antibody U. The manufacture of a semi-Ig binding protein produces a homogenous single major product of the desired living' without any sequence of G-decorations or chemical modifications of any kind. Other previously described methods of producing "semi-antibody" do not produce I-major production (9). Rather, previous attempts to produce "half antibodies" have resulted in intracellular production or secretion of monomeric and dimeric "half" 160877.doc 201247704 antibody cocktails, or monomeric antibodies that do not have an Fc domain (eg Fab or F (ab) 2 fragment), or a "half" antibody having a substantially truncated CH3 domain (ie, a CH3 domain sequence of less than about 30°, 25%, 20%, 15%). The compositions and methods also provide multispecific semi-Ig binding proteins for binding to a variety of different antigens. At least 30. /. At least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97°/. At least 98% or at least 99% of the assembled and expressed half Ig binding protein is not dimerized via the CH3 domain. The percentage of semi-Ig binding protein present in a sample can be determined using any of the methods provided in this application (eg, in Examples 3 and 4). In a preferred embodiment, by size exclusion chromatography (SEC) The percentage of half ig binding protein present was determined. In some embodiments, other methods can be used to determine the percentage of half of the bound protein present. This aspect particularly enhances the commercial utility of the present invention. Thus, the invention encompasses a method of expressing a specific antigen binding domain having an Fc region to promote effector function' wherein the peptide (i.e., the semi-Ig binding protein) is not dimerized via the CH3 domain. In a preferred embodiment, the semi-Ig binding protein is expressed in a single cell, producing a single major product of a "semi-Ig" binding protein. A semi-Ig binding protein comprises a single polypeptide having at least one heavy chain antigen binding domain and a non-dimerized CH3 domain. In another embodiment, the semi-Ig binding protein comprises two polymorphisms. The first polypeptide comprises at least one heavy chain antigen binding domain and a non-dimerized CH3 domain, and the second polypeptide comprises at least one light chain antigen binding domain, wherein the first and second polypeptides form a single half Ig binding protein. Since the second polypeptide does not include the CH3 domain, the interaction between the first and second polypeptides does not include CH3-CH3 dimerization. The present invention provides a method for expressing a semi-Ig binding protein in a single cell to produce a "major product" of "semi-Ig binding protein", wherein the "main product" is at least 30%, at least 40%, at least 5%, at least μ%, At least 嶋, at least 65%, at least 70%, at least 75%, at least 8%, at least 85 〇/〇, at least 90%, at least 91%, at least 92%, at least %%, at least 94. /. At least 95%, at least 96%, at least 97%, at least 9 hearts or up to 99. /. All of the assembled proteins comprise a non-CH3_dimerized protein. The amount of CH3 dimerized protein can be determined, for example, using any of the methods provided herein (see, e.g., Example 3). In one embodiment, the amount of dimerized protein is determined by a quantitative method. In another embodiment, the amount of dimerized protein is determined by size exclusion chromatography. In one embodiment, the amount of dimerized protein is determined by analytical ultracentrifugation. It will be understood that the dimerized protein does not include non-specific aggregates, i.e., by conventional denatured SDS_PAGE, does not dissociate into aggregates of monomers. F• Derivatized Semi-Ig Binding Proteins One example provides a labeled binding protein in which a binding protein of the invention is derivatized or linked to another work: a knife (e.g., another peptide or protein). For example, a labeled binding protein of the invention can be obtained by functionally linking a binding protein of the invention to one or more other molecular entities (eg, by chemical coupling, gene fusion, non-covalent association, or other means). The one or more knives are, for example, another antibody (eg, a bispecific antibody or a bipotent b antibody), a detectable agent, a cytotoxic agent, a medicinal agent, and/or can be mediated by 160877.doc 201247704, a protein or peptide in which the σ 5 protein is associated with another molecule such as a streptavidin core region or a polyhistidine tag. Suitable detectable agents that can be used to derivatize the binding proteins of the invention include fluorescent compounds. Exemplary fluorescent detectable agents include luciferin, fluorescein isothiocyanate, rhodamine, 5-dimethylamine naphthalenesulfonate, phycoerythrin, and the like. The binding protein can also be derivatized with a detectable enzyme such as alkaline phosphatase, horseradish peroxidase, glucose oxidase and the like. When a binding protein is derivatized with a detectable enzyme, it is produced by adding other reagents (the enzyme uses the reagents)

The reaction product can be detected and detected. For example, when a detectable agent, horseradish peroxidase, is present, the addition of hydrogen peroxide and diaminobenzidine produces a detectable colored reaction product. Biotin-derived binding to protein can also be detected by indirect measurement of avidin or streptavidin binding. Another embodiment of the invention provides a crystalline binding protein and formulations and compositions comprising the same. In one embodiment, the crystallized binding protein has an in vivo half-life that is longer than the soluble counterpart of the binding protein. In another embodiment, the binding protein retains biological activity after crystallization. The crystallized binding protein of the present invention can be produced according to the method disclosed in the art and as disclosed in the publication of WO 02/072636. Another embodiment of the invention provides a glycosylated binding protein (such as an antibody) wherein the antibody or antigen binding portion thereof comprises one or more carbohydrate residues. Initial in vivo protein production can undergo further processing known as post-translational modification. In particular, the sugar (glycosyl) residue can be added by enzymatic treatment. The resulting (4) side bond with a covalently bonded (4) side bond (59. 160877.doc 201247704 protein is called a glycosylated protein or glycoprotein. The antibody has one or more carbohydrate residues in the domain and variable domains. Glycoprotein. The residue of the hydrated water compound in the Fc domain has an important effect on the effector function of the Fc domain, but has little effect on the antigen binding or half-life of the antibody (jefferis, r·丨2〇〇5) Biotechnol. Prog· 21 : 11-16). In contrast, glycosylation of the variable domain can have an effect on the antigen binding activity of the antibody. Glycosylation in the variable domain may have a negative impact on antibody binding affinity, which may be caused by steric effects (Co, MS et al, (1993) Mol. Immunol. 30: 1361-1367) or lead to affinity for antigen Increase (Wallick, SC et al., (1988) Εχρ

Med. 168:1099-1 109; Wright, Α· et al., (1991) EMBO J. 10:2717 2723). One aspect of the invention is directed to a glycosylation site mutant in which a conjugated or n-linked glycosylation site that produces a binding protein has been mutated. Those skilled in the art can generate such mutants using standard well-known techniques. A glycosylation site mutant that retains biological activity but has enhanced or attenuated binding activity is another object of the invention. In another embodiment, the glycosylation of a binding protein (e.g., an antibody) or antigen binding portion of the invention is modified. For example, aglycosylated antibodies can be prepared (i.e., the antibody lacks glycosylation). Glycosylation can be altered, for example, to increase the affinity of the binding protein for the antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more glycosylation sites in the binding protein sequence. By way of example, one or more amino acid substitutions can be made which result in the elimination of one or more variable region glycosylation sites, thereby eliminating glycosylation at the site. This glycosylation increases the affinity of the binding protein for the antigen. Such a method is described in further detail in the pCT publications, which are hereby incorporated by reference in its entirety. Alternatively or additionally, a low-fucosylated antibody having a reduced amount of a modified binding protein of the invention having an altered glycosylation type, such as a trehalyl residue, can be prepared (see Kanda et al., (2007) J. Biotechnol. 130(3): 300-3 3.) or an antibody that increases the structure of the GlcNAc. These altered glycosylation patterns have been shown to enhance the ADCC ability of antibodies. Such carbohydrate modifications can be achieved, for example, by expressing the binding protein in a host cell altered by a glycosylation machinery. Cells altered by glycosylation machinery have been described in the art and can be used as host cells for the expression of the recombinant binding proteins of the invention thereby producing a glycosylation-binding binding protein. See, for example, Shields, R丄 et al.

(2002) J. Biol. Chem. 277: 26733-26740; Umana et al., (1999) Nat. Biotech. 17: 176-1, and European Patent EP

1,176,195; and PCT Publication Nos. WO 03/035835 and WO 99/54342 80 ° Protein glycosylation depends on the amino acid sequence of the relevant protein and the host cell expressing the protein. Different organisms can produce different glycosylation enzymes (e.g., glycosyltransferases and glycosidases) and have different available substrates (nucleoside sugars). Because of these factors, the protein glycosylation pattern and the composition of the glycosyl residues may vary depending on the host system that represents the particular protein. Glycosyl residues suitable for use in the present invention may include, but are not limited to, glucose, galactose, mannose, trehalose, eta-acetylglucosamine, and sialic acid. In one embodiment, the glycosylated binding protein comprises a glycosyl residue such that the glycosylation pattern is a human glycosylation pattern. 160877.doc -171 - 201247704 It is known to those skilled in the art that different protein glycosylation can produce different protein characteristics. For example, the efficacy of a therapeutic protein produced in a microbial host such as yeast and utilizing glycosylation of the yeast endogenous pathway may be comparable to the efficacy of the same protein expressed in mammalian cells such as CHO cell lines. low. These glycoproteins may also be immunogenic in humans and exhibit reduced in vivo half-life after administration. Special stimuli in humans and other animals recognize specific glycosyl residues and promote rapid clearance of proteins from the bloodstream. Other adverse effects may include protein folding, solubility, sensitivity to protease, transport, transport, compartmentalization, secretion, recognition by other proteins or factors, antigenicity or allergenicity. Thus, a physician may select a therapeutic protein having a particular glycosylation composition and pattern (e.g., a glycosylation composition and pattern that is identical or at least similar to that produced in a human cell or a species-specific cell of a predetermined individual animal). A glycosylated protein that behaves differently from a glycosylated protein of a host cell can be achieved by genetically modifying the host cell to express a heterologous glycosylation enzyme. Using techniques known in the art, physicians can produce antibodies or antigen binding portions thereof that exhibit glycosylation of human proteins. For example, yeast strains have been genetically modified to represent non-naturally occurring glycosylation enzymes such that glycosylated proteins (glycoproteins) produced in such yeast strains are expressed with animal cells (especially human cells) Protein glycosylation is the same protein glycosylation (U.S. Patent Nos. 7,449,308 and 7,029,872 and pCT Publication No. 2005/100584). In addition to binding proteins, the invention also targets anti-idiotypic (anti-Id) antibodies specific for the binding proteins of the invention. The anti-Id antibody is an antibody that recognizes the unique determinant associated with the antigen binding region of another binding protein, such as an antibody, 160877.doc • 172· 201247704. Anti-Id can be prepared by immunizing an animal with a binding protein or a CDR-containing region thereof. The immunized animal will recognize and respond to the individual genotype determinant of the immunological binding protein and produce an anti-Id antibody. It is apparent that it may be easier to produce anti-idiotypic antibodies that bind to two or more parental binding proteins in a DVD-Ig molecule; and that binding is confirmed by methods well-recognized in the art (eg, BiAC0re, ELISA) Studies to verify that an individual genotype antibody specific for an individual genotype of each parent binding protein also recognizes an individual genotype (eg, an antigen binding site) in the case of a semi-Ig binding protein. Anti-individual genotype antibodies specific for each of the more antigen binding sites of the semi-Ig binding protein (especially when more than one antigen binding site is present) provide half of the human semi-Ig binding protein in the patient's serum The ideal reagent for Ig binding protein concentration; the semi-binding protein concentration assay can be established using the "sandwich assay ELISA format" in which antibodies against the antigen binding region are applied to a solid phase (eg, BIAcore® wafer, eusa disk, etc.). Rinse with wash buffer, incubated with serum sample to 'wash again, and when a second antigen binding site is present, finally incubated with another anti-idiotypic antibody against another antigen binding site, the other antibody The individual genotype antibody itself is enzymatically labeled to bind to the reaction. In one embodiment, for a half 1 g of binding protein having more than two different binding sites, anti-individual genotype antibodies directed against the two outermost binding sites (located at the most distal and proximal ends of the constant region) will not only have It helps to determine the concentration of the semi-Ig binding protein in human serum and also helps to prove the integrity of the molecule in vivo. Each anti-Id antibody can also be used as an "immunogen" for inducing an immune response in another animal 160877.doc-173-201247704, thereby producing a so-called anti-antigen. Furthermore, it will be appreciated by those skilled in the art that a library of host cells genetically engineered to express various glycosylation enzymes can be used to express a related protein' such that a host cell member of the library produces a variant glycosylation pattern. Related proteins. The physician can then select and isolate related proteins with specific novel glycosylation patterns. In an embodiment, the protein f having a specifically selected novel glycosylation pattern exhibits improved or altered biological properties.

Use of π·half-ig-binding protein In view of the ability of the binding protein of the invention to bind to one or more antigens, such binding proteins can be used to use conventional immunoassays such as ELISA, radioimmunoassay (ria) or tissue immunohistochemistry. Predicted antigens (eg, in biological samples, such as antigens in serum or plasma). The semi-Ig binding protein is directly or indirectly labeled with a detectable substance to facilitate detection of bound or unbound antibodies.

Suitable for detectable substances include various enzymes, prosthetic groups, fluorescent substances, luminescent substances and radioactive substances. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, and acetylcholinesterase; examples of suitable prosthetic complexes include streptavidin/biotin and avidin Examples of proteins/biotin's suitable for fluorescent substances include umbelHferone, luciferin, luciferin isothiocyanate, rhodamine, dichlorotriazinylamine luciferin, tannin gas (dansyl chl0ride) And phycoerythrin; examples of luminescent substances include luminol; and examples of suitable radioactive materials include 3H, 35 s Ύ 99

Tc 11 】Ιη 125ι 131 177

Lu, 166Ηο, and 153

Sm 160877.doc -174- 201247704 In one implementation, the binding proteins of the invention are active in vitro and in vivo and antigenically. Thus, such semi-binding proteins may inhibit antigenic activity in, for example, a cell culture containing antigen, in a human subject or other mammalian subject having an antigen that is reactive with the binding protein of the present invention. In another embodiment, the invention provides a method of reducing the antigenic activity of an individual suffering from a disease or condition that is deleterious to antigenic activity. A binding protein of the invention can be administered to a human subject for therapeutic purposes.

As used herein, the term "disease that is harmful to antigenic activity" is intended to include the following diseases and other conditions in which the presence of an antigen in an individual suffering from the condition has been shown or suspected to be the cause of the pathophysiology of the condition, or The cause of the deterioration of the condition. Thus, a condition which is detrimental to antigenic activity is a condition which is expected to reduce the activity of the antigen and alleviate the symptoms and/or progression of the disease. Such conditions can be confirmed by, for example, an increase in the concentration of the antigen in the biological fluid of the individual suffering from the condition (e.g., an increase in the concentration of the antigen in the serum, plasma, synovial fluid, etc. of the individual). Non-limiting examples of disorders which may also be treated with the binding proteins of the invention include those disorders discussed below and in the section on pharmaceutical compositions of the binding proteins of the invention. The semi-Ig binding protein of the present invention can bind to one antigen or multiple antigens. Such antigens include, but are not limited to, the targets listed in the following databases. These target databases include the following list: Treatment targets (乂111. 23.111113.6 (111.3 Lv/Lü 1:0叩/. Plus (!"1(1.&8卩); cytokines and cytokines) Receptors (www.cytokinewebfacts.com, www.copewithcytokmes.de/cope.cgi and cmbi.bjmu.edu.cn/ cmbidata/cgf/CGF_Database/cytokine.medic.kumamoto-u.ac.jp/ -175- 160877. Doc 201247704 CFC/indexR.html); Chemokine (cytokine.medic.kumamoto-u.acjp/CFC/CK/Chemokine.html); Chemokine Receptor and GPCR (csp.medic.kumamoto-u_ac.jp/ CSP/Recep tor .html and www.gpcr.org/7tm/); Olfactory Receptor (senselab.med.yale.edu/senselab/ORDB/default.asp); Receptor (www.iuphar-db .org/iuphar-rdnist/index.htm);

Cancer target (cged.hgc.jp/cgi-bin/input.cgi); secreted protein as a potential antibody target (spd.cbi.pku.edu.cn/); protein kinase (spd.cbi.pku.edu.cn /), and the human CD mark (content.labvelocity.eom/tools/6/1226/CD_ table_final_locked.pdf) and (Zola Η (2005) Blood 106: 3123-6). A half Ig binding protein comprising two functional antigen binding sites is useful as a therapeutic to simultaneously block two different targets, thereby enhancing efficacy/safety and/or increasing patient coverage. Such targets may include soluble targets (e.g., TNF) and cell surface receptor targets (e.g., VEGFR and EGFR). It can also be used to induce tumor cells and T cells (such as Her2 and CD3) (for cancer therapy), or between autoreactive cells and effector cells (for autoimmune diseases or transplants), or any target cells. Reorientation of cytotoxicity with effector cells to eliminate cells that cause disease in any given disease. Furthermore, when a semi-Ig binding protein is designed to dictate two different epitopes on the same receptor, it can be used to initiate receptor clustering and activation. This has 160877.doc -176- 201247704 for the preparation of agonistic and antagonistic anti-GPCR therapeutics. In this case, a semi-Ig binding protein can be used to target two different epitopes on one cell (including epitopes on both the loop and extracellular domains) to achieve clustering/signaling (two cell surface molecules) ) or signal conduction (on a molecule). Similarly, a semi-Ig binding protein can be designed to elicit immunity by targeting two different epitopes of the CTLA-4 extracellular domain (or two copies of the same epitope) to initiate CTLA-4 junction and negative signals. The reaction is lowered. CTLA-4 is a clinically proven target for the therapeutic treatment of a variety of immunological disorders. CTLA-4/B7 interaction negatively regulates T cell activation by impairing cell cycle progression, IL-2 production, and T cell proliferation after activation, and CTLA-4 (CD 152) engagement down-regulates T cell activation and promotes immunity Induction of tolerance. However, the strategy of attenuating T cell activation by the agonistic antibody engagement of CTLA-4 was not successful because CTLA-4 activation requires conjugation. As confirmed by crystal structure analysis, the molecular interaction of CTLA-4/B7 is in the form of a "slanted zipper" array (Stamper (2001) Nature 410: 608). However, currently available CTLA-4 binding reagents, including anti-CTLA-4 mAbs, do not have binding properties. Several attempts have been made to resolve this issue. In one case, a single-chain antibody that binds to a member of the cell is produced, which significantly inhibits allogeneic rejection in mice (Hwang (2002) J. Immunol. 169:633). In another case, an artificial APC surface-linked single-chain antibody against CTLA-4 was produced and proved to attenuate the T cell response (Griffin (2000) J. Immunol. 164:4433). In both cases, CTLA-4 engagement is achieved by tightly localizing antibodies bound to members in an artificial system. Although these experiments provide a concept of immune down-regulation by triggering CTLA-4 negative signaling, I60877.doc -177- 201247704 verifies that the reagents used in these reports are not suitable for therapeutic use. To this end, CTLA-4 conjugation can be achieved by using a semi-Ig binding protein that targets two different antigenic determinants of the CTLA-4 extracellular domain. The rationale is that the distance (about 150-170 A) across the two binding sites of IgG is too large to achieve active ligation of CTLA-4 (30-50 A between 2 CTLA-4 homodimers). However, the distance between the two binding sites on the semi-Ig binding protein (one arm) is much shorter (also in the range of 30-50 A), allowing for proper engagement of CTLA_4. The semi-Ig binding proteins of the invention can be used to pair antigens without clustering. Two copies of the same antigen binding site can be incorporated in a single half Ig to facilitate conjugation of the two copies containing the antigen of interest without clustering. Similarly, two different target antigens can be joined without clustering. A half Ig binding protein with three (or more) binding sites can be used to similarly bind a small number of targets without promoting clustering. Similarly, a semi-Ig binding protein having at least two antigen-binding sites can target two different members of the cell surface receptor complex (eg, m" and 13). In addition, the hemi-(tetra)-protein can be non-directional to CR1 and soluble. The protein "pathogen' thus drives rapid clearance of the target soluble protein/pathogen. Furthermore, the semi-binding protein of the invention having at least two antigen-binding sites can be used for tissue-specific delivery of taste-to-tissue markers and disease mediators to enhance local PK 'Therefore, it has higher efficacy and/or lower toxicity, including intracellular transmission of axial internalization receptors and intracellular molecules) and transmission to the transferrin receptor and CNS disease, and disease. The body can also pass through the blood-brain barrier, and can also be used as a carrier protein to transfer the antigen to a specific location via a non-neutralizing antigen to the antigen, and to extend the half-life of the antigen. ^ 160877.doc • 178. 201247704 The external 'semi-Ig binding protein can be designed to be physically linked to or targeted to a medical device implanted in a patient (see Burke, SE et al. (2006) Adv. Drug Deliv. Rev. 58(3) : 437 -446; Hildebrand, HF et al., (2006) 511 also. 6 311 (1 (3〇 has 1^11§3丁6.1111〇1.200(22-23): 6318-6324,Wu,Ρ· et al. (2006) Biomaterials 27(11): 2450-2467; Marques, AP fA, (2005) Biodegrad. Syst· Tissue Eng.. and Regen· Med. 377-397). Briefly, guide cells of the appropriate type To the medical implant site to promote healing and repair of normal tissue function. Or, also mediators released by the semi-Ig binding protein coupled to or targeted to the device when implanted (including (but not Limited to) cytokines) provide inhibition. For example, stents have been used in interventional cardiology for years to clear obstructed arteries and improve blood to myocardial flow. However, traditional bare metal stents are known in some patients. Causes restenosis (re- narrowing the arteries in the treated area) and produces blood clots. Recently, stents coated with anti-CD34 antibodies have been described which capture endothelial progenitor cells (EPC) circulating throughout the blood. To reduce restenosis and prevent blood clots. Endothelial cells The cells lined inside the blood vessels to allow smooth flow of blood. EPC adheres to the hard surface of the stent to form a smooth layer that not only promotes healing but also prevents restenosis and blood clots previously associated with stent use (Aoji Et al. (2005) J. Am. Coll. Cardiol. 45(10): 1574-9). In addition to improving the outcome of patients requiring stents, there is also a sense for patients requiring cardiovascular bypass surgery. For example, repairing a vascular tube (artificial artery) coated with an anti-EPC antibody will eliminate the need to use an artery from the patient's leg or arm for bypass surgery. 160877.doc .179· 201247704 This will reduce surgery and anesthesia: the number of sputum, ❹ will reduce the crown (four) pulse surgery dead = ° designed by -^ multi-specific semi-(10) protein, which makes it bind to cell surface markers ( Such as coffee 4) and proteins that have been applied to implanted devices to promote cell recruitment (or any type of epitope, including but not limited to proteins, lipids, and polysaccharides).豸 and other methods can also be used for other medical objects. Alternatively, a half 1 § binding protein can be applied to a medical device and after the device is implanted and all semi-匕 binding proteins are released from the device (or any other requirement that may require other fresh hemi-(tetra) protein, including the loaded half Ig binding protein aging and denaturation), the device can be reloaded by systemically administering fresh semi-Ig binding protein to the patient, and the middle half 1 § combines the egg. 10% in groups, · Ό σ sites bind to related targets (cytokines, cell surface markers (such as CD34), etc.) and bind to other targets (including proteins and any species) Antigenic determinants, including but not limited to lipid 'polysaccharides and polymers. This technique has the advantage of extending the applicability of coated implants. A. Use of a semi-Ig binding protein in various diseases The semi-Ig binding protein of the present invention is also useful as a therapeutic molecule for treating various diseases. The semi-Ig binding proteins can bind to one or more of the targets involved in a particular disease. Examples of such targets in various diseases are described below. 1. Human autoimmune and inflammatory responses Generally, many proteins are involved in autoimmune and inflammatory reactions, including C5, CCL 1 (1-309), CCL11 (eotaxin), CCL13 ( Mcp-4), CCL15 (MIP-Id), CCL16 (HCC-4), CCL17 (TARC), CCL18 (PARC), CCL19, 160877.doc -180- 201247704

CCL2 (mcp-l), CCL20 (MIP-3a), CCL21 (MIP-2) 'CCL23 (MPIF-1), CCL24 (MPIF-2/eosinophil chemotactic factor-2), CCL25 (TECK), CCL26, CCL3 (MIP-la), CCL4 (MIP-lb), CCL5 (RANTES), CCL7 (mcp-3), CCL8 (mcp-2), CXCL1, CXCLIO (IP-IO), CXCLll (I-TAC/ IP-9), CXCL12 (SDF1), CXCL13, CXCL14, CXCL2, CXCL3, CXCL5 (ENA-78/LIX), CXCL6 (GCP-2), CXCL9, IL13, IL8, CCL13 (mcp-4), CCR1, CCR2 , CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CX3CR1, IL8RA, XCR1 (CCXCR1), IFNA2, IL10, IL13, IL17C, ILIA, IL1B, IL1F10, IL1F5, IL1F6, IL1F7, IL1F8, IL1F9, IL22, IL5, IL8, IL9, LTA, LTB, MIF, SCYE1 (endothelial mononuclear cell cytokine), SPP1, TNF, TNFSF5, IFNA2, ILIORA, IL10RB, IL13, IL13RA1, IL5RA, IL9, IL9R, ABCF1, BCL6, C3 , C4A, CEBPB, CRP, ICEBERG, IL1R1, IL1RN, IL8RB, LTB4R, TOLLIP, FADD, IRAKI, IRAK2, MYD88, NCK2, TNFAIP3, TRADD, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6, ACVR1, ACVR1B, ACVR2 , ACVR2B, ACVRL1, CD28 CD3E, CD3G, CD3Z, CD69, CD80, CD86, CNR1, CTLA4, CYSLTR1, FCER1A, FCER2, FCGR3A, GPR44, HAVCR2, OPRD1, P2RX7, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, BLR1, CCL1, CCL2, -181 - 160877.doc 201247704 CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL13, CCL15, CCL16, CCL17, CCL18, CCL19, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCR1 , CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CX3CL1, CX3CR1, CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL10, CXCL11, CXCL12, CXCL13, CXCR4, GPR2, SCYE1, SDF2, XCL1, XCL2 , XCR1, AMH, AMHR2, BMPR1A, BMPR1B,

BMPR2, C19orflO (IL27w), CER1, CSF1, CSF2, CSF3, DKFZp451J0118, FGF2, GFI1, IFNA1, IFNB1, IFNG, IGF1, ILIA, IL1B, IL1R1, IL1R2, IL2, IL2RA, IL2RB, IL2RG, IL3, IL4, IL4R , IL5, IL5RA, IL6, IL6R, IL6ST, IL7, IL8, IL8RA, IL8RB, IL9, IL9R, IL10, IL10RA, IL10RB, IL11, IL11RA, IL12A, IL12B, IL12RB1, IL12RB2, IL13, IL13RA1, IL13RA2, IL15, IL15RA , IL16, IL17,

IL17R, IL18, IL18R1, IL19, IL20, KITLG, LEP, LTA, LTB, LTB4R, LTB4R2, LTBR, MIF, NPPB, PDGFB, TBX21, TDGF1, TGFA, TGFB1, TGFB1I1, TGFB2, TGFB3, TGFBI, TGFBR1, TGFBR2 TGFBR3, TH1L, TNF, TNFRSF1A, TNFRSF1B, TNFRSF7, TNFRSF8, TNFRSF9, TNFRSF11A, TNFRSF21, TNFSF4, TNFSF5, TNFSF6, TNFSF11, VEGF, ZFPM2 and RNF110 (ZNF144). In one aspect, a half-ig binding protein that binds one or more of the targets listed herein is provided. 2. Asthma Allergic asthma is characterized by eosinophilia, goblet cell metaplasia, epithelial cell changes, tracheal overreaction (AHr), and Th2 and Th1 cytokine expression and elevated serum IgE levels. At present, tracheal inflammation is widely recognized as a potential key factor in the pathogenesis of asthma, involving inflammatory cells such as T cells, B cells, eosinophils, mast cells and macrophages and their secreted mediators (including cytokines and chemokines). The complex interactions between the two. Corticosteroids are today's most important anti-inflammatory therapeutics for asthma, but their mechanisms of action are not specific and have safety issues, especially in the juvenile patient population. It is therefore necessary to develop more specific and dry-type therapies. There is growing evidence that IL·丨3 in mice mimics many of the characteristics of asthma, including AHR, excessive mucus secretion, and tracheal fibrosis, but not with eosinophilic inflammatory inflammation (Fin〇tt〇 et al., (2005) Internat.

Immunol. 17(8): 993-1007; Padilla et al, (2005) J. Immunol. 174(12): 8097-8105). Yu has suggested that IL·13 plays a key role in causing pathological reactions associated with asthma. The development of anti-IL_13 mAb therapy that reduces the effects of IL-13 in the lungs is an inspiring new approach that has considerable promise as a novel treatment for asthma. However, other mediators of different immune pathways are involved in the pathogenesis of asthma, and blocking these mediators in addition to IL-13 provides additional therapeutic benefits. Such target pairs include, but are not limited to, IL_丨3 and pro-inflammatory cytokines such as tumor necrosis factor-a (TNF_a). TNF-a enhances the inflammatory response of asthma' and may be associated with disease severity (McDonnell et al, 160877. doc-183. 201247704 (2001) Progr. Respir. Res 31: 247-250). This suggests that blocking both IL-13 and TNF-[alpha] may have beneficial effects, especially in severe airway diseases. In another embodiment, a half Ig binding protein of the invention binds to a target IL-13 and/or TNFa and is used to treat asthma. Animal models for assessing both inflammation and AHR (such as mouse models of OVA-induced asthma) are known in the art and can be used to determine the ability of various semi-Ig binding proteins to treat asthma. Animal models for studying asthma. Rev. Coffman et al., (2005) J. Exp. Med. 201(12): 1875-1879; Lloyd et al., (2001) Adv. Immunol. 77: 263-295; Boyce et al., (2005) J. Exp. Med. 201(12): 1869-1873; and Snibson et al., (2005) J. Brit. Soc. Allerg. Clin. Immunol. 35(2): 146-52. In addition to the routine safety assessment of this target, it may be necessary to conduct specific tests for the degree of immunosuppression and these tests help to select the best target pair (see Luster et al., (1994) Toxicology 92 (1-3) ): 229-43, Descotes et al., (1992) Devel. Biol. Stand. 77: 99· 102, Hart et al., (2001) J. Allerg. Clin. Immunoh 108(2).... 250-257). Based on the basic principles disclosed herein and using the same s-estimation model of efficacy and safety, it is possible to determine that other semi-Ig binding proteins can be combined and can be adapted to treat other targets or target pairs of asthma. In one embodiment, such targets include, but are not limited to, IL-13 and IL-Ιβ, since IL-ip is also involved in the inflammatory response of asthma; IL-13 and cytokines and chemotaxis involved in inflammation Factors such as IL-13 and IL-9; 11^13 vs. _4; ; and -25; IL-13 and TARC; IL-13 and MDC; IL-13 and MIF; IL-13 and 160877 .doc •184- 201247704

TGF-β; IL-13 and LHR agonists; IL-13 and CL25; IL-13 and SPRR2a; IL-13 and SPRR2b; and IL-13 and ADAM8. The present invention also provides a semi-Ig binding protein which can bind to one or more targets selected from the group consisting of CSF1 (MCSF), CSF2 (GM-CSF), CSF3 (GCSF), FGF2, IFNA1, IFNB1. , IFNG, histamine and histamine receptors, ILIA, IL1B, IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL9, IL10, IL11, IL12A, IL12B, IL13, IL14, IL15, IL16, IL17, IL18 , IL19, KITLG, PDGFB, IL2RA, IL4R, IL5RA, IL8RA, IL8RB, IL12RB1, IL12RB2, IL13RA1, IL13RA2, IL18R1, TSLP, CCL1, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL13, CCL17, CCL18, CCL19 , CCL20, CCL22, CCL24, CX3CL1, CXCL1, CXCL2, CXCL3, XCL1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CX3CR1, GPR2, XCR1, FOS, GATA3, JAK1, JAK3, STAT6, TBX21, TGFB1 , TNF, TNFSF6 ' YY1, CYSLTR1, FCER1A, FCER2, LTB4R, TB4R2, LTBR and chitinase. 3. Rheumatoid arthritis Rheumatoid arthritis (RA) is a systemic disease characterized by a chronic inflammatory response in the synovial membrane and is associated with cartilage degradation and proximal joint bone erosion. Many pro-inflammatory cytokines, including TNF, chemokines and growth factors, are present in diseased joints. Systemic administration of an anti-TNF antibody or sTNFR fusion protein to a RA mouse model has been shown to have anti-inflammatory and joint protection as 160877.doc-185-201247704. Intravenous administration of infliximab (Harriman, G et al, (1999) Ann. Rheum. Dis. 58 (suppl. 1): 161-4) (-the anti-TNF mAb) blocks RA patients Clinical studies of TNF activity have provided evidence that TNF regulates IL-6, IL-8, MCP-1 and VEGF production, immunization and recruitment of inflammatory cells into the joint, angiogenesis and decreased blood levels of matrix metalloproteinases 1 and 3. . A better understanding of the inflammatory pathway of rheumatoid arthritis has led to the identification of other therapeutic targets involved in rheumatoid arthritis. Promising treatments have been tested in randomized controlled trials, such as the interleukin-6 antagonist (IL-6 receptor antibody MRA developed by Chugai, Roche), see Nishimoto, N. et al., (2004) Arthrit. Rheum. 50(6): 1761-1769)), CTLA4Ig (abatacept, Genovese, Μ· et al., (2005) N. Engl. J. Med. 353:1 114-23.) and anti-B cells Therapy (Rituximab, Okamoto, Η. and Kamatani, N. (2004) N. Engl. J. Med. 351:1909). Other cytokines have been identified and have been shown to have benefits in animal models, including interleukin-15 (therapeutic antibodies HuMax-IL_15, AMG 714 (see Baslund, B. et al., (2005) Arthrit. Rheum. 52(9): 2686-2692)), interleukin-17 and interleukin-18, and clinical trials of these agents. Dual-specific antibody therapy combined with anti-TNF and another mediator has great potential for improving clinical efficacy and/or patient coverage. For example, blocking both TNF and VEGF may eradicate inflammation and angiogenesis, all of which are involved in the pathophysiology of RA. Other target pairs involved in RA are also expected to be blocked by one or more specific semi-Ig, including but not limited to TNF and IL-18; TNF and IL-12; TNF and IL-23; TNF and IL-Ιβ; TNF and MIF; TNF and IL-17; TNF with IL-15, TNF and 160877.doc -186 - 201247704 SOST. In addition to routine safety assessments of these goals or objectives, it may be necessary to conduct specific tests for the degree of immunosuppression and these tests may help to select the best target pair (see Luster et al., (1994) Toxicol. 92 ( 1-3): 229-43; Descotes et al., (1992) Devel. Biol. Stand- 77·· 99-102; Hart et al., (2001) J. Allerg. Clin. Immunol. 108(2)·· 250-257) The pre-clinical animal RA model (such as a mouse model of collagen-induced arthritis) can be used to assess whether a half-binding protein will be suitable for the treatment of rheumatoid arthritis. Other applicable models are also well known in the art (see Brand, D. D. (2005) Comp. Med. 55(2): 1 14-22). Based on the reactivity of the parent antibody to human and mouse orthologues (eg, reactivity with human and mouse TNF, human and mouse IL-15, etc.), the half-Ig produced by "matching the replacement antibody" can be Binding proteins for validation studies in mouse CIA models; in short, semi-Ig binding proteins based on one (or more) mouse target-specific antibodies can be matched to possible extent for human semi-Ig binding protein construction The characteristics of the parent binding protein (eg, human antibody or humanized antibody) (similar affinity, similar neutralizing potency, similar half-life, etc.). In one embodiment, a binding protein of the invention binds to a combination of three targets 'such as: NGF, TNF, and PGE2; and IL-la, Jiang_113, and PGE2. 4. Systemic lupus erythematosus (SlE) The immunogenic pathogen of SLE is characterized by multiple B cell activation, which leads to hyperglobulinemia, autoantibodies and immune complexes. The basic abnormality seems to be that T cells cannot suppress the b-line pure line due to systemic tau cell dysfunction. In addition, several cytokines (such as Jiang-1〇) and co-stimulatory molecules (such as CD40, CD40L, B7, CD28, CTLA-4) that initiate the second 160877.doc 201247704 signal promote B cell interaction with T cells. These interactions, together with the phagocytic clearance abnormalities of the immune complexes and apoptotic substances, contribute to the immune response and the resulting tissue damage. The following targets may be involved in SLE and potentially used in semi-Ig binding protein methods for therapeutic intervention: B cell targeted therapy: CD-20, CD-22, CD-19, CD28, CD4, CD80, HLA -DRA, IL10, IL2, IL4, TNFRSF5, TNFRSF6, TNFSF5, TNFSF6, BLR1, HDAC4, HDAC5, HDAC7A, HDAC9, ICOSL, IGBP1, MS4A1, RGS1, SLA2, CD81, IFNB1, IL10, TNFRSF5, TNFRSF7, TNFSF5, AICDA , BLNK, GALNAC4S-6ST, HDAC4, HDAC5, HDAC7A, HDAC9, IL10, IL11, IL4, INHA, INHBA, KLF6, TNFRSF7, CD28 'CD38, CD69, CD80, CD83, CD86, DPP4, FCER2, IL2RA, TNFRSF8, TNFSF7 , CD24, CD37, CD40, CD72, CD74, CD79A, CD79B, CR2, IL1R2, ITGA2, ITGA3, MS4A1, ST6GAL1, CD1C, CHST10, HLA-A, HLA-DRA and NT5E; costimulatory signals: CTLA4 or B7.1 /B7.2; Inhibition of B cell survival: BlyS or BAFF; Complement inactivation: C5; Cytokine regulation: The key principle is that the net biological response in any tissue is a balance between pro-inflammatory cytokines or local levels of anti-inflammatory cytokines The result (see Sjf Ikakis, P.P. et al., (2005) Curr. Opin. Rheumatol. 17: 550-7). It is believed that SLE is a Th-2 driven disease, and it has been confirmed that serum IL-4, IL-6 and IL-10 are elevated, and it is also expected to be combined with one or more selected from the group consisting of 160877.doc -188 - 201247704 .IL- 4. The half-ig binding protein and TNF-a of the target of IL-6, IL-10, and iFN-a. The combination of goals described herein will enhance the therapeutic efficacy against the disease's ability to test in a variety of pre-clinical models of lupus (see 1, S.L. (2_) Meth() ds version fine m: 227 72). Based on the interaction of the parent antibody with human and mouse orthologues (eg, (4) humans and matched mice, CD20, human and + mouse interference, etc.), the half of the "antibody" can be produced. (10) Protein binding in mouse lupus model

Xun research. Briefly, a semi-Ig binding protein based on one (or more) mouse target-specific antibodies can match, to the extent possible, a parental binding protein (eg, human antibody or humanization) for human half 1 § binding protein construction. The characteristics of the antibody (similar to affinity) are similar to neutralizing potency, similar half-life, etc.). 5 · Multiple Sclerosis Multiple sclerosis (MS) is a complex human autoimmune disease whose cause is largely unknown. The immunological destruction of myelin-detecting protein (MBp) throughout the nervous system is the major pathology of multiple sclerosis. MS is a disease having a complex pathology involving CD4+ and CD8+ T cell infiltration and a response in the central nervous system. The performance of cytokines, reactive nitrogen species and costimulatory molecules in CN S has been described in MS. The primary consideration is the immune mechanism that promotes autoimmunity. In particular, antigenic expression, cytokines interact with white blood cells, and regulatory T cells that help balance/modulate other T cells, such as Th 1 and Th2 cells, are important for the identification of therapeutic targets. IL-12 is a pro-inflammatory cytokine produced by APC and promoting the differentiation of Th1 effector cells. IL-12 is produced in lesions in patients with MS and in animals with EAE in 877160877.doc -189- 201247704 "Previously demonstrated interference with IL-12 pathway to effectively prevent experimental autoimmunity in rodents Encephalomyelitis (EAE), and the use of an anti-IL-12 mAb to neutralize IL-12p40 in vivo has a beneficial effect in the myelin-induced EAE model of common sputum. TWEAK is a member of the TNF family and is constitutively expressed in the central nervous system (CNS), which has pro-inflammatory, proliferative or apoptotic effects depending on the cell type. Its receptor Fnl 4 is expressed in the CNS by endothelial cells, reactive astrocytes, and neurons. During experimental autoimmune encephalomyelitis (EAE), TWEAK and Fnl4 mRNA expression was increased in the spinal cord. Anti-TWEAK antibody treatment of myelin oligodendrocyte glycoprotein (MOG)-induced sputum in C57BL/6 mice caused disease severity and decreased white blood cell infiltration after treatment in mice after the pre-sensitization phase. One aspect of the invention pertains to a semi-Ig binding protein which binds to one or more (e.g., two) targets selected from the group consisting of IL-1 2, TWEAK, IL-23, CXCL13, CD40, CD40L. , IL-18, VEGF, VLA-4, TNF, CD45RB, CD200, IFNy, GM-CSF, FGF, C5, CD52 and CCR2. One embodiment includes a dual specific anti-IL-12/TWEAK semi-DVD Ig binding protein as a therapeutic agent beneficial for MS therapy. Several animal models for assessing the suitability of semi-Ig binding protein treatments for MS are known in the art (see Steinman. L. et al., (2005) Trends Immunol. 26(1 1): 565-71; Lublin , FD et al., (1985) Springer Semin. Immunopathol. 8(3): 197-208; Genain, CP et al., (1997) J. Mol. Med. 75(3): 187-97; Tuohy, VK 160877 .doc -190· 201247704 et al., (1999) J. Exp. Med. 189(7): 1033-42; Owens, T. et al., (1995) Neurol. Clin.l3(l): 51-73; And Hart, BA et al, (2005) J. Immunol. 175(7): 4761-8). The half-Ig binding produced by "matching replacement antibodies" based on the reactivity of parental antibodies of human and animal species orthologs (eg, human and mouse IL-12, human and mouse TWEAK, etc.) The protein was validated in a mouse EAE model. Briefly, a half Ig binding protein based on one (or more) mouse target-specific antibodies can match, to the extent possible, a parental binding protein (eg, a human antibody or a humanized antibody) for human semi-Ig binding protein construction. Characteristics (similar to affinity, similar neutralization efficacy, similar half-life, etc.) ^ The same concept applies to animal models of other non-rodent species, where a half-DVD-Ig binding protein produced by "matching replacement antibodies" will be selected for the intended pharmacology Learn and possibly use for safety research. In addition to these objectives for routine safety assessments, it is necessary to conduct specific tests for the degree of immunosuppression and these tests help to select the best target pair (see Luster et al., 〇 994).

Toxicol. 92(1-3): 229-43; Descotes et al., (1992) 〇 6乂61·

Biol. Stand. 77: 99-102; Jones, R. (2000) IDrugs 3(4): 442-6). 6. The pathophysiology of septicemia is caused by the outer membrane components of gram-negative organisms (lipopolysaccharide (LPS), lipid A, endotoxin) and membrane components of gram-positive organisms (fat) Start with ipoteeichoic acid, peptidoglycan. These outer membrane components are capable of binding to the surface of monocytes. 〇14 receptor. With the help of the recently described toll-like receptor, the signal is then transmitted to the cell, resulting in the production of the pro-inflammatory cytokine tumor necrosis factor-a (TNF-a) and interleukin-1 (IL-) at the final 160877.doc -191 · 201247704. 1). Severe inflammatory and immune responses are essential features of septic shock' and play a major role in the pathogenesis of sepsis-induced tissue damage, multiple organ failure, and death. Cytokines (especially TNF and IL-1) have been shown to be key mediators of septic shock. These cytokines have a direct toxic effect on tissues; they also activate phospholipase A2. These and other effects result in increased platelet activating factor concentrations, promotion of nitric oxide synthase activity, promotion of tissue infiltration of neutrophils, and promotion of neutrophil activity. The treatment of sepsis and septic shock remains a clinical challenge, and recent prospective trials with biological response modifiers for inflammatory responses (i.e., anti-TNF and anti-MIF) show only modest clinical benefits. Recently, the focus of attention has been on reversing the treatment with the immunosuppressive phase. Studies in experimental animals and critically ill patients have shown cells in the lymphoid organs and some parenchymal tissues; increased peripheral death contributes to this immunosuppression, non-reactivity and organ system dysfunction. During septicemia, apoptosis of lymphocytes can be triggered by the absence of il_2 or by the release of glucocorticoids, granzymes or so-called "death" cytokines (ie, tumor necrosis factor a or Fas ligand). Apoptosis is continued by autoactivation of intracellular and/or mitochondrial caspase, which can be affected by pro-apoptotic and anti-apoptotic members of the Bcl-2 family. In experimental animals, treatment with an apoptosis inhibitor not only prevents apoptosis of lymphocytes, but it also improves the results. Although clinical trials using anti-apoptotic agents are largely attributable to technical difficulties associated with their administration and tissue targeting, inhibition of apoptosis of lymphocytes represents attractiveness for patients with sepsis. The goal of treatment. As with 160877.doc • 192·201247704, dual-specific agents that target both inflammatory mediators and apoptotic mediators may have other benefits. One aspect of the present invention relates to a binding protein that binds to one or more targets (in the embodiment, two targets) selected from the group consisting of sepsis: TNF, IL-1, MIF, IL,

6. IL-8, IL-18, IL-12, IL-23, FasL, LPS, Toll-like receptor, TLR-4, tissue factor, MIP-2, AD0RA2A, CASP1, CASP4, IL-10, IL- 1B, NFKB1, PR0C, TNFRSF1A, CSF3, CCR3, IL1RN, MIF, NFKB1, PTAFR, TLR2, TLR4, GPR44, HMOX1, midkine, IRAKI, NFKB2, SERPINA1, SERPINE1 and TREM1. In one embodiment, the binding protein binds to a combination of three targets, such as: HMGB1, VEGF and TNF (eg, TNFa); RAGE, VEGF and TNF (eg, TNFa); NGF, TNF (eg, TNFa) and PGE2; IL-la , IL-lb and PGE2; A IL-1 a, IL-1 b and NGF. The efficacy of such semi-Ig binding proteins for sepsis can be assessed in preclinical animal models known in the art (see Buras, JA et al., (2005) Nat. Rev. Drug Discov. 4(10): 854- 65; and Calandra, T. et al., (2000) Nat. Med. 6(2): 164-70). 7. Neurological disorders 7.1. Neurodegenerative diseases Chronic neurodegenerative diseases are generally age-related diseases characterized by progressive loss of neuronal function (neuronal cell death, demyelination), loss of mobility, and loss of memory. Emerging knowledge about the mechanisms underlying chronic neurodegenerative diseases such as Alzheimer's disease (AD) shows complex etiology and has been recognized for its many genesis and progress, for example, year 160877.doc -193· 201247704 Age, blood glucose status, amyloid production and multimerization, advanced glycation end product (AGE) (which binds to its receptor RAGE (the receptor for AGE)) increases brain oxidative stress, reduces cerebral blood flow, including Inflammation of inflammatory cytokines and chemokines, neuroinflammation, neuronal dysfunction, and microglial activation. Thus, such chronic neurodegenerative diseases represent complex interactions between multiple cell types and mediators. Therapeutic strategies for these diseases are limited and primarily use non-specific anti-inflammatory agents (e.g., corticosteroids, COX inhibitors) to block the inflammatory process or to use agents that prevent neuronal loss and/or synaptic function. These treatments cannot stop the progression of the disease. Studies have shown that more targeted therapies, such as antibodies against soluble AP peptides (including Af3 oligomeric forms), can not only help stop disease progression, but can also help maintain memory. These preliminary observations indicate that specific therapies targeting more than one disease mediator (eg, Aβ and pro-inflammatory cytokines (such as TNF)) provide therapeutic efficacy for chronic neurodegenerative diseases even more than targeting a single disease mechanism ( For example, the therapeutic effect observed by soluble Αβ alone is better. (See Nelson, R_B. (2〇〇5) Curr pharm — ιι: 3335 ; Klein. W. (2002) Neurochem. Int. 41: 345 ;

Janelsins, M.C., (2005) j. Neuroinflamm. 2: 23;

Soloman, B. (2004) Curr. Alzheimer Res. 1: 149; Klyubin, I. Specialist' (2005) Nat. Med. 11: 556-61; Bornemann, KD et al., (2001) Am. J. Pathol. 158: 63; Deane, R., et al., (2003) Nat. Med. 9·· 907-13; and Masliah, E. et al., (2005) Neuron. 46: 857) 半 semi-Ig binding protein of the present invention One or more targets involved in chronic neurodegenerative diseases such as Az 160877.doc 201247704 Helm's disease may be combined. Such targets include, but are not limited to, any soluble or cell surface mediator involved in the pathogenesis of AD, such as AGE (S100 A, amphotericin), pro-inflammatory cytokines (eg, IL-1), chemokines (eg MCP 1), molecules that inhibit nerve regeneration (eg Nogo, RGM A) and molecules that enhance neurite outgrowth (neurotrophin). The efficacy of a semi-Ig binding protein can be verified in preclinical animal models of transgenic mice such as overexpressing starch-like precursor proteins or RAGE and forming Alzheimer's disease-like symptoms. In addition, a half Ig, ''° 0 protein can be constructed and tested for its efficacy in animal models, and the best therapeutic semi-Ig binding protein can be selected for testing in human patients. Semi-W binding proteins can also be used to treat other neurodegenerative diseases such as Parkinson's disease. Alpha-Synuciein is involved in the pathology of Parkinson's disease. The half 1 § binding protein that can target ... synaptophysin and inflammatory mediators (such as TNF, 丨-丨, Mcp-i) may prove to be an effective therapy for Parkinson's disease and is encompassed by the present invention. 7.2 Neuronal regeneration and spinal cord injury Although the understanding of pathological mechanisms has increased, spinal cord injury (SCI) remains a destructive condition and represents a medical indication characterized by high medical needs. Most spinal cord injuries are contused or crushed and the primary injury is usually a secondary injury mechanism that exacerbates the initial injury and causes a significant enlargement (sometimes more than 10 times) of the lesion (inflammatory mediators such as cytokines and Chemokine). These primary and secondary mechanisms in SCI are very similar to those of brain damage caused by other means such as stroke. There is no satisfactory treatment, and high-dose rapid injections of methylprednisone (methylprednis() 1(10)e; 160877.doc-195-201247704 MP) are the only treatments that are used within a short period of 8 hours after injury. However, this treatment is only intended to prevent secondary damage without causing any significant functional recovery. It is severely criticized for its lack of definitive efficacy and its adverse effects (such as immunosuppression with subsequent infections and severe histopathological muscle changes). No other drugs, biologics or small molecules that stimulate endogenous regenerative potential have been approved, but promising therapeutic principles and drug candidates have shown efficacy in SCI animal models in recent years. To a large extent, the lack of recovery of human SCI function is caused by factors that inhibit neurite outgrowth at the lesion site, in scar tissue, in myelin, and on injury-associated cells. These factors are myelin-associated proteins NogoA, OMgp and MAG, RGM A, scar-related CSPG (chondroitin sulfate proteoglycan) and inhibitory factors on reactive astrocytes (some semaph〇rin and Ephemerin (ephHn). However, not only growth inhibitory molecules are found at the lesion site, but also neurite outgrowth stimulating factors (such as neurotrophins, laminin, L1 and other factors) are found. This combination of growth promoting molecules may explain that blocking a single factor such as NogoA or RGM A will cause significant functional recovery in the rodent SCI model, as reducing the inhibitory effect may shift the balance from growth inhibition to growth promotion. Incomplete recovery is observed when blocking a single extra-sexual growth inhibitory molecule. To achieve faster and more significant recovery, it may be necessary to block two neurite outgrowth inhibitory molecules (eg Nogo and RGM A), Or blocking neurite outgrowth inhibitory molecules and enhancing the function of neurite outgrowth enhancing molecules (eg, N〇g〇 and neurotrophins), or A neurite outgrowth inhibitory molecule (eg, N〇g〇) and a pro-inflammatory molecule (eg, TNF) (see McGee, AW et al. 160877. doc-196-201247704 (2003) Trends Neurosci. 26: 193; Domeniconi, Μ. et al., (2005) J. Neurol. Sci. 233: 43; Makwanal, M. et al., (2005) FEBS J. 272: 2628; Dickson, BJ (2002) Science 298: 1959; Yu, F. And Teng, H. et al., (2005) J. Neurosci. Res. 79: 273; Karnezis, T. et al., (2004) Nature Neurosci. 7: 736; Xu, G. et al., (2004) J. Neurochem. 91: 1018). In one aspect, a half Ig binding protein that binds to a single target or target pair is provided: such as NgR and RGM A; NogoA and RGM A; MAG and RGM A; OMGp and RGM A; RGM A and RGM B; CSPG and RGM A; aggrecan, midkine, neuroglycan, versican, phosphoprotein, Te3 8 and TNFa; and antibodies that promote dendritic and axon sprouting Combined Αβ globulomer-specific antibodies. Dendritic lesions are very early signs of AD, and NOGO 已知 is known to limit dendritic growth. One type of Αβ can be used. Any SCI- candidate (myelin - proteins) Ab combination. Other semi-Ig binding protein targets may include any combination of NgR-p75, NgR-Troy, NgR-Nogo66 (Nogo), NgR-Lingo, Lingo-Troy, Lingo-p75, MAG, and Omgp. In addition, the target may also include inhibition of any soluble or cell surface mediator involved in neurites, such as Nogo, Ompg, MAG, RGM A, brachialin, pterin, soluble Aβ, pro-inflammatory cytokines (eg IL- 1) Chemokines (such as MIP la) and molecules that inhibit nerve regeneration. The efficacy of anti-Nogo/anti-RGM(R) or similar semi-Ig binding proteins can be demonstrated in preclinical animal models of spinal cord injury. In addition, these semi-Ig binding proteins can be constructed and tested for their efficacy in animal models, and the optimal therapeutic semi-Ig binding protein can be selected for testing in human 160877.doc-197-201247704 patients. In addition, two different ligand binding sites on a single receptor (eg, a Nogo receptor that binds three ligands N〇g〇, Ompg, and MAG, and a RAGE that binds Ab and S 100 A) can be constructed. Semi-Ig binding protein. Furthermore, in immunological diseases such as multiple sclerosis, inhibitors of neurite outgrowth such as Nogo and Nogo receptors also play a role in preventing nerve regeneration. It has been shown in animal models of multiple sclerosis that inhibition of Nogo-Nog(R) receptor interaction enhances recovery. Thus, a semi-"binding protein that blocks the function of an immune mediator (such as a cytokine such as IL_12) and a neurite outgrowth inhibitor (such as Nogo or RGM) can provide greater than inhibition of immune or neurite outgrowth inhibition alone. Molecular faster and greater efficacy. 8. Tumor disease monoclonal antibody therapy has emerged as an important therapeutic modality for cancer (v〇n

Mehren et al. (2003) Annu. Rev. Med. 54: 343-69). Antibodies can exert anti-tumor effects by inducing apoptosis, redirecting to cytotoxicity, interfering with ligand-receptor interactions, or preventing protein expression critical to the neoplastic phenotype. In addition, antibodies can target components of the tumor microenvironment, disrupting important structures, such as the formation of tumor-associated vascular structures. Antibodies can also target ligands as receptors for growth factors, such as the epidermal growth factor receptor. The antibody thus inhibits the binding of the natural ligand that stimulates cell growth to the targeted tumor cell. Alternatively, antibodies can induce anti-idiotypic network, complement-mediated cytotoxicity or antibody-dependent cellular cytotoxicity (ADCC). The use of dual-specific antibodies that are dry to two separate tumor mediators provides additional benefits over monospecific therapy. However, monospecific therapy is still applicable in many situations. It is also expected that the following targets can be combined with either or both to treat tumors I60877.doc •198· 201247704 Ig of Ig: IGF1 and IGF2; IGF1/2 and HER-2; VEGFR and EGFR; CD20 and CD3; CD138 CD20; CD38 and CD20; CD38 and CD138; CD40 and CD20; CD138 and CD40; CD38 and CD40; CD-20 and CD-19; CD-20 and EGFR; CD-20 and CD-80; CD-20 and CD -22; CD-3 and HER-2; CD-3 and CD_19; EGFR and HER-2; EGFR and CD-3; EGFR and IGF1, 2; EGFR and IGF1R; EGFR and RON; EGFR and HGF; EGFR and c -MET; HER-2 and IGF1,2; HER-2 and IGF1R; RON and HGF; VEGF and EGFR; VEGF and HER-2; VEGF and CD-20; VEGF and IGF1,2; VEGF and DLL4; VEGF and HGF VEGF and RON; VEGF and NRP1; CD20 and CD3; VEGF and PLGF; DLL4 and PLGF; ErbB3 and EGFR; HGF and ErbB3; HER-2 and ErbB3; c-Met and ErbB3; HER-2 and PLGF; With HER-2; TNF with SOST. Other target combinations include one or more members of the EGF/erb-2/erb-3 family. Other target(s) to which the semi-Ig binding protein involved in a neoplastic disease may bind include, but are not limited to, a group selected from the group consisting of: CD52, CD20, CD19, CD3, CD4, CD8, BMP6, IL12A, ILIA, IL1B, IL2, IL24, INHA, TNF, TNFSF10, BMP6, EGF, FGF1, FGF10, FGF11, FGF12, FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF2, FGF20, FGF21, FGF22, FGF23, FGF3, FGF4, FGF5, FGF6, F0F7, FGF8, FGF9, GRP, IGF1, IGF2, IL12A, ILIA, IL1B, IL2, INHA, TGFA, TGFB1, TGFB2, TGFB3, VEGF, CDK2, 160877.doc -199· 201247704 FGF10, FGF18 , FGF2, FGF4, FGF7, IGF1R, IL2, BCL2, CD164, CDKN1A, CDKN1B, CDKN1C, CDKN2A, CDKN2B, CDKN2C, CDKN3, GNRH1, IGFBP6, ILIA, IL1B, ODZ1, PAWR, PLG, TGFB1I1, AR, BRCA1, CDK3 , CDK4, CDK5, CDK6, CDK7, CDK9, E2F1, EGFR, ENOl, ERBB2, ESR1, ESR2, IGFBP3, IGFBP6, IL2, INSL4, MYC, NOX5, NR6A1, PAP, PCNA, PRKCQ, PRK D1, PRL, TP53, FGF22, FGF23,

FGF9, IGFBP3, IL2, INHA, KLK6, TP53, CHGB, GNRH1, IGF1, IGF2, INHA, INSL3, INSL4, PRL, KLK6, SHBG, NR1D1, NR1H3, NR1I3, NR2F6, NR4A3, ESR1, ESR2, NR0B1, NR0B2, NR1D2, NR1H2, NR1H4, NR1I2, NR2C1, NR2C2, NR2E1, NR2E3, NR2F1, NR2F2, NR3C1, NR3C2, NR4A1, NR4A2, NR5A1, NR5A2, NR6A1, PGR, RARB, FGF1, FGF2, FGF6, KLK3, KRT1, APOC1 BRCA1

CHGA, CHGB, CLU, COL1A1, COL6A1, EGF, ERBB2, ERK8, FGF1, FGF10, FGF11, FGF13, FGF14, FGF16, FGF17, FGF18, FGF2, FGF20, FGF21, FGF22, FGF23, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, GNRH1, IGF1, IGF2, IGFBP3, IGFBP6, IL12A, ILIA, IL1B, IL2, IL24, INHA, INSL3, INSL4, KLK10, KLK12, KLK13, KLK14, KLK15, KLK3, KLK4, KLK5, KLK6, KLK9, MMP2, 160877.doc -200- 201247704

MMP9, MSMB, NTN4, ODZ1, PAP, PLAU, PRL, PSAP, SERPINA3, SHBG, TGFA, TIMP3, CD44, CDH1, CDH10, CDH19, CDH20, CDH7, CDH9, CDH1, CDH10, CDH13, CDH18, CDH19, CDH20, CDH7, CDH8, CDH9, R0B02, CD44, ILK, ITGA1, APC, CD164, COL6A1, MTSS1, PAP, TGFB1I1, AGR2, AIG1, AKAP1, AKAP2, CANT1, CAV1, CDH12, CLDN3, CLN3, CYB5, CYC1, DAB2IP, DES, DNCL1, ELAC2, EN02, EN03, FASN, FLJ12584, FLJ25530, GAGEB1, GAGEC1, GGT1, GSTP1, HIP1, HUMCYT2A, IL29, K6HF, KAI1, KRT2A, MIB1, PARTI, PATE, PCA3, PIAS2, PIK3CG 'PPID, PR1, PSCA, SLC2A2, SLC33A1, SLC43A1, STEAP, STEAP2, TPM1, TPM2, TRPC6, ANGPT1, ANGPT2, ANPEP, ECGF1, EREG, FGF1, FGF2, FIGF, FLT1, JAG1, KDR, LAMA5, NRP1, NRP2, PGF, PLXDC1, STAB1, VEGF, VEGFC, ANGPTL3, BAI1, COL4A3, IL8, LAMA5, NRP1, NRP2, STAB1, ANGPTL4, PECAM1, PF4, PROK2, SERPINF1, TNFAIP2, CCL11, CCL2, CXCL1 CXCL10, CXCL3, CXCL5, CXCL6, CXCL9, IFNA1, IFNB1, IFNG, IL1B, IL6, MDK, EDG1, EFNA1, EFNA3, EFNB2, EGF, EPHB4, FGFR3, HGF, IGF1, ITGB3, PDGFA, TEK, TGFA, TGFB1, TGFB2, 160877.doc -201 - 201247704 TGFBR1, (: CL2, CDH5, COL18A1, EDG1, ENG, ITGAV, ITGB3, THBS1, THBS2, BAD, BAG1, BCL2, CCNA1, CCNA2, CCND1, CCNE1, CCNE2, CDH1 (E -Cadherin), CDKNlB(p27Kipl) ' CDKN2A(pl6INK4a) ' COL6A1, CTNNBl (b-sodanes), CTSB (Cathepsin B), ERBB2 (Her-2), ESR1, ESR2, F3 (TF), FOSLl (FRA-l), GATA3, GSN (Plastin), IGFBP2, IL2RA, IL6, IL6R, IL6ST (glycoprotein 130), ITGA6 (a6 integrin), JUN, KLK5, KRT19, MAP2K7 (c-Jun) , MKI67 (Ki-67), NGFB (NGF), NGFR, NME1 (NM23A), PGR, PLAU (uPA), PTEN, SERPINB5 (mammostatin), SERPINEl (PAI-1), TGFA, THBS1 (platelet reaction Protein-1), TIE (Tie-1), TNFRSF6 (Fas), TNFSF6 (FasL), TOP2A (topoisomerase Iia), TP53, AZGP1 (word-a-glycoprotein), BPAG1 (net protein), CDKNl A (p21Wapl/ Cipl), CLDN7 (Claudin-7), CLU (Clotin), ERBB2 (Her-2), FGF1, FLRT1 (Fiber Binding Protein), GABRP (GABAa), GNAS1, ID2, ITGA6 (a6 Integrin), ITGB4 (b4 integrin), KLF5 (GC box BP), KRT19 (keratin 19), KRTHB6 (hair-specific type II keratin), MACMARCKS, MT3 (metallothionein-III), MUC1 (sticky) Protein), PTGS2 (COX-2), RAC2 (p21Rac2), S100A2, SCGB1D2 (lipophilic B), SCGB2A1 (mammon globin 2), SCGB2A2 (mammon globin 1), SPRRlB (Sprl), THBS1, THBS2 THBS4, and TNFAIP2 (B94), RON, c-Met, CD64, DLL4, PLGF, CTLA4, phospholipid lysine, R0B04, 160877.doc -202- 201247704 CD80, CD22, CD40, CD23, CD28, CD80, CD55 , CD38, CD70, CD74, CD30, CD138, CD56, CD33, CD2, CD137, DR4, DR5, RANKL, VEGFR2, PDGFR, VEGFR1, MTSP1, MSP, EPHB2, EPHA1, EPHA2, EpCAM, PGE2, NKG2D 'LPA, SIP , APRIL, BCMA, MAPG, FLT3, PDGFRa, PDGFRp, ROR1, PSMA, PSCA, SCD1 and CD59 » Other target combinations include specific targeting of tumors The cells and the immune effect are closely related to the tumor to initiate and/or enhance the three targets of the immune response to the tumor. In one embodiment, a binding protein of the invention binds to two different cell surface molecules present on CD3 and tumor heterologous cells (e.g., tumors having a mixture of cell types). In another embodiment, the binding protein of the invention binds to two different cell surface molecules present on an immune cell receptor (such as NKG2D or Fey receptor) and tumor heterologous cells (e.g., a tumor having a mixture of cell types). 9. Other diseases, conditions and conditions • Brain natriuretic peptide (BNP) has been implicated in cardiac function. Among other diseases, BNP semi-Ig binding proteins are potentially useful in the treatment of cardiovascular diseases, including various clinical diseases, disorders or conditions involving the heart, blood vessels or circulation. Such diseases, disorders or conditions may be caused by atherosclerotic lesions of the coronary arteries, cerebral arteries or peripheral arteries. The potentially treatable cardiovascular diseases include, but are not limited to, coronary artery disease, peripheral vascular disease, hypertension, Myocardial infarction, heart failure and similar diseases. HIV semi-Ig binding proteins are potentially useful for treating AIDS or AIDS symptoms. 160877.doc •203· 201247704 IL-18 has been identified as a marker for a variety of conditions or diseases, including but not limited to inflammatory conditions such as allergies and autoimmune diseases (Kawashima et al., (1997) J Educ. Inform· Rheumatol. 26(2): 77), Acute Kidney Injury (Parikh et al., (2005) J. Am. Soc. Nephrol. 16: 3046-3052; and Parikh et al., (2006) Kidney Int 'l. 70: 199-203), chronic kidney disease (such as when used as part of a panel assay), minimally pathological renal disease syndrome (MCNS) (Matsumoto et al., (2001) Nephron 88: 334-339 ), adult-onset Sterling's disease (Kawaguchi et al., (2001) Arthrit Rheum. 44(7): 1716-1717), adolescent atopic dermatitis (Hon et al., (2004) Ped. Derm. 21(6): 619-622), hemophagocytic lymphohistiocytosis (HLH) (Takeda et al., (1999) Brit. J. Haematol. 106(1): 182-189), adolescent idiopathic joints Inflammation (Lotito et al., (2007) J. Rheumatol. 34(4): 823-830), ovarian cancer (Le Page et al., (20060 Int'l J. Cancer 1 18: 1750-1758), systemic Lupus (Amerio et al. (2002) Clin. Exp. Rheum. 20(4): 535-538) and future cardiovascular events (Blankenberg et al., (2003) Circul. 108(20): 2453-2459) » Neutrophilated gelatinase-associated lipocalin (NGAL) is an early marker of acute kidney injury or disease. In addition to secretion from specific particles that activate human neutrophils, NGAL is also produced by renal cells in response to tubular epithelial damage. It is also a marker of tubulointerstitial (TI) injury. Due to ischemia or nephrotoxicity, NGAL levels in acute tubular necrosis (ATN) are elevated even after mild "subclinical" renal ischemia. In addition, it is known to be slow in 160877.doc -204 - 201247704 nephropathy (CKD) and acute kidney injury ((ΑΚΙ); see, for example, Devarajan et al. (2008) Amer. J. Kidn. Dis. 52(3): 395 -399 and Bolignano et al. (2008) Amer. J_ Kidn_ Dis. 52(3): 595-605) NGAL is manifested by the kidney. High urine NGAL levels have been suggested as precursors to progressive renal failure. It has previously been demonstrated that in animal and human models, NGAL is manifested by the renal tubules very early after ischemia or nephrotoxic injury. NGAL is rapidly secreted into the urine, and NGAL can be easily detected and measured in urine and precedes the appearance of any other known sputum or serum markers of ischemic injury. The protein is resistant to proteases, suggesting that it can be recovered in urine as a reliable marker of NGAL expression in the renal tubules. In addition, NGAL (eg, filtered from the blood) from outside the kidney does not appear in the urine, but is quantitatively absorbed by the proximal tubules. NGAL is also a diagnosis and/or prognosis for a variety of other diseases (see, for example, Xu et al., (2000) Biochim. et Biophys. Acta 1482: 298-307), signs of conditions and conditions, including inflammation, such as inflammation associated with infection. . In particular, it is a marker of the following conditions: large intestine urgency (see, for example, U.S. Patent Publication Nos. 2008/0166719 and 2008/0085524); renal dysfunction, nephropathy and kidney damage (see, for example, U.S. Patent Publication No. 2008/0090304) , 2008/0014644, 2008/0014604, 2007/0254370, and 2007/0037232); systemic inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, and multiple organ dysfunction syndrome (MODS) (see, for example, U.S. Patent Publication Nos. 2008/0050832 and 2007/0092911; see also U.S. Patent No. 6,136,526); periodontal disease (see, e.g., U.S. Patent No. 5,866,432); and venous thromboembolicity 160877 .doc - 205 - 201247704 Disease (see, for example, U.S. Patent Publication No. 2007/0269836). When in the free, non-complexed form, it is a marker for ovarian cancer, invasive and non-invasive breast cancer, and atypical ductal hyperplasia, which is a major risk factor for breast cancer (see, for example, US Patent Publication No. 2007/0196876) See also U.S. Patent Nos. 5,627,034 and 5,846,739 for the evaluation of the proliferative state of cancer. It is also a marker for colon cancer (Nielsen et al. (1996) Gut 38: 414-420), pancreatic cancer (Furutani et al., (1998) Cane. Lett. 122: 209-214) and esophageal cancer. It is also a marker of the condition associated with tissue remodeling when it is complexed with rivers < _9 (see, for example, U.S. Patent Nos. 7,432,066 and 7,153,660). High NGAL levels (eg, about 350 pg/L (Xu et al., (1995) Scand. J. Clin. Lab. Invest. 55: 125-13 1)) may also indicate bacterial infection (as opposed to viral infection) (see For example, U.S. Patent No. 7,056,702). In other diseases, IL-18 and NGAL semi-Ig binding proteins are potentially useful in the treatment of kidney disease, including any disease, condition or damage or injury to the kidney, including, for example, acute renal failure, acute renal disease, analgesic nephropathy, porridge Embolism nephropathy, chronic renal failure, chronic nephritis, congenital renal syndrome, end-stage renal disease, Gubus Dessert syndrome (G〇〇dpasture: - called, interstitial nephritis, kidney cancer, kidney damage, kidney infection, kidney Injury, kidney stones, lupus nephritis, membrane proliferative QN, membrane proliferative GN II, membranous nephropathy, minimally pathological nephropathy # disease, necrotic spheroid nephritis, f blastoma, renal deposition disease, Renal diabetes insipidus, nephropathy-IgA, nephropathy (kidney syndrome), polycystic kidney disease, post-Keyococcus infection (10), reflux nephropathy, renal artery examination 160877.doc 201247704 Plug, renal artery stenosis, kidney stagnation疋 τ τ 孔 硕 坏死 、 第 第 第 第 第 第 τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ . It is known that nerve growth factor (NGF) affects inflammatory pain and neuralgia, and anti-NGF therapy has been shown to improve both diseases. Therefore, in the disease, NGF can be used for household travel - %,. Treat sepsis, rheumatoid arthritis, bone inflammation and pain. Other factors shown in pain include, for example, TNF, :L-la, IL_lb, IL_6, CGRp, substance p, and Prostate 2 (PGE2). Thus, in one embodiment, the binding protein of the present invention is selected from the group consisting of =, c - rw - a combination of targets: IL_la, IL_n and NGF, IL-1 a, IL-1 b Pgt?1? . ττ 1 x ” PGE2, IL-Ia 'NGF and substance p IL_la, NGF and CGRP 〇 In addition, the NGF and IL-lb of the ancient master * 曰 also b between the performance of osteoarthritis pain Therefore, in a sinus sinus, in the example, the binding protein of the present invention binds to three groups selected from the group consisting of p. j Η铩. IL-la, jiang la, IL - lb and PGE 2. HI. Pharmaceutical Composition The present invention also provides a pharmaceutical composition comprising the binding protein of the present invention and a pharmaceutically acceptable carrier. The inclusion of taiji. A pharmaceutical composition for use in, but not limited to, diagnosis, detection, or monitoring of a condition; prevention (eg, inhibition or delay of a disease, condition, or its immortality, occupation, elderness, condition, treatment, management, or improvement) a condition or one or more symptoms thereof; and/or and/or research. In a particular embodiment, the composition comprises a In another embodiment, the pharmaceutical composition comprises - 3 or more of the binding proteins of the present invention and a binding protein other than the inventive protein of 160877.doc-207-201247704 One or more prophylactic or therapeutic agents for treating a condition. In one embodiment, a prophylactic or therapeutic agent is known to be or has been or is currently being used for prophylaxis (eg, inhibiting or delaying a disease, disorder, or other A prophylactic or therapeutic agent for the treatment, management or amelioration of a condition or one or more symptoms thereof. According to such embodiments, the composition may further comprise a carrier, diluent or excipient. The protein may be incorporated into a medicinal composition comprising a binding protein of the invention and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable" The carrier includes any and all solvent dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, which are physiologically compatible. Examples of pharmaceutically acceptable carriers include water, physiological saline, phosphate buffered physiological saline, sulphate dextrose, glycerol, ethanol, and the like: or evenings, and combinations thereof. In some embodiments, a special agent is included in the composition, such as 撼. - ρ ^ , , ,, such as mannitol, sorbitol; == pharmaceutically acceptable carrier can be further included A small amount of auxiliary: a diluent or emulsifier, preservative or buffer that enhances the shelf life or effectiveness of the antibody or antibody portion. It is known that various delivery lines are white or heart (four) - or a plurality of combinations of the present invention combined with egg delay # & ^ protein and for use in prevention (such as inhibition or or the onset of other conditions), management of 'treatment or improvement of the package can be packaged _ as = A combination of prophylactic or therapeutic agents. Delivery of binding protein microparticles, microcapsules; recombinant cells that can express slaves; receptor-mediated internal beverages for use in 160877.doc • 208-201247704 (see, for example, Wu and Wu (1987) J. Biol. Chem. 262: 4429-4432); and construction of nucleic acids, etc. as part of a retrovirus or other vector. Methods of administering a prophylactic or therapeutic agent of the invention include, but are not limited to, parenteral administration (eg, intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous), epidural administration, intratumoral administration And mucosal administration (eg intranasal and oral route). Further, the administration of the lung can be carried out, for example, by using an inhaler or a nebulizer and a formulation having an aerosol. See, for example, U.S. Patent No. 6,019,968; 5,985,320; 5,985,309; 5,934,272; 5,874,064; 5,855,913; 5,290,540; and 4,880,078; and PCT Publication No. WO 92/19244; No. 97/32572; WO 97/44013; WO 98/31346; and WO 99/66903. In one embodiment, the binding protein, combination therapy, or composition of the invention β of the invention is administered in a particular embodiment using Alkermes AIR® pulmonary music delivery technology (Alkermes, Inc., Cambridge, ΜΑ). The prophylactic or therapeutic agent of the present invention is administered intramuscularly, intravenously, intratumorally, orally, intranasally, pulmonaryly or subcutaneously. The prophylactic or therapeutic agent can be administered by any suitable route, for example by infusion or rapid injection, by absorption through the epithelial or cutaneous inner layer of the skin (eg, oral mucosa, rectal and intestinal mucosa, etc.) and can be combined with other bioactive agents. Give it together. The administration can be administered systemically or locally. In a particular embodiment, it may be desirable to topically administer a prophylactic or therapeutic agent of the invention to a region in need of treatment; this may be by, for example, but not limited to, local infusion, injection or by means of an implant (implant) It is achieved for porous or non-porous materials 'including membranes and matrices' such as sialastic membranes, 160877.doc • 209-201247704 poly. fibrous matrix (eg Tissuel®) or collagen matrix. & ^In the embodiment, one of the effective amounts of one or more of the gentlemen of the present invention is administered locally to the diseased area of the individual! &,, ° 〇 protein to prevent, treat, manage and / or improve the disease or, symptoms. In another embodiment, an effective amount of the drug-containing region of the individual is administered - or more - a binding protein of the invention and an effective amount - or a plurality of therapies other than the binding protein of the invention (eg, - or a plurality of prophylactic agents or treatments) A combination of agents to prevent, treat, and/or ameliorate a condition or one or more symptoms thereof. In another embodiment, the prophylactic or therapeutic agent can be delivered by controlled release or sustained release systems. In one embodiment, a pump can be used to achieve controlled release or sustained release (see Langer, supra; Sefton (1987) CRC Crit. Ref. Biomed. Eng. 14: 20; Buchwald et al., (1980)

Surgery 88: 507; Saudek et al., (1989) N. Engl. J. Med. 321: 574). In another embodiment, a polymeric material can be used to achieve controlled release or sustained release of the therapy of the invention (see, for example, Medical Applications of Controlled Release, Langer and Wise (ed.), CRC Pres., Boca Raton, Fla. (1974). Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (ed.), Wiley, New York (1984); Ranger and Peppas (1983) J., Macromol. Sci. Rev. Macromol. Chem. 23:61; See Levy et al., (1985) Science 228: 190; During et al, (1989) Ann. Neurol. 25: 351; Howard et al., (1989) J. Neurosurg. 71: 105); U.S. Patent No. 5,679,377; Nos. 5, 916, 597; 5, 912, 015; 5, 989, 463; and 5, 128, 326; and PCT Publication No. 160877.doc-210-201247704, No. WO 99/15154; WO 99/20253, "Continuous Release Formulation" Examples of polymers used include, but are not limited to, poly(2-hydroxyethyl methacrylate), poly(methyl methacrylate), poly(acrylic acid), poly(ethylene-co-vinyl acetate), poly (mercaptoacrylic acid), Glycolide (PLG), polyanhydride, poly(N-vinylpyrrolidone), poly(vinyl alcohol), polyacrylamide, poly(ethylene glycol), polylactide (PLA), poly(propylene) Ester-co-glycolide (PLGA) and polyorthoester. In one embodiment, the polymer used in the sustained release formulation is inert, free of leachable impurities, storage stable, sterile and biodegradable In another embodiment, the controlled release or sustained release system can be placed adjacent to the prophylactic or therapeutic target, thus requiring only one portion of the systemic dose (see, for example, Goodson, Medical Applications of Controlled Release, supra, Vol. 2, No. 115). Page - page 138 (1984)) The controlled release system is discussed in the review by Langer (1990) Science 249: 1527-1533). The preparation of one or more therapeutic agents of the invention can be prepared using any technique known to those skilled in the art. Continuous release of the formulation. See, e.g., U.S. Patent No. 4,526,938; PCT Publication No. WO 91/05548; WO 96/20698, Ning et al., (1996) Radiotherap. Oncol 39: 179-189; Song et al., (1995) PDA J. Pharma. Sci. Tech. 50:372-397; Cleek #,, (1997) Pro.Int, l.Symp. Control. Rel. Bioact· Matter. 24:853-854; and Lam et al., (1997) Proc. Int'l. Symp. Control Rel. Bioact. Matter. 24: 759-760 〇 In the embodiment of the present invention, the composition of the nucleic acid encoding the prophylactic or therapeutic agent is 160877.doc 211 · 201247704, By constructing the 醆 . . 4 4 4 4 4 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 之一 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且 且Directly left-handed or by using particle bombardment (eg, gene robbing; Bwiistic, Dup〇m) or; M-month enamel or cell surface receptor or transfection agent coated ' or by discuring its disc It is known that the homeobox-like peptide-linked peptides entering the nucleus (see, for example, Joliot et al. /] on n D, tracing, (199!) Proc. Nath % usa 88.1864 1868) It is administered such that it becomes administered intracellularly to the nucleic acid to promote the performance of the prophylactic or therapeutic agent it encodes. Alternatively, the nucleic acid can be introduced into a cell and incorporated into the host cell DNA for expression by homologous recombination. The pharmaceutical compositions of the present invention are formulated to be compatible with their intended route of administration. Examples of routes of administration include, but are not limited to, parenteral, such as intravenous, intradermal, subcutaneous, oral, intranasal (e.g., inhalation), transdermal (e.g., topical), transmucosal; and rectal administration. In a particular embodiment, the composition is formulated for intravenous, subcutaneous, intramuscular, oral, intranasal or local according to conventional procedures. It is also a pharmaceutical composition with humans. Typically, the composition for intravenous administration is a solution in sterile isotonic aqueous buffer. The composition may also include a solubilizing agent and a local anesthetic (such as Ugnocaine) to reduce the pain at the injection site, if desired. If the composition of the present invention is to be administered topically, the composition can be formulated as an ointment, a cream, a 'skin patch, a lotion, a gel, a shampoo, a spray, an aerosol, a solution, an emulsion. Forms or other forms familiar to those skilled in the art. See, for example, Remington's Pharmaceutical Sciences and

Introduction to Pharmaceutical Dosage Forms, 19th Edition 160877.doc • 212- 201247704

Mack Pub. Co., Easton, Pa. (1995). In one embodiment, for non-sprayable topical dosage forms, a carrier comprising one or more excipients compatible with topical application and having a dynamic viscosity greater than that of water to a semi-solid or solid form is used. Suitable formulations include, but are not limited to, solutions, suspensions, lotions, creams, ointments, powders, liniments, ointments and the like, if necessary sterilized or with additives which affect various properties such as osmotic pressure Mix (eg preservatives, stabilizers, wetting agents, buffers or salts). Other suitable topical dosage forms include sprayable aerosol formulations wherein, in one embodiment, the active ingredient is combined with a solid or liquid inert carrier with a pressurized volatile material (eg, a gas propellant such as Ion (freon) The mixture is packaged or encapsulated in a squeeze bottle. If necessary, a moisturizer or a moisturizer may be added to the pharmaceutical composition and the dosage form. Examples of such other ingredients are well known in the art. If the method of the invention comprises intranasal administration of the composition, the composition may be formulated in the form of an aerosol, spray, mist or drops. In particular, the prophylactic or therapeutic agent used according to the invention may be formulated with a propellant (for example, dichloroethane gas, trichlorofluorodecane, dioxotetrafluoroethane, carbon dioxide or other suitable gas) as an aerosol. The spray is presented in a form that is conveniently delivered from a pressurized pack or spray. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount of capsules and cartridges containing a powder mixture of the compound and a suitable powder base such as lactose or starch (for example by Gelatin is used for use in an inhaler or insufflator. If the method of the invention comprises oral administration, the composition can be formulated as an oral lozenge, capsule, cachet, gelcap, solution, suspension and 160877.doc • 213-201247704 form. Tablets or capsules may be prepared by conventional means using pharmaceutically acceptable excipients such as binders (for example, pregelatinized corn powder, polyethylene to ketone or ketone) a methylcellulose); a filler (such as lactose, microcrystalline cellulose or hydrogen peroxide); a lubricant (such as magnesium stearate, talc or vermiculite); a disintegrant (such as potato starch or glycolic acid starch) Sodium); or a wetting agent (such as sodium lauryl sulfate). Tablets can be coated by methods well known in the art. The liquid preparation for oral administration can be in the form of, but not limited to, a solution, syrup or suspension, or it can be provided as a dry product which is reconstituted with water or other suitable vehicle before use. Such liquid preparations can be prepared by conventional means using pharmaceutically acceptable additives such as such additives such as suspending agents (for example, sorbitol glycoside, cellulose derivatives or hydrogenated edible fats); emulsifiers (eg eggs) "Dish grease or gum arabic" 'non-aqueous vehicle (eg almond oil, oily ester, ethanol or vegetable oil); and preservatives (for example methyl p-hydroxybenzoate or propyl p-hydroxybenzoate or sorbic acid). The formulation may also contain buffer salts, flavoring agents, coloring agents, and sweetening agents, as appropriate. Formulations for oral administration may be suitably formulated for slow release, controlled release or sustained release prophylactic or therapeutic agents. The method of the invention may comprise administering a composition formulated with an aerosolizing agent, for example, by using an inhaler or a nebulizer. See, for example, U.S. Patent No. 6,019,968; U.S. Patent No. 5,985,320; U.S. Patent No. 5,985,309; 5,934,272; 5,874,064; 5,855,913; 5,290,540; and 4,880,078; and PCT Publication No. WO 92/19244; No. 97/32572; WO 97/44013; WO 98/3 1346; and WO 99/66903. In a specific embodiment, the binding proteins, combination therapies, and/or compositions of the invention of the invention are administered using Alkermes 160877.doc -214.201247704 AIR® Lung Drug Delivery Technology (Alkemes, Inc, cambridge, ma). The S method of the present invention may comprise administering a composition formulated for parenteral administration by injection (example &# by rapid injection or continuous infusion). Formulations for injection can be provided in unit dosage form with a preservative (for example, in ampoules or in multi-dose containers). The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain such compositions as suspending agents, stabilizing agents and/or dispersing agents. Alternatively, the active ingredient may be in the form of a powder which is suitable for reconstitution (e.g., sterile (4) water) prior to use. The method of the present invention may additionally comprise a composition of a genital formulation formulated as a depot preparation. Such long acting formulations may be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the composition may be formulated with a suitable dispersing substance (for example, an emulsion formulated as an acceptable oil) or with an ion exchange resin; or formulated as a sparingly soluble derivative (for example, formulated as a sparingly soluble salt) .

The method of the invention encompasses the administration of a composition formulated in a neutral or salt form. Pharmaceutically acceptable salts include salts formed with anions such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, and tartaric acid; and salts formed with cations, which are derived from sodium hydroxide, potassium hydroxide, Salts of ammonium oxyhydroxide, hydrogen hydroxide, iron hydroxide, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, and procaine. In general, the ingredients of the composition are supplied separately or mixed together in unit dosage form, e.g., in the form of a dry, ortho-dried powder or anhydrous concentrate, in a sealed cereal such as an ampoule or sachet indicating the amount of active agent. When the mode of administration is 160877.doc -215- 201247704, the drug can be dispensed with aseptic pharmaceutical grade water or saline. "The drug mode is injection" can provide sterile water for injection or saline. Ampoule so that the ingredients can be mixed before administration.

Further, the present invention also provides a prophylactic or therapeutic agent or a pharmaceutical composition of the present invention which is encapsulated in a sealed container (such as an ampoule or a capsule) in an amount indicative of the pharmaceutical agent. In one embodiment, the prophylactic or therapeutic agent or the pharmaceutical composition of the present invention is provided in a sealed container in the form of (d) no (iv) dry powder or anhydrous concentrate and is reversible (eg, with water or saline). Appropriate concentrations are available for administration to individuals. In one embodiment, one or more of the prophylactic or therapeutic agent or the pharmaceutical composition of the invention is in a dry sterile lyophilized form at least 5 mg, at least 10 mg, at least 15 mg, at least

A unit dose of at least 35 mg, at least 45 mg, at least 50 mg, at least 75 mg, or at least 100 mg of mg is provided in a sealed container. The lyophilized prophylactic or therapeutic agent or the pharmaceutical composition of the present invention should be stored in an initial container between 8 c and the prophylactic or therapeutic agent or the pharmaceutical composition of the present invention should be within 1 week after recovery, for example 5 Within days, within 72 hours, within 48 hours, within 24 hours, within 12 hours, within 6 hours, within 5 hours, within 3 hours, or within hours of work. In an alternative embodiment, one or more of the prophylactic or therapeutic agent or the inventors of the present invention are provided in liquid form in a sealed container indicating the amount and concentration of the agent, The liquid form of administration of I and the compound is at least 2525 mg/ml, at least 0.5 mg/ml, at least 1 mg/ml, at least 2.5 mg/nU, at least 5 mg/ml, at least 8 mg/mi, A concentration of at least 1 mg/ml, at least 15 mg/kg, at least 25 mg/ml, at least 50 mg/ml, to 75 mg/ml or at least 100 mg/ml is provided in a sealed container. 160877.doc • 216 · 201247704 The liquid form should be stored between 2 ° C and 8. 在 in its original container. The binding protein of the invention can be incorporated into a pharmaceutical composition suitable for parenteral administration. In a consistent application, The protein or binding protein portion will be prepared as an injectable solution containing 0.1-250 mg/ml binding protein. The injectable solution may be comprised of a liquid or lyophilized dosage form in a vermiculite or amber vial, ampule or pre-filled syringe. L-Histamine (ΐ·5〇mM), optimal 5-10 mM, pH 5.0 to 7.0 (optimally pH 6,0). Agents include, but are not limited to, sodium succinate, sodium citrate, sodium phosphate or potassium phosphate. The solution can be adjusted using sodium chloride at a concentration of 0-300 mM (15 mM optimal for liquid dosage forms). Toxicity. For lyophilized formulations, cryoprotectants may be included 'mainly 〇-1〇% sucrose (best 〇·5%_1〇%^ other suitable cryoprotectants including trehalose and lactose. For lyophilized formulations) It may include a bulking agent, mainly mannitol (optimally 2% _4%). Stabilizers can be used in both liquid and lyophilized formulations, mainly S50 mM L-methionine (best 5 - 10 mM). Other suitable bulking agents include glycine and arginine (any of which can be included in a concentration of 0-0.05%), and polysorbate _ 8〇 (best 〇.〇〇5 %-〇.〇1% concentration is included. Other surfactants include, but are not limited to, polysorbate 2〇 and Brjj® surfactants. Prepared for injectable solutions for parenteral administration. A pharmaceutical composition comprising a binding protein of the invention may further comprise an agent suitable for use as an adjuvant, such as for increasing a therapeutic protein (eg, An agent that absorbs or disperses proteins. Particularly suitable adjuvants are hyaluronidases such as Hylenex® (recombinant human hyaluronics & L per). Addition of hyaluronan to injectable solutions after modified parenteral administration 'especially Human bioavailability after subcutaneous administration. It also allows for higher injection volume (ie greater than 1 ml) and less pain and discomfort, as well as the lowest incidence of injection site reactions (see PCT disclosure). Case No. 4〇4/〇7814〇 and US Patent Publication No. 2006/104968). The compositions of the invention may take a wide variety of forms. Such forms include, for example, liquid, semi-solid, and solid dosage forms such as liquid solutions (e.g., injectable solutions and infusible solutions), dispersions or suspensions, lozenges, pills, powders, liposomes, and suppositories. The form chosen will depend on the intended mode of administration and therapeutic application. A typical composition is in the form of an injectable or infusible solution, such as a composition similar to that used for passive immunization of humans with other binding proteins (e.g., antibodies). The mode of administration selected is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular;). In one embodiment, the binding protein is administered by intravenous infusion or injection. In another embodiment the binding protein is administered by intramuscular or subcutaneous injection. Therapeutic compositions are generally sterile and stable under the conditions of manufacture and storage. Group. The materials may be formulated as solutions, microemulsions, dispersions, liposomes or other ordered structures suitable for high drug concentrations. Sterile injectable solutions can be prepared by incorporating the active compound (i.e., antibody or antibody portion) in a desired amount, together with one of the ingredients listed herein, or such a guillotine, into a suitable solvent, followed by filtration sterilization as desired. In general, dispersions are prepared by incorporating the active compound into a liquid vehicle containing a basic carrier medium and other desired ingredients from the ingredients enumerated herein. In the case of a sterile, toluene powder for the preparation of a sterile, main spray solution, the preparation is vacuum drying and spray drying to produce a live!·Production plus any other desired ingredients from its prior sterile filtration solution. The powder can be maintained, for example, by using a material such as (iv) fat, 160877.doc 201247704, by maintaining the desired particle size (in the case of dispersions), and by using a surfactant to maintain proper fluidity of the solution. Prolonged absorption of the injectable compositions can be brought about by the inclusion of agents which delay absorption, such as monostearate and gelatin, in the compositions. The conjugated proteins of the invention can be administered by a variety of methods known in the art, but in one embodiment, for many therapeutic applications, the route/mode of administration is subcutaneous, intravenous or infusion. As will be appreciated by those skilled in the art, the route and/or mode of administration will vary depending on the desired result. In certain embodiments, active sigma, such as controlled release formulations, including implants, transdermal patches, and microencapsulated delivery systems, can be prepared with carriers that prevent rapid release of the compound. Biodegradable, biocompatible polymers such as ethyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Many of the methods for preparing such formulations are patented or generally known to those skilled in the art. See for example

Sustained and Controlled Release Drug Delivery Systems, edited by J.R. Robinson, Marcel Dekker, Inc, New Y〇rk, i978. In certain embodiments, a binding protein of the invention can be administered orally, for example, with an inert carrier 3 and a deteriorating edible carrier. The compound, if necessary in combination with other ingredients, may also be enclosed in a hard or soft shell gelatin capsule, compressed into a lozenge or incorporated directly into the individual's diet. For oral therapeutic administration, the compounds may be combined with excipients and used in the form of ingestible lozenges, buccal tablets, dragees, capsules, elixirs, suspensions, syrups, wafers and Similar form. In order to administer a compound of this month by means other than parenteral administration, it may be necessary to coat the compound with a material that prevents its inactivation, or to co-inject the compound with a material that prevents its inactivation. versus. Supplementary active compounds can also be incorporated into the compositions. In certain embodiments, a binding protein of the invention is co-administered and/or co-administered with one or more additional therapeutic agents suitable for treating a condition with a binding protein of the invention. For example, a binding protein of the invention can be co-administered and/or co-administered with one or more other antibodies that bind to other targets (e.g., antibodies that bind to other cytokines or bind to cell surface molecules). In addition, one or more of the binding proteins of the invention may be used in combination with two or more of the above therapeutic agents. 'The combination therapy is preferably administered with a lower dose of the therapeutic agent, thereby avoiding possible toxicity associated with various monotherapies or complication. In certain embodiments, the binding protein is linked to a half-life extending agent known in the art. Such vehicles include, but are not limited to, domains, polyethylene glycols, and dextran. Such vehicles are described, for example, in U.S. Patent No. 6,66, 843, issued to PCT Publication No. WO 99/25044. In a particular embodiment, a nucleic acid sequence encoding a binding protein of the invention or another prophylactic or therapeutic agent of the invention is administered to treat, prevent, manage or ameliorate a condition or one or more symptoms thereof by means of gene therapy. Gene therapy refers to a therapy performed by administering to a subject an already expressed or expressible nucleic acid. In this embodiment of the invention, the nucleic acid produces a binding protein or prophylactic or therapeutic agent of the invention encoded by it that mediates a prophylactic or therapeutic effect. Any method for gene therapy that can be utilized in the art can be used in accordance with the present invention. For a general review of the methods of gene therapy, see

Goldspiel et al., (1993) Clin. Pharm. 12: 488-505; Wu and Wu (1991) Biotherapy 3: 87-95; Tolstoshev (1993) Ann. Rev 160877.doc • 220· 201247704

Pharmacol. Toxicol. 32:573-596; Mulligan (1993) Science 260:926-932; and Morgan and Anderson (1993) Ann. Rev. Biochem. 62:191-217; May (1993) TIBTECH 11(5) :155-215. Methods commonly known in the art for recombinant DNA techniques that can be used are described in Ausubel et al. (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); and Kriegler, Gene.

Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990). A detailed description of various methods of gene therapy is disclosed in U.S. Patent Publication No. 20090297514. The binding proteins of the present invention are useful for treating various diseases which are harmful to the targets recognized by such binding proteins. Such diseases include, but are not limited to, rheumatoid arthritis, osteoarthritis, adolescent chronic arthritis, septic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic Lupus erythematosus, Crohn's disease, ulcerative colitis, inflammatory bowel disease, insulin-dependent diabetes mellitus, verrucous, asthma, allergic disease, psoriasis, inflammatory scleroderma, graft versus host disease , organ transplant rejection, acute or chronic immune disease associated with organ transplantation, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki's disease, Gracie's disease, renal syndrome, chronic fatigue syndrome, Wegner Granulomatosis, Henry-Siegen Lai purpura, renal microscopic vasculitis, spastic active hepatitis, (4) membranous inflammation, septic shock, toxic shock: syndrome, depleted syndrome, cachexia m disease, Parasitic diseases, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington's disease, Parkinson's disease, _axis disease n original bile liver 160877.doc -221· 201247704 Sclerosis, hemolytic anemia, malignant disease, heart failure, myocardial infarction, Addison's disease, sporadic type I polygland secretion deficiency and π-type polygland secretion deficiency, Schmidt's syndrome Adult (acute) respiratory distress syndrome, alopecia, plaque alopecia, seronegative joint disease, joint disease, Lyttle's disease, psoriasis arthropathy, ulcerative colitis, joint disease, enteric synovitis, and cloak Bacterial, Yarrowia and Salmonella-associated joint diseases, spondyloarthropathy, atherosclerosis/arteriosclerosis, atopic allergy, autoimmune bullous disease, pemphigus vulgaris, peony pemphigus, pemphigus , linear IgA disease, autoimmune hemolytic anemia, Comm's positive hemolytic anemia, acquired pernicious anemia, adolescent pernicious anemia, myalgia encephalitis / royal free disease, chronic skin mucosal candidiasis, giant cell arteritis , primary sclerosing hepatitis, unexplained autoimmune hepatitis, acquired immunodeficiency syndrome, acquired immunodeficiency-related diseases, hepatitis B, hepatitis C, general Variant immunodeficiency (common variant hypogammaglobulinemia), dilated cardiomyopathy, female infertility, ovarian failure, premature ovarian failure, fibrotic lung disease, unexplained fibrotic alveolitis, post-inflammatory interstitial Pulmonary lung disease, interstitial pneumonia, connective tissue disease-associated interstitial lung disease, mixed, 'dung tissue disease-related lung disease, systemic sclerosis-related interstitial lung disease, rheumatoid arthritis-related interstitial Lung disease, systemic lupus erythematosus-associated lung disease, dermatomyositis/polymyositis-associated lung disease, Hugh's disease-related lung disease, ankylosing spondylitis-related lung disease, vasculitis, diffuse lung disease, 3 hemacysis Related lung diseases, drug-induced interstitial lung disease, fibrosis, radiation fibrosis, obstructive bronchiolitis, chronic eosinophilic pneumonia, lymphocytic infiltrating lung disease, post-infection interstitial lung 160877.doc 201247704 disease, Gouty arthritis, autoimmune hepatitis, sputum autoimmune hepatitis (typical autoimmune or lupus-like hepatitis), type 2 autoimmune liver (anti-LKM antibody hepatitis), autoimmune-mediated hypoglycemia, type B insulin resistance with acanthosis nigricans, parathyroid hypoenergy, acute immune diseases associated with organ transplantation, chronic immune diseases associated with organ transplantation, Osteoarthrosis, primary sclerosing cholangitis, spastic psoriasis, type 2 psoriasis, idiopathic leukopenia, autoimmune neutropenia, nephropathy, spheroid nephritis, renal microscopic Vasculitis, Lyme disease, discoid red • Lupus erythematosus, idiopathic *N0S male infertility, sperm autoimmune, multiple sclerosis (all subtypes), sympathetic ophthalmia, connective tissue disease secondary Pulmonary hypertension, Gubus Deer syndrome, pulmonary manifestations of nodular polyarteritis, acute rheumatic fever, rheumatoid spondylitis, Still's disease, systemic sclerosis, Hugh's syndrome, high Inn's disease / Arteritis, autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmune verrucous disease, hyperthyroidism, goiter, autoimmune verrucous dysfunction (Hashimoto ), atrophic autoimmune thyroid hypoenergy, pre-lumilous mucinous edema, lens-like uveitis, primary vasculitis, leukoplakia acute liver disease, chronic liver disease, alcoholic cirrhosis, alcohol-induced liver injury , bile stasis, characteristic liver disease, drug-induced hepatitis, nonalcoholic steatosis hepatitis, allergies and asthma, group B streptococcus (GBS) infection, mental disorders (such as depression and schizophrenia), Th2 and Sputum-mediated diseases, acute and chronic pain (different forms of pain), and cancers such as lung cancer, breast cancer, stomach cancer, bladder cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer, and rectal cancer, and hematopoietic system malignancy Disease (leukemia and leaching 160877.doc -223- 201247704 barnoma), no beta lipoproteinemia, hand and foot cyanosis, acute and chronic parasitic or infection process, acute leukemia, acute lymphoblastic leukemia (ALL), acute bone marrow leukemia (AML), acute or chronic bacterial infection, acute pancreatitis, acute renal failure 'adenocarcinoma, atrial ectopic beat, AIDS dementia complex, wine Induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergic rhinitis, allograft rejection, α-^antitrypsin deficiency, amyotrophic lateral sclerosis, anemia, angina pectoris, anterior horn cell degeneration, Anti-cd3 therapy, antiphospholipid syndrome, anti-receptor allergic reaction, aortic and peripheral aneurysms, aortic dissection, arterial hypertension, atherosclerosis, arteriovenous fistula, ataxia, atrial fibrillation (continuous or Paroxysmal), atrial flutter, atrioventricular block, B-cell lymphoma, bone graft rejection, bone marrow transplantation (BMT) rejection, bundle branch block, Burkitt's lymphoma, burn, arrhythmia , cardiac suppressor syndrome, cardiac tumor, cardiomyopathy, cardiopulmonary bypass inflammatory response, cartilage transplant rejection, cerebellar cortical degeneration, cerebellar disorder, turbulent or multifocal atrial tachycardia, chemotherapy-related disorders, chronic myeloid cells Leukemia (CML), chronic alcoholism, chronic inflammatory disease, chronic lymphocytic leukemia (CLL), chronic obstructive pulmonary disease Disease (c〇PD), chronic salicylic acid poisoning, colorectal cancer, congestive heart failure, conjunctivitis, contact dermatitis, pulmonary heart disease, coronary artery disease, Cui's disease, culture-negative sepsis, cystic disease Fibrosis, cytokine therapy-related disorders, boxer dementia, demyelinating disease, hemorrhagic dengue, dermatitis, dermatological conditions, diabetes, diabetic arteriosclerosis, diffuse Lewy body disease, dilated congestive myocardium Disease, basal ganglia disease, middle-aged Down's syndrome 160877.doc -224· 201247704 candidate, drug-induced dyskinesia induced by drugs that block CNS dopamine receptors, drug sensitivity, eczema, encephalomyelitis, heart Endometritis, endocrine disease, epiglottis, EB virus infection, acromegaly, extrapyramidal and cerebellar disorders, sputum hemolymphatic histiocytoma, fetal thymus transplant rejection, Friedreich Arrhythmia, functional peripheral arterial disease, fungal sepsis, gas gangrene, gastric ulcer, nephron nephritis, graft rejection of any organ or tissue Gram-negative sepsis, Gram-positive sepsis, granuloma caused by intracellular organisms, hairy cell leukemia, Harlowden-Spartz's disease, Hashimoto's disease, hay fever, heart transplant rejection , hemochromatosis, hemodialysis, hemolytic uremic syndrome/thrombotic thrombocytopenic purpura, hemorrhage, hepatitis (A), His bundle of arrhythmia, HIV infection/HIV neuropathy, Hodgkin's disease, hyperactive motor disorder , allergic reactions, allergic pneumonia, high pressure, slow movement

Dynamic condition, hypothalamic-pituitary-adrenal axis assessment, idiopathic Addison's disease, idiopathic pulmonary fibrosis, antibody-mediated cytotoxicity, debilitation, infant spinal muscular atrophy, aortic inflammation, type A Influenza, ionizing light exposure, iridocyclitis/uvitis/opic neuritis, ischemia-reperfusion injury: ischemic stroke, juvenile rheumatoid arthritis, adolescent spinal muscular atrophy, Kaposi's sarcoma 'Kidney transplant rejection, Legionnaires' disease, Leishman's disease, leprosy, corticospinal disorders, fat edema, liver transplant rejection, lymphedema, malaria, malignant lymphoma, subpopular apoptotic, malignant melanin Tumor, meningitis, meningococcal disease: syndrome, metabolic/idiopathic disease, migraine, mitochondrial multiple "disease, mixed connective tissue disease, gamma globulin disease, multiple bone tumor, multiple I60877. Doc .225- 201247704 System degradation (also cut, Dynay_Thomas, Spreg and Machado 'Joseph) myasthenia gravis, mycobacterial avian mycobacteria, Mycobacterium tuberculosis, bone marrow development Adverse syndrome, myocardial infarction, myocardial ischemic disease, nasopharyngeal carcinoma, neonatal chronic lung disease, tendinitis, nephropathy, neurodegenerative diseases, neuromuscular atrophy, neutrophilic fever, non-Hodgkin's lymph Tumor, abdominal aorta and its branch blockage, obstructive arterial disease, 〇ω therapy, Pillillary / epididymitis, testicular vas deferens, organ enlargement, osteoporosis, ligature rejection, sputum cancer , paraneoplastic syndrome / malignant hypercalcemia, parathyroid transplant rejection, pelvic inflammatory disease, perennial rhinitis, pericardial disease, peripheral atherosclerotic disease, peripheral vascular disease, peritonitis, pernicious anemia, Pneumocystis carinii Pneumonia pneumonia, POEMS syndrome (polyneuropathy, organ enlargement, endocrine disease, gamma globulin disease and skin variability syndrome), post-perfusion syndrome, post-pump syndrome, post-cardiac syndrome, pre-eclampsia, progressive Nuclear paralysis, primary pulmonary hypertension 'radiotherapy, Raynaud's phenomenon and disease, Raynaud's disease, Lefsum's disease, regular narrow QRS, e tachycardia, renal vascular blood pressure, reperfusion injury, restrictive cardiomyopathy, sarcoma, scleroderma, senile chorea, Lewy body dementia, seronegative joint disease, shock, sickle cell anemia Skin allograft rejection 'skin change syndrome, small bowel transplant rejection, solid tumor, specific arrhythmia, spinal ataxia, spinocerebellar degeneration, streptococcal myositis, cerebellar structural lesions, subacute sclerosing panencephalitis , fainting, cardiovascular system syphilis, systemic allergy 'systemic inflammatory response syndrome, systemic adolescent rheumatoid arthritis, T-cell or FAB ALL, telangiectasia, 160877.doc •226· 201247704 thromboangiitis obliterans, Thrombocytopenia, poisoning, transplantation, trauma/bleeding, type III allergic reaction, type Iv allergy, unstable angina, uremia, urinary septicemia, urticaria, valvular heart disease, varicose veins, vasculitis, venous disease, Venous thrombosis, ventricular fibrillation, viral and fungal infections, viral encephalitis/aseptic meningitis , virus-associated hemophagocytic syndrome, Wernicke-Korsakov syndrome, Wilson's disease, and xenograft rejection of any organ or tissue (see PCT Publication No. WO 2002/097048; WO 95/24918) ; and W0 • 00/56772). The binding protein of the present invention can be used for treating humans suffering from autoimmune diseases, especially related to inflammation, including rheumatoid arthritis, spondylitis, allergy, autoimmune diabetes and autologous Immunological uveitis. In one embodiment, a binding protein of the invention or antigen binding portion thereof is for use in the treatment of rheumatoid arthritis, Crohn's disease, multiple sclerosis, insulin dependent diabetes, and psoriasis. In one embodiment, the diseases or treatments that can be treated or diagnosed by the compositions and methods of the invention include, but are not limited to, primary and metastatic cancers, including breast cancer, colon cancer, rectal cancer, lung cancer, oropharyngeal cancer, Pharyngeal cancer, esophageal cancer, gastric cancer, pancreatic cancer, liver cancer, gallbladder cancer and cholangiocarcinoma, small bowel cancer, urinary tract cancer (including kidney cancer, bladder cancer and urothelial cancer), female genital cancer (including sub-S cancer, child Lung cancer and India's nest cancer, as well as choriocarcinoma and genital tract trophoblastic disease), male genital tract cancer (including prostate cancer, seminal vesicle cancer, testicular cancer and germ cell tumor), endocrine adenocarcinoma (including thyroid cancer, adenocarcinoma and Pituitary adenocarcinoma) and skin cancer, as well as hemangioma 'melanoma, sarcoma (including bone 160877.doc -227· 201247704 and sarcoma produced by soft tissue and Kaposi's sarcoma), brain tumor, neurological tumor, eye tumor and meninges Tumors (including astrocytoma, glioma, glioblastoma, retinoblastoma, neuroma, neuroblastoma, schwannomas, meningioma), by such as leukemia Solid malignant tumor disease caused by blood 'and lymphoma (Hodgkin and non-Hodgkin lymphoma). In one embodiment, a binding protein of the invention, or an antigen binding portion thereof, can be used to treat cancer or inhibit tumor metastasis as described herein when used alone or in combination with radiation therapy and/or other chemotherapeutic agents. The binding protein of the present invention or antigen-binding portion thereof may be combined with agents including, but not limited to, anti-neoplastic agents, radiation therapy, chemotherapeutic agents, such as DNA priming agents, cispiatin, and card bots. (carboplatin), anti-tubulin, paclitaxel, docetaxel, paclitaxel, doxorubicin, gemcitabine, gemzar, anthracycline, ade Adriamycin, topoisomerase! Inhibitors, topoisomerase 11 inhibitors, 5-fluorouracil (5-FU), leucovorin, irinotecan, and receptor tyrosine kinase inhibitors (eg, Ero Erlotinib, gefitinib, C〇X-2 inhibitor (eg, cereac〇xib), kinase inhibitor, and siRNA. The binding proteins of the invention may also be administered with one or more other therapeutic agents suitable for the treatment of various diseases. The binding proteins of the present invention may be used alone or in combination to treat such diseases. 160877.doc 201247704::, the broad binding protein may be used alone or in combination with other agents (eg, ', η' and '' It is selected by those who are familiar with the technology. 蛊You 丨& ^ ^, 八用w~士举^ " 'This other drug is recognized in the industry as appropriate / alpha treatment of the disease or condition treated by the antibody of the present invention The therapeutic agent may also be an agent that imparts a beneficial property to the therapeutic composition, such as a drug viscosity of the composition.

It is to be understood that the combinations to be included in the present invention are combinations thereof suitable for their intended purpose. The agents described below are illustrative and are not intended to be limiting. The combination of the parts of the present invention may be a binding protein of the present invention and at least one other agent selected from the list below. If the combination is such that the formed group = object can perform the (four) function, the secret ingredient includes more than one other agent, such as two or three other agents. The combination of autoimmune and inflammatory diseases is a non-steroidal anti-inflammatory drug (also known as NSAIDS) which includes a drug such as ibuprofen. Other combinations are corticoids, including prednisolone; the well-known side effects of steroid use can be reduced or even eliminated by gradually reducing the amount of steroid required to treat a patient in combination with the semi-Ig of the present invention. Non-limiting examples of therapeutic agents for rheumatoid arthritis that may be combined with or in combination with the present invention include the following: cytokine inhibitory anti-inflammatory drugs (CSAID); antibodies against other human cytokines or growth factors Or an antagonist, such other human cytokines or growth factors are, for example, 1^1?, 1/1, ratio_1, 11^2, _3 > IL-4 ^ IL-5 ^ IL-6 > IL-7 > IL-8 ^ IL-15 > IL-16 ^ IL-18, IL-21, IL-23, interferon, EMAp_n, GM CSF, FGF and PDGF. The binding protein of the present invention or antigen-binding portion thereof can be combined with an antibody against a cell surface molecule of fine I60877.doc-229-201247704, such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD80 (B7.1), CD86 (B7.2), CD90 and CTLA or their ligands, including CD154 (gp39 or CD40L). Combinations of therapeutic agents can interfere with autoimmune and subsequent inflammatory cascades at different points; examples include TNF antagonists (eg, chimeric, humanized or human TNF antibodies), adalimumab (ADALIMUMAB) (PCT Publication No. WO No. 97/29131), CA2 (RemicadeTM), CDP 571 and soluble p55 or p75 TNF receptor, its derivatives (p75TNFRlgG (EnbrelTM) or p55TNFRlgG (anazepa); and TNFa invertase (TACE) inhibitor; similar In fact, IL-1 inhibitors (interleukin-1 converting enzyme inhibitors, IL-1RA, etc.) may be effective for the same reason. Other combinations include interleukin 11. Another combination includes function with IL-12 A key player in autoimmune response that acts in parallel, depends on IL-12 function or synergizes with IL-12 function; especially IL-18 antagonists, including IL-18 antibodies, soluble IL-18 receptors And IL-18 binding proteins. IL-12 and IL-18 have been shown to have overlapping but different functions, and combinations of antagonists of both may be most effective. Another combination is a non-consumptive anti-CD4 inhibitor. Other combinations include Antagonism of the co-stimulatory pathway CD80 (B7.1) or CD86 (B7.2) Including antibodies, soluble receptors, and antagonistic ligands. The binding proteins of the invention may also be combined with agents such as amidoxime, 6-MP, sulfur, sputum, sulphate, and bite (bite) Sulphasalazine), mesalazine, olsalazine chloroquinine/base sputum, pencillamine, thiomalate 160877.doc -230- 201247704 (aurothiomalate) (intramuscular) And oral), sulfur, sitting, colchicine (cochicine), corticosteroids (oral, inhalation and topical injection), beta-2 adrenergic receptor agonist (salbutamol, terbutaline) (terbutaline), salmeteral, scutellaria (theophylline, aminophylline), cromoglycate, nedocromil, ketotifen , ipratropium and oxitropium, cyclosporine, FK5 06, rapamycin, mycophenolate mofetil, leflunomide, NSAID (eg ibuprofen), cortex Steroids (such as pour nylon), phosphodiesterases Inhibitors, adenosine agonists, anti-suppressants, complement inhibitors, adrenergic agents, agents that interfere with pro-inflammatory cytokines (such as TNF-α or IL-1) signaling (eg IRAK, NIK, IKK) , p38 or] VIAP kinase inhibitor), IL-Ιβ converting enzyme inhibitor, TNFa converting enzyme (TACE) inhibitor, T cell signaling inhibitor (such as kinase inhibitor), metalloproteinase inhibitor, sulfasalazine Azathioprine, 6-mercaptopurine, angiotensin-converting enzyme inhibitor, soluble cytokine receptor and its derivatives (eg soluble p55 or p75 TNF receptor and derivative p75 TNFR IgG (EnbrelTM & p55 TNFR IgG) ), sIL-1RI, sIL-lRII and sIL-6R), anti-inflammatory cytokines (eg 11^4, 11^-10, IL-11, IL-13 and TGFP), senecab, folic acid, hydroxyl sulfate Hydroxychloroquine sulfate, rofecoxib, etanercept, infliximab, naproxen (napr〇Xen), valdecoxib, sulfasalazine, sputum nylon, merlot Meloxicam, methylprednisolone acetate, thiomalate (Gold 160877.doc -231 - 201247704

Sodium thiomalate), aspirin, triamcinolone acetonide, propoxyphene napsylate/apap, folate, nabumetone, dipivoxil Diclofenac), ° piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone hcl, hydrocodone tartaric acid Hydrocodone bitartrate/apap, diisoprostol/misoprostol, fentanyl, human recombinant anakinra (humankin), tramadol hydrochloride (tramadol hcl) ), salsalate, sulindac, cyanocobalamin/fa/° ratio pyridoxine 'acetaminophen, alendronate (alendronate sodium), splashed nylon, morphine sulfate, lidocaine hydrochloride, indomethacin, glucosamine sulf/chondroitin, amitriptyline hydrochloride Hcl), amine, bite Adiazine), oxycodone hydrochloride / acetaminophen phenol, olopatadine hydrochloride (olopatadine HC1), misoprostol, naproxen, omeprazole. Omeprazole, cyclophosphamide, rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18 BP, anti-IL-18, anti-IL15, BIRB-796, SCIO-469 VX-702, AMG-548, VX-740, Roflumilast, IC-485, CDC-801 and Mesopram. Combinations include amidoxime or leflunomide, and cyclosporine in the case of moderate or severe rheumatoid arthritis. Non-limiting for the treatment of rheumatoid arthritis in combination with binding proteins 160877.doc -232- 201247704

Other agents include, but are not limited to, the following: non-steroidal anti-inflammatory drugs (NSAID); cytokine inhibitory anti-inflammatory drugs (CSAID); CDP-571/BAY-10-3356 (humanized anti-TNFa antibody; Celltech/Bayer); cA2 / Infliximab (chimeric anti-TNFa antibody; Centocor); 75 kdTNFR-IgG/etanercept (75 kD TNF receptor-IgG fusion protein; Immunex; see eg (1994) Arthr. Rheum. 37: S295; (1996) J. Invest· Med. 44: 235A); 55 kdTNF-IgG (55 kD TNF receptor-IgG fusion protein; Hoffmann-LaRoche); IDEC-CE9.1/SB 210396 (non-expendable primate anti-antibody) CD4 antibody; IDEC/SmithKline; see for example (1995) Arthr. Rheum. 38: S185); DAB 486-IL-2 and/or DAB 389-IL-2 (IL-2 fusion protein; Seragen; see eg (1993) Arthrit. Rheum. 36: 1223); anti-Tac (humanized anti-IL-2Ra; Protein Design Labs/Roche); IL-4 (anti-inflammatory cytokine; DNAX/Schering); IL-10 (SCH 52000; recombinant IL- 10 anti-inflammatory cytokines; DNAX/Schering); IL-4; IL-10 and / or IL-4 agonists (such as agonist antibodies); IL-1RA (IL-1 receptor antagonist; Synergen / Amgen ); Anakinin ( Kineret®/Amgen); TNF-bp/s-TNF (soluble TNF-binding protein; see for example (1996) Arthr. Rheum. 39 (9 (suppl)): S284; (1995) Amer. J. Physiol. -Heart and Circ. Physiol. 268: 37-42); R973401 (type IV phosphodiesterase inhibitor; see eg (1996) Arthr. Rheum. 39 (9 (suppl): S282); MK-966 (COX-2 inhibitor) See, for example, (1996) Arthr Rheum. 39 (9 (suppl): S81); Iloprost (see for example (1996) Arthr. Rheum. 39 (9 (suppl): S82); methotrexate I60877.doc -233 - 201247704 呤'thalidomide (see for example (1996) Arthr. Rheum. 39 (9 (suppl): S282) and thalidomide-related drugs (eg Celgen) ), leflunomide (anti-inflammatory and cytokine inhibitors; see for example (1996) Arthr. Rheum. 39 (9 (suppl): S131; (1996) Inflamm.

Res· 45: 103-107); tranexamic acid (plasminogen activation inhibitor; see for example (1996) Arthr Rheum 39 (9 (Supp.). S284) 'T-614 (cytokine) Inhibitors; see, for example, (1996) Arthr. Rheum. 39 (9 (suppl): s282); prostaglandin £1 (see for example (1996) Arthr. Rheum. 39 (9 (suppl): S282); Tinidap (Tenidap) (non-steroidal anti-inflammatory drugs; see for example (1996) Arthr Rheum. 39 (9 (suppl): S280); naproxen (non-steroidal anti-inflammatory drugs; see eg (1996) Neuro. Report 7: 1209-1213) ; meloxicam (non-steroidal anti-inflammatory drugs); ibuprofen (non-steroidal anti-inflammatory drugs); piroxicam (non-steroidal anti-inflammatory drugs); diclofenac (non-steroidal anti-inflammatory drugs); indomethacin (non-steroidal anti-inflammatory drugs) Sulfasalazine 11 pyridine (see, for example, (1996) Anhr Rheum. 39 (9 (suppl): S281); azathioprine (see for example (1996)

Arthr·Rheum. 39 (9 (suppl): S281); ICE inhibitor (inhibitor of enzyme interleukin-1 beta-convertase); zap_7〇 and/or lck inhibitor (tyrosine kinase zap-70 or lck) Inhibitors; VEgf inhibitors and / or VEGF-R inhibitors (jk official endothelial growth factor or inhibitor of endothelial cell growth factor receptor; angiogenesis inhibitors); corticosteroid anti-inflammatory drugs (eg SB203 5 80 TNF-converting enzyme inhibitor; anti-antibody; anti-ns antibody, interleukin-11 (see, for example, (1996) Arthr. Rheum. 39 (9 (suppl) _ S296); interleukin-13 (see for example ( 1996) Arthr. Rheum. 39 (9 (Added 160877.doc -234 - 201247704): S308); Interleukin-17 inhibitor (see for example (1996) Arthr Rheum 39 (9 (suppl): S120) ; gold; penicillamine; gas quinquin; chlorambucil; chlorhexidine; cyclopurine; ring guanamine; systemic lymphoid tissue irradiation; antithymocyte globulin; anti-CD4 antibody; CD5 toxin; Oral administration of peptides and collagen; lobenzarit disodium; cytokine modulators (CRA) HP228 and HP466 (Houghten P Harmaceuticals, Inc.; ICAM-1 antisense thioacetate oligodeoxynucleotide (ISIS 2302; Isis Pharmaceuticals, Inc.); Soluble Complement Receptor 1 (TP10; T Cell Sciences, Inc.); Prednisone; orgotein; glycosaminoglycan polysulphate; minocycline; anti-IL2R antibody; marine organisms and plant lipids (fish and plant seed fatty acids; See, for example, DeLuca et al., (1995) Rheum. Dis. Clin. North Am. 21: 759-777); auranofin; phenylbutazone; meclofenamic acid Flufenamic acid; intravenous immunoglobulin; zileuton; azaribine; mycophenolic acid (RS-61443); tacrolimus ( Tacrolimus) (FK-506); sirolimus (rapamycin); amyprilose (therafectin); cladribine (2-chloro Deoxyadenosine); methotrexate; bcl-2 inhibitor (see Bruncko, Μ et al, (2007) J. Med. Chem. 5 0(4): 641-662 ); and antiviral agents and immunomodulators. In one embodiment, the binding protein or antigen binding portion thereof is administered in combination with one of the following agents to treat rheumatoid arthritis: a small molecule of KDR inhibitor 160877.doc-235-201247704; a small molecule inhibitor of Tie.2 Methotrexate; prednisone; seneoxib; folic acid; sulfuric acid by base gas; rofecoxib; etanercept; infliximab mt#; zeppson; valdecoxib; ratio. ; methylprednisolone; ibuprofen; beauty, and then electricity. ^ „ Wu Luo Xi Kang, methylprednisolone acetate; sodium thiomalate; aspirin L 吟; triamcinolone acetonide; Propoxyphene/apap; folate; nabumetone; difenfen; piroxicam; etodolac; diclofenac sodium; oxaprozil; hydrochloric acid test _; nitrogen can be tartrate hydrogen salt / apa Shuangqifen sodium/misoprostol; fentanyl; human recombinant anakinra, tramadol hydrochloride; disalicylate; sulindac; cyanocobalamin/fa/pyridoxine; Amine phenol; alendronate sodium; palladium; morphine sulfate; lidocaine hydrochloride; indomethacin; glucosamine sulfate/chondroitin; cyclosporine, amitriptyline hydrochloride; Coxone/Butylamine benzoate olothetamine hydrochloride; misoprostol; naproxen sodium; omeprazole d; mycophenolate mofetil; cyclophosphamide; rituximab; TRA"TRAp;

MRA; CTLA4-IG; IL-18 BP; IL-12/23; anti-IL 18; anti-IL 15, BIRB-796, SCIO-469; VX-702; AMG-548; VX 740; roflumilast; -485; CDC-801; and Mesopan. Non-limiting examples of therapeutic agents for inflammatory bowel disease that can be combined with the binding proteins of the invention include the following: budesonide; epidermal growth factor; corticosteroids, cyclosporine; sulfasalazine; Acid salt; 6-mercaptopurine; azathioprine; sputum nitrate; sitting; lipid oxygenase inhibitor; mesalazine; olsalazine; balsalazide; antioxidant; thromboxane inhibitor; IL-1 Body antagonist; anti-IL-Ιβ mAb; anti-IL-6 mAb; growth factor; elastase inhibitor; pyridyl-imidazole compound; and other humans 160877.doc • 236 · 201247704 cytokines or growth factors (eg TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-17, IL-18, EMAP-II, GM-CSF, FGF and PDGF) An antibody or antagonist. The binding protein of the invention or antigen binding portion thereof can be combined with antibodies to cell surface molecules such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69 and CD90, or any of their ligands. The binding protein of the present invention or antigen-binding portion thereof may also be combined with an agent such as amidoxime; cyclosporine; FK506; rapamycin; mycophenolate mofetil; defluzamide; NSAID, Such as ibuprofen; corticosteroids, such as prednisolone; phosphodiesterase inhibitors; adenosine agonists; antithrombotic agents; complement inhibitors; adrenaline stimulants; interference with pro-inflammatory cytokines such as TNFa or IL -1) a signaling agent (eg IRAK, NIK, IKK, p38 or MAP kinase inhibitor); an IL-1 beta converting enzyme inhibitor; a TNFa converting enzyme inhibitor; a T cell signaling inhibitor, such as a kinase inhibitor; Metalloproteinase inhibitor; sulfasalazine; azathioprine; 6-mercaptopurine; angiotensin-converting enzyme inhibitor; soluble cytokine receptor and its derivatives (eg, φ soluble ρ55 or p75 TNF receptor, sIL -lRI, sIL-lRII and SIL-6R); anti-inflammatory cytokines (eg IL-4, IL-10, IL-11, IL-13 and TGFp); and bcl-2 inhibitors. Examples of therapeutic agents for Crohn's disease that can be combined with binding proteins include the following: TNF antagonists (e.g., anti-TNF antibodies), adalimumab (PCT Publication No. WO 97/29131; HUMIRA®), CA2 (REMICADE®), CDP 571, TNFR-Ig constructs (p75 TNFR IgG (ENBREL®) and p55 TNFR IgG (ennespril 8)) inhibitors and PDE4 inhibitors. The binding protein of the present invention 160877.doc-237-201247704 or an antigen binding portion thereof can be combined with a corticosteroid such as budesonide or dexamethasone, the binding protein of the present invention or an antigen binding portion thereof, and the following agent Combinations: such as sulfasalazine, 5-aminosalicylic acid, and olsalazine, as well as agents that interfere with the synthesis or action of pro-inflammatory cytokines such as ICP (eg, IL_丨β-converting enzyme inhibitors and IL-lra). The binding protein of the present invention or antigen-binding portion thereof can also be used together with a butyl cell signaling inhibitor such as the tyrosine kinase inhibitor 6-mercaptopurine. The binding protein of the present invention or an antigen binding portion thereof can be combined with IL_i. The binding protein of the present invention or antigen-binding portion thereof can be combined with the following agents: mesalazine, prednisone, azathioprine, guanidinopurine, infliximab, sputum nylon glass station sodium, diphenoxy (diphenoxylate)/atrop sulfate, i〇peramide hydrochloride, amidoxime, omeprazole, folate, ciprofloxacin/dextrose-water Hydrocodone, hydrogen azide/apap, tetracycline hydrochloride, fluocinonide, sulphate, thimerosal, succinic acid, cholestyramine/sucrose, hydrochloric acid ring Proscyamine sulfate, meperidine hydrochloride, midazolam hydrochloride, ketamine hydrochloride, acetaminophen hydrochloride, and Pumin Taiding Promethazine hydrochloride), sodium sulphate, cans of sulfamethoxazole / methoxybenzylamine. Trimethoprim, celecoxib, polycarbophil (P〇lycarb〇phil), propofol propionate, hydrocortisone, multivitamin 'Baliu nitrogen> dinars, Codeine acidate (codeine 160877.doc - 238 - 201247704 phosphate)/apap, colesevelam hcl, cyanocyanine, folic acid levofloxacin, methylprednisolone, natalizumab And interferon-γ. Non-limiting examples of therapeutic agents that can be used in combination with the binding proteins of the invention for multiple sclerosis include the following: corticosteroids; splashed nylon; strontium nylon; azathioprine; cyclophosphamide; cyclosporine; methylamine喋呤; 4-aminopyridine; tizanidine; interferon-pia (AVONEX®; Biogen); interferon-plb (BETASERON®; Chiron/Berlex); interferon a-n3 (Interferon Sciences/ Fujimoto); interferon-a (Alfa Wassermann/J&J); interferon plA-IF (Serono/Inhale Therapeutics); pegylated interferon a 2b (Enzon®/Schering-Plough); copolymer l (Cop -l; COPAXONE®; Teva Pharmaceutical Industries, Inc.; hyperbaric oxygen; intravenous immunoglobulin; clabribine; against other human cytokines or growth factors and their receptors (eg TNF) , LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-23, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF and PDGF ) an antibody or antagonist. The binding protein of the present invention can be associated with an antibody against a cell surface molecule such as CD2, CD3, CD4, CD8, CD19, CD20, CD25, CD28, CD30, CD40, CD45, CD69, CD80, CD86, CD90 or a ligand thereof combination. The binding proteins of the invention may also be combined with agents such as amidoxime, cyclosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAID (eg, ibuprofen), Corticosteroids (such as prednisolone), phosphodiesterase inhibitors, adenosine agonists, antithrombotic agents, complement inhibitors, adrenergic agonists, interfering pro-inflammatory cytokines 160877.doc -239- 201247704 TNFa or IL-1) signaling agents (eg IRAK, NIK, IKK, p38 or MAP kinase inhibitors), IL-ΐβ converting enzyme inhibitors, TACE inhibitors, T cell signaling inhibitors (eg kinase inhibitors) , metalloproteinase inhibitors, sulfasalazine, azathioprine, 6-mercaptopurine, angiotensin-converting enzyme inhibitors, soluble cytokine receptors and their derivatives (eg soluble P55 or P75 TNF receptor, 51!^ -1RI, sIL-lRII and SIL-6R), anti-inflammatory cytokines (eg IL-4, IL-10 'IL-13 and TGFp) and beb 2 inhibitors. Examples of therapeutic agents for multiple sclerosis which can be combined with the binding proteins of the present invention include interferon 邛 (e.g., 11?>^1& and 11?1'^113); kepasone, corticosteroids, cass Protease inhibitors (eg, caspase-1 inhibitors), IL-1 inhibitors, TNF inhibitors, and antibodies against CD40 ligands and CD80. The binding proteins of the invention may also be combined with the following agents; such as alemtuzumab, dronabinol, Unimed, dalizumab, mitoxantrone, zalibin hydrochloride (xalipr) 〇den hydrochloride), amine 0 fluffridine 'glatiramer acetate, natalizumab, sinnabidol, a-imonokin NNS03 (a-immunokine NNS03), ABR -215062, AnergiX.MS®, chemokine receptor antagonist, BBR-2778, calagualine, CPI-1189, LEM (liposome encapsulated mitoxantrone), THC.CBD (classigenic marijuana Cannabinoid agonist), MBP-8298, mesoprofen (PDE4 inhibitor), MNA-715, anti-IL-6 receptor antibody, neurovax, 0 fenfenidone ), atropipin 1258 (allotrap 160877.doc -240- 201247704 1258) (RDP-1258), sTNF-Rl, talampanel, teriflunomide, TGF-P2 'Teli Tiplimotide, VLA-4 antagonist (eg TR-14035, VLA4 Ultrahaler®, Antegran®-ELAN/Biogen), interferon gamma Agents and anti-IL-4 agonist.

Non-limiting examples of therapeutic agents for angina pectoris that can be combined with the binding proteins of the invention include the following: aspirin, nitroglycerin, isosorbide mononitrate, metoprolol succinate (metoprolol) Succinate), atenolol, metoprolol tartrate, amlodipine besylate, diltiazem hydrochloride, isosorbide dinitrate, clopidogrel Bisulfate), indeed 'nifedipine, atorvastatin, atorvastatin calcium, potassium chloride, furosemide, simvastatin, verapamil hydrochloride Verapamil hcl), digoxin, propranolol hydrochloride, carvedilol, lisinopril, spironolactone, hydrochlorothiazide, butadiene Elaprilril maleate, nadolol, ramirril, enoxaparin sodium, heparin sodium, vasartan, Sotalol hydrochloride, fenofibrate, ezetimibe, bumetanide, losartan potassium, lisinopril/hydrogen Benzopyrazine, felodipine, captopru and bisoprolol fumarate 160877.doc -241 - 201247704 (bisoprolol fumarate). Non-limiting examples of therapeutic agents for ankylosing spondylitis that can be combined with the binding proteins of the invention include the following: ibuprofen, difenfen and misoprostol, naproxen, mezlocine, and ami Xin, difenfen, seneoxib, rofecoxib, sulfasalazine, methotrexate, azathioprine, diammine tetracycline, prednisone, etanerxi popular infliximab. Non-limiting European examples of therapeutic agents for asthma that can be combined with the binding proteins of the invention include the following: albuterol, salmetero1/fluticasone, montelukast sodium , cecasin propionate, budesonide (bU (jesonide), prednisone, salmeterol hydroxynaphtholate, levobutamol hydrochloride (levalbuter 〇 1 hcl), salbutamol sulfate / ipratropium, pep) Sodium, triamcinolone acetonide, beclomethasone dipropionate, desertified ipratropium, azithromycin, pirbuterol acetate, splashed nylon, anhydrous theophylline, methylprednisolone Sodium diacid, clarithromycin, zafirlukast, formoterol fumarate, influenza virus vaccine, methylprednisolone, amoxicillin trihydrate , flunisolide (flunisolide), allergy injection, sodium cromoglycate, fexofenadine hydrochloride, oxynipines / menthol, amoxicil Clavulanate, levofloxacin, inhaler aid, guaifenesin, dexamethasone sodium, moxifloxacin hcl, doxycycline hydrochloride Doxycycline hyclate), guaifenesin/d-methorphan, ephedrine 160877.doc -242- 201247704 /cod/chlorpheniramine (chi〇rphenir), gatifloxacin (gatifl〇) Xacin), cetirizine hydr〇chl〇ride, mometasone furoate, salmeterol hydroxynaphthoate, benzalnatate, cephalexin Cephalexin), pe/hydrocodone / fentanamine, cetirizine hydrochloride/pseud ephedrine, phenylephrine/cod/Pultin, dynasty/Pultin Cefpr〇zil, dexamethasone, guaifenesin/pseudoephedrine, chlorpheniramine/hydrocodone, nedocromil sodium, terbutaline sulfate, adrenaline, Metaproterenol sulfate between methylprednisolone and sulfuric acid. Non-limiting examples of therapeutic agents for c〇pD that can be combined with the binding proteins of the invention include the following: salbutamol sulfate/isopropylidonium bromide, ipratropium bromide, salmeterol/fluticasone, salbutamol, hydroxynaphthylquinone Salmeterol, fluticasone propionate, prednisone, anhydrous theophylline, methylprednisolone sodium succinate, montelukast sodium, budesonide, fumarate fumarate, triamcinolone acetonide, levofloxacin , guaifenesin, azithromycin, dipropionic acid bismuth • rice pine, l-salbutamol hydrochloride, flunisolide, ceftrixone sodium, amoxicillin trihydrate, gatifloxacin, zafirlukast, Moxilin / clavulanate, flunisolide / menthol, chlorpheniramine / ketone, hydroxyisoproterenol, methylprednisolone, mometasone citrate, ephedrine / C 〇d/ fenfenamine, guanbuterol acetate, ephedrine/loratadine, terbutaline sulphate, tiotropium bromide, (R,R)·Fumo Tro, TgAAT, cilomilast and roflumilast. 160877.doc • 243 · 201247704 Non-limiting examples of therapeutic agents for HCV that can be combined with the binding proteins of the invention include the following: interferon a-2a, interferon a 2b, interferon alpha c〇nl, interferon a -nl, pegylated interferon a_2a, pegylated interferon a-2b, ribavirin, pegylated interferon a_2b + viral saliva, ursodeoxycholic acid, glycyrrhizic acid (glycyrrhizic acid), thymalfasin, maxamine, VX-497, and any compound used to treat hcv by interfering with the following targets: HCV polymerase, HCV protease, HCV helicase and HCV IRES (internal ribosome entry site). Non-limiting examples of therapeutic agents for idiopathic pulmonary fibrosis that can be combined with the binding proteins of the invention include the following: prednisone, azathioprine, albuterol, colchicine, salbutamol sulfate, digoxin , gamma interferon, methylprednisolone sodium succinate, lorazepam (lorazepam), flubenzidine, lisinopril, nitroglycerin, spironolactone, cyclophosphamide, ipratropium bromide Bacteriocin d, alteplase, fluticasone propionate, levofloxacin, isoproterenol sulfate, morphine sulfate, oxycodone hydrochloride, potassium hydride, triamcinolone acetonide, anhydrous tacrolimus Division, fishing, interferon alpha, methotrexate, mycophenolate mofetil and interferon gamma-1β. Non-limiting examples of therapeutic agents for myocardial infarction that can be combined with the binding proteins of the invention include the following: aspirin, nitroglycerin, metoprolol tartrate, enoxaparin sodium, heparin sodium, gaspirite hydrogen sulfate Salt, carvedilol, atenolol, morphine sulfate, metoprolol succinate, warfarin sodium 'lisinopril, isosorbide mononitrate, digoxin, furosemide Acid, simvastatin, ramipril, tenecteplase 160877.doc • 244- 201247704

(tenecteplase), erbium dibutyl succinate, tosemide, retavase, losartan, quinapril hcl/mag carb, bume Nitalactide, alteplase, enalaprilat, amiodarone hydrochloride, tirofiban hcl m-hydrate, diltiazem hydrochloride, captopril , irbesartan (irbesartan), saponin, propranolol hydrochloride, fosinopril sodium, lidocaine hydrochloride, eptifibatide, head hug. Cefazolin sodium, atropine sulfate, aminocaproic acid, spironolactone, interferon, sotalol hydrochloride, gasification _, docusate sodium, dopaamine hydrochloride (dobutamine hcl), Apu. Alprazolam, pravastatin sodium, atorvastat, j弼, hydrochloric acid. Mide "Spirulina, pethidine hydrochloride, isosorbide dinitrate, adrenaline, dopamine hydrochloride, bivalirudin, rosuvastatin, ezetimibe / simvastatin, ar Non-limiting examples of therapeutic agents for psoriasis that can be combined with the binding proteins of the present invention include: KDR small molecule inhibitors, small Tie-2 Molecular inhibitors, calcipotriene, clobetasol propionate, triamcinolone acetonide, halobetasol propionate, tazarotene, methaqualin , fluocinolone acetonide, betamethasone diprop augmented, fluocinolone acetonide, 160877.doc -245- 201247704 acitretin, tar shampoo, Betamethasone valerate, mometasone citrate, and sylvestre. Ketoconazole, pramoxine / fluocinolone, hydrocortisone valerate, flurandrenolide, urea, betamethasone, dexamethasone propionate / emollient, propionic acid Fluticasone, azithromycin, hydrocortisone, moisturizing formula, folic acid, desonide, pimecrolimus, coal tar, diflorasone diacetate ), etanercept folate, lactic acid, methoxsalen, he/ bismuth subgal / znox/resor, methylprednisolone acetate, prednisone, sunscreen, Hasina Halcinonide, salicylic acid, anthralin, clocortolone pivalate, coal extract, coal tar/salicylic acid, coal tar/salicylic acid/sulfur, gossip Rice (des〇ximetasone), diazepam, emollient, fluocinolone acetonide/emollient, mineral oil/E sesame oil/na lact, mineral oil/peanut oil, petroleum/isopropyl myristate, bone Psoralen, salicylic acid, soap/tribromsaian, Thimerosal / boric acid, celecoxib stuffed inside, infliximab, cyclosporine, A special Tour de France, efalizumab, tacrolimus. Bimemus, PUVA, UVB and sulfasalazine. Non-limiting examples of therapeutic agents for psoriatic arthritis that can be combined with the binding proteins of the invention include the following: methotrexate, etanercept, rofecoxib, senecab, folic acid, sulfasalamide比bipyridine, naproxen, leflunomide, methylprednisolone acetate, indomethacin, hydroxyquine sulfate, prednisone, sulindac, intensive betamethasone dipropionate, infliximab, A Amine, 160877.doc •246· 201247704 Folic acid, triamcinolone acetonide, difenfen acid, disulfoxide, „beroxicon, sodium bisphenolate, ketoprofen, merlot Xikang, methylprednisolone, nabumetone, tolmetin sodium, barbiturate, cyclosporine, diclofenac sodium/misoprostol, fluocinolone acetonide, glucosamine sulfate, thiomalate Gold sodium, hydrocodone bitartrate/apap, ibuprofen, risedronate sodium, reductive amine bite, thioguanine, valdecoxib, afaset, legal liberia Anti-bcl-2 inhibitor. Non-therapeutic agents for restenosis that can be combined with the binding proteins of the present invention Restrictive examples include the following: sirolimus, paclitaxel, everolimus, tacrolimus, zotatolimus, and acetaminophen. It can be used in combination with the binding proteins of the present invention. Non-limiting examples of therapeutic agents for sciatica include the following: hydrocodone bitartrate/apap, rofecoxib, CyCl〇benzaprine hcl, methylprednisolone, naproxen, ibuprofen , oxycodone hydrochloride / acetaminophen hydroxy, seneoxib, valdecoxib, acetaminophen acetate, prednisone, codeine phosphate / apap, tramadol hydrochloride / acetaminophen, metaxalone (metaxalone ), meloxicam, mesocarbamol, lidocaine hydrochloride, sodium bisphenolate, gabapentin, dexamethasone, carisoprodol, ketosole ketorolac Tromethamine), 0 丨 0 meimeixin, acetaminophen phenol, diazepam, nabumetone, oxycodone hydrochloride, tizanidine hydrochloride, sodium bisphenolate/misoprostol, propane oxygenated naphthalene sulfonate Fen / apap, asa / oxycod / test 鲷 ter (asa / oxycod / o Xycodone ter), ibuprofen / 160877.doc -247- 201247704 Hydrogen steel tartrate, tramadol hydrochloride, etodolac, propoxyphene hydrochloride, amitriptyline hydrochloride, muscle tranquilization / codeine phosphate /asa , morphine sulfate, multivitamins, naproxen sodium, orphenadrine citrate and temazepam. Non-limiting examples of therapeutic agents for systemic lupus erythematosus (SLE) that can be combined with the binding proteins of the invention include the following: NSAIDs, such as diclofenac, naproxen, ibuprofen, piroxicam, ami Xin; COX2 inhibitors, such as senebutr's rofecoxib, valdecoxib; antimalarial agents, such as hydroxyquinoquine; steroids, such as prednisone, prednisolone, budesonide, dexamethasone; cytotoxic agents, For example, azathioprine, cyclomethamine, mycophenolate mofetil, methotrexate; and PDE4 inhibitors or purine synthesis inhibitors, such as Cellcept. The binding protein of the present invention may also be combined with an agent such as sulfasalazine, 5-aminosalicylic acid, olsalazine, Imuran, and pro-inflammatory cytokine synthesis, production or Acting agents (eg, caspase inhibitors, such as IL-Ιβ converting enzyme inhibitors, the binding proteins of the invention may also be associated with tau cell signaling inhibitors (eg, tyrosine kinase inhibitors) or targeted tau cell activating molecules Molecules (eg, CTLA_4_IgG or anti-B7 family antibodies and anti-PD-1 family antibodies) are used together. The binding proteins of the invention may be combined with or anti-cytokine antibodies (eg, aryltozumab (f〇n〇t〇Uzumab) (anti- IFNg antibody)) or an anti-receptor receptor antibody (eg, an anti-receptor antibody) and an antibody combination against a B cell surface molecule. The antibody of the present invention or an antigen-binding portion thereof can also be combined with LJP 394 (Abemex ( Abetimus)) an agent that depletes or does not activate B cells, such as rituximab (anti-cd2〇 antibody); 160877.doc •248- 201247704 lymphostat-B (anti-BlyS antibody), TNF antagonist (eg anti-TNF antibody) Ada Wood monoclonal antibody (PCT Publication No. WO 97/29131; HUMIRA), CA2 (REMICADE), CDP 571, TNFR-Ig construct (p75 TNFR IgG (ENBREL) and p55 TNFR IgG (LENERCEPT)) and bcl The -2 inhibitor was used together, since the overexpression of bcl-2 in transgenic mice has been shown to cause a lupus-like phenotype (see Marquina, R et al., (2004) J. Immunol. 172(11): 7177 -7185), therefore, it is expected that inhibition will produce a therapeutic effect. The pharmaceutical composition of the present invention may comprise a "therapeutically effective amount" or a "prophylactically effective amount" of a binding protein of the present invention. "Therapeutically effective amount" means at a necessary dose and is necessary for a period of time. The effective amount of the binding protein is determined by the skilled artisan and may be based on factors such as the disease condition, the age, sex and weight of the individual, and the ability of the binding protein to elicit the desired response in the individual. A therapeutically effective amount is also one in which the therapeutically beneficial effect of the binding protein or binding protein moiety exceeds any toxic or detrimental effect. "Prophylactically effective amount" means a period of time necessary and necessary for a period of time Effectively achieve the amount of result to be prevented. Usually, because the prophylactic dose is administered to an individual before the early stage of the disease or at an early stage of the disease, the prophylactically effective amount will be less than the therapeutically effective amount. The prophylactically effective amount does not require prevention of the disease or condition. For example, a prophylactically effective amount can delay the onset of a disease or condition. The dosage regimen can be adjusted to provide the optimal desired response (eg, a therapeutic or prophylactic response). For example, a single large dose can be administered, which can be administered over time. With a number of open doses, or as indicated by the urgency of treatment, a reduction of 160877.doc -249· 201247704 or an increase in dose may be made. It is especially preferred to formulate parenteral compositions as unit dosage forms for ease of administration and uniformity of dosage. Unit dosage form and suitable beta as used herein are used as a unitary dosage for the physical individual unit of the individual to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the desired pharmaceutical carrier . The specification of the dosage unit form of the present invention is directly dependent on the following factors: (4) the unique characteristics of the active compound and the particular therapeutic or prophylactic effect to be achieved; and (b) the tailing of the active compound for treatment of the individual Limitations inherent in sensitive technologies. An exemplary, non-limiting range of therapeutically or prophylactically effective amounts of a binding protein of the invention is from 0.1 to 20 mg/kg, for example! · i G mg/kg. It should be noted that the dose value may vary depending on the type and severity of the condition to be alleviated. It should be further appreciated that for any particular individual, the particular dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the individual administering or supervising the administration of the composition, and the dosage ranges described herein are for illustrative purposes only and are not It is intended to limit the scope or implementation of the claimed compositions. Ιν·Diagnostic Applications The present invention also provides diagnostic applications. This will be further described below. Diagnostic Methods The invention also provides for the use of at least one half as described herein. Binding Protein A method of determining the presence, amount or concentration of an analyte (or a fragment thereof) in a test sample. Any suitable assay known in the art can be used in the method. Examples include, but are not limited to, immunoassays, such as sandwich immunoassays (eg, single, multiple, and/or semi-binding protein sandwich immunoassays or any of their variations of 160877.doc-250·201247704 (eg, single plant) /Semi-Ig binding protein, semi-Ig binding protein/multiple strains, etc.), including radioisotope detection (Radioimmunoassay (RIA)) and enzyme detection (EIA) or enzyme-linked immunosorbent assay (ELISA) ELISA) (eg Quantikine ELISA assay, R & D Systems, Minneapolis, MN), competitive inhibition immunoassay (eg forward and reverse), fluorescence polarization immunoassay (FPIA), enzyme doubling immunoassay (EMIT), Bioluminescence Resonance Energy Transfer (BRET), and homogeneous chemiluminescence analysis, etc. In a SELDI-based immunoassay, a capture reagent that specifically binds a related analyte (or a fragment thereof) is ligated to a mass spectrometer probe ( Surface such as a pre-activated protein wafer array. The analyte (or a fragment thereof) is then specifically captured on a biochip and the captured analyte is detected by mass spectrometry. Or a fragment thereof. Alternatively, the analyte (or a fragment thereof) may be eluted from the capture reagent and detected by conventional MALDI (matrix-assisted laser desorption/ionization) or by SELDI. Chemiluminescence microparticle immunoassay ( In particular, the use of ARCHITECT® automated analyzers (Abbott Laboratories, Abbott Park, IL) for chemiluminescent microparticle immunoassay) is an example of a preferred immunoassay. For example, a semi-Ig binding protein as described herein is used as For immunodiagnostic reagents and/or for use in analyte immunoassay kits, the methods well known in the art for collecting, processing and processing urine, ash, gold and commercial and other body fluids are used. The test sample may contain other moieties in addition to the relevant analytes, such as antibodies, antigens, haptens, hormones, drugs, enzymes, receptors, proteins, peptides, polypeptides, oligonucleotides and/or polynucleotides. For example, the sample can be a whole blood sample obtained from an individual 160877.doc-251 - 201247704. It may be necessary or desirable to pre-treat, for example, prior to the immunoassay as described herein. Treatment of test samples (especially whole blood) ^ Pre-treatments can be performed as appropriate (eg as part of a commercial platform-based approach) even where pre-treatment is not necessary (eg most urine samples). Any reagent suitable for use with the immunoassay and kit of the present invention. The pretreatment comprises, as appropriate: (a) one or more solvents (eg, methanol and ethylene glycol) and optionally a salt, (b) ) one or more solvents and salts and optionally detergents, (c) detergents, or (d) detergents and salts. Pretreatment reagents are known in the art and can be, for example, as described in the literature (see, for example, Yatscoff et al., (1990) Clin. Chem. 36: 1969-1973 and Wallemacq et al. (1999) Clin. Chem. 45. : 432-435) The form used in the analysis performed on Abbott TDx, AxSYM® and ARCHITECT8 analyzers (Abb〇tt Laboratories, Abbott Park, IL), and/or commercially available forms. Further, the pretreatment can be carried out as described in U.S. Patent No. 5,135,875, European Patent Publication No. eu 471 293, U.S. Patent No. 6,660,843, and U.S. Patent Application No. 2__2. The pretreatment reagent can be a (tetra) reagent or a homogeneous reagent. The pretreatment reagent precipitates an analyte binding protein (e.g., a protein that binds to the analyte or a fragment thereof) present in the sample using a heterogeneous pretreatment reagent. The pretreatment step comprises removing any analyte binding protein by isolating the supernatant of the mixture formed by adding the pretreatment agent to the sample to the analyte binding protein of the sink. In this assay, the supernatant of the mixture without any binding protein is analyzed (4) and directly subjected to the antibody capture step. 160877.doc •252· 201247704 Under the use of homogeneous pretreatment reagents, there is no complete (tetra) compound and analytical procedure for this sample and pretreatment reagent. The test marker-specific binding partner (such as labeled: /, fragment) will be used before or during the capture of a white-like antibody (or its antigen-reactive "sediment-binding egg-specific binding partner"; The t:r agent is a pretreatment reagent that still exists (or remains) in the mixture of diuradine I(tetra) in the pretreated material.

Can the conjugate be a half Ig binding protein body? Β-specific segment). ^ ^A fragment of a variant or variant thereof In a heterogeneous format, after obtaining a test sample from an individual, preparing a first mixture: the mixture contains a test sample for evaluation of the analyte (or a fragment thereof) and the first or only specificity A binding partner, wherein the specific binding partner forms a specific binding partner cleavage complex with any analyte contained in the test sample. The specific binding partner is preferably an anti-analyte annex protein (e.g., an antibody) or a fragment thereof. The specific binding partner may be a half Ig binding protein (or a fragment thereof, a variant or a variant fragment) as described herein. The order in which test samples are added and the specific binding partners are combined to form a mixture is not critical. Preferably, the specific binding partner is immobilized on a solid phase. The solid phase used in immunoassays (for specific binding partners and optionally for the first specific binding partner) can be used in this technology. Know any solid phase, such as (but not limited to) magnetic particles, beads, test tubes, microtiter plates, cuvettes, membranes, backbone molecules, membranes, filter paper, discs, and wafers. 160877.doc •253 · 201247704 After the first or only specific binding of the conjugate-analyte complex, any of the 7 unbound analytes are removed from the complex using any technique known in the art. For example, unbound analytes can be removed by silk. However, it is desirable to have the specific binding partner present in excess of any analyte present in the test sample so that all of the analytes present in the test sample are combined with the specific binding partner. After removal of any unbound analyte, if a second specific binding partner is present, a second specific binding partner can be added to the mixture to form a specific binding partner - analyte - second specific binding Matching compound. The second specific binding partner is preferably an anti-analyte binding protein that binds to an antigen on the analyte that is different from the antigenic determinant of the epitope bound to the analyte by the first specific binding partner. In addition, the second specific binding partner is preferably labeled with a detectable label as described above or contains a detectable label as described above. The second specific binding partner can be a semi-Ig binding protein (or variant thereof) as described herein. Any suitable detectable label known in the art can be used. For example, the detectable label can be a radioactive label (such as 3H, 125I, 35s, 14C, 32P, and 33p), an enzyme label (such as horseradish peroxidase 'alkaline peroxidase, glucose 6-phosphate dehydrogenase) And its analogs), chemiluminescent labels (such as acridine rust, thioester or sulfonamide; luminol, isoluminol, phenanthridinium ester and its analogues), fluorescein Light labeling (such as luciferin (eg 5-fluorescein, rebel leucine, 3·6-carboxy luciferin, 5(6)-carboxyfluorescein, 6-hexa-luciferin, 6 - four-gas luciferin, luciferase isothiocyanate and its analogues)), rhodamine, 160877.doc • 254 · 201247704

Phycobiliprotein, R-phycoerythrin, quantum dots (e.g., cadmium selenide sulphide), thermolabeling or immunopolymerase chain reaction labeling. Introduction of markers, labeling procedures and detection of markers can be found in Polak and Van Noorden, Introduction to Immunocy to chemistry, 2nd edition, Springer Verlag, Ν·Υ. (1997) and Haugland, Handbook of Fluorescent Probes and Research Chemicals ( 1996) (which is a combined manual and catalogue published by Molecular Probes, Inc., Eugene, Oregon). Fluorescent markers can be used in the FPIA (see, for example, U.S. Patent Nos. 5,593,896; 5,573,904; 5,496,925; 5,3,59,093 and 5,3,52,803). Acridine compounds can be used as detectable labels in homogeneous or heterogeneous chemiluminescence assays (see, for example, Adamczyk et al. (2006) Bioorg. Med. Chem. Lett. 16: 1324-1328; Adamczyk et al., ( 2004) Bioorg. Med. Chem. Lett. 4: 2313-2317; Adamczyk et al., (2004) Biorg. Med. Chem. Lett 14: 3917-3921; and Adamczyk et al., (2003) Org. Lett. 5 : 3779-3782). Preferably, the acridinium compound is acridinium-9-nonylamine. A method for the preparation of acridinium 9-nonylamine is described in Mattingly (1991) J. Biolumin. Chemilumin. 6: 1 07-114; Adamczyk et al., (1998) J. Org. Chem. 63: 5636-5639; Adamczyk Et al., (1999) Tetrahedron 55: 10899-10914; Adamczyk et al., (1999) Org. Lett. 1: 779-781; Adamczyk et al., (2000) Biocon. Chem. 11: 714-724; Mattingly et al. , In Luminescence Biotechnology: Instruments and Applications; Dyke, KV; CRC Press: 160877.doc -255 - 201247704

Boca Raton, pp. 77-105 (2002); Adamczyk et al., (2003) Org. Lett. 5: 3779-3782; and U.S. Patent Nos. 5,468,646; 5,543,524; and 5,783,699. Another preferred acridinium compound is acridine rust-9-aryl aryl ester. An example of an acridine rust-9-aryl aryl ester is 10 0-methyl-9-(phenoxycarbonyl)acridine rust fluorosulfonate (available from Cayman Chemical, Ann Arbor, MI). A method for the preparation of acridinium 9-carboxylic acid aryl esters is described in McCarra et al. (1965) Photochem. Photobiol. 4: 1111-21; Razavi et al., (2000) Luminescence 15: 245-249; Razavi et al., (2000) Luminescence 15: 239-244; and U.S. Patent No. 5,241,070. Further details regarding acridine rust-9-decanoic acid aryl ester and its use are set forth in U.S. Patent Publication No. 20080248493. Chemiluminescence analysis can be carried out according to the method described in Adamczyk et al. (2006) Anal. Chim. Acta 579(1): 61-67 (for example using acridine rust or other chemiluminescent agents as described above). Any suitable analytical format can be used, but a microplate chemiluminometer (Mithras LB-940, Berthold

Technologies U.S.A., LLC, Oak Ridge, TN) enable rapid analysis of multiple small volume samples. The order in which test samples and specific binding partners are added to form a mixture for chemiluminescence analysis is not critical. If the first specific binding partner is detectable by a chemiluminescent agent such as an acridine rust compound, a first specific binding partner-analyte complex with a detectable label is formed. Or 'if a second specific binding partner is used and the second specific binding partner is detectable by a chemiluminescent agent such as an acridine rust compound, a first specific binding partner of the detectable label is formed - Analyte - Second Special 160877.doc -256- 201247704 Heteroconjugate binding complex. Any labeled or unlabeled unbound specific binding partner can be removed from the mixture using any technique known in the art, such as washing. Hydrogen peroxide may be generated in situ in the mixture prior to, simultaneously with or after the addition of the acridine compound described above, or hydrogen peroxide may be supplied or supplied to the mixture (eg, the source of hydrogen peroxide is one or more known to contain hydrogen peroxide) The buffer or other solution, hydrogen peroxide, can be generated in situ in a variety of ways, such as in a manner well known to those skilled in the art. • After at least one alkaline solution is added to the sample simultaneously or subsequently, a non-analytical analysis is produced. A detectable signal, i.e., a chemiluminescent signal, present. An alkaline solution having at least one base and having a pH greater than or equal to 1 Torr, preferably greater than or equal to 12, includes, but is not limited to, Sodium hydroxide, potassium oxyhydroxide, calcium hydroxide, ammonium hydroxide, magnesium hydroxide, sodium carbonate, sodium hydrogencarbonate, calcium hydroxide, calcium carbonate and calcium hydrogencarbonate. The amount of alkaline solution added to the sample depends on the alkali The concentration of the solution depends on the concentration of the alkaline solution used, and those skilled in the art can readily determine the amount of the alkali solution to be added to the sample. Conventional techniques known to those skilled in the art detect the resulting chemiluminescent signal. Based on the resulting signal intensity, the amount of analyte in the sample can be quantified. In particular, the amount of analyte in the sample and the resulting signal Strength Scale can be used to quantify the amount of analyte present by comparing the amount of light produced to the standard curve of the analyte or by comparison to a reference standard. Mass spectrometry, gravimetric analysis, and Other techniques known in the art use serial dilutions or solutions of known concentrations of analytes to generate the standard 160877.doc • 257·201247704 compounds as chemical statements to use other chemical quasi-curves. Although the above description focuses on the use of acridine rust Light agent', but the general technologist can easily adapt this illuminant. Analyte immunoassay can generally use this arbitrage

The formula (such as, but not limited to, a sandwich format) is performed. In particular, at least two binding proteins (e. g., antibodies) are employed in an immunoassay format to: isolate and quantify an analyte in a sample, such as a human analyte or fragment thereof. More specifically, at least two antibodies bind to different epitopes on the analyte (or a fragment thereof) to form an immune complex, which is referred to as a "sandwich." In general, in immunoassay, one or more antibodies can be used to capture the analyte (or a fragment thereof) in the sample (these antibodies are often referred to as "capture" antibodies) and one or more antibodies can be used. To allow detectable (ie, quantitative) labeling to be incorporated into the sandwich (these antibodies are often referred to as "detecting antibodies" or "conjugates J". Thus, 'in the context of a sandwich immunoassay format, as described herein. A binding protein or a semi-匕 binding protein (or a variant thereof) can be used as a capture antibody detection antibody or both. For example, for a half 1 § binding protein containing at least two complete antigen binding sites, one can be used. A binding protein or a semi-Ig binding protein having a domain that binds to a first epitope on an analyte (or a fragment thereof) as a capture reagent and/or may be used on another having a bindable analyte (or a fragment thereof) a binding protein or a semi-Ig binding protein of a domain of a second epitope as a detecting agent. In this regard, having a first domain capable of binding to a first epitope on an analyte (or a fragment thereof)The beta protein or semi-Ig binding protein of the second domain of the second epitope on the analyte (or a fragment thereof) can be used as a capture reagent and/or a primary test agent. Or I60877.doc -258- 201247704 a binding protein having a first domain that binds to an antigenic thiol group on a first analyte (or a fragment thereof) and a second domain that binds to an epitope on a second analyte (or a fragment thereof) or The semi-Ig binding protein acts as a capture and/or detection agent to detect and optionally quantify two or more analytes. The analyte may be in more than one form (such as monomeric form and dimeric/multimeric form, Where it may be homomeric or hetero meric in the sample, a binding egg having a domain that binds to an epitope that is only exposed to the monomeric form can be used. Or a half-binding protein and another binding protein or semi-binding protein having a binding to a dimeric/multimeric form, a domain of an epitope on the P-knife, as a scavenger and/or a detecting agent, thereby Capable of detecting and depending on the situation Different forms of established analytes. In addition, binding proteins or semi-Ig binding proteins with different affinities between single-binding or semi-Ig binding proteins and/or between binding proteins or semi-Ig binding proteins can provide an affinity advantage. In the case of immunoassays as described herein, it may be helpful or desirable to incorporate one or more linkers into the structure of the binding protein or semi-Ig binding protein. If a linker is present, it is optimal. The linker should be of sufficient length and structural flexibility to enable the internal domain to bind to the epitope and the external domain to bind to another epitope. In this regard, when the binding protein or the semi-Ig binding protein can bind two different When the analyte is one greater than the other, it is desirable to incorporate a larger analyte from the external domain. In general, a test (eg, suspected of containing) a sample of an analyte (or a fragment thereof) can be simultaneously or sequentially and in any order with at least one capture agent and at least 160877.doc -259.201247704 a detection agent (eg, In the case where the cockroach is/the smear and/or the detection agent contains a plurality of reagents, it may be the first-predicate apricot-Ή丨ate丨4+J and the second detection agent or even serial numbering Agent) contact. For example, you can: _ 丨 便利 Convenience 6 The pattern βσ is first contacted with at least one of the traps' and then (sequentially) in contact with at least one of the detectors. Alternatively, the test sample can be first contacted with at least one detector and then (sequentially) contacted with at least one capture agent. In another alternative, the test sample can be brought into contact with the capture agent and the detection agent at the same time.

The sample suspected of containing the analyte (or a fragment thereof) is first contacted with at least one of the first capture reagents under conditions which permit formation of the first reagent/analyte complex in the form of the assay. If more than one type of capture agent is used, a first capture/analyte complex comprising two or more capture agents is formed. In the decentering assay, the reagent (i.e., preferably at least one capture reagent) is used in an amount greater than the maximum molar excess of the analyte (or its tablet) expected in the test sample. For example, from about 5 micrograms to about 1 milligram of reagent per milliliter of buffer (e.g., microparticle coating buffer) can be used.

Since only one type of binding protein and/or a half Ig, '' is required in the context of the present invention. The combination of prion protein is often used to measure the competitive inhibition of small analytes. *Including sequential and typical lattices. In the sequential competitive inhibition immunoassay, the capture agent of the relevant analyte is applied to the well of a microtiter plate or other solid support. When a sample containing the relevant analyte is added to the sputum, the relevant analyte binds to the capture reagent. After washing, a known amount of labeled (e.g., biotin or horseradish peroxidase (HRP)) knife extract capable of binding to the binding protein is added to the well. Enzyme-labeled receptors are necessary for generating signals. An example of a suitable substrate for HRP is 3,3',5,5'-tetradecylbenzidine 160877.doc •260· 201247704 (TMB). After washing, the signal produced by the labeled analyte is measured and the signal is inversely proportional to the amount of analyte in the sample. In a typical competitive inhibition immunoassay, in the context of the present invention, a binding protein of a relevant analyte and/or a semi-Ig binding protein (e.g., an antibody) is typically applied to a solid support (e.g., a well of a microtiter plate). However, unlike sequential competitive inhibition immunoassays, both the sample and the labeled analyte are added to the well. Any analyte in the sample competes with the labeled analyte for binding to the capture reagent. After washing, the signal produced by the labeled analyte is measured and the signal is inversely proportional to the amount of analyte in the sample. Of course, there are many variations of these formats (for example, when a combination with a solid substrate occurs, the format is one step, two steps, two steps of delay, and the like), and the average technician will recognize this change. Body format. Depending on the situation, at least one capture agent can be bound to the solid support prior to contacting the test sample with at least one capture agent (eg, the first capture agent), which facilitates the first reagent/analyte (or fragment thereof) The complex is separated from the test sample. The substrate to which the capture agent is combined can be any suitable solid support or solid phase that facilitates separation of the capture agent-analyte complex from the sample. Examples include wells of trays (such as microtiter plates), test tubes, porous gels (such as silicone, agarose, dextran or gelatin), polymeric films (such as polyacrylamide), beads (such as polystyrene beads). Strips or particles of filter paper/film (eg nitrocellulose or nylon), particles (eg latex particles, magnetizable particles (eg having iron oxide or chromium oxide cores and homo- or heteropoly coatings) The semi-active particles are about 1-10 micrometers. The substrate may comprise a suitable porous material having a surface affinity suitable for binding to the antigen and a porosity sufficient to allow detection of antibody entry and exit. Although a gel state in the hydrated state may be used. Materials, but micro 160877.doc -261- 201247704 Pore materials are generally preferred. The porous substrates are preferably in the form of flakes having a thickness of from about 0.01 mm to about 0.5 mm, preferably about 〇 1 mm. It may vary widely, but preferably has a pore size of from about 0.025 to about 15 micrometers, more preferably from about 0.15 to about 15 micrometers. The surface of the substrates may be chemically bonded to the substrate by covalent bonding of the antibody to the substrate. Process passivation coating or activation. Generally borrowed The antigen or antibody is irreversibly bound to the substrate via hydrophobic force adsorption; alternatively, the antibody may be covalently bound to the substrate using a chemical coupling agent or other means as long as the binding does not interfere with the ability of the antibody to bind the analyte. Alternatively, the antibody (In the context of the present invention, ie, a binding protein and/or a semi-匕 binding protein) can be bound to a microparticle, which is pre-coated with a streptavidin (eg DYNAL® magnetic bead, invitrogen, Carlsbad, CA). Or biotin (for example, using Power-BindTM-SA-MP streptavidin coated microparticles (Seradyn, Indianapolis, IN)) or anti-species specific monoclonal antibodies (in the context of the present invention' Protein and/or DVD-Ig). If necessary or desired, the substrate (eg for labeling) can be derivatized to various types of antibodies (in the context of the present invention, ie binding proteins or semi-binding proteins) The functional group is reactive. The derivatization requires the use of certain coupling agents, examples of which include, but are not limited to, maleic anhydride, N-hydroxybutanediimide, and oxime ethyl _3_ (3·2曱Aminopropyl propyl Carbodiimide. If desired, one or more capture agents (such as antibodies (or fragments thereof), each of which may be specific for the analyte (in the context of the present invention, ie, a binding protein and/or a semi-Ig binding protein) )) is attached to a different physical or addressable location of the solid phase (e.g., in a biochip configuration) (see, e.g., U.S. Patent No. 6,225, No. 47; pCT Publication No. WO 99/51773, U.S. Patent No. 6>329>; 2 () 9 said; ρ (: τ & % _ $ 160877.doc 201247704 WO 00/56934, and US Patent No. 5,242, 828). If the capture reagent is attached to a mass spectrometer probe as a solid support, the amount of analyte bound to the probe can be detected by laser desorption ionization mass spectrometry. Alternatively, a single column can be filled with different beads derivatized with one or more capture agents, thereby capturing the analyte at a single location (see techniques based on antibody-derivatized beads, such as LUminex (AUStin, TX)) χΜΑΡTechnology). After the test sample used to analyze the analyte (or a fragment thereof) is contacted with at least one capture agent (eg, the first capture agent), the mixture is grown to allow formation

The first capture agent (or the first capture agent)-analyte (or a fragment thereof) complex. The incubation can be at about 2 at a value of about 4.5 to about 1 11. (: to about 45. (: at a temperature of at least about - (1) minutes to about ten (18) hours, preferably about 1 minute to about 24 minutes, preferably about 4 minutes to about 18 minutes) The immunoassay described herein can be carried out in one step (meaning that the test sample, at least one of the capture agents, and at least one of the four (four) agents are added to the reaction vessel sequentially or simultaneously) or in more than one step, such as Two steps, three steps, etc. - w (di- or a few) capture agent / analyte (or a fragment thereof) complex formation ^ then make the complex with at least a formulation in the form of allowing (the - or :) Capture agent/analyte (or its month) / second detection agent complex = touch. Although for the sake of clarity, the name "second - detection agent" is captured, and the reagent is captured. And / / 一种 V a detection agent can be used in immunoassay "two: detection agent, third agent, fourth detection agent, etc.. If the capture agent (or its fragment) complex and more than one Contact with the detector to form a 16 〇 877.doc 201247704 (first or first) capture agent/analyte (or Fragment) / (several) detector complex. Like a capture reagent (eg, a first capture reagent), when at least one species (eg, a second and any subsequent number) is detected with a capture reagent/analyte (or sheet

Section) When the complex is contacted, it is necessary to incubate for a period of time under conditions similar to those described above to form (first or first) capture agent/analyte (or fragment thereof) / (second or first) Detector complex. Preferably, at least one of the detectors contains a detectable label. The detectable label can bind to at least one detection agent before, simultaneously or after (first or a plurality of) capture agent/analyte (or a fragment thereof) / (second or a plurality of) detection agent complexes ( For example, the second detection agent). Any detectable label known in the art can be used (see above for details, including Polak and Van Noorden (1997) and Haugland (1996) references). The detectable label can be bound to the reagent either directly or via a coupling agent. An example of a coupling agent that can be used is EDAC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride), which is commercially available from Sigma Aldrich, St. Louis, M. Other coupling agents that can be used are known in the art. Methods for binding detectable labels to binding proteins are known in the art. In addition, a variety of detectable labels that have been promoted to couple detectable labels to the end groups of the reagent are commercially available or synthesized, such as CPSP• acridinium ester (ie, 9-[N-toluene bromide N (3) -carboxypropyl)]·1〇_(3_sulfopropyl) acridine rust carbamide) or spsp_ acridine rust (ie Νΐ〇·(3-sulfopropyl)_N_(3 sulfopropyl) _ acridine rust _9-nonylamine). Prior to the 疋1 labeling, (but not necessarily) (first or first) trapping knife extract / (second or a number of) detector complexes and test samples 160877.doc -264- 201247704 Partially isolated "For example, if at least one capture agent (eg, a first capture agent, such as a binding protein of the invention and/or a semi-binding protein), a gentleman in a solid support (such as a well or a bead), then The separation can be achieved by removing (testing the sample) fluid, no longer contacting the solid support, or if at least the first capture agent is bound to the solid support, it can simultaneously be combined with the sample containing the analyte and at least one The second detection agent contacts to form the first plurality of reagents/analyte/second (several) reagent complex, and then the removal fluid (test sample) is no longer in contact with the solid support. If at least one first capture agent Not bonded to a solid support, no need

The test sample is removed (first or a number of) of capture agent/analyte/(second or a number of) detector complex to quantify the amount of label. After the labeled capture agent/analyte/detector complex (eg, the first capture agent/analyte/second detection agent complex) is formed, the complex is quantified using techniques known in the art; ^ Tickets are the amount of their own. In the case of I, if the enzyme label S is used, the labeled complex is reacted with a standard that produces a quantifiable reaction (such as color development). If labeled as a radioactive label, the label is quantified using a suitable method, such as a scintillation counter. If marked as a fluorescent marker, the marker is quantified by exciting the marker with monochromatic light (referred to as the "excitation wavelength") and detecting another color emitted by the marker response excitation (which is referred to as the "emission wavelength"). . If the mark is a chemiluminescent mark, the amount of the mark is quantified by visual inspection or by using a photometer, an X-ray film, a high-speed photographic film, a CCE camera, or the like to detect the emitted light. Thereafter, the concentration of the analyte in the test sample or the concentration of the fragment of 160877.doc-265 · 201247704 is determined by a suitable method, such as by using a standard curve generated using serial dilutions of analytes of known concentrations or fragments thereof. . In addition to the use of serial dilutions of the analyte or fragments thereof, standard curves can be generated gravimetrically, by mass spectrometry, and by other techniques known in the art. In the chemiluminescence particle analysis method using the ARCHITECT® analyzer, the pH of the conjugate diluent should be about 6.0 +/- 0.2, and the particle coating buffer should be maintained at about room temperature (ie, at about 17 ° C to At about 27 ° C), the particle coating buffer should have a pH of about 6.5 +/- 0.2 ' and the particulate diluent pH should be about 7 8+/_ 〇·2. The solid is preferably less than about 0.2°/. For example, less than about 0.15% and less than about 0.14. /. Less than about 0.13%, less than about 〇.12% or less than about %11%, such as about 0.10%. FPIA is based on the principle of competitive binding immunoassays. Fluorescent cursors emit fluorescence that is inversely proportional to the rate of rotation of the compound when excited by linearly polarized light. When the fluorescently labeled tracer-antibody complex is excited by linearly polarized light, the emitted light remains highly polarized because the rotation of the fluorophore between the time of absorbing light and the time of emitting light is limited. When a "free" tracer compound (ie, an unbound: antibody-based compound) is excited by linearly polarized light, its rotation ratio is competitively combined with the corresponding agent-antibody conjugate produced in the immunoassay (or In the present invention, the vertical agent - any octa protein and / or tracer · hemi (tetra) protein) is much faster. FPIA is superior to ria because of the radioactivity of special handling and disposal. In addition, it is a homogeneous analysis method that can be easily and quickly performed. A method of determining the presence, amount, or concentration of an analyte (or a fragment thereof) in a test sample. The τ ° χ method includes the following methods: 四 Α Α Α ( ( 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 四 ' ' ' ' ' ' ' ' ' ' ' ' ' ' ' ' ' ' ' ' 201247704 A antibody, an antibody fragment, an antibody that binds to an analyte, a fragment of an antibody variant that binds to an analyte, a binding protein as disclosed herein, and an analyte that binds to the analyte At least one of the DVD-Ig binding protein (or a fragment thereof or a fragment of the variant), and () to the y detectable label; and (ii) the inclusion of the detectable marker as an assay Direct or indirect indication of the presence, amount or concentration of the substance (or a fragment thereof) in the test sample and the presence, amount or concentration of the analyte (or fragment thereof) in the control or control standard directly or Indirect indicator numbers are compared. The control standard is a series of control standard knives as appropriate, with each control standard having a different analyte concentration than the other control standards. The method can comprise (1) contacting a test sample with at least one first specific binding partner selected from the group consisting of: (or a fragment thereof) to form a first specific binding partner/analyte (or fragment thereof) a complex: an antibody, an antibody fragment that binds to an analyte; a variant of an antibody that binds to an analyte, a tablet that binds to an antibody variant of the analyte; a binding protein as disclosed herein; Binding to a half of the analyte binding protein (or a fragment, variant or variant thereof (ii) such that the first specific binding partner/analyte (or fragment thereof) complex and at least one selected from the group consisting of The second specific binding partner of the analyte (or a fragment thereof) is contacted to form a first-specific binding partner/analyte (or a fragment thereof) / a second specific binding partner complex: can be combined with the analysis The detectably labeled anti-analyte antibody, the detectably labeled anti-analyte antibody fragment can bind to the detectable labeled anti-analyte antibody variant of the analyte. I60877.doc •267- 201247704 A detectable labeled anti-analyte antibody variant fragment that binds to an analyte, can be bound to an analyte of a detectable marker, a purine protein, and is detectable as disclosed herein. a half-marked DVD-Ig binding protein (or a fragment thereof, a variant or a variant thereof); and (iii) a specific binding partner/analyte formed by detecting or measuring (...) Or a fragment thereof / a first specific binding partner that complexes the detectable marker calving signal to determine the presence, amount or concentration of the analyte in the test sample. At least one first-specific binding partner for the analyte (or a fragment thereof) and/or at least one second specific binding partner for the analyte (or a fragment thereof) is a binding protein as described herein or as A method of the semi-DVD-Ig binding protein (or a fragment, variant or variant thereof) described herein may be preferred, the method comprising the step of bringing the test sample to at least two analytes (or fragments thereof) The first-specific binding collocation (4) is the second antibody to the analyte, the antibody fragment; the antibody variant, which can be bound to the phase separation, -τα A 1 , such as, can be - the antibody variant of the knife a fragment; a binding protein as disclosed herein, and a fragment of a half 1 § allogeneic or variant); and simultaneously or sequentially 5, and the sample and at least the genus of the genus in any order The second specific partner is contacted, and the one of the first ligands can be competitively bound to the analyte (or a fragment thereof). The first heterosexual binding partner can be combined with the group selected from the following: · The specificity of the combined binding of the Hungarian, can be detected浐4 焱 Detectable marker analysis 贝 軚 己 分析 分析 分析 分析 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析 分析A detectably labeled analyte variant fragment that specifically binds to the conjugate at 160877.doc 201247704. Any analyte (or a fragment thereof) present in the test sample competes with at least one first specific binding partner to form a first specific binding partner/analyte (or fragment thereof) complex and a first specificity, respectively Sex binding conjugate/second specific binding partner complex. The method further comprises determining an analyte by detecting or measuring a signal generated by the detectable label in the first specific binding partner/second specific binding partner complex formed in (ii) The presence, amount, or concentration in the test sample, wherein the signal produced by the detectable label in the first specific binding partner/second specific binding partner complex and the amount or concentration of the analyte in the test sample In inverse proportion. The above method may further comprise diagnosing, prognosing or assessing the efficacy of the therapeutic/prophylactic treatment of the patient obtaining the test sample. If the method further comprises assessing the efficacy of the therapeutic/prophylactic treatment of the patient obtaining the test sample, the method further comprises altering the therapeutic/prophylactic treatment of the patient as needed to improve efficacy. This method can be modified for use in a self-rotating system or a semi-automated system. Regarding the analytical method (and its kit), a commercially available anti-analyte antibody or a method of producing an anti-analyte as described in the literature can be employed. Suppliers of various antibodies include, but are not limited to, Santa Cruz Biotechnology Inc. (Santa Cruz, CA), GenWay Biotech, Inc. (San Diego, CA), and R&D Systems (RDS; Minneapolis, MN). Generally, 'a predetermined amount can be used as a knot obtained when evaluating an analyte or a fragment thereof (for example, for detecting a disease or a disease risk) of a test sample, 160877.doc • 269-201247704 benchmark & In comparison, a predetermined amount is obtained by performing a specific analysis of a sufficient number of times under suitable conditions to:: present the analyte, its content or its concentration, and the characteristic stage or end point of the disease, disorder or condition or Relevance or relevance of specific clinical indicators. Typically, an analyte obtained by analyzing a reference individual (or a population of individuals) can be assayed to include its tablet, its degradation products, and/or its enzymatic cleavage product.

In particular, the amount or concentration of an analyte or a fragment thereof may be "unchanged", "favorable" (ancient or "favorably altered") or "unfavorable" with respect to the predetermined amount used to monitor disease progression and/or treatment ( “Unfavorable change” “two-door” or “if plus” means that the amount or concentration in the test sample is higher than the typical or normal content or range (eg, predetermined content), or higher than another reference content or range (eg, initial or baseline) Sample). The term "reduce" or "reduce" the heart test: the amount or concentration in the sample is lower than the typical or normal content or range (eg: pre-set) or below another reference content or range (eg initial Or baseline, "surgical improvement" means that the amount or concentration change (increase or decrease) in the sample exceeds the typical or JL constant content or range (eg & predetermined content) or exceeds another reference content or range (eg, initial or baseline sample) The typical or normal content or range of analytes is defined according to standard operating methods. Since the analyte content is extremely low in some cases, it is compared to typical or normal content or range or reference content or range. When any net change cannot be explained by experimental error or sample difference, it can be considered that the content or change of the so-called change has occurred. Because &, the measured content in a specific sample is measured in a similar sample from a so-called normal individual. Content or content range 160877.doc • 270- 201247704 compared. In this context, “individuals who are not associated with the disease, and for example “just have no detectable disease, often” (sometimes referred to as “control”) Patients or groups are patients or groups that do not have a debtable disease. In addition, high levels of analytes are generally not found in most human populations, and "normal =" can be considered as the amount or concentration of the analyte. (d) an increase or a "normal" (sometimes referred to as "control") patient or group of patients or groups whose scallops have no substantial increase or increase in the amount or concentration of the knife. A normal individual is an individual who has not yet evaluated the analyte or gone before = assessing the analyte. When the analyte is usually undetectable (eg, the normal content is zero, or is about 25% to about 75% of the normal population) Internal) 'But when the analyte is detected in the test sample, and the analyte content in the # test sample is higher than the normal content, the analyte content is considered to be "increased". Therefore, the present invention particularly provides screening for specific diseases. , a disease or condition, or a method of treating an individual at risk of a particular face, disorder, or condition. The analytical method may also involve the analysis of other markers and their analogs. Thus, the methods described herein may also be used to determine whether an individual is diarrhea or not. The disease, disorder or condition is at risk of developing a given disease, disorder or condition. In particular, the method may comprise the following steps: (4) determining the indifference of an analyte (or a fragment thereof) from a test sample from an individual. Or amount (for example, using the methods described herein or methods known in the art); and (b) comparing step (a) to the concentration or amount of the analyte (or fragment thereof) as measured by a predetermined amount. 'If the concentration or amount of the analyte measured in step (4) is a favorable result with respect to the content, then it is confirmed that the individual is not -27. I60S77.doc 201247704 Disorder or condition or no risk of a given disease, disorder or condition of the risk. However, when the concentration or amount of the analyte measured in the step (4) is unfavorable with respect to the predetermined amount, it is determined that the individual suffers from the established disease, disorder or condition or is suffering from a predetermined disease, disorder or condition. risk. In addition, this article provides a way to monitor the health of the money show. Preferably, the method comprises the steps of: (4) determining the concentration or amount of the analyte in the test sample from the individual; (8) determining the concentration of the analyte in the post-hypox test sample from the individual; and (4) measuring as in step (b) The concentration or amount of the analyte is compared with the concentration or amount of the analyte measured in (4) (4), wherein the concentration or amount measured in step (b) is in the concentration or amount of the analyte measured in step (4). When there is no change or an unfavorable result at the time of comparison, it is determined that the disease of the individual continues, progresses or deteriorates. In contrast, if the concentration or amount of the analyte measured in step (b) is a favorable result when compared to the concentration or amount of the analyte measured in step (4), then the disease of the individual has been determined Suspend, subside or improve. The method further comprises, as the case may be, comprising comparing the concentration or amount of the analyte measured in step (b) to, for example, a predetermined amount [further, the method optionally comprises: if comparing the analysis measured in step (b) If the concentration or amount of the substance is adversely altered, for example, relative to the predetermined amount, the individual is treated with the composition of the drug for a period of time. In addition, such methods can be used to monitor treatment of an individual receiving treatment with __ or a plurality of medical remedies. In particular, the methods involve providing a first test from an individual 160877.doc • 272. 201247704 before administering to the individual - or a plurality of pharmaceutical compositions. Subsequently, the concentration or amount of the analyte in the first test sample from the individual (eg, after determining the concentration or amount of the analyte using methods described herein or as known in the art), then the concentration of the analyte is then taken as appropriate ^ Or the amount is compared with the predetermined content. If the concentration or amount of the divided substance measured in the first test sample is lower than the predetermined content, the individual is not treated with - or a plurality of pharmaceutical compositions. However, if in the first test sample When the concentration or amount of the analyte is higher than the predetermined amount, one or more pharmaceutical compositions (4) (iv) are used for a period of time. The time period for treating the individual with - or a plurality of pharmaceutical compositions may be determined by those skilled in the art (eg, the time period) It may be from about seven (7) days to about 2 years, preferably about fourteen (14) days to about one year. In the course of treatment with one or more pharmaceutical compositions, then the second and subsequent test samples are obtained from the individual. The number of test samples and the time from which the individual obtains the test samples is not critical. For example, one or more (7) days after the first administration of one or more medical groups to the individual The second test sample 'can be administered to the individual for the first time—the second test sample is obtained two (2) weeks after the Gexi seed composition, and the first person can be administered one or more medical treatments to the individual. (7) The fourth test sample is obtained weekly, and the fifth test sample can be obtained four (4) weeks after the first time to the individual = or a plurality of pharmaceutical compositions, and so on. The second or subsequent sowing * spray test is obtained from the individual. After the sample, determine the analyte red 'mountain morning or amount in the second or subsequent test sample (eg, using the squares known to be described herein or such a technique, π, ) ° then will be measured in each second and subsequent The degree, quantity or amount is compared with the concentration or amount of the analyte in the first test sample (for example, the test sample which is initially compared with the predetermined content phase α tt). If the step is compared, the concentration or amount of the precipitate is compared. The concentration or amount of the analyte measured in (4) is compared with the concentration or amount of the analyte measured in step (4): when the result is profitable, 'the individual's money has been stolen, subsided or improved and should continue to be The individual is administered to step (8) - or a combination of drugs However, if the concentration or amount measured in step (c) is unchanged or unfavorable when compared with the concentration or amount of the analyte measured in step (a), then the individual's disease is confirmed. Sustained, progressed, or worsened, and the individual should be treated with a higher concentration of one or more of the pharmaceutical compositions administered to the individual in step (b) or should be administered to the individual in one or more different steps (b) A pharmaceutical composition of one or more pharmaceutical compositions for treating the individual. In particular, one or more pharmaceutical compositions different from one or more of the pharmaceutical compositions previously received by the individual can be used to treat the individual to reduce or reduce the individual. Analyte content. Generally, for an assay that may be subjected to repeated testing (eg, monitoring disease progression and/or response to treatment), a second or subsequent test is obtained for a period of time after the first test sample has been obtained from the individual. sample. In particular, the second test sample can be obtained from the individual several minutes, hours, days, weeks or years after the first test sample has been obtained from the individual. For example, about 1 minute, about 5 minutes, about 10 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 60 minutes, about 2 hours, about 3 hours, after obtaining the first test sample from the individual, About 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 1 hour, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 5 hours, About 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 Day, about 6 160877.doc -274·201247704 days, about 7 days, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks 'about 1 week, about 丨 week, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 17 weeks, about "week, about ^ weeks, about 20 weeks, about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, about 25 weeks, about 26 weeks, about 27 weeks, about 28 weeks, about 29 weeks, about 30 weeks, about 31 weeks, about 32 weeks, about 33 weeks, about 34 weeks , about 35 weeks, about 36 weeks, about 37 weeks, about % weeks, about 39 weeks, about 40 weeks About 41 weeks, about 42 weeks, about 43 weeks, about 44 weeks, about 45 weeks, about 46 weeks, about 47 weeks, about 48 weeks, about 49 weeks, about 5 weeks, weeks, about 51 weeks, about 52 weeks, About two years, about two years, about 2.5 years, about three years, about 3.5 years, about 4.0 years, about 4.5 years, about five years, about 55 years, about 6.0 years, about 6.5 years, about 7. The second test sample is obtained from the individual during the leap year, about 7.5 years, about 8 years, about 85 years, about 9.0 years, about 9.5 years, or about 10.0 years. When the above analysis is used to monitor disease progression, Use of the above assays to monitor disease progression in individuals with acute conditions. Acute conditions are also referred to as critical care conditions and refer to life-threatening acute illnesses or other serious medical conditions, such as the cardiovascular system or the excretory system. Critical care conditions usually refer to the need for acute medical intervention in hospital facilities (including but not limited to emergency rooms, intensive care units, trauma centers, or other emergency care facilities) or by medical personnel in paramedics or other fields. Symptoms. For critical care conditions, generally in a shorter time frame Repeated monitoring, ie minutes, hours or days (eg about 丨 minutes, about 5 minutes, about 1 〇 minutes, about 15 minutes, about 30 minutes, about 45 minutes, about 6 minutes, about 2 hours, about 3 Hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, 160877.doc -275-201247704 about Μ, hr, about 9 hours, about 10 hours, about u hours, about η hours, about = hours, About 14 hours, about 15 hours, about 16 hours, about 17 hours, about hours, about 19 hours, about 2G hours, about, about, about 22 hours, about 23 hours, about 24 hours, about 2 days, about 3 Days, about 4 days, about $day, about 6 = 7 days)' and the initial analysis is also generally performed within the heart time range of the onset of the disease or condition, such as in about minutes, hours or days. These assays can also be used to monitor disease progression in individuals with chronic or non-acute conditions. Non-emergency care or non-acute disease (4) refers to conditions other than life-threatening acute illnesses and other critical medical conditions involving, for example, the cardiovascular system and/or the genital system. Non-acute conditions usually include conditions with longer or chronic duration. For non-acute conditions, repeated monitoring is typically performed over a longer period of time 'eg, hours, days, weeks, months, or years (eg, about hours, about 2 hours, about 3 hours, about 4 hours, about $ Hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about (7) hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about..., time of week, week, week, week, week, about 3 about 10 about 16 about 22 about 28 about 34 about 40 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 2 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, About 2, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 2 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 21 weeks, about 27 weeks, about 33 weeks, about 39 weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 2 weeks, about 23 weeks. About 24 weeks, about 25 weeks, about 26 weeks, about 29 weeks, about 3 weeks, about 31 weeks, about 32 weeks, about 35 weeks, about 36 weeks, about 37 weeks, about 38 weeks. 160877.doc •276· 201247704 Week, about 41 weeks, about 42 weeks, about 43 weeks, about About weeks, about 46 weeks, about 47 weeks, about 48 weeks, about 49 weeks, about 5 weeks, about "week, about 52 weeks, about 1.5 years, about 2 years, about 2.5 years, about 3 years. About "year, about * year, about 4_5 years, about 5.G years, about 5.5 years, about 6 years, about 6 rides, about 7.0 years, about 7.5 years, about 8 years, about 8 5 Years, about 9 years, about 9 years or about 1 year. The silk analysis method is also generally in the longer time range of a disease or a large episode of a disease, such as about hours, days, months or numbers. 0 年 During the year, the above assay can be performed using the first test sample obtained from the individual, wherein the first test sample is obtained from a source such as urine, serum or plasma. The second test sample obtained from the individual can then be used as the case may be repeated. The second test sample is obtained from another source. For example. If the first test sample is obtained from urine, the second test sample can be obtained from serum or plasma. The results obtained by the analysis using the first test sample and the second test sample can be compared. This comparison can be used to assess the status of a disease or condition in an individual. • In addition, the present invention is also directed to methods of determining whether an individual susceptible to or suffering from a given disease, disorder, or condition will benefit from treatment. In particular, the present invention relates to analyte-associated diagnostic methods and products. Thus, the method of "monitoring the treatment of a disease of an individual as described herein preferably further encompasses candidates for selection or differential therapy. Thus, in a particular embodiment, the invention also provides a method of determining whether an individual suffering from a given disease, disorder, or condition or having a risk of developing a given disease, disorder, or condition is treated; - generally, the individual is already 160877.doc •277- 201247704 Some of the symptoms of a given money, illness or condition or have actually been diagnosed as having a given disease, disorder or condition or suffering from a given disease, disorder or disease An individual at risk; and/or an individual who exhibits an unfavorable concentration or amount of analyte or fragment thereof as described herein. The method optionally comprises an assay as described herein, wherein the treatment is performed with one or more pharmaceutical compositions (e.g., particularly with a drug involved in the mechanism of action of the analyte), with immunosuppressive therapy, or by immunoadsorption therapy. The analyte is assessed before and after the individual, or where the analyte is assessed after the treatment and the concentration or amount of the analyte is compared to a predetermined amount. An unfavorable concentration or amount of analyte observed after treatment determines that the individual does not benefit from further or continued treatment, and an advantageous concentration or amount of analyte is observed after treatment to determine that the individual will benefit from further or Continue treatment. This does help to manage clinical research and provide improved patient care. It will be understood that while certain embodiments herein are suitable for assessing a given disease, disorder, or condition as described herein, such assays and kits can be employed to assess other diseases, disorders, and An analyte in a condition. Analytical methods can also involve the analysis of other markers and their analogs. Analytical methods can also be used to identify compounds that improve a given disease, disorder, or condition. For example, cells expressing an analyte can be contacted with a candidate compound. The amount of analyte in the cells contacted with the compound can be compared to the amount of analyte in the control cells using the analytical methods described herein. Β·Sets A kit for analyzing the presence, amount, or concentration of an analyte (or a fragment thereof) in a test sample for a test sample is also provided. The kit contains at least one test for analysis 160877.doc -278- 201247704. The composition of the analyte (or a fragment thereof) of the model and the instructions for analyzing the sample (or a fragment thereof) of the test sample. At least one component of the analyte (or a fragment thereof) for use in analyzing the test sample can comprise a binding protein and/or an anti-analyte half-binding protein (or a fragment thereof) comprising the invention disclosed herein, which is optionally immobilized on a solid phase. A composition of a variant or a fragment of a variant). The kit may comprise at least one component for analyzing the analyte of the test sample by immunoassay (eg, chemiluminescent microparticle immunoassay) and analyzing the test sample by immunoassay (eg, chemiluminescent microparticle immunoassay) Instructions for the analyte. For example, a kit can comprise at least one species-specific binding partner of an analyte, such as an analyte-resistant individual that binds to a fragment of the analyte, which can bind to an analyte variant, = a variant fragment of the analyte Any of the binding proteins or anti-analyte recombination proteins disclosed herein (or fragments, variants or variant fragments thereof) may be labeled. Alternatively or additionally, the kit may comprise _ = votes: Analyte (or a monoclonal antibody/multiple antibody that binds to an anti-analyte, : a fragment of the disclosed binding protein or anti-analyte hemi-protein (or a fragment thereof; or a fragment of the variant)) Compatible with the =precipitate in the test sample to bind the anti-analyte single/multiple antibodies (or a fragment thereof that can bind to the variant of the analyte), Revealing the binding protein 2, "/ knife-dissociated protein (or its fragment, variant 5, a few centimeters, half Ig, is fixed on the solid support). The package can be isolated or purified. Things, sets can be packaged == 照物' example points 妍 夕 夕 交 匕 至 至 至 至 至 至(eg tubes, microtiter plates or strips, which may for example be 160877.doc - 279 - 201247704 have been coated with a first-specific binding partner); and / or buffers, such as assay buffers or sachet buffers, Anyone can concentrate Mm# supply; detect the labeling solution (such as enzyme label); or stop the solution. The kit preferably contains all the components necessary for the analysis, ie reagents, standards, buffers Liquid, diluent, etc. The instructions may be in paper form or computer readable form, such as a magnetic disk, CD, DVD, or the like. More specifically, a set of antigens (or fragments thereof) for analyzing the test sample is provided. The kit includes at least one component for analyzing an antigen (or a fragment thereof) of the test sample and an antigen for analyzing the test sample (or a fragment thereof), wherein the at least one component includes at least one of the present inventions Composition and instructions for use. Another set of antigens (or fragments thereof) for analysis of test samples is provided. The kit contains at least one component and fraction of the antigen (or sheet k) used to analyze the test sample. A description of an antigen (or a fragment thereof) of a test sample, wherein the at least one component comprises at least one composition comprising a binding protein, the binding protein (i.) comprising a polypeptide chain comprising Vdi_xi_x2, wherein: VD1 comprises a first heavy chain a variable domain; χι comprises a heavy chain constant 1 (cm) domain; and X2 comprises at least a portion of a CH3 domain, wherein X2 comprises at least one mutation at a residue selected from the group consisting of C220, C226, C229, T366, L3 68, P3 95, F405 'Y407 and K409, thereby inhibiting CH3-CH3 dimerization' wherein the binding protein forms a functional antigen binding site and wherein the binding protein is detectably labeled as appropriate. In certain embodiments, The polypeptide chain further comprises a second polypeptide chain 'which comprises ¥〇1-(parent 1)]^, wherein 乂〇1 comprises a light bond antigen binding domain; X1 comprises a light chain domain; and N is 〇 or 1. 160877.doc -280· 201247704 Another set of antigens (or fragments thereof) for analysis of test samples is provided. The kit comprises at least one component for analyzing an antigen (or a fragment thereof) of a test sample and an assay for analyzing an antigen (or a fragment thereof) of the test sample, wherein the at least one component comprises at least one composition comprising a binding protein, The binding protein (i,) comprises the polypeptide chain VD1-(X1)N-VD2_(X2)N-X3, wherein .VD1 comprises a first heavy chain antigen binding domain; XI is a linker; vd2 comprises a second heavy chain antigen binding Domain; X2 comprises a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain, and a linker; N is deuterium or 1; and X3 comprises a polypeptide having at least a portion of the CH3 domain, wherein X3 comprises at least one mutation at the residue in the CH3/CH3 contact region, thereby inhibiting CH3-CH3 dimerization, wherein the binding protein forms a functional antigen binding site; and wherein the binding protein is detectably labeled as appropriate. In certain embodiments, the polypeptide chain further comprises a second polypeptide chain comprising VD1_(X1)N' wherein VD1 comprises a light chain antigen binding domain; X1 comprises a light chain constant domain; and N is deuterium or 1. Another set of antigens (or fragments thereof) for analysis of the test sample is provided. The kit comprises at least one component for analyzing an antigen (or a fragment thereof) of the test sample and instructions for analyzing the antigen (or a fragment thereof) of the test sample, wherein the at least one component comprises at least one of the semi-匕 binding proteins. combination. Any antibody (such as an anti-analyte antibody), any binding protein as disclosed herein, any anti-analyte half-Ig binding protein or tracer can have a detectable label such as a fluorophore, a radioactive moiety as described herein. The enzyme, biotin/avidin label, chromophore, chemiluminescent label or analog thereof' or kit can include reagents for performing a detectable label. Anti-160877.doc -281 · 201247704 =:! The quasi- and/or control may be provided in separate containers or in advance in an analytical format, such as pre-dispensed in a microtiter plate. The kit includes quality control components (eg, sensitivity group, control standard positive control) as appropriate. The preparation of quality control reagents is described in the art on the inserts of various immunodiagnostic products. Use the bar as appropriate; r VIII and the members to establish the analytical performance characteristics, and the step-by-step situation is the standard of the r-assay reagent set and the standardization of the analytical method. Other reagents, such as buffers, salts, enzymes, enzyme cofactors, enzymes, and other analogs, which are required for diagnostic analysis or for quality control evaluation, may also be included. Other components may also be included in the kit, such as buffers and solutions for separating and/or processing test samples (eg, pretreatment reagents) kits may additionally include one or more other controls. In this case, the kit may further comprise reagents suitable for reconstituting the lyophilized component. The various components of the kit are optionally provided in a suitable container, such as a microtiter tray, as needed. A container for holding or storing a sample (for example, a container or a filter cartridge for a urine sample) may be further included. Where appropriate, the kit may optionally contain a reaction container, a mixing container, and a reagent or test sample. Other components. The kit may also include one or more instruments for assisting in obtaining a test sample, such as a syringe, pipette, forceps, measured spoon, or the like. If the detectable marker is at least one t> bite rust compound, the kit may comprise at least one acridine rust-9-guanamine, at least one acridine rust _9_decanoic acid aryl 酽160877.doc -282- 201247704 or any combination thereof. Detective Marked as at least one acridine rust compound, the kit may also comprise a source of hydrogen peroxide, such as a buffer, solution and/or to >, an assay solution. If necessary, the kit may contain a solid phase such as magnetic particles. , beads, tubes, microtiter plates, cuvettes, membranes, matrix molecules, membranes, filter paper, discs or wafers. c. Adaptation of kits and methods by analytical methods (such as the immunoassay described herein) The kit (or component thereof) and method for determining the presence, amount or concentration of an analyte in a test sample can be adapted for use in, for example, U.S. Patent No. 5, No. 89,424 and No. 5, No. 6, 3 A variety of automated and semi-automated systems (including systems containing solid phase containing particulates), such as those sold by Abbott Laboratories (Abbott Park, IL) as ARCHITECT®, are described in 〇9 waves. Some differences between ELISA) include ligation of a first specific binding partner (eg, an anti-analyte, a single/multiple antibody (or a fragment thereof, a variant thereof, or a variant thereof thereof), a binding protein as disclosed herein or A substrate for an anti-analyte half-binding protein (or a fragment thereof, a variant thereof, or a variant thereof) that may affect sandwich formation and analyte reactivity, as well as affect capture, speculation, and/or any visual The duration and timing of the washing step in the case. Although non-automated formats (such as ELISA) may require the sample to be incubated with the capture reagent for a relatively long period of time (eg, about 2 hours) 'automatic or semi-automatic format (eg ARCHITECT®, Abbott Laboratories) ) may have a relatively short incubation time (eg, about 18 minutes for ARCHITECT®). Similarly 'although non-automated formats (such as ELISA) can be used to incubated detection antibodies (such as knot 160877.doc -283 - 201247704 reagents) ) A relatively long incubation time (eg, about 2 hours), but an automatic or semi-automatic format (eg, ARCHITECT®) may have a relatively short incubation time (eg, about 4 minutes for ARCHITECT®). Other platforms available from Abbott Laboratories include, but are not limited to, AxSYM®, IMx® (see, e.g., U.S. Patent No. 5,294,404), PRISM®, EIA (beads), and QuantumTM II, among others. In addition, assays, kits, and kit components can be used in other formats, such as on electrochemical or other handheld or point-of-care analysis systems. The present invention is, for example, applicable to a commercial Abbott Point of Care (i-STAT®, Abbott Laboratories) electrochemical immunoassay system that performs a sandwich immunoassay. An immunosensor, a method of manufacturing the same, and a method of operating in a single use test device are described, for example, in U.S. Patent Nos. 5,063,081; 7,419,821; and 7,682,833; and U.S. Patent Publication No. 20040018577 and 2006/0160164 No. In detail, the following configuration is preferred for improving analyte analysis to fit the I-STAT® system. A micromachined tantalum wafer having a pair of gold current analysis working electrodes and a silver-vaporized silver reference electrode was fabricated. On one working electrode, there will be a fixed anti-analyte monoclonal/multiple antibody (or a fragment thereof, a variant thereof or a variant thereof thereof), a binding protein as disclosed herein or an anti-analyte semi-Ig binding protein ( Polystyrene beads (0.2 mm diameter) of or a fragment thereof, a variant thereof or a variant thereof thereof are adhered to the polymer coating of the patterned polyvinyl alcohol on the electrode. This wafer was assembled into an I_S T A T ® cartridge having a fluid format suitable for immunoassays. On a portion of the wall of the chamber in which the cartridge contains the sample, there is a layer comprising a 160877.doc •284·201247704 conjugate that is specific for the analyte, such as an anti-analyte single/multibody antibody (or a fragment thereof, a variant thereof, or a variant thereof thereof, which binds to a binding protein or an anti-analyte half-binding protein as disclosed herein (or a fragment thereof, or a variant thereof, or a variant thereof) Variant fragments), any of which can be detected by a detectable label. In the fluid capsule of the filter cartridge is an aqueous reagent comprising a pair of amino benzene vinegar. In operation +, a sample suspected of containing an analyte is suspected. Add to the containment chamber of the test cartridge and insert the filter cartridge into the i_STAT8 reader command. After the analytical object-specific binding partner dissolves in the sample, the pump element in the filter cartridge forces the sample into In the tube containing the wafer. At this point, it is oscillated to promote the formation of a sandwich. In the penultimate step of the analysis, the fluid is forced out of the capsule and into the tube to wash the sample from the wafer and into the waste. In the chamber, in the final step of the assay, the alkaline luciferase label reacts with the aminophenol phenol phosphate to cleave the phosphate group and allows the released aminophenol to be electrochemically oxidized on the working electrode. The measured current, the reader is able to calculate the amount of analyte in the Lu sample by means of an embedded algorithm and a factory-determined calibration curve. In addition, it is apparent that the methods and kits as described herein necessarily involve the use of immunoassays. Other reagents and methods. For example, various buffers are contemplated, such as are known in the art and/or can be readily prepared or optimized for use, for example, in washing, as a binder diluent, as a particulate diluent, and / or used as a buffer for the control standard diluent. Exemplary conjugate diluents are used in certain kits (Abbott Laboratories, Abbott Park, IL) and contain 2-(N-Merlinyl) ethyl benzoate (MES), salt, protein blocker, anti-micro 160877.doc -285- 201247704 biological agent and detergent architecture® conjugate thinner ◊ exemplary control standard diluent for certain kits (Abbott Laborator Ies, ARCHITECT® Human Control Standard Diluent for use in Abb (m Park, IL), which comprises a buffer containing MES, other salts, protein blockers, and antimicrobial agents. In addition, as disclosed in U.S. Patent Publication No. As described, improved signal generation can be obtained, for example, in a I-Stat cartridge format using a nucleic acid sequence coupled to a signal antibody as a signal amplifier. It will be apparent to those skilled in the art that other methods described herein are suitable. The modifications and improvements are obvious and can be made using suitable equivalents without departing from the scope of the claimed invention or the embodiments disclosed herein. The present invention will now be described in detail, and the invention It is to be understood that the examples are included for purposes of illustration only and are not intended to limit the invention. EXAMPLES Example 1: Molecular selection of semi-Ig binding proteins Semi-immunoglobulin (semi-Ig) binding proteins were designed based on dual variable domain immunoglobulin molecules (DVD_IgTM). The semi-Ig also binds to the antigen instead of the bivalent binding target antigen (Fig. 1B). This concept applies to all immunoglobulin-like molecules involved in the ch3 contact region in dimerization. Such molecules include, but are not limited to, immunoglobulin, dual variable domain immunoglobulin (DVD_IgrM), protein, triple variable domain or triple variable domain immunoglobulin (TVDjgTM) protein, and receptor antibody (RAb). Example 1.1. Molecular Colonization of Anti-C-Met Semi-Ig Binding Proteins Hepatocyte growth factor (HGF)/c-Met pathway has been found to be associated with cancer progression due to driving proliferation, 160877.doc 201247704 motility, invasion, and angiogenesis. (See Nakamura et al., (1989) Nature 342: 440-3; Lokker et al., (1992) EMBO J. 1 1: 2503-10; Naka et al., (1992) J. Biol. Chem. 267: 201 14 -9; and Peruzzi et al., (2006) Clin. Cancer Res. 12:3 657_60, each of which is incorporated herein by reference. It is expected that targeting this pathway will inhibit cancer growth and metastasis. However, conventional c-Met antibodies are inherently efficacious due to the promotion of c-Met dimerization on the cell surface (see Prat et al., (1998) J. Cell Science 1 1 1: 237-247; Ohashi et al. (2000) Nature Med. 6: 327-331, each of which is incorporated herein by reference. The semi-Ig binding protein includes a heavy chain and a light chain which are linked to each other via a disulfide bond. As demonstrated herein, with this feature, the anti-c_Met semi-Ig-binding protein is a pure antagonist because of the monovalent binding to c-Met. Example 1.1.1: Production of anti-C-Met Ig heavy chain (HC) and light bond (LC) constructs Mouse fusion tumor HB-1 1895 (5D5.11·6) was purchased from the American Type Culture Center (American Type) Culture Collection ; ATCC » Manassas, VA). The Vh and vl cdNA sequences were cloned using methods well known in the art. The cDNA sequences and translated amino acid sequences are shown in Tables 8 and 9. Table 8: Anti-c-Met variable domain cDNA sequence

Domain Sequence identifier Cloned cDNA sequence 12345678901234567890 Anti-C-Met VH SEQ ID ΝΟ:ϊ --- CAGGTCCAACTGCAGCAGTC TGGGCCTGAGCTGGTGAGGC CTGGGGCTTCAGTGAAGATG TCCTGCAGGGCTTCGGGCTA TACCTTCACCAGCTACTGGT TGCACTGGGTTAAACAGAGG CCTGGACAAGGCCTTGAGTG GATTGGCATGATTGATCCTT CCAATAGTGACACTAGGTTT •287· 160877.doc 201247704

AATCCGAACTTCAAGGACAA GGCCACATTGAATGTAGACA GATCTTCCAACACAGCCTAC ATGCTGCTCAGCAGCCTGAC ATCTGCTGACTCTGCAGTCT ATTACTGTGCCACATATGGT AGCTACGTTTCCCCTCTGGA CTACTGGGGTCAAGGAACCT CAGTCACCGTCTCCTCA anti-c-Met VL SEQ ID NO: 2 GACTTTATGATGTCACAGTC TCCATCCTCCCTAACTGTGT CAGTTGGAGAGAAGGTTACT GTGAGCTGCAAGTCCAGTCA GTCCCTTTTATATACTAGCA GTCAGAAGAACTACTTGGCC TGGTACCAGCAGAAACCAGG TCAGTCTCCTAAACTGCTGA TTTACTGGGCATCCACTAGG GAATCTGGGGTCCCTGATCG CTTCACAGGCAGTGGATCTG GGACAGATTTCACTCTCACC ATCACCAGTGTGAAGGCTGA CGACCTGGCAGTTTATTACT GTCAGCAATATTATGCCTAT CCGTGGACGTTCGGTGGAGG CACCAAGTTGGAGCTCAAAC GG Table 9: Anti variable domain amino acid C-Met sequence

Protein amino acid sequence region sequence identifier 12345678901234567890 anti-c-Met VH SEQ ID NO: 3 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSS anti-C-Met VL SEQ ID NO: 4 DFMMSQSPSSLTVSVGEKVT VSCKSSQSLLYTSSQKNYLA WYQQKPGQSPKLLIYWASTR ESGVPDRFTGSGSGTDFTLT IT SVKADDLAVYYCQQYYAY PWTFGGGTKLELKR light and heavy chain cDNA sequences PCR amplification was performed using Platinum PCR SuperMix High Fidelity (Invitrogen, Carlsbad, CA). The PCR products were separately selected to pHyBE-hCk and pHybE-hCgl, z, non-a vector (Abbott Laboratories) " after preparation, using the ABI 3730S gene analyzer by the dideoxy bond termination method (Applied Biosystesm, Foster City, CA) Confirm the plastid sequence. Table 10 shows the full length light and heavy chain sequences. 288 -

160877.doc 201247704 Table 10: Full-length anti-c-Met HC and LC sequences

Protein amino acid sequence region sequence identifier 12345678901234567890 anti-c-Met HC SEQ ID NO: 5 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK anti-c-Met LC SEQ ID NO: 6 DFMMSQSPSSLTVSVGEKVT VSCKSSQSLLYTSSQKNYLA WYQQKPGQSPKLLIYWASTR ESGVPDRFTGSGSGTDFTLT ITSVKADDLAVYYCQQYYAY PWTFGGGTKLELKRTVAAPS VFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYS LSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC

Example 1.1.2: Generation of anti-c-Met HC chain constructs with Fc mutations Two factors play an important role in maintaining intact antibodies: disulfide bonds between the two hinge regions and non-covalent proteins between the CH3 domains - Protein interactions (see Deisenhofer (1981) Biochem. . 20: 2361-2370, which is incorporated herein by reference). The disulfide bond in the chain region is destroyed by a mutation in the cysteine in the hinge region to, for example, serine. Non-covalent protein-protein interactions between the CH3 domains can be prevented by a variety of methods. For example, random mutagenesis of a particular region of the Fc can be used to subsequently identify which mutation -289-160877.doc 201247704 or a combination of mutations produces a semi-Ig molecule for screening. Alternatively, molecular modeling assisted mutagenesis in the Fc/CH3 domain can be used to identify and mutate specific residues critical for CH3 domain interaction and dimerization. Individually exchange and test 16 interdomain contact residues of single-stranded CH3 dimers (Q347, Y349, T350, L351, T366, L368, K370, K392, T394, P395, v397, L398, D399, F405, Y407 and The alanine substitution study of each of K409) identified six contact residues (T3 66, L368, P395, F405, Y407, and K409) critical to the stability of the single-stranded CH3 domain dimer (see Dall'). Acqua et al. (1998) Biochem. 37: 9266-9273). Twenty heavy chain constructs having heavy chain variable domains from an anti-cMet antibody ligated in-frame with an Fc region comprising a combination of different mutations were prepared to investigate how to eliminate Fc dimerization. In addition to the mutation at the position of the disulfide bond in the hinge region, 1 to 6 CH3 mutations were introduced at residues important for the dimerization of the CH3 domain (see Table 11 and Table 12). Table 11: Fc mutations in the anti-c-Met HC sequence

HC CH3 mutation hinge mutation cMetHC-Mutl no mutation C226S > C229S cMetHC-Mut2 T366F, L368F, P395A, F405R, Y407R, no mutation K409D cMetHC-Mut3 T366F, L368F, P395A, F405R, Y407R, C226S, C229S K409D cMetHC-Mut4 P395A, F405R, Y407R, K409D No mutation cMetHC-Mut5 P395A, F405R, Y407R, K409D C226S 'C229S cMetHC-Mut6 P395A, F405R, Y407R, K409D C220S ' C226S cMetHC-Mut7 P395A, F405R, Y407R C226S, C229S cMetHC-Mut8 F405R , Y407R, K409D C226S, C229S 160877.doc -290- 201247704

cMetHC-Mut9 P395A, Y407R, K409D C226S, C229S cMetHC-MutlO P395A, F405R, K409D C226S 'C229S cMetHC-Mutll P395A, F405R C226S, C229S cMetHC-Mutl2 P395A, Y407R C226S > C229S cMetHC-Mutl3 P395A, K409D C226S, C229S cMetHC-Mutl4 F405R, Y407R C226S ' C229S cMetHC-Mutl5 F405R, K409D C226S ' C229S cMetHC-Mutl6 Y407R, K409D C226S, C229S cMetHC-Mutl7 P395A C226S, C229S cMetHC-Mutl8 F405R C226S ' C229S cMetHC-Mutl9 Y407R C226S, C229S cMetHC- Mut20 K409D C226S, C229S Table 12: Full-length anti-c-MetHC sequences with Fc mutations

Protein amino acid sequence region sequence identifier 12345678901234567890 cMetHC-Mutl SEQ ID NO: 7 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut2 SEQ ID NO: 8 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY 160877.doc -29Bu 201247704

ICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLFCEVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSFRLRSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut3 SEQ ID NO: 9 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLFCFVKGFYPSDIA VEWESNGQPENNYKTTPAVL DS DG S FRLRSDLTV DK SRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC -Mut4 SEQ ID NO: 10 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQ TY ICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DS DGS FRLRSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut5 SEQ ID NO: 11 QVQLQQSGPELVRPGASVKM

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SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DS DGS FRLRSDLTVDKSRWQ QGNVFSCS ^ HEALHNHYTQ KSLSLSPGK cMetHC-Mut6 SEQ ID NO: 12 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SSDKTHTSPPCPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSFRLRSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut7 SEQ ID NO: 13 QVQLQQSGPELVRPGASVKM SCR ASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS 160877.doc -293- 201247704

TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSraLRSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut8 SEQ ID NO: 14 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVS PLDYWGQGTSVTVS SA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFRLRSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut9 SEQ ID NO: 15 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TY RVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSFFLRSDLTVDKSRWQ QGNVFSCS ^ HEALHNHYTQ KSLSLSPGK cMetHC-MutlO SEQ ID NO: 16 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA

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STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSFRLYSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutll SEQ ID NO: 17 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPP cloud Pk5tLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSFRLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl2 SEQ ID NO: 18 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSS A STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL 160877.doc -295 - 201247704

DSDGSFFLRSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl3 SEQ ID NO: 19 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPAVL DSDGSFFLYSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl4 SEQ ID NO: 20 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSMKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DS ! DGSFRLRSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut 5 SEQ ID NO: 21 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK -296- 160877.doc 201247704

SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFRLYSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl6 SEQ ID NO: 22 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVS PLDYWGQGTSVTVS SA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFFLRSDLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl7 SEQ ID NO: 23 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSWTVPSSSLGTQTY ICNVNHKNTNTVVDKKKEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl8 SEQ ID NO: 24 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR 160877.doc • 297 · 201247704

PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVE PK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFRLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mutl9 SEQ ID NO: 25 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY station CNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEM TKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVL DSDGSFFLRSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPGK cMetHC-Mut20 SEQ ID NO: 26 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVK QR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPK SCDKTHTSPPSPAPELLGGP SVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKE

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YKCKVSNKALPAPIEKTISK

AKGQPREPQVYTLPPSREEM

TKNQVSLTCLVKGFYPSDIA

VEWESNGQPENNYKTTPPVL

DSDGSFFLYSDLTVDKSRWQ

QGNVFSCSVMHEALHNHYTQ —---s_____I KSLSLSPGK_ Example 1.2: Molecular selection of anti-CD28 semi-Ig binding protein Although not the only co-stimulatory receptor on T lymphocytes, cd28 was first discovered (see Hara et al., (1985) j Exp_ Med. 161:1513-24' is incorporated herein by reference and has been shown to be the most important sputum cell receptor for the initial activation of native tau cells. CD28 is a 44 kDa homodimer with an extracellular domain attached to the transmembrane region via a stalk and a cell-rich domain rich in cell-conducting motifs. CD28 is a prototype of the homodimeric receptor family, which includes inducible costimulatory factor (ICOS) and cytotoxic T lymphocyte antigen-4 (CTLA-4, CD152) (see Sharpe et al., (2002) Nat. Rev Immunol. 2: 116-26, which is incorporated herein by reference. Conventional CD28-specific mAbs have agonistic or super-potentiating effects due to their role in promoting CD28 dimerization or stabilizing lattice formation (see Hunig et al., (2005) Immunol. Lett. 100: 21-28, The manner of reference is incorporated herein). To investigate whether the monovalent half Ig binding protein is antagonistic, anti-CD28 antibody 9.3 was selected as a parental antibody constructed against the antimony molecule. Example 1.2.1: Generation of anti-CD28 Ig light and heavy chain constructs After codon optimization, the variable domain sequence of anti-CD28 antibody 9.3 was synthesized (see Damle et al., (1988) J. Immunol. 140:1753 -1761) (See Table 13 and Table 14) and used as a PCR template for subsequent steps. 160877.doc -299- 201247704 Table 13: Anti-CD28 9.3 variable domain amino acid sequence

Protein amino acid sequence region sequence identifier 12345678901234567890 anti-CD28 VH SEQ ID NO: 27 QVKLQQSGAELVKPGASVRL SCKASGYTFTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DFSGYDALPYWGQGTMVTVS S anti-CD28 VL SEQ ID NO: 14 28 DIQMTQSPASLSVSVGETVT ITCRTNENIYSNLAWYQQKQ GKSPQLLIYAATHLVEGVPS RFSGSGSGTQYSLKITSLQS EDFGNYYCQHFWGTPCTFGG GTKLEIKR Table: CD28 variable domain cDNA sequences of anti

Protein region sequence identifier codonoptimized CDNA sequence 12345678901234567890 anti-CD28 VH SEQ ID NO: 29 CAGGTGAAGCTGCAGCAGAG TGGCGCTGAGCTCGTGAAGC CTGGCGCCTCAGTCCGGCTG AGTTGTAAGGCATCAGGTTA CACGTTTACCGAGTATATTA TCCATTGGATTAAACTCAGG TCTGGGCAAGGATTGGAATG GATAGGCTGGTTCTATCCTG GATCAAATGACATCCAGTAC AATGCTAAGTTCAAGGGGAA GGCCACACTGACCGCAGACA AGTCCTCCTCTACAGTGTAT ATGGAACTCACTGGGCTGAC CAGCGAAGACAGTGCAGTGT ATTTCTGCGCGAGGAGGGAC GATTTCTCCGGTTATGACGC TCTGCCATATTGGGGGCAGG GCACGATGGTTACCGTGTCT AGC anti-CD28 VL SEQ ID NO: 30 GACATACAGATGACCCAAAG CCCGGCCTCCCTCTCCGTCT CAGTAGGGGAGACTGTAACA ATCACATGTAGGACTAATGA GAATATCTACTCTAATCTGG CGTGGTACCAGCAAAAGCAG GGCAAATCCCCCCAGCTGCT CATCTATGCTGCCACCCATC TTGTAGAAGGAGTCCCCTCT CGCTTCAGCGGCTCCGGGTC CGGGACACAATATTCTCTGA AAATTACCAGCCTCCAATCA GAAGACTTCGGGAACTACTA TTGCCAGCACTTTTGGGGAA CCCCCTGTACCTTTGGAGGC •300· 160877.doc 201247704

GGCACAAAGCTCGAGATAAA __GCGG_ PCR products were separately selected into pHyBE-hCk and pHybE-hCgl, z, non-a vectors (Abbott Laboratories). After preparation, the plastid sequence was confirmed by the dideoxy chain termination method using an ABI 3730S Genetic Analyzer (Applied Biosystesm, Foster City, CA). Table 15 shows the full length light and heavy chain sequences. Table 15: Full length anti-CD28 HC and LC sequences

Protein amino acid sequence region sequence identifier 12345678901234567890 anti-CD28 HC SEQ ID NO: 31 QVKLQQSGAELVKPGASVRL SCKASGYTFTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DFSGYDALPYWGQGTMVTVS SASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQS SGLYSLSSWTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRT PEVTCVWDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHY TQKSLSLSPGK anti-CD28 LC SEQ ID NO: 32 DIQMTQSPASLSVSVGETVTIT CRTNENIY SNLAWYQQKQGKS P QLLIYAATHLVEGVPSRFSGSG SGTQYSLKITSLQSEDFGNYYC QHFWGTPCTFGGGTKLEIKRTV AAPSVFIFPPSDEQLKSGTASV VCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTH QGLS S PVTKS FNRGEC example 1.2.2: Fc mutations generating a key anti-CD28 construct produced a weight having 5 -Fc heavy chains of different combinations of mutations in the Fc domain construct. All constructs contained two mutations (C220S, C226S) in the hinge region. The constructs pCD28HC-Mut2 and pCD28HC-Mut3 contain 4 -301 - 160877.doc 201247704 other mutations in the CH3 domain. The constructs pCD28HC-Mut4 and pCD28HC-Mut5 contain six other mutations in the CH3 domain (see Tables 16 and 17). Table 16: Fc mutations in the anti-c-Met HC sequence

HC CH3 mutation 狡 bond mutation CD28HC-Mut21 no mutation C220S, C226S CD28HC-Mut22 P395A, F405A, Y407A, K409D C220S, C226S CD28HC-Mut23 P395A, F405R, Y407R, K409D C220S ' C226S CD28HC-Mut24 T366F, L368F, P395A, F405A , Y407A ' K409D C220S, C226S CD28HC-Mut25 T366F, L368F, P395A, F405R, Y407R, K409D C220S ' C226S Table 17: Full-length anti-CD28HC sequence with Fc mutation

Protein amino acid sequence region sequence identifier 12345678901234567890 CD28HC-Mut21 SEQ station D NO: 33 QVKLQQSGAELVKPGASVRL SCKASGYT FTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DF SGYDAL PYWGQGTMVTV S SASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV SWNS GALT S GVHT F PAVLQS SGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSSDKTHTSPPCPAPELLG GPSVFLFPPKPKDTLMISRT PEVTCVWDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSR WQQGNV FS C SVMHEALHNHY TQKSLSLSPGK CD28HC-Mut22 SEQ ID NO: 34 QVKLQQSGAELVKPGASVRL SCKASGYTFTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DFSGYDALPYWGQGTMVTVS SASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV • 302 · 160877.doc 201247704

SWNSGALTSGVHTFPAVLQS SGLYSLSSWTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSSDKTHTSPPCPAPELLG GPSVFLFPPKPKDTLMISRT PEVTCVWDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTAV LDSDGSFALASDLTVDKSRW QQGNVFSCSVMHEALHNHYT QKSLSLSPGK CD28HC-Mut23 SEQ ID NO: 35 QVKLQQSGAELVKPGASVRL SCKASGYTFTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DFSGYDALPYWGQGTMVTVS SASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQS SGLYSLSSWTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSSDKTHTSPPCPAPELLG GPSVFLFPPKPKDTLMISRT PEVTCVWDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTAV LDSDGSE ^ LRSDLTVDKSRW QQGNVFSCSVMHEALHNHYT QKSLSLSPGK CD28HC- Mut24 SEQ ID NO: 36 QVKLQQSGAELVKPGASVRL SCKASGYTFTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DFSGYDALPYWGQGTMVTVS SASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV SW NSGALTSGVHTFPAVLQS SGLYSLSSWTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSSDKTHTSPPCPAPELLG GPSVFLFPPKPKDTLMISRT PEVTCVWDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRE EMTKNQVSLFCByKGFYPSD IAVEWESNGQPENNYKTTAV LDSDGSFALASDLTVDKSRW QQGNVFSCSVMHEALHNHYT 160877.doc • 303 · 201247704

QKSLSLSPGK CD28HC-Mut25 SEQ ID NO: 37 QVKLQQSGAELVKPGASVRL SCKASGYTFTEYIIHWIKLR SGQGLEWIGWFYPGSNDIQY NAKFKGKATLTADKSSSTVY MELTGLTSEDSAVYFCARRD DFSGYDALPYWGQGTMVTVS SASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVE PKSSDKTHTSPPCPAPELLG GPSVFLFPPKPKDTLMISRT PEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRE EMTKNQVSLFC | VKGFYPSD IAVEWESNGQPENNYKTTAV LDSDGSEWiRSDLTVDKSRW QQGNVFSCSVMHEALHNHYT QKSLSLSPGK Example 1.3: an anti-CD3 / CD19 half DVD-IgTM * son molecule selected from The DVD-IgTM molecular technology enables immunoglobulins to bind simultaneously to two targets or epitopes. This is particularly useful in the treatment of complex diseases in which multiple mediators contribute to disease pathogenesis through unique or redundant mechanisms. Simultaneous blockade of multiple targets may result in better therapeutic efficacy than inhibition of a single target (see Wu et al. (2007) Nature Biotech 25: 1290-1297). Similar to the antibody, the DVD-Ig assay consists of two heavy chains (HC) and two light chains (LC), and the Fc-Fc interaction plays an important role in keeping the two HC-LC heterodimers together. . To generate a monovalent semi-DVD-Ig binding protein, anti-CD3/CD19 DVD-Ig was selected as the parent molecule. Example 1.3.1: hc and LC constructs producing anti-CD3/CD19 DVD-Ig Anti-CD3 and CD19 variable domain sequences (Table 18) were obtained from U.S. Patent Application Serial No. 20070123479. After codon optimization (sequences are shown in Table 19;), • 304-160877.doc 201247704 The variable domain DNA sequences are constructed into heavy and light chains according to the method described in PCT Publication No. WO2008024188. A body with a short linker or a long linker between the two variable domains (Table 2's sequence is shown in bold). Full length DVD-IgHC and LC amino acid sequences are shown in Table 21. Table 18: Anti-CD3 and anti-CD19 variable domain amino acid sequences

Protein amino acid sequence region sequence identifier 12345678901234567890 anti-CD3 VH SEQ ID NO: 38 DIKLQQSGAELARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQLSSLTSEDSAVYYCARYY DDHYCLDYWGQGTTLTVSS anti-CD3 VL SEQ ID NO: 39 DIQLTQS PAIMSAS PGEKVT MTCRASSSVSYMNWYQQKSG TSPKRWIYDTSKVASGVPYR FSGSGSGTSYSLTISSMEAE DAATYYCQQWSSNPLTFGAG TKLELK anti-CD19 VH SEQ ID NO: 40 QVQLQQSGAELVRPGSSVKI SCKASGYAFSSYWMNWVKQR PGQGLEWIGQIWPGDGDTNY NGKFKGKATLTADESSSTAY MQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTV TVSS Anti-CD19 VL SEQ ID NO: 41 DIQLTQSPASLAVSLGQRAT ISCKASQSVDYDGDSYLNWY QQIPGQPPKLLIYDASNLVS GIPPRFSGSGSGTDFTLNIH PVEKVDAATYHCQQSTEDPW TFGGGTKLEIK Table 19: Anti-CD3 and anti-CD19 variable domain DNA sequences

Protein region sequence identifier codonoptimized CDNA sequence 12345678901234567890 anti-CD3 VH SEQ ID NO: 42 GATATCAAACTGCAGCAGTC AGGGGCTGAACTGGCAAGAC CTGGGGCCTCAGTGAAGATG TCCTGCAAGACTTCTGGCTA CACCTTTACTAGGTACACGA TGCACTGGGTAAAACAGAGG CCTGGACAGGGTCTGGAATG GATTGGATACATTAATCCTA GCCGTGGTTATACTAATTAC AATCAGAAGTTCAAGGACAA GGCCACATTGACTACAGACA AATCCTCCAGCACAGCCTAC 160877.doc -305- 201247704

ATGCAACTGAGCAGCCTGAC ATCTGAGGACTCTGCAGTCT ATTACTGTGCAAGATATTAT GATGATCATTACTGCCTTGA CTACTGGGGCCAAGGCACCA CTCTCACAGTCTCCTCA anti-CD3 VL SEQ ID NO: 43 GACATTCAGCTGACCCAGTC TCCAGCAATCATGTCTGCAT CTCCAGGGGAGAAGGTCACC ATGACCTGCAGAGCCAGTTC AAGTGTAAGTTACATGAACT GGTACCAGCAGAAGTCAGGC ACCTCCCCCAAAAGATGGAT TTATGACACATCCAAAGTGG CTTCTGGAGTCCCTTATCGC TTCAGTGGCAGTGGGTCTGG GACCTCATACTCTCTCACAA TCAGCAGCATGGAGGCTGAA GATGCTGCCACTTATTACTG CCAACAGTGGAGTAGTAACC CGCTCACGTTCGGTGCTGGG ACCAAGCTGGAGCTGAAA anti-CD19 VH SEQ ID NO: 44 CAGGTGCAGCTGCAGCAGTC TGGGGCTGAGCTGGTGAGGC CTGGGTCCTCAGTGAAGATT TCCTGCAAGGCTTCTGGCTA TGCATTCAGTAGCTACTGGA TGAACTGGGTGAAGCAGAGG CCTGGACAGGGTCTTGAGTG GATTGGACAGATTTGGCCTG GAGATGGTGATACTAACTAC AATGGAAAGTTCAAGGGTAA AGCCACTCTGACTGCAGACG AATCCTCCAGCACAGCCTAC ATGCAACTCAGCAGCCTAGC ATCTGAGGACTCTGCGGTCT ATTTCTGTGCAAGACGGGAG ACTACGACGGTAGGCCGTTA TT AC TAT GCT AT GGAC T AC T GGGGCCAAGGGACCACGGTC ACCGTCTCCTCC Anti-CD19 VL SEQ ID NO: 45 GATATCCAGCTGACCCAGTC TCCAGCTTCTTTGGCTGTGT CTCTAGGGCAGAGGGCCACC ATC TCCTGCAAGGCCAGCCA AAGTGTTGATTATGATGGTG ATAGTTATTTGAACTGGTAC CAACAGATTCCAGGACAGCC ACCCAAACTCCTCATCTATG ATGCATCCAATCTAGTTTCT GGGATCCCACCCAGGTTTAG TGGCAGTGGGTCTGGGACAG ACTTCACCCTCAACATCCAT CCTGTGGAGAAGGTGGATGC TGCAACCTATCACTGTCAGC AAAGTACTGAGGATCCGTGG ACGTTCGGTGGAGGGACCAA GCTCGAGATCAAA

160877.doc •306- 201247704 Table 20: Linker Sequences

Table 21: Full length CD3/CD19 HC and LC amino acid sequences

Protein amino acid sequence region sequence identifier 12345678901234567890 anti-CD3 / CD19-LL VH SEQ ID NO: 54 DIKLQQSGAELARPGASVKMSC KTSGYTFTRYTMHWVKQRPGQG LEWIGYINPSRGYTNYNQKFKD KATLTTDKSSSTAYMQLSSLTS EDSAVYYCARYYDDHYCLDYWG QG TT LT VSS ASTKGPSVFPLAP QVQLQQSGAELVRPGSSVKISC KASGYAFSSYWMNWVKQRPGQG LEWIGQIWPGDGDTNYNGKFKG KATLT ADE SS ST AYMQLS S LAS EDSAVYFCARRETTTVGRYYYA MDYWGQGTTVTVSSASTKGPS VFPLAPSSKSTSGGTAALGC LVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPR EPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLS PGK anti-CD3 / CD19-LL Vl SEQ ID NO: 55 DIQLTQSPAIMSASPGEKVTMT CRASSSVSYMNWYQQKSGTSPK RWIYDTSKVASGVPYRFSGSGS GTSYSLTISSMEAEDAATYYCQ QWSSNPLTFGAGTKLELKTVAA

Linker sequence identifier amino acid sequence 12345678901234567890 VH long linker amino acid sequence SEQ ID NO: 46 ASTKGPSVFPLAP νΗ long linker DNA sequence SEQ ID NO: 47 GCGTCGACCAAGGGCCCATC GGTCTTCCCCCTGGCACCC vH short linker amino acid sequence SEQ ID NO: 48 ASTKGP Vh short linker DN A sequence SEQ ID NO: 49 GCGTCGACCAAGGGCCCA VL long linker amino acid sequence SEQ ID NO: 50 TVAAPSVFIFPP VL long linker DNA sequence SEQ ID NO: 51 ACGGTGGCTGCACCATCTGT CTTCATCTTCCCGCCA vL short linker amino acid sequence SEQ ID NO: 52 TVAAP VL short linker DNA sequence SEQ ID NO: 53 ACGGTGGCTGCACCA -307· 160877.doc 201247704

PSVFIFPPDIQLTQSPASLAVS LGQRATISCKASQSVDYDGDSY LNWYQQIPGQPPKLLIYDASNL VSGIPPRFSGSGSGTDFTLNIH PVEKVDAATYHCQQSTEDPWTF GGGTKLEIKTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSF NRGEC anti-CD3 / CD19-SS VH SEQ ID NO: 56 DIKLQQSGAELARPGASVKMSC KTSGYTFTRYTMHWVKQRPGQG LEWIGYINPSRGYTNYNQKFKD KATLTTDKSSSTAYMQLSSLTS EDSAVYYCARYYDDHYCLDYWG QGTTLTVSSASTKGPQVQLQQS GAELVRPGSSVKISCKASGYAF SSYWMNWVKQRPGQGLEWIGQI WPGDGDTNYNGKFKGKATLTAD ESSSTAYMQLSSLASEDSAVYF CARRETTTVGRYYYAMDYWGQG TTVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPS SSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCP APELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHED PEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK Anti-CD3/CD19-SS VL SEQ ID NO: 57 DIQLTQSPAIMSASPGEKVTMT CRASSSVSYMNWYQQKSGTSPK RWIYDTSKVASGVPYRFSGSGS GTSYSLTISSMEAEDAATYYCQ QWSSNPLTFGAGTKLELKTVAA PDIQLTQSPASLAVSLGQRATI SCKASQSVDYDGDSYLNWYQQI PGQPPKLLIYDASNLVSGIPPR FSGSGSGTDFTLNIHPVEKVDA ATYHCQQSTEDPWTFGGGTKLE IKTVAAPSVFIFPPSDEQLKSG TASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC Example 1.3.2: generating a Fc mutations anti-CD3 / CD19 HC construct produce two heavy chains have different combinations of mutations of the construct. The two constructed 308 · 160877.doc 201247704 constructs have two mutations (C220S and C226S) in the hinge region and four mutations (P395A, F405R, Y407R and K409D) in the CH3 region. The construct pCD3/CD19HC-LL-Mut26 has a long linker between the two VH domains and the two VL domains. The construct pCD3/CD19HC-SS-Mut27 has a short linker between the two VH domains and the two VL domains (Tables 22 and 23). Four heavy chain-Fc region constructs with different combinations of mutations in the Fc region were generated. All four constructs produced had two mutations (C226S and C229S) in the hinge region and long linkages between the two VH domains and the two VL domains (Tables 22 and 23). Table 22: Fc mutations in the anti-c-Met HC sequence

HC CH3 Mutant 狡 Chain Mutation CD3/CD19HC-LL-Mut26 P395A ' F405R ' Y407R ' K409D C220S, C226S CD3/CD19HC-SS-Mut27 P395A, F405R, Y407R, K409D C220S ' C226S CD3/CD19HC-LL-Mut28 P395A, F405R , Y407R, K409D C226S > C229S CD3/CD19HC-LL-Mut29 F405R, Y407R, K409D C226S, C229S CD3/CD19HC-LL-Mut30 P395A ' K409D C226S, C229S CD3/CD19HC-LL-Mut31 F405R C226S ' C229S

Table 23: Full length anti-CD3/CD19 DVD-Ig sequences with Fc mutations

Protein amino acid sequence region sequence identifier 12345678901234567890 CD3 / CD19HC-LL-Mut26 SEQ ID NO: 58 DIKLQQSGAELARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQLSSLTSEDSAVYYCARYY DDHYCLDYWGQGTTLTVSSA STKGPSVFPLAPQVQLQQSG AELVRPGSSVKISCKASGYA FSSYWMNWVKQRPGQGLEWI GQIWPGDGDTNYNGKFKGKA T LTADE S S S TAYMQLS S LAS EDSAVYFCARRETTTVGRYY YAMDYWGQGTTVTVS SAS TK GPSVFPLAPSSKSTSGGTAA 160877.doc -309- 201247704

LGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSSD KTHTSPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPAVLDSD GSF ^ LRSDLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSL SLSPGK CD3 / CD19HC-SS-Mut27 SEQ ID NO: 59 DIKLQQSGAEIARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQLSSLTSEDSAVYYCARYY DDHYCLDYWGQGTTLTVSSA STKGPQVQLQQSGAELVRPG SSVKISCKASGYAFSSYWMN WVKQRPGQGLEWIGQIWPGD GDTNYNGKFKGKATLTADES SSTAYMQLSSLASEDSAVYF CARRETTTVGRYYYAMDYWG QGTTVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSN TKVDKKVEPKSSDKTHTSPP CPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQV YTLPPSREEMTKNQVSLTCL VKGFYPSDIAVEWESNGQPE NNYKTT PAVLDS DGS FRLRS DLTVDKSRWQQGNVFSCSVM HEALHNHYTQKSLSLSPGK CD3/CD19HC-LL-Mut28 SEQ ID NO: 60 DIKLQQSGAELARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQLSSLTSEDSAVYYCARYY DDHYCLDYWGQGTTLTVSSA STKGPSVFPIiAPQVQLQQ S G AELVRPGSSVKISCKASGYA FSSYWMNWVKQRPGQGLEWI GQIWPGDGDTNYNGKFKGKA TLTADESSSTAYMQLSSLAS EDSAVYFCARRETTTVGRYY YAMDYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSG

160877.doc •310· 201247704

ALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCD KTHTSPPSPAPELLGGPSVF LFTPiPK ^ rLMI SRTPEVTC VVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPAVLDSD GS FRLRSDLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSL SLSPGK CD3 / CD19HC-LL-Mut29 SEQ ID NO: 61 DIKLQQSGAELARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQL SS LT SE DSAVYYCARYY DDHYCLDYWGQGTTLTVSSA STKGPSVPPLAPQVQLQQSG AELVRPGSSVK station SCKASGYA FSSYWMNWVKQRPGQGLEWI GQIWPGDGDTNYNGKFKGKA TLTADESSSTAYMQLSSLAS EDSAVYFCARRETTTVGRYY YAMDYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCD KTHTSPPSPAPELLGGPSVF LFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSD GSFRLRSDLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSL SLSPGK CD3 / CD19HC-LL-Mut30 SEQ ID NO: 62 DIK LQQSGAELARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQLSSLTSEDSAVYYCARYY DDHYCLDYWGQGTTLTVSSA STKGPSVFPLAPQVQLQQSG AELVRPGSSVK work SCKASGYA FSSYWMNWVKQRPGQGLEWI GQIWPGDGDTNYNGKFKGKA TLTADESSSTAYMQLSSLAS EDSAVYFCARRETTTVGRYY YAMDYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSG 160877.doc -311 · 201247704

ALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCD KTHTSPPSPAPELLGGPSVF LFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPAVLDSD GSFFLYSDLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSL SLSPGK CD3 / CD19HC-LL-Mut31 SEQ ID NO: 63 DIKLQQSGAELARPGASVKM SCKTSGYTFTRYTMHWVKQR PGQGLEWIGYINPSRGYTNY NQKFKDKATLTTDKSSSTAY MQLSSLTSEDSAVYYCARYY DDHYCLDYWGQGTTLTVSSA STKGPSVFPLAPQVQLQQSG AELVRPGSSVKISCKASGYA FSSYWM and VKQRPGQGLEWI GQIWPGDGDTNYNGKFKGKA TLTADESSSTAYMQLSSLAS EDSAVYFCARRETTTVGRYY YAMDYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCD KTHTSPPSPAPELLGGPSVF LFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKN QVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSD GSFRLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSL SLSPGK

Example 2: Performance and Purification of Semi-Ig Binding Proteins Example 2.1: Expression and Purification of Anti-C-Met Semi-Ig Binding Proteins by Transfection Using Polyethylideneimine (Sigma, St. Louis, MO) A plastid paired from the common light chain of anti-C-Met Ab for expression of the plastids linked to the anti-c-Met heavy chain of the Fc domain including the mutations listed above to human fetal kidney 293-6E cells (US Culture Center, -312- 160877.doc 201247704

Manassas, V A) is used to transiently express semi-Ig molecules. Cell culture media were harvested 6 to 7 days after transient transfection and antibodies were purified using Protein G stratification (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. The data in Table 24 shows that, in addition to the constructs with six mutations in the CH3 domain, the amount of anti-c-Met half Ig with Fc mutations is comparable to that of the parental full-length antibody, indicating that these semi-Ig Binding proteins are effective in mammalian cells. Table 24: Anti-C-Met Ig expression bp with Fc mutation in 293 cells bg/ml) cMet Ig 22.6 cMet-Mutl 17.4 cMet-Mut2 2.0 cMet-Mut3 3.8 cMet-Mut4 12.0 cMet-Mut5 18.0 cMet-Mut6 15.0 cMet-Mut7 5.6 cMet-Mut8 20.0 cMet-Mut9 25.0 cMet-MutlO 24.0 cMet-Mutl 1 17.9 cMet-Mutl2 18.5 cMet-Mutl 3 19.5 cMet-Mutl 4 15.7 cMet-Mutl 5 19.4 cMet-Mutl 6 10.2 cMet-Mutl 7 21.2 cMet-Mutl 8 18.9 cMet-Mutl 9 30.3 cMet-Mut20 16.2

-313-160877.doc 201247704 Example 2.2: Performance and Purification of Anti-CD28 Semi-Ig Binding Proteins Transfection with Polyethylideneimine (Sigma, St. Louis, MO) will be associated with the coding from anti-CD28 Ab The plastid pairing of the light chain is used to express the plastids of the anti-CD28 heavy chain plastids linked to the Fc domain including the mutations listed above to human fetal kidney 293-6E cells (American Type Culture Collection, Manassas, VA) Used for short-term performance of half Ig. Cell culture media were harvested 6 to 7 days after transient transfection and antibodies were purified using protein G chromatography (In vitro, Carlsbad, CA) according to the manufacturer's instructions. The data in Table 25 shows that the amount of the anti-CD28 half Ig binding protein having the Fc mutation is comparable to or superior to that of the parent full length antibody, indicating that these semi-Ig binding proteins can be effectively expressed in mammalian cells. Table 25: Anti-CD28 Ig expression in 293 cells with Fc mutation (pg/ml) CD28 Ig 11.0 CD28-Mut21 18.0 CD28-Mut22 14.0 CD28-Mut23 60.0 CD28-Mut24 20.0 CD28-Mut25 10.0 Example 2.3: Anti-CD3 /CD19 Semi-DVD-Ig Binding Protein Performance and Purification Paired with plastids encoding common corresponding light chains from anti-CD3/CD19 DVD-Ig using polyethylenimine (Sigma, St. Louis, MO) The linker is paired with a long linker, and the short linker is paired with a short linker) to express the plastid of the anti-CD3/CD19 DVD-Ig heavy chain linked to the Fc domain including the mutations listed above to the human fetal kidney 293-6E cells (American strain preservation 160877.doc-314-201247704 heart, Manassas, VA) for transient expression of semi-DVD-Ig binding protein 0 Cell culture medium was collected 6 to 7 days after transient transfection, and according to manufacturing The manufacturer's instructions used protein G chromatography (Invitrogen, Carlsbad, CA) to purify the antibody. The data in Table 26 shows that the amount of anti-CD3/CD19 semi-DVD-Ig binding protein with Fc mutation is relatively lower than that of the parental full-length DVD-Ig binding protein. Table 26: Anti-CD3/CD19 DVD-Ig binding protein expression (pg/ml) showing Fc mutation in 293 cells CD3/CD19 DVD-Ig 12.4 CD3/CD19HC-LL-Mut26 1.2 CD3/CD19HC-SS-Mut27 2.5 Example 3: Characterization of physicochemical properties of semi-Ig binding proteins Example 3.1: SDS-PAGE analysis of semi-Ig binding proteins Example 3.1.1: SDS-PAGE analysis of anti-C-Met semi-Ig binding proteins under non-reducing and reducing conditions The anti-c-Met semi-Ig binding protein having the above Fc mutation was analyzed by SDS-PAGE. Under non-reducing conditions, each protein sample showed a single band. The anti-c-Met semi-Ig binding protein with Fc mutation migrates faster than the parental wild type antibody, which is consistent with the molecular weight of the semi-Ig. Under reducing conditions, each protein sample produced two bands, one corresponding to the heavy chain and the other corresponding to the light chain. The light chain from the parent antibody and the half Ig binding protein having the Fc mutation is the same. However, the heavy chain specific gravity chain wild type Fc peptide having the Fc domain peptide including the mutation migrated slightly more slowly. Without being bound by the mechanism, changes in the migration of the letter are caused by changes in configuration and/or post-translational modifications. SDS-PAGE showed that each half of the Ig binding protein was produced as a single substance, and the heavy chain and the light 160877.doc-315-201247704 chain were effectively paired to form a half-IgG binding protein-like molecule. Example 3.1.2: SDS-PAGE analysis of anti-CD28 half Ig binding protein An anti-CD28 semi-Ig binding protein having the above Fc mutation was analyzed by SDS-PAGE under non-reducing and reducing conditions. Under non-reducing conditions, Mut21 with only 2 hinge mutations (C220S and C226S) is electrophoresed with multiple bands, of which 2 major slow-moving bands are close to each other. LC dissociation. All anti-CD28 half Ig binding proteins with CH3 mutations migrate faster than the parental anti-purine or control antibodies, indicating that their CH3 mutations disrupt Fc/Fc dimerization. Under reducing conditions, each protein sample produced two bands, one corresponding to the heavy chain and the other corresponding to the light chain. The heavy and light chains of the semi-Ig binding protein have the same mobility as the parent antibody. SDS-PAGE shows that the selected CH3 mutation combination coupled to the hinge mutation produces a half IgG binding protein. Example 3.1.3: SDS-PAGE analysis of anti-CD3/CD19 half Ig binding protein An anti-CD3/CD19 semi-DVD-Ig binding protein having the above Fc mutations was analyzed by SDS-PAGE under non-reducing and reducing conditions. Under non-reducing conditions, Mut26 and Mut27 semi-DVD-Ig binding proteins with long and short linkers with two hinge mutations (C220S and C226S) and four CH3 mutations (P395A, F405R, Y407R, K409D) migrated respectively. Faster than its parent DVD-Ig, indicating that the mutation disrupts DVD-Ig dimerization. Under reducing conditions, each protein sample produced two bands, one corresponding to the heavy chain and the other corresponding to the light chain. The heavy and light chains of Mut26 and Mut27 semi-DVD-Ig binding proteins have the same mobility as the parent antibody. SDS-PAGE indicated that the selected CH3 mutation combination coupled with a hinge mutation produced a semi-Ig binding protein. 160877.doc -316- 201247704 Example 3.2: Size Exclusion Chromatography (SEC) Analysis of Semi-Ig Binding Proteins Example 3.2.1: Size Exclusion Chromatography (SEC) Analysis of Anti-c-Met Semi-Ig Binding Proteins Chromatographic (SEC) analysis was performed to confirm the semi-Ig state of the various anti-c-Met half Ig binding proteins listed above. For SEC analysis, phosphate buffered saline (PBS) containing purified parental anti-c-Met semi-Ig binding protein with Fc mutation was applied to Superdex® 200, 300 x 10 mm column (GE® Healthcare, Piscataway) , NJ). SEC was performed using a HPLC instrument 'Model 10A (Shimadzu®, Columbia, MD). All proteins were tested at 280 nm and 214 nm using UV light. Isocratic dissolution at a flow rate of 0.5 mL/min. Size exclusion chromatography demonstrated that the individual hinge mutations (Mutl, C226S and C229S) were insufficient to dissociate the antibody dimer/prevent the formation of antibody dimers. However, CH3 mutations alone (Mut2 and Mut4) are sufficient to prevent dimerization. Interestingly, Mut4 (with 4 CH3 mutations) was more effective than Mut2 (with 6 CH3 mutations) to prevent dimerization as determined by the percentage of half Ig. These results indicate that the number of mutated amino acids is not necessarily related to the half Ig state. This is additionally evidenced by other combinations of mutations in the CH3 domain. For example, a half Ig state can be achieved to varying degrees by combining 1-6 mutations in the CH3 region by hinge mutation. Mut6 with two different hinge mutations (C220S and C226S) and four CH3 mutations (P395A, F405R, Y407R, K409D) was also confirmed by the SEC as a half Ig. These data indicate that the disulfide bond between the two HCs is less important than the non-covalent interaction in the CH3 domain when the antibody is dimerized. The results are shown in Table 27 below. 160877.doc -317- 201247704 Table 27: Half Ig Percentage Constructed by SEC Half Ig% Wild Type 0 Mutl 0 Mut2 56.0 Mut3 82.4 Mut4 91.6 Mut5 97.2 Mut6 100 Mut7 98.8 Mut8 97.6 Mut9 73.5 MutlO 97.5 Mutll 94.8 Mutl 2 96.5 Mutl 3 94.9 Mutl 4 96.1 Mutl 5 97.7 Mutl 6 58.9 Mutl 7 90.1 Mutl 8 94.5 Mutl 9 94.4 Mut20 91.7

Example 3.2.2: Size Exclusion Chromatography (SEC) Analysis of Anti-CD28 Semi-Ig Binding Protein Size exclusion chromatography analysis was performed to confirm the dimerization state of the anti-CD28 half Ig binding protein. For SEC analysis, PBS containing purified anti-CD28 Ig with Fc mutation was applied to a Superdex® 200, 300 x 10 mm column (GE® Healthcare, Piscataway, NJ). SEC was performed using a HPLC instrument, Model 10A (Shimadzu®, Columbia, MD). All proteins were extracted at 280 nm and 214 nm using UV light. Isocratic dissolution at a flow rate of 160 877.doc -318 - 201247704 at 0.5 mL/min. The SEC results showed that individual hinge mutations (Mum, C22〇S and C226S) were not sufficient to prevent CH3 dimerization. Hinge mutations (C220S and C226S) combined with 4 or 6 CH3 mutations are sufficient to prevent Fc dimerization. The results are shown in Table 28 below: Table 28: Half Ig Percentage Determined by SEC Half Ig% Wild Type 0 Mut21 84.1 Mut22 0 Mut23 92.7 Mut24 98.3 Mut25 84.7 These results confirm the half shown in the previous SDS-PAGE analysis Ig status. Example 3.2.3: Size exclusion stratification (SEC) analysis of anti-CD3/CD19 semi-DVD-Ig binding proteins

Size exclusion chromatography analysis was performed to confirm the dimerization state of the anti-CD3/CD19 half Ig. For SEC analysis, PBS containing purified Mut26 and Mut27 was applied to a Superdex® 200, 300 x 10 mm column (GE® Healthcare, Piscataway, NJ). SEC was performed using a HPLC instrument, Model 10A (Shimadzu, Columbia, MD). All proteins were detected at 280 nm and 214 nm using UV light. Isocratic dissolution at a flow rate of mL 5 mL/min. SEC results showed that Mut26 with two hinge mutations (C220S and C226S) and four CH3 mutations (P395A, F405R, Y407R, K409D) and 160877.doc -319- 201247704

Mut27 migrates in the form of a molecule of about 100 kDa. The results are shown in Table 29 below: Table 29: Percent Ig as determined by SEC Constructs Half Ig% Wild type 0 Mut26 96.0 Mut27 98.5 This confirms the half Ig state revealed in the SDS-PAGE analysis. Example 3.3: Size exclusion chromatography against anti-c-Met semi-Ig binding protein - Multi-angle laser light scattering (SEC-MALLS) analysis Size exclusion chromatography - multi-angle laser light scattering (SEC-MALLS) to further characterize the resistance c-Met semi-Ig binding protein. Protein samples were separated using a Superdex 200 HR10/30 size exclusion column (GE® Healthcare, Piscataway, NJ) by HPLC 1100 quaternary pump (Agilent Technologies, Santa Clara, CA). A 50 μί sample volume was injected at 0.6 mL/min into a phosphate buffered saline mobile phase containing 1 mM sodium azide. Data were generated using a Dawn® HELEOS multi-angle light scattering detector (Wyatt Technology), a variable wavelength U\M detector (Agilent Technologies, Santa Clara, CA), and an Optilab rEX refractive index detector (Wyatt Technology). Data collection and analysis was performed using Astra v5.3.4.14 (Wyatt Technology, Santa Barbara, CA) and plotted using Origin v6.0 (Microcal Software Inc. Piscataway, NJ). The absorption coefficient was calculated from the mono-amino acid sequence of each construct using GPMAW v8.0 (ChemSW Inc.), and a refractive index increment (dn/dc) of 0.180 ml/g was used in the molecular weight calculation. 160877.doc - 320- 201247704

The wild-type c-Met antibody has a major peak with a calculated molecular weight of about 154 kDa (78%) > this is consistent with a typical IgG consisting of two light chains and two heavy chains with other slower migration peaks. The 〇-Met-Mut5, Mutll, Mutl2, Mutl4, Mutl8 and Mutl9 with different combinations of CH3 mutations showed a main peak of the experimental molecular weight and anti-c-Met semi-Ig. c-Met-Mut20 and Mutl3, each having a single mutation (K409D) and a double mutation combination (P395A, K409D) in the CH3 domain, respectively, appear to have a half-Ig binding protein and a wild-type antibody as evidenced by the slope in the calculated molecular weight. The main peak of the variable molecular weight material. Without being bound by the mechanism, the binding may reflect residual weak non-covalent interactions between the light chain/heavy chain subunits. The sample c-Met-Mutl6 with a double mutation combination (Y407R, K409D) in the CH3 domain has two main peaks (43% and 52%) consistent with the complete IgG and half Ig molecular weight. The data is summarized in Table 30 below: Table 30: Half Ig Percentage as determined by SEC-MALLS Half Ig% of the construct Wild type 0 Mut5 95 Mutll 94 Mutl2 95 Mutl3 94 Mutl4 95 Mutl6 52 Mutl8 96 Mutl9 93 Mut20 91.5 Example 3.4: Analytical ultracentrifugation (AUC) analysis of CD28-Mut23 semi-Ig binding protein 160877.doc -321 - 201247704 Analytical ultracentrifugation (AUC) was performed to further characterize the molecular weight of the solution of the CD28-Mut23 semi-Ig binding protein. Dilute all samples so that the absorbance at 280 nm in the 2.2 cm path length unit is approximately ^2 ^ sample solution and reference blank are loaded to have a 1.2 cm path length, sapphire window and carbon epoxy center cup Standard two-part unit. All samples were simultaneously inspected using a 4-well (AN-60Ti) rotor in a Beckman Pr〇te〇meLab XL-Ι analytical ultracentrifuge (sequence pli〇6C01). Allow the sample (each 435 μm) and the rotor to heat equilibrate for more than 1 hour after the temperature set point (20.0 ± 〇.rc) is reached before starting the speed operation. The sample was centrifuged at 45 rpm (168 〇〇〇>^) for 6 hours at 2 (TC). Data were collected using a 280 μηι UV source and a cm·〇〇 3 cm radial step. Use as implemented in SEDFIT The c(s) method (v 118 〇, see "Heart and Peter, Biophys. J. 2000, 78: 1606-1619, which is incorporated herein by reference) analyzes the initial settling velocity data. This method uses 1 ^ 〇 1 claw equation (1) direct fitting while simulating the influence of diffusion on the data to improve the resolution 'self-settling velocity data to derive information. dc dt \d_ r dr dc rD^.~Sc〇W dr

To avoid noise, all data sets use a maximum entropy and a 1 σ dimension adjustment fit. The result of the C(s) analysis is the distribution of the sinking coefficient, in which the area under the curve of each substance can be integrated to obtain quantitative information. After measuring the sedimentation coefficient and the diffusion coefficient (D), the molar mass can be calculated using j§vedberg equation (2). 160877.doc •322· 201247704

_ MD{\ - Up) S = ~~E~ (2) where M is the molar mass of the protein ''partial specific volume, p is the solvent density' R is the general gas constant and T is the absolute temperature. CD28- The sedimentation rate of Mut23 semi-Ig binding protein in pBS was characterized by the main factor (89.7%), which is consistent with the molecule formed by a light chain and a heavy chain bonded by disulfide bond.

The sub-quantity conversion yields a calculated value of 73 kDa for the main substance. This is within 6% of the theoretical value of the anti-(: 半 28 half Ig. The difference between these values may be the result of trying to fit a c(s) program of multiple substances with a single coefficient of friction, and there is 8.8% The polymer and i · 5% high molecular weight material will logically affect the accuracy of the calculation. The second most abundant (8.8%) material has a sedimentation coefficient consistent with the dimer of the main substance, and with two chains and two The size of the fully sized antibody consisting of the heavy chains was approximately the same. The high molecular weight material # (丨5(6) was detected in each of the three replicates and exhibited an oligomeric state distribution in the size range of 225 to 550 kDa. ^Mo (4) The average fit of the number (1.445) is close to the value usually measured for intact antibodies (1.4 to 1.6). This indicates that the half 1 § binding protein is similar to the intact antibody because it has a spherical shape with a typical coefficient of friction of about 12. The asymmetric configuration of the egg. This data indicates that the substance present in the anti-CD-2eIg binding protein sample consists of a light chain and a heavy chain < protein. 160877.doc •323· 201247704 Example 3.5: Using SEC Fractionated CD28-Mut23 Semi-Ig Binding Protein Analytical centrifugation In order to more accurately measure the molecular weight of the CD28-Mut23 semi-Ig binding protein solution and further study its solution stability, the monomer Mut23 was purified after SEC fractionation. Using TSK G300WXL column (Toso, Tokyo, Japan) at 1200 The material was fractionated on a Quaternary HPLC system (Agilent, Santa Clara, CA). The purified material was reanalyzed by SEC and the data showed successful purification. The separated monomer was partially loaded into a single centrifuge unit at 45,000 rpm (168,000 xg) Settling for 6 hours. The SEDFIT analysis was used to fit the data to a single major substance, which is the same as the monomeric anti-CD28. The calculated molecular weight of the c(M) conversion based on the sedimentation rate data of this material is 74.3 kDa, which is in the primary amino acid. The theoretical value of the sequence prediction is within 5% of 77.5 kDa. About 2% dimer and 1% high molecular weight oligomer were detected. Although these values are slightly higher than the values observed by SEC of fractionated monomers, It is significantly lower than the dimer and high molecular weight oligomer content detected in the unfractionated CD28-Mut half Ig (about 8% for the AUC, about 1% for the dimer and about 1% for the high molecular weight oligomer). In the top 6 After the settling speed was operated, the sample was allowed to stand in an ultracentrifuge for 4 days at 20 ° C. The sample unit was then removed from the rotor and mixed to redistribute the protein solution, returned to the rotor, and subjected to a second 6 hour settling. Speed operation. The results of the second operation demonstrated that the dimer or high molecular weight oligomers hardly increased after initial settling speed operation and 4 days of incubation. 160877.doc •324- 201247704 After the second 6-hour settling speed operation, the sample was allowed to stand in the ultracentrifuge at 2 〇t and the sample was dispensed for a third time. There was no significant increase in dimer or high molecular weight oligomers after 7 days at 2 ° C with two additional settling speeds. However, low molecular weight substances were detected in the velocity data. Based on the sedimentation system, the number can represent the fracture and/or single bond Fc ° absorbance data overlap from the first three scans of the sedimentation velocity operation i-3 and is displayed at 0·85 to 0ϋ28. Randomly distributed between them, which is within the noise level of the super-central optical system. This data indicates that there is no detectable loss of protein due to irreversible protein. After the third settling speed operation and 7 days under the machine, the protein solution was recovered from the ultracentrifugation and analyzed by size (4) (4) (4) to confirm the presence of the fragment material.

The protein recovered from the centrifugation unit is made up of the pre-operated weaving part of the pool (4). S(10)(4) towel clearly exists _ material speed: the fragment detected by the tearing off... Bu, no semi-Ig binding protein-Hg-I or molecular weight oligomer is detected in the sample recovered from the centrifugation unit Significantly increased, this is consistent with the results of the third settling speed operation. These data indicate that anti-CD28 hemimorphin is present as a relatively stable monomer under the conditions tested during these experiments. Although some breaks were observed after the day and three 6 hour settling speeds, there was no indication of increased dimerization, formation of high molecular weight oligomers or formation of insoluble aggregates. Example 3.6: Mass spectrometric analysis of CD28-MUt23 semi-Ig binding protein 160877.doc • 325- 201247704 To measure the complete molecular weight of the CD28-Mut23 semi-Ig binding protein, 2 pL half Ig (0.8 pg/pL) was injected into Poroshell 300 SB - C3 column (1·〇χ75 mm, 5 μπι, Agilent Technologies Inc., Pala Alto, CA). LC/MS analysis was performed on an Agilent HP1100 capillary HPLC (Agilent Technologies Inc., Pala Alto, CA) coupled to a mass spectrometer Agilent 6510 Q-Tof LC/MS system. Buffer A is 0.1 ° /. Water of formic acid, and buffer B is acetonitrile containing 0.1% formic acid. The flow rate is 50 pL/min. The separation gradient was maintained at 5% B for the first 5 minutes, increased to 95% B in 0.5 minutes and remained at 95% B for the next 9.5 minutes, then changed to 5% B in 0.5 minutes and maintained at 5% B for 4.5 minute. The mass spectrometer operates at a 5 kV spray voltage and has a scan range of 600 to 3200 mass to charge ratio. To measure the molecular weight (MW) of the light and heavy chains of the protein sample, a 10 μΐ protein sample (08 pg/pl) was reduced with a 0.2 μί 1 M DTT solution for 30 minutes at 37 °C. Light and heavy chains were separated using a porosheii 300SB-C3 column, 1.0 x 75 mm, 5 pm (Agilent Technologies Inc., Pala Alto, CA). LC/MS analysis was performed on an Agilent HP1100 capillary HPLC (Agilent Technologies Inc., Pala Alto, CA) coupled to a mass spectrometer Agilent 6510 Q-Tof LC/MS system. Buffer A is water containing 〇 1 〇 / 曱 citric acid, and buffer B is acetonitrile containing 〇 1% formic acid. The flow rate was 50 μΐ/min and the sample injection volume was 2 . The column temperature was set to 60 °C. The separation gradient starts at 5% Β; increases to 35% in 5 minutes, then increases to 65% B in 15 minutes; at! Increase to 95% B in minutes; and keep %% for 4 minutes, and drop to 5% B within 丨 minutes and keep 5% β for 5 minutes. The mass spectrometer operates at a 5 kV spray voltage and has a scan range of 6〇〇 to 160877.doc 201247704 3200 mass-to-charge ratio. The molecular mass of the experimental determination of CD28-Mut23 semi-Ig including light bonds, heavy chains and full-length proteins as shown in Table 31 was in good agreement with the predicted values. Table M· CD28-Mut23 Hemimorphin Expression and Molecular Weight Analysis Light Chain 23,478 (23,477) By Mass Spectrometry 49,228 (49,: Full Length ί! 72,706 (72,707), Light Chain, Heavy Chain and Full Length Half Ig Molecular mass

Example 4: Characterization of antigen binding properties of semi-Ig binding proteins Example 4.1: Characterization of anti-c_Met semi-Ig binding proteins by ELISA A c-Met binding ELISA was performed to characterize the antigen binding properties of anti-c-Met semi-Ig. A 96-well ELISA plate was coated overnight at 4 C with PBS containing 5 pg/mi anti-mouse IgG Fc per well. After washing 3 times with pBS/〇〇5% Tween®20, 'block the plate with 5% milk in PBS for 1 hour at room temperature' and then wash it 3 times with PBS/0-05% Tween®2〇. 100 μl of c_Met antibody or half Ig was added to each well, followed by incubation at room temperature for 1 hour. Biotin-labeled c-Met was detected by streptavidin-horseradish peroxidase (KPL, Gaithersburg, MD) and tetramethylbenzidine (Thermo Scientific, ROCkf0rd, IL). After stopping the reaction with 2 mol/L H2S04, the final absorbance was obtained at 450 nm. The data show that the cMet-Mut6 half Ig binding protein binds to its antigen with 0.46 nM of EC5Q, which is comparable to the parental c_Met antibody (EC5g = 0.36 nM) or slightly less potent than the parental c_Met antibody. Example 4·2: FACS analysis of anti-CD28 semi-Ig binding protein Jurkat E6-1 cells (ATCC, Manassas, VA) were collected and washed with PBS 160877.doc-327-201247704 and resuspended at 2 x 107 cells per ml. In ice-cold PBS with 10% FCS (GIBCO®). Anti-CD28 antibody or semi-Ig binding protein (1 μM) was added to a tube containing 100 μM of cell suspension, followed by incubation on ice for 30 minutes. The cells were washed 3 times and resuspended in ice-cold PBS, followed by staining with tritiated anti-human IgGl antibody (Southern Biotech, Birmingham, AL) according to the manufacturer's instructions. After secondary staining, the cells were washed and resuspended in ice-cold PBS containing 10% FCS. FACS analysis was performed on a FACScan (Becton Dickinson, Franklin Lakes, NJ). The data indicate that the binding of the hinge region mutation and the anti-CD28 half Ig with 4 or 6 CH3 mutations to the target antigen cell surface CD28 is comparable to the corresponding parent antibody. Example 4.3: FACS Analysis of Anti-CD3/CD19 Semi-DVD-Ig Binding Proteins Anti-CD3/CD19 Semi-DVDs were analyzed by fluorescence activated cell sorting (FACS) using conventional methods in the art, such as the methods provided herein. Binding of the Ig binding protein to the ligands CD3 and CD19. A self-organized culture flask was used to collect a stable cell strain or a tumor cell strain which excessively expresses a cell surface antigen (e.g., CD3, CD 19), and was resuspended in PBS containing 5% fetal bovine serum (PBS/FBS). Prior to staining, cells were blocked on ice using (100 μΐ) PBS/FCS containing 5 pg/ml human IgG. On ice, cells (1-5 χ 105) were incubated with PBS/FBS containing antibody or DVD-Ig or semi-Ig (2 pg/mL) for 30-60 minutes. Wash the cells twice and add the appropriate secondary antibody, eg 100 μΐ F(ab') 2 goat anti-human IgG, Fey-phycoerythrin (1:200 fold diluted in PBS) (Jackson ImmunoResearch®, West Grove, PA) ). After incubation on ice for 30 minutes, the cells were washed twice and resuspended in PBS/FBS. Use Becton Dickinson FACSCaliburTM (Becton Dickinson, 160877.doc -328- 201247704

San Jose, CA) measures fluorescence. It will be appreciated that cells that naturally or heterologously express two cell surface antigens bound by a semi-DVD-Ig binding protein can be similarly tested for half-DVD-Ig binding proteins directed against different cell surface antigens. Example 5: In vitro characterization of biological activity of semi-Ig binding proteins Example 5.1: Characterization of the elbow in the c-Met phosphonium assay half of the binding protein A549 cells (ATCC, Manassas, VA) were seeded in 24-well plates (c 〇ming, Lowell, ΜΑ), and grow until it reaches 9〇% to 100% confluence. The cells were serum starved for 18 to 20 hours. C_Met was induced to phosphorylate with 200 ng/ml human growth factor (HGF) (R&D Systems, Minneapolis, MN) for 10 minutes. To test for antibody inhibition, the cells were pre-incubated with the parental 5D5 anti-cMet antibody or anti-cMet mut6 semi-Ig binding protein for an hour, followed by the addition of HGF. Immediately after incubation with HGF, the cells were placed on ice and dissolved. The fill, c-Met, was measured using a sandwich ELISA (R&D Systems, Minneapolis, MN). The degree of phosphorylation of c-Met is low in the absence of HGF stimulation. As expected, HGF induced c-Met phosphorylation via the HGF/c-Met signaling pathway. The parental 5D5 c-Met antibody acts as an agonist due to bivalent binding to c-Met and induces c-Met phosphorylation comparable to HGF alone. Co-administration of 5D5 antibody with HGF also stimulated phosphorylation of c-Met. However, it is interesting to note that cMet-Mut6 semi-Ig binding protein, which is administered alone or in combination with HGF-administered monovalent c-Met, acts as a CMet-Mut6 half Ig binding protein administered alone or in combination with HGF, 5D5 antibody or HGF and 5D5 antibodies. Antagonists and inhibit c-Met squamation. Example 5.2: Characterization of anti-CD3/CD19 semi-DVD-Ig binding protein in a redirected cytotoxicity assay (rCTL) 160877.doc - 329- 201247704

Human CD3+ T cells were isolated from PBMC by negative selection of a thickened column (R&D Systems, Minneapolis, MN). T cells were plated in flasks coated with 10 pg/mL OKT-3 anti-CD3 antibody (BD, Franklin Lakes, NJ) and 2 pg/ml CD28.2 anti-CD28 antibody (Abeam, Cambridge, MA) in complete RPMI medium. Stimulate for 4 days. T cells were allowed to stand overnight in 301;/1111^11^2 (Peprotech, Rocky Hill, NJ) and subsequently used in the assay. Raji B lymphocytes were labeled with the cell membrane dye PKH26 (Sigma, St. Louis, MO) according to the manufacturer's instructions. Phenol-free RPMI 1640 medium (Invitrogen®, Carlsbad, CA) containing 1% L-face prosamine (Invitrogen®, Carlsbad, CA) and 10% FBS (Hyclone) was used throughout the rCTL assay. Effector T cells (E) and target Raji cells (T) were seeded in 96-well plates (Corning®, Lowell, sputum) at 105 and 104 cells per well, respectively. The anti-CD3/CD19 DVD-Ig binding protein and the semi-DVD-Ig binding protein were appropriately diluted to obtain a concentration-dependent titration curve. After overnight incubation, the cells were pelleted and washed once with PBS, then resuspended in 0.1% BSA (Invitrogen®, Carlsbad, CA) and 0.5 pg/mL moth-activated (BD, ·

Franklin Lakes, NJ) in PBS. FACS data were collected on a FACSCantoTM machine (BD, Franklin Lakes, NJ) and analyzed in Flowjo software (Treestar, Ashland, OR) to detect redirection of cytotoxic cytolytic products. The data indicate that both Mut26 and Mut27 semi-DVD-Ig binding proteins play a role in redirecting cytotoxic cytolysis, while Mut27 (IC50 = 52.4 pM) with a long linker combination between variable domains has a shorter linker combination. 160877.doc •330· 201247704

Mut26 is more effective in redirecting cytotoxic cytolysis. Mut26 and Mut27 semi-DVD-Ig binding proteins were less potent in redirecting cytotoxic cytolysis than the parental DVD-Ig (IC50 = 4.3 pM). Example 5.3: Characterization of semi-DVD-Ig binding proteins in a redirected cytotoxicity assay (rCTL) such as FACS-based assays (Dreier, T. et al., 2002.

Cawcer 100: 690-697) and impedance-based analysis (Zhu, J. et al., 2006. J Immunological Methods 309: 25-33) i%# t X. Φ] I® Toxicity Assay β For FACS-based Analytical method, human CD3+ T cells were isolated from previously frozen PBMCs by negative selection of a thickened column (R&D catalog number HTCC-525). T cells were coated with complete RPMI medium (L-glutamine, 55 mM β-ΜΕ) containing 10 pg/mL anti-CD3 (OKT-3, BD) and 2 pg/mL anti-CD28 (CD28.2, Abeam) , Pen/Strep, 10% FCS) was stimulated for 4 days in a flask. T cells were allowed to stand overnight in 30 U/mL IL-2 (Peprotech) and subsequently used in the analysis. DoHH2 or Raji target cells were labeled with PKH26 (Sigma) according to the manufacturer's instructions. RPMI 1640 medium containing L-glutamine indole and 10% FBS (Hyclone) (Invitrogen®) was used for the entire rCTL analysis. Effect T cells (E) and target (T) were 105 cells per well. And 104 cells per well were seeded in 96-well plates (Costar® #3799) to produce a 10:1 E:T ratio. The semi-DVD-Ig binding protein was appropriately diluted to obtain a concentration-dependent titration curve. After overnight incubation, cells were pelleted and washed once with PBS and subsequently resuspended in PBS containing 0.1% BSA (Invitrogen®) and 0.5 160877.doc -331 - 201247704 pg/mL propidium iodide (BD). FACS data was collected on a FACSCanto machine (BD) and analyzed in Flowjo (Treestar). The percentage of live targets in the semi-DVD-Ig binding protein treated samples was calculated by dividing the total target percentage (control 'untreated) to determine the specific lysis percentage. The data was plotted and the IC50 was calculated in Prism (Graphpad). The target cells expressing EGFR were adhered to an ACEART-CES96 well plate (ACEA Bio, San Diego) overnight for impedance-based assays such as the preparation of tau cells as described above. The effector T cells (E) and target (T) were then seeded at 2 χ 105 cells per well and 2 x 104 cells per well to produce an e:T ratio of 1〇:1. The DVD-Ig molecule was appropriately diluted to obtain a concentration-dependent titration curve. The cell index of the target in the sample treated with the semi-DVD-Ig binding protein was divided by the cell index of the control target (untreated) to calculate the specific dissolved percentage. The data was plotted and the IC50 was calculated in Prism (GraPhPad). The data for this experiment is shown in Table 32 and indicates that the semi-DVD-Ig binding protein plays a role in redirecting cytotoxic cytolysis. Table 32. Restricted cytotoxicity (rCTL) with half Ig binding protein DVD-Ig ID N-terminal variable domain (VD) C-terminal variable domain (VO) Tumor target cell rCTL IC50 (pM) assay */2〇¥ 0857 CD3 CD19 DoHH2 (DSMZ ACC47) 52 FACS /-0859 CD3 EGFR A431 (ATCC CRL-1555) 7.5 Impedance • ΛϋνϋδόΟ EGFR CD3 A431 (ATCC CRL-1555) 1240 Impedance '/2DVD018 CD3 EGFR A431(ATCC CRL-1555) 33 Impedance ViOvmn HER-2 CD3 N87 (ATCC CRL-5822) 62221 Impedance 160877.doc -332- 1 Native Human PBMC Effector Example 5.4: Cytokine Release Assay 201247704 Ability to cause cytokine release from parent antibody or DVD-Ig And the ability of the semi-DVD-Ig binding protein to prevent the release of cytokines. Peripheral ash is drawn from a healthy donor (e.g., at least 3) to a heparin-treated vacuum blood collection tube by venipuncture. #rPMI-1640 medium 1: 5 times diluted whole blood' and placed in a 24-well tissue culture dish at 0.5 mL per well. An anti-cytokine antibody (eg, anti-IL-4) was diluted in RPMI-1640 and placed in a culture dish at 0.5 ml per well to obtain 2〇〇, 1〇〇, 5〇, 10, and 1叩/... Final concentration. The final dilution of whole blood in the culture dish was 1:1 〇β lipopolysaccharide (LPS) and polyhydroxyalkanoate (ΡΗΑ) were added to each well at a final concentration of 2 gg/mL and 5 gg/mL. A positive control for cytokine release. Multiple human IgGs were used as negative control antibodies. The experiment was performed in duplicate. The plates were incubated at 37 C at 5% C〇2. After 24 hours, the contents of each well were transferred to a test tube, and centrifuged at 1200 rpm for 5 minutes (35 〇 Xg) to collect the cell-free supernatant and freeze. The cells remaining on the culture plate and in the test tube were dissolved in a 血5 blood-dissolved solution, and placed at -20 ° C and thawed. Add medium (〇5 balances the volume of cells and non-cell samples and lysates, and cold-bundle samples. Thaw cell-free supernatants and cell lysates and analyze their cells by ELISA using conventional methods (eg, commercially available kits) Hormone content, for example, content of -8, IL-6, IL-Ιβ, IL-1RA or TNF-α. Example 6: Characterization of Fc function of a half-ig-binding protein Example 6.1: Anti-battering The neonatal Fc receptor (FcRn) is present in the small intestine and on vascular endothelial cells. The neonatal Fc receptor is located in the acidic endosome, in which the neonatal & receptor binds to the Fc portion, ie, has been borrowed Absorbed by the pinocytosis 1 § (5. . ^ ^ followed by 160877.doc -333 · 201247704

FcRn is released back to the cell surface. This process allows control of IgG transport through a single layer of epithelial barrier and protects IgG molecules from catabolism, thereby affecting IgG half-life (see Blumberg RS and Lencer WI. Nature Biotechnology, 2005, 23: 1232-1234, by way of citation Incorporated herein). CHO-FcRnGPI cells (stabilized FcRn receptor expression) and CHO-pBudll cells (non-FcRn representation) were aliquoted into 96-well culture dishes at 1 x 10 cells per well. The cells were resuspended in 150 μM FACS buffer. Anti-c-Met Ab and semi-Ig binding proteins were diluted to 100 gg/ml in FACS buffer pH 6.4 and pH 7.4. 30 μM of diluted antibody or half Ig binding protein was added to each well and incubated on ice for 1 hour. After washing twice with FACS buffer, add 50 μΐ secondary antibody R-phycoerythrin-conjugated AfiniPure F(ab') 2 fragment goat anti-human IgG diluted in FACS buffer pH 6.4 or ρΗ7·4 (Jackson ImmunoResearch) ®, West Grove, PA). The mixture was incubated on ice for 30 minutes. The cells were resuspended in 100 μl of PBS containing 1% FBS (Invitrogen®, Carlsbad, CA) and 2 pg/ml propofol (Invitrogen®, Carlsbad, CA) with corresponding pH. Final FACS data was obtained on a FACScan (Becton Dickinson, Franklin Lakes, NJ). The wild-type c-Met antibody showed strong binding to the FcRn receptor. Interestingly, cMet-Mutl with only two hinge mutations (C226S, C229S) showed extensive binding profiles, suggesting that hinge mutations may have a minor impact on FcRn binding capacity. For all constructs with 3-6 mutations in the CH3 domain, the FcRn receptor binding function is completely or almost completely lost. However, cMet-Mutl 1 and cMet-Mutl 8 with two CH3 mutations (P395A, F405R) and a single CH3 mutation (F405R), respectively, showed relatively strong FcRn receptor binding. 160877.doc • 334· 201247704 Two fronts were detected from the binding profiles of cMet-Mutl3 and cMet-Mut20, which may indicate the presence of two or more populations in such purified antibody/semi-Ig binding proteins. Four constructs (Mutl2, Mutl4, Mutl5, Mutl9) showed weak binding to the FcRn receptor. In certain instances, the presence of a strong FcRn-IgG interaction in the therapeutic composition is preferred because the higher affinity FcRn-IgG interaction extends the half-life of the IgG and Fc coupling drugs in serum. These improved pharmacokinetics will reduce the frequency of administration of individual antibodies and reduce patient risk and discomfort.

Table 33 · FcRn binding analysis of cMet semi-Ig binding protein. EC50〇iM) at molecular pH 6.4 cMet-Mutl 1,088 cMet-Mut3 1,129 cMet-Mut5 3,286,000 cMet-MutlO 292,804 cMet-Mutl 1 4,697,000 cMet-Mutl3 630 cMet-Mutl 4 1,445 cMet-Mutl 8 1,977,000 cMet-Mutl 9 2,981 cMet-Mut20 1,227 Control mAb 50.37 Example 6.2: Fcy binding of anti-c-Met semi-Ig binding protein The human FcyR system is regulated by activation (FCyRi, FcyRiia, FcyRiHa) and inhibition (FqRIIb) Body composition. FqRI is a high-affinity IgGi receptor expressed on monocytes and macrophages and a variety of monocyte cell lines (including ΤΉι> 1). FcyRIIa is a more widely expressed low affinity IgG receptor and is also present on K562 cells. Both FcyRI and FcyRIIa are activating molecules. FeYRlIb is widely expressed on effector cells and inhibits IgG receptors with low affinity. 0 at 160877.doc •335· 201247704

When FcyRIIb binds to IgG complexes, calcium-dependent processes such as degranulation, swallowing, ADCC, cytokine release, and pro-inflammatory activation are blocked (see Ravetch JV et al., Annu. Rev. Immunol. 2001, 19: 275-290, which is incorporated herein by reference. THP-1 cells, K562 cells or CHO-FcyRII-b-1 cells were aliquoted into 96-well culture plates at a ratio of lxlO5 cells per well for FcyRI, FcyRIIa or FcyRIIb binding assays. Diluted c-Met antibody or semi-Ig binding protein (30 μM) was added to each well, followed by incubation on ice for 1-2 hours. The cells were washed twice with 150 μΐ FACS buffer. After staining for 30 minutes on ice with the secondary antibody R-phycoerythrin-conjugated AHniPure F(ab')2 fragment goat anti-human IgG (Jackson ImmunoResearch®, West Grove, PA), add 50 to each well in each well. ί PBS containing 4% trimeric (Electron Microscopy Sciences, Hatfield, PA). After final strip washing, cells were resuspended in 100 μί PBS 1% FBS for analysis and analyzed on a FACScanTM (Becton Dickinson, Franklin Lakes, NJ). Example 6.3: Clq binding of anti-c-Met semi-Ig binding protein The complement complex component C1 q binding of a semi-Ig binding protein can be performed using conventional methods to identify half or Ig of activated or non-activated complement. Briefly, antibodies were diluted to various concentrations (eg, 8, 4, 2, 1, 0.5, and 0.125 g/mL in PBS, total volume 325 μΐ). Diluted half Ig and appropriate controls (e. g., parent antibody, non-specific antibody) were aliquoted (100 μM each) into wells. The plates were incubated overnight at 4 °C. 200 μί TBS with SuperBlockTM Blocking Buffer in each well (Thermo Fisher Scientific, 160877.doc -336- 201247704

Waltham, ΜΑ) washed 4 times. Add Cql to HBSS (Invitrogen®, Carlsbad, CA) '0.1% BSA (Sigma, St. Louis, MO) '0.02% Tween® 20 (JT Baker, Phillipsburg NJ) up to 2 pg/mL and to each well Add 50 gL of diluted Clq. The plates were incubated for 3 hours at room temperature and then washed 4 times with TTBS. Cql protein was detected using 50 μΐ of 1:2000 HBSS, 0.1% BSA, 0.02% Tween® 20 buffer containing anti-huClq-HRP (Biogenesis, Mill Creek, WA). The plates were incubated for 1 hour at room temperature with shaking and then washed 4 times with TTBS. The plates were developed for 10 minutes using a TMB substrate kit (Thermo Fisher Scientific, Waltham, MA). The reaction was stopped with 100 gL of 1 N NaS〇4 (J.T. Baker, Phillipsburg NJ), and the reaction product was detected by reading the sample OD45G.65Q using a plate reader. Example 7: Pharmacokinetic analysis of semi-Ig binding proteins Pharmacokinetic studies were performed in Sprague-Dawley rats and Balb/c mice. Male and female rats and mice are administered intravenously in a single dose, for example 4 mg/kg for rats and 5 mg/kg for mice. Semi-Ig binding proteins and samples were analyzed using antigen capture ELISA and pharmacokinetic parameters were determined by non-compartmental analysis. Briefly, ELISA plates were coated with goat anti-biotin antibody (5 mg/m b 4 ° C, overnight), blocked with SuperBlockTM (Pierce, Rockford, IL) and at 50 ng/ml at room temperature 10% SuperBlockTM TTBS of biotinylated human antigen was incubated for 2 hours. Serum samples were serially diluted (0.5° in TTBS, serum, 10% SuperBlockTM) and incubated on the plate for 30 minutes at room temperature for detection by HRP-labeled goat anti-human antibodies, and by standard The curve was determined using a four parameter logistic fit of 160877.doc -337 - 201247704. Pharmacokinetic parameter values were determined by a non-compartment model using WinNonlin® software (Pharsight Corporation, Mountain View, CA). The pharmacokinetic profiles of cMet semi-Ig in SD rats and Balb/c mice are similar. In all half Ig, cMet semi-Ig mutlO has a longest half-life (Τι/2)' of 5-6 days similar to the parent cMet mAb. Mutl 1, mutl 4 and mutl8 have a Tm shorter than 1 day. All cMet half Ig showed a rapid clearance. Without being bound by the mechanism, this is due, at least in part, to the reduced ability of the FcRn binding of the mutants reported in Example 6. Table 34: Pharmacokinetic profile of cMet semi-Ig binding protein in rats in Spogdogi *Tm (days) Vss (mL/Kg) CL (mL/hr/kg) Cmax (pg/mL) AUCinf (hr *mg/mL) cMet mAb 7.7 82.9 0.3 111.4 11.8 cMet semi-Ig mutlO 5.8 423.3 9.0 95.1 0.5 cMet semi-Ig mutl 1 0.7 61.8 12.2 97.5 0.3 cMet semi-Ig mutl4 0.6 64.8 17.0 83.2 0.2 cMet semi-Ig mutl 8 0.8 73.5 12.7 94.1 0.3 * The harmonic mean and pseudo standard deviation used. Table 35: Pharmacokinetic profile of cMet semi-Ig binding protein in Balb/c mice *T1/2 Vss CL Cmax AUCinf (days) (mL/Kg) (mL/hr/kg) (pg/mL) ( Hr*mg/mL) cMet semi-Ig mutlO 6.1 183.6 6.70 50.1 0.74 cMet semi-Ig mutl 1 0.8 32.5 8.38 44.2 0.61 cMet semi-Ig mutl4 0.3 18.1 9.40 41.5 0.54 cMet semi-Ig mutl 8 0.6 30.1 8.45 44.5 0.60 * Harmonic average used And pseudo standard deviation. 160877.doc -338 - 201247704 Example 8: Other Semi-Ig Binding Proteins The semi-Ig technique described in the previous examples can be applied to many other formats of protein molecules to produce novel semi-Ig binding proteins as provided herein. Specific examples of semi-Ig binding proteins are provided below. Example 8.1 Molecular Construction of Semi-Ig Binding Proteins The same method as described in the previous examples was used to generate other semi-Ig binding proteins, such as: semi-triple variable domain immunoglobulin (1/2 TVD) binding protein, semi-scFv-Fc Binding protein, half domain antibody (DA)-Fc binding protein and semi-φ receptor (R)-Fc binding protein. Briefly, the PCR products of the relevant protein binding domains were separately colonized into the appropriate pHyBE vector (Abbott Laboratories). The plastid sequence was confirmed by the dideoxy bond termination method using an ABI 3730S Genetic Analyzer (Applied Biosystesm, Foster City, CA). The cDNA sequences of the respective domains (constructed blocks) of the final molecule and the translated amino acid sequences are shown in Table 36 and Table 37. The structural information of the semi-Ig binding protein and the control protein is shown in Table 38. Table 36: Binding sequences of the half-Ig binding protein of various formats, constant or Fc region 瘫 cDNA sequence

Protein region / domain sequence identifier cDNA sequence 12345678901234567890 IL12 / IL18 / PGE2 TVD VH SEQ ID NO: 64 GAGGTCACCTTGAGGGAGTC TGGTCCTGCGCTGGTGAAAC CCACACAGACCCTCACACTG ACCTGCACCTTCTCTGGGTT CTCACTCAGCAAATCTGTTA TGGGTGTGAGCTGGATCCGT CAGCCCCCAGGGAAGGCCCT GGAGTGGCTTGCACACATTT ACTGGGATGATGACAAGTAC TATAATCCATCCCTAAAGAG CAGGCTCACCATCTCCAAGG ACACCTCCAAAAACCAGGTG GTCCTTACAATGACCAACAT GGACCCTGTGGACACAGCCA -339- 160877.doc 201247704

CGTATTATTGTGCACGGAGA GGGATACGAAGTGCTATGGA CTATTGGGGGCAAGGGACCA CGGTCACCGTCTCCTCAGCG TCGACCAAGGGCCCAGAGGT GCAGCTGGTGCAGTCTGGAA CAGAGGTGAAAAAACCCGGG GAGTCTCTGAAGATCTCCTG TAAGGGTTCTGGATACACTG TTACCAGTTACTGGATCGGC TGGGTGCGCCAGATGCCCGG GAAAGGCCTGGAGTGGATGG GATTCATCTATCCTGGTGAC TCTGAAACCAGATACAGTCC GACCTTCCAAGGCCAGGTCA CCATCTCAGCCGACAAGTCC TTCAATACCGCCTTCCTGCA GTGGAGCAGTCTAAAGGCCT CGGACACCGCCATGTATTAC TGTGCGCGAGTCGGCAGTGG CTGGTACCCTTATACTTTTG ATATCTGGGGCCAAGGGACA ATGGTCACCGTCTCTTCAGC GTCGACCAAGGGCCCAGAGG TGCAGCTGGTGCAGAGCGGC GCCGAGGTGAAGAAGCCCGG CGCCAGCGTGAAGGTGAGCT GCAAGGCCAGCGGCTACACC TTCACCAAGTACTGGCTGGG CTGGGTGCGGCAGGCCCCCG GCCAGGGCCTGGAGTGGATG GGCGACATCTACCCCGGCTA CGACTACACCCACTACAACG AGAAGTTCAAGGACCGGGTG ACCCTGACCACCGACACCAG CACCAGCACCGCCTACATGG AGCTGCGGAGCCTGCGGAGC GACGACACCGCCGTGTACTA CTGCGCCCGGAGCGACGGCA GCAGCACCTACTGGGGCCAG GGCACCCTGGTGACCGTGAG CAGC hCgl-Mutl8 SEQ ID NO: 65 GCGTCGACCAAGGGCCCATC GGTCTTCCCCCTGGCACCCT CCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTG CCTGGTCAAGGACTACTTCC CCGA ACCGGTGACGGTGTCG TGGAACTCAGGCGCCCTGAC CAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCA GGACTCTACTCCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCA CAAGCCCAGCAACACCAAGG TGGACAAGAAAGTTGAGCCC AAATCTTGTGACAAAACTCA CACATCACCACCGTCTCCAG CACCTGAACTCCTGGGGGGG •340- 160877.doc 201247704

CCGTCAGTCTTCCTCTTCCC CCCAAAACCCAAGGACACCC TCATGATCTCCCGGACCCCT GAGGTCACATGCGTGGTGGT GGACGTGAGCCACGAAGACC CTGAGGTCAAGTTCAACTGG TACGTGGACGGCGTGGAGGT GCATAATGCCAAGACAAAGC CGCGGGAGGAGCAGTACAAC AGCACGTACCGTGTGGTCAG CGTCCTCACCGTCCTGCACC AGGACTGGCTGAATGGCAAG GAGTACAAGTGCAAGGTCTC CAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCG AGAACCACAGGTGTACACCC TGCCCCCATCCCGCGAGGAG ATGACCAAGAACCAGGTCAG CCTGACCTGCCTGGTCAAAG GCTTCTATCCCAGCGACATC GCCGTGGAGTGGGAGAGCAA TGGGCAGCCGGAGAACAACT ACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTT CCGGCTCTACAGCAAGCTCA CCGTGGACAAGAGCAGGTGG CAGCAGGGGAACGTCTTCTC ATGCTCCGTGATGCATGAGG CTCTGCACAACCACTACACG CAGAAGAGCCTCTCCCTGTC TCCGGGTAAATGA hCgl SEQ ID NO: 66 GCGTCGACCAAGGGCCCATC GGTCTTCCCCCTGGCACCCT CCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTG CCTGGTCAAGGACTACTTCC CCGAACCGGTGACGGTGTCG TGGAACTCAGGCGCCCTGAC CAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCA GGACTCTACTCCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCA CAAGCCCAGCAACACCAAGG TGGACAAGAAAGTTGAGCCC A AATCTTGTGACAAAACTCA CACATGCCCACCGTGCCCAG CACCTGAACTCCTGGGGGGA CCGTCAGTCTTCCTCTTCCC CCCAAAACCCAAGGACACCC TCATGATCTCCCGGACCCCT GAGGTCACATGCGTGGTGGT GGACGTGAGCCACGAAGACC CTGAGGTCAAGTTCAACTGG TACGTGGACGGCGTGGAGGT GCATAATGCCAAGACAAAGC CGCGGGAGGAGCAGTACAAC AGCACGTACCGTGTGGTCAG 160877.doc -341 - 201247704

CGTCCTCACCGTCCTGCACC AGGACTGGCTGAATGGCAAG GAGTACAAGTGCAAGGTCTC CAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCG AGAACCACAGGTGTACACCC TGCCCCCATCCCGCGAGGAG ATGACCAAGAACCAGGTCAG CCTGACCTGCCTGGTCAAAG GCTTCTATCCCAGCGACATC GCCGTGGAGTGGGAGAGCAA TGGGCAGCCGGAGAACAACT ACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTT CTTCCTCTACAGCAAGCTCA CCGTGGACAAGAGCAGGTGG CAGCAGGGGAACGTCTTCTC ATGCTCCGTGATGCATGAGG CTCTGCACAACCACTACACG CAGAAGAGCCTCTCCCTGTC TCCGGGTAAATGA IL12 / IL18 / PGE2 TVD VL SEQ ID NO: 67 GACATCGTGATGACCCAGTC TCCAGACTCCCTGGCTGTGT CTCTGGGCGAGAGGGCCACC ATCAACTGCAAGGCCAGTCA GAGTGTGAGTAATGATGTAG CTTGGTACCAGCAGAAACCA GGACAGCCTCCTAAGCTGCT CATTTACTATGCATCCAATC GCTACACTGGGGTCCCTGAC CGATTCAGTGGCAGCGGGTC TGGGACAGATTTCACTCTCA CCATCAGCAGCCTGCAGGCT GAAGATGTGGCAGTTTATTA CTGTCAGCAGGATTATAACT CTCCGTGGACGTTCGGCGGA GGGACCAAGGTGGAGATCAA ACGTACGGTGGCTGCACCAG AAATAGTGATGACGCAGTCT CCAGCCACCCTGTCTGTGTC TCCAGGGGAAAGAGCCACCC TCTCCTGCAGGGCCAGTGAG AGTATTAGCAGCAACTTAGC CTGGTACCAGCAGAAACCTG GCCAGGCTCCCAGGCTCTTC ATCTA TACTGCATCCACCAG GGCCACTGATATCCCAGCCA GGTTCAGTGGCAGTGGGTCT GGGACAGAGTTCACTCTCAC CATCAGCAGCCTGCAGTCTG AAGATTTTGCAGTTTATTAC TGTCAGCAGTATAATAACTG GCCTTCGATCACCTTCGGCC AAGGGACACGACTGGAGATT AAACGAACGGTGGCTGCACC AGACGTGCTGATGACCCAGA CCCCCCTGAGCCTGCCCGTG ACCCCCGGCGAGCCCGCCAG CATCAGCTGCACCAGCAGCC

160877.doc -342 · 201247704

AGAACATCGTGCACAGCAAC GGCAACACCTACCTGGAGTG GTACCTGCAGAAGCCCGGCC AGAGCCCCCAGCTGCTGATC TACAAGGTGAGCAACCGGTT CAGCGGCGTGCCCGACCGGT TCAGCGGCAGCGGCAGCGGC ACCGACTTCACCCTGAAGAT CAGCCGGGTGGAGGCCGAGG ACGTGGGCGTGTACTACTGC TTCCAGGTGAGCCACGTGCC CTACACCTTCGGCGGCGGCA CCAAGGTGGAGATCAAGCGG hCk SEQ ID NO: 68 ACGGTGGCTGCACCATCTGT CTTCATCTTCCCGCCATCTG ATGAGCAGTTGAAATCTGGA ACTGCCTCTGTTGTGTGCCT GCTGAATAACTTCTATCCCA GAGAGGCCAAAGTACAGTGG AAGGTGGATAACGCCCTCCA ATCGGGTAACTCCCAGGAGA GTGTCACAGAGCAGGACAGC AAGG AC AGC ACC TAC AGC CT CAGCAGCACCCTGACGCTGA GCAAAGCAGACTACGAGAAA CACAAAGTCTACGCCTGCGA AGTCACCCATCAGGGCCTGA GCTCGCCCGTCACAAAGAGC TTCAACAGGGGAGAGTGTTG A CD3 scFv SEQ ID NO: 69 GATATCAAACTGCAGCAGTC AGGGGCTGAACTGGCAAGAC CTGGGGCCTCAGTGAAGATG TCCTGCAAGACTTCTGGCTA CACCTTTACTAGGTACACGA TGCACTGGGTAAAACAGAGG CCTGGACAGGGTCTGGAATG GATTGGATACATTAATCCTA GCCGTGGTTATACTAATTAC AATCAGAAGTTCAAGGACAA GGCCACATTGACTACAGACA AATCCTCCAGCACAGCCTAC ATGCAACTGAGCAGCCTGAC ATCTGAGGACTCTGCAGTCT ATTACTGTGCAAGATATTAT GATGATCATTACTGCCTTGA CTACT GGGGCCAAGGCACCA CTCTCACAGTCTCCTCAGTC GAAGGTGGAAGTGGAGGTTC TGGTGGAAGTGGAGGTTCAG GTGGAGTCGACGACATTCAG CTGACCCAGTCTCCAGCAAT CATGTCTGCATCTCCAGGGG AGAAGGTCACCATGACCTGC AGAGCCAGTTCAAGTGTAAG TTACATGAACTGGTACCAGC AGAAGTCAGGCACCTCCCCC AAAAGATGGATTTATGACAC ATCCAAAGTGGCTTCTGGAG TCCCTTATCGCTTCAGTGGC 160877.doc • 343 - 201247704

• AGTGGGTCTGGGACCTCATA CTCTCTCACAATCAGCAGCA TGGAGGCTGAAGATGCTGCC ACTTATTACTGCCAACAGTG GAGTAGTAACCCGCTCACGT TCGGTGCTGGGACCAAGCTG GAGCTGAAA ILip DAI SEQ ID NO: 70 GAAGTTCAGCTGTTGGAAAG CGGCGGAGGTTTGGTGCAGC CTGGAGGGTCTCTCCGGCTC TCTTGTGCCGCATCAGGGTT TACCTTCGCTGATGAGGGAA TGATGTGGGTTCGGCAGGCC CCAGGAAAGGGACTGGAGTG GGTGTCACGAATCACCTATA GCGGCAAGAATACCTACTAT GCCGACTCCGTGAAAGGGCG GTTTACCATTTCACGCGACA ACAGTAAGAACACCCTGTAC CTGCAAATGAATTCACTCCG CGCGGAAGACACTGCGGTGT ACTACTGCGCGAAATATACA GGTCGGATTCTGGGACACCA TCTGTTCGACTACTGGGGAC AAGGCACCTTGGTCACAGTC TCTTCA IL1P DA2 SEQ ID NO: 71 GAGGTGCAACTGCTCGAATC TGGCGGGGGACTGGTACAAC CTGGGGGTAGCCTTCGACTC AGCTGCGCCGCCTCCGGATT TACCTTCGCCGAGGAAAGCT GGATGTGGGTGAGACAGGCG CCTGGAAAAGGGCTGGAGTG GGTGAGTCGGATTGGTCAGG ATGGAAAAAACACCTATTAC CGAGAGGATGTGAAGGGACG ATTCACCATATCCAGGGATA ATAGTAAAAACACTCTGTAT CTCCAGATGAACAGCCTTCG CGCGGAAGACACCGCCGTCT ACTATTGTGCTAAGTACACT GGACGGATCATGGGCCATCA TCTGTTTGATTACTGGGGCC AGGGGACATTGGTGACCGTT TCCTCA hTNFaR SEQ ID NO:72 ATGGCCCCTGTTGCCGTCTG GG CTGCGTTGGCTGTGGGGC TGGAACTGTGGGCCGCAGCG CACGCTCTGCCTGCTCAGGT AGCATTTACTCCTTACGCAC CCGAGCCCGGATCCACGTGC AGACTCAGGGAGTATTATGA TCAAACCGCACAGATGTGTT GTAGCAAATGTAGCCCAGGC CAGCATGCCAAGGTGTTCTG CACCAAGACCTCCGATACAG TCTGTGATAGCTGTGAGGAT TCTACCTACACACAGCTCTG GAACTGGGTTCCGGAGTGCT TGTCCTGTGGTTCTCGGTGC • 344- 160877.doc 201247704

AGCAGCGATCAAGTCGAAAC TCAGGCCTGCACCCGAGAGC AGAATCGCATCTGTACCTGC AGACCGGGTTGGTACTGCGC ACTGTCAAAACAGGAAGGTT GCCGACTGTGCGCCCCACTC AGAAAGTGCAGGCCCGGATT CGGAGTTGCTAGACCCGGAA CAGAAACCAGTGATGTGGTT TGTAAACCTTGTGCTCCGGG GACCTTTAGTAACACAACTA GCAGCACGGACATCTGCAGA CCCCACCAGATCTGCAATGT TGTGGCAATTCCAGGAAACG CCTCCATGGATGCCGTCTGC ACTTCAACCTCTCCCACGAG AAGTATGGCTCCCGGCGCCG TGCATCTCCCGCAGCCGGTG TCAACTCGGTCCCAGCATAC CCAGCCCACACCAGAGCCAA GCACCGCCCCTTCAACTTCA TTTTTGCTGCCAATGGGACC CTCCCCACCAGCCGAAGGAA GCACTGGCGAC hCTLA4ECD SEQ ID NO: 73 ATGGGCGTTCTGCTGACACA GAGGACGCTGCTTTCACTCG TGTTGGCTCTCCTCTTCCCG TCTATGGCGTCTATGGCAAT GCACGTGGCCCAGCCGGCGG TCGTTCTGGCTTCCTCAAGA GGAATCGCATCTTTTGTTTG CGAATATGCCTCTCCAGGCA AGGCAACAGAAGTCCGAGTA ACGGTCCTCAGACAGGCCGA TTCCCAGGTGACAGAGGTCT GTGCCGCTACTTACATGATG GGCAATGAACTGACATTTCT GGATGATTCAATCTGCACCG GCACCTCCAGCGGTAATCAA GTGAATCTTACCATCCAGGG CCTTCGCGCCATGGATACAG GACTGTATATCTGCAAAGTG GAACTGATGTATCCGCCTCC CTACTATCTGGGAATCGGAA ACGGGACACAAATATATGTG ATCGATCCCGAACCGTGTCC CGATAGCGAC Fc-Mutl 8 SEQ ID NO: 74 GAGCCCAAATCTAGCGACAA AACTCACACATCACCACCGT CTCCAGCACCTGAACTCCTG GGGGGGCCGTCAGTCTTCCT CTTCCCCCCAAAACCCAAGG ACACCCTCATGATCTCCCGG ACCCCTGAGGTCACATGCGT GGTGGTGGACGTGAGCCACG AAGACCCTGAGGTCAAGTTC AACTGGTACGTGGACGGCGT GGAGGTGCATAATGCCAAGA CAAAGCCGCGGGAGGAGCAG TACAACAGCACGTACCGTGT 160877.doc • 345 · 201247704

GGTCAGCGTCCTCACCGTCC TGCACCAGGACTGGCTGAAT GGCAAGGAGTACAAGTGCAA GGTCTCCAACAAAGCCCTCC CAGCCCCCATCGAGAAAACC ATCTCCAAAGCCAAAGGGCA GCCCCGAGAACCACAGGTGT ACACCCTGCCCCCATCCCGC GAGGAGATGACCAAGAACCA GGTCAGCCTGACCTGCCTGG TCAAAGGCTTCTATCCCAGC GACATCGCCGTGGAGTGGGA GAGCAATGGGCAGCCGGAGA ACAACTACAAGACCACGCCT CCCGTGCTGGACTCCGACGG CTCCTTCCGGCTCTACAGCA AGCTCACCGTGGACAAGAGC AGGTGGCAGCAGGGGAACGT CTTCTCATGCTCCGTGATGC ATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTC CCTGTCTCCGGGTAAATGA Fc SEQ ID NO: 75 GAGCCCAAATCTTGTGACAA AACTCACACATGCCCACCGT GCCCAGCACCTGAACTCCTG GGGGGACCGTCAGTCTTCCT CTTCCCCCCAAAACCCAAGG ACACCCTCATGATCTCCCGG ACCCCTGAGGTCACATGCGT GGTGGTGGACGTGAGCCACG AAGACCCTGAGGTCAAGTTC AACTGGTACGTGGACGGCGT GGAGGTGCATAATGCCAAGA CAAAGCCGCGGGAGGAGCAG TACAACAGCACGTACCGTGT GGTCAGCGTCCTCACCGTCC TGCACCAGGACTGGCTGAAT GGCAAGGAGTACAAGTGCAA GGTCTCCAACAAAGCCCTCC CAGCCCCCATCGAGAAAACC ATCTCCAAAGCCAAAGGGCA GCCCCGAGAACCACAGGTGT ACACCCTGCCCCCATCCCGC GAGGAGATGACCAAGAACCA GGTCAGCCTGACCTGCCTGG TCAAAGGCTTCTATCCCAGC GACATCGCCGTGGAGTGG GA GAGCAATGGGCAGCCGGAGA ACAACTACAAGACCACGCCT CCCGTGCTGGACTCCGACGG CTCCTTCTTCCTCTACAGCA AGCTCACCGTGGACAAGAGC AGGTGGCAGCAGGGGAACGT CTTCTCATGCTCCGTGATGC ATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTC CCTGTCTCCGGGTAAATGA

160877.doc 346- 201247704 Table 37: Amino acid sequence of the binding domain, constant or Fc region of a semi-Ig binding protein

Protein region / domain of the amino acid sequence a sequence identifier 12345678901234567890 IL12 / IL18 / PGE2 TVD VH SEQ ID NO: 76 EVTLRESGPALVKPTQTLTL TCTFSGFSLSKSVMGVSWIR QPPGKALEWL7VHIYWDDDKY YNPSLKSRLTISKDTSKNQV VLTMTNMDPVDTATYYCARR GIRSAMDYWGQGTTVTVSSA STKGPEVQLVQSGTEVKKPG ESLKISCKGSGYTVTSYWIG WVRQMPGKGLEWMGFIYPGD S ETRYS PT FQGQVTISADKS FNTAFLQWS S LKAS DTAMYY CARVGSGWYPYTFDIWGQGT MVTVSSASTKGPEVQLVQSG AEVKKPGASVKVSCKASGYT FTKYWLGWVRQAPGQGLEWM GDIYPGYDYTHYNEKFKDRV TLTTDTSTSTAYMELRSLRS DDTAVYYCARSDGSSTYWGQ GTLVTVSS hCgl -Mutl8 SEQ ID NO: 77 ASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEP KSCDKTHTSPPSPAPELLGG PSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPV LDSDGSE ^ LYSKLTVDKSRW QQGNVFSCSVMHEALHNHYT QKSLSLSPGK hCgl SEQ ID NO: 78 ASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEP K SCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYT QKSLSLSPGK IL12 / IL18 / PGE2 TVD VL SEQ ID NO: 79 DIVMTQSPDSLAVSLGERAT -347 · 160877.doc 201247704

INCKASQSVSNDVAWYQQKP GQPPKLLIYYASNRYTGVPD RFSGSGSGTDFTLTISSLQA EDVAVYYCQQDYNSPWTFGG GTKVEIKRTVAAPEIVMTQS PATLSVSPGERATLSCRASE SISSNLAWYQQKPGQAPRLF IYTASTRATDIPARFSGSGS GTEFTLTISSLQSEDFAVYY CQQYNNWPSITFGQGTRLEI KRTVAAPDVLMTQTPLSLPV TPGEPASISCTSSQNIVHSN GNTYLEWYLQKPGQS PQLLI YKVSNRFSGVPDRFSGSGSG TDFTLKISRVEAEDVGVYYC FQVSHVPYTFGGGTKVEIKR hCk SEQ station D NO: 80 TVAAPSVFIFPPSDEQLKSG TASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLS S PVTKS FNRGEC CD3 scFv SEQ ID NO: 81 MEFGLSWLFLVAILKGVQCD IKLQQSGAELARPGASVKMS CKTSGYTFTRYTMHWVKQRP GQGLEWIGYINPSRGYTNYN QKFKDKATLTTDKS S STAYM QLSSLTSEDSAVYYCARYYD DHYCLDYWGQGTTLTVSSVE GGSGGSGGSGGSGGVDDIQL TQSPAIMSASPGEKVTMTCR ASSSVSYMNWYQQKSGTSPK RWIYDTSKVASGVPYRFSGS GSGTSYSLTISSMEAEDAAT YYCQQWSSNPLTFGAGTKLE LK ILip DAI SEQ ID NO: 82 MEFGLSWLFLVAILKGVQCE VQLLESGGGLVQPGGSLRLS CAASGFTFADEGMMWVRQAP GKGLEWVSRITYSGKNTYYA DSVKGRFTISRDNSKNTLYL QMNSLRAEDTAVYYCAKYTG RILGHHLFDYWGQGTLVTVS S IL13 DA2 SEQ ID NO: 83 MEFGLSWLFLVAILKGVQCE VQLLESGGGLVQ PGGSLRLS CAASGFTFAEESWMWVRQAP GKGLEWVSRIGQDGKNTYYR EDVKGRFTISRDNSKNTLYL QMNSLRAEDTAVYYCAKYTG RIMGHHLFDYWGQGTLVTVS S hTNFaR SEQ ID NO: 84 MAPVAVWAALAVGLELWAAA HALPAQVAFTPYAPEPGSTC RLREYYDQTAQMCCSKCSPG QHAKVFCTKTSDTVCDSCED STYTQLWNWVPECLSCGSRC SSDQVETQACTREQNRICTC RPGWYCALSKQEGCRLCAPL RKCRPGFGVARPGTETSDVV

160877.doc -348 - 201247704

CKPCAPGTFSNTTSSTDICR PHQICNWAIPGNASMDAVC T ST S PTRSMAPGAVHLPQPV STRSQHTQPTPEPSTAPSTS FLLPMGPSPPAEGSTGD hCTLA4ECD SEQ ID NO: 85 MGVLLTQRTLLSLVLALLFP SMASMAMHVAQPAVVLAS SR GI AS FVCEYAS PGKATEVRV TVLRQADSQVTEVCAATYMM GNELTFLDDSICTGTSSGNQ VNLTIQGLRAMDTGLYICKV ELMYPPPYYLGIGNGTQIYV IDPEPCPDSD Fc-Mutl8 SEQ ID NO: 86 EPKSSDKTHTSPPSPAPELL GGPSVFLFPPKPKDTLMISR TPEVTCVWDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQ YNSTYRWSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTP PVLDSDGSE ^ LYSKLTVDKS RWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK Fc SEQ ID NO: 87 EPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISR TPEVTCVWDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQ YNSTYRWSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKS RWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK table 38: semi-Ig binding protein and the structural sequence the parent molecule of the protein molecules IL12 / IL18 / PGE2 TVD heavy chain: SEQ ID NO: 76 + SEQ ID NO:78 light chain: SEQ ID NO: 79 + SEQ ID NO: 80 '/jILn/ILlS/PG ElTVD heavy chain: SEQ ID NO: 76 + SEQ ID NO: 77 light chain: SEQ ID NO: 79 + SEQ ID NO: 80 CD3 scFv Fc SEQ ID NO: 81 + SEQ ID NO: 87 V2 CD3 scFv Fc SEQ ID NO :81 + SE〇ID NO:86 ILlbDAl Fc SEQ ID NO:82 + SEQ ID NO:87 »/2 ILlb DAI Fc SE〇ID NO:82 + SEQ ID NO:86 ILlb DA2 Fc SE〇ID NO:83 + SE〇ID NO:87 »/2 ILlb DA2 Fc SE〇ID NO:83 + SE〇ID NO:86 hTNFaRFc SEQ ID NO:84 + SEQ ID NO:87 160877.doc •349· 201247704 */2 hTNFaR Fc SE 〇ID NO:84 + SE〇ID NO:86 hCTLA4ECD Fc SEQ ID NO:85 + SEQ ID NO:87 '/2 hCTLA4ECD Fc SEQ ID NO:85 + SEQ ID NO:86 Example 8.2 Performance and Purification Semi-TVD, Half The performance and purification scheme of scFv-Fc, semi-DA-Fc and semi-R-Fc binding protein half Ig binding proteins are described in Example 2. Performance data is shown in Table 39 below. Table 39. Transient expression of semi-Ig binding protein in 293 cells. Protein molecule expression amount hg/ml) IL12/IL18/PGE2TVD 10.5 */2 IL12/IL18/PGE2 TVD 0.3 CD3 scFv Fc 1.5 '/2 CD3 scFv Fc 10.0 ILlb DAI Fc 109 '/2 ILlb DAI Fc 107 ILlb DA2 Fc 278 */2 ILlb DA2 Fc 100 hTNFaR Fc N/A •/2 hTNFaR Fc 40 hCTLA4ECD Fc N/A */2 hCTLA4ECD Fc 87 Example 8.3 Half Ig TVD, half Characterization of physicochemical properties of Ig scFv-Fc, semi-Ig DA-Fc and semi-Ig R-Fc binding proteins All semi-Ig binding proteins were subjected to SDS-PAGE analysis and SEC analysis according to the same protocol as in Example 3. All molecules showed the expected molecular weight in both PAGE gel analysis and SEC analysis. The percentage of monomers is shown in Table 40 〇 160877.doc • 350· 201247704 Table 40: Half Ig Percentage Determined by SEC Half Ig% IL12/IL18/PGE2TVD 78 */2 IL12/IL18/PGE2 TVD 78 CD3 scFv Fc 77 */2 CD3 scFv Fc 58 ILlb DAI Fc 93 */2 ILlb DAI Fc 82 ILlb DA2 Fc 89 */2 ILlb DA2 Fc 83 hTNFaRFc 100 */2 hTNFaR Fc 55 hCTLA4ECD Fc 98 '/2 hCTLA4ECD Fc 98 Example 8.4 Characterization of the semi-Ig binding protein CD3scFvFc by FACS The binding properties of the semi-CD3 scFv Fc were characterized by FACS analysis using Jurkat cells. The detailed procedure is described in Example 4.2. The binding properties of the semi-CD3 scFv Fc are similar to the parental molecule CD3 scFv Fc. Both show an effective combination of singular digits in the nM range.

Table 41. FACS analysis of half hCTLA4 ECD Fc CD3 scFv Fc */2 CD3 scFv Fc EC50 (nM) 2.00 6.45 Example 8.5 Characterization of half-hCTLA4ECDFc by ELISA Analysis of semi-Ig binding protein hCTLA4 ECD Fc compared to human IgG in a sandwich ELISA assay (Jackson Immunoresearch®, West Grove, PA) or CTLA4-Fc (R&D Systems, Minneapolis, MN) in combination with human B7.1. EIA plates (Corning®, Lowell) coated with 1 pg/ml human gamma globulin, hfTLA4ECD Fc 160877.doc -351 - 201247704 or semi-Ig hCTLA4ECD Fc carbonate-argon carbonate buffer (Pierce, Rockford, IL) , ΜΑ) » The plate was then blocked with SuperBlock® (Pierce, Rockford, IL) and hB7.1-Fc titrated in PBST. Biotin-labeled mouse anti-hB7.1 (R&D Systems, Minneapolis, MN) was used for indifference binding and developed using HRP-conjugated streptavidin (Pierce, Rockford, IL). OD45 was read on a 190 (Molecular Devices, Sunnyvale, CA). And use Graphpad Prizm software to analyze data and plots. The data show that the semi-Ig hCTLA4ECD Fc effectively binds to its ligand, human B7.1, however, the binding of the semi-Ig to its ligand is weaker than the parental molecule hCTLA4ECD Fc.

Table 42. Direct binding ELISA of the semi-Ig binding protein hCTLA4 ECD Fc hCTLA4ECD Fc */2 hCTLA4ECD Fc EC50 (pg/ml) 0.036 0.53

Example 8.6: Characterization of the semi-Ig binding protein IL12/IL18/PGE2 TVD using cell-based bioassay Example 8.6.1 PGE2 Bioassay To determine the potency of TVD semi-Ig binding protein against PGE2, EP4 HEKG al6#2 cells were used. FLIPR analysis. EP4 HEKG al6#2 cells were seeded in black/transparent poly-D-lysate plates (Corning®) at 3 x 104 cells per well. The cells were incubated for 15 minutes at room temperature to allow them to evenly settle. The plates were incubated overnight at 37 ° C, 5% C02. The FLIPR is turned on 30 minutes before use. A FLIPR buffer consisting of lxHBSS (Invitrogen®), 20 mM HEPES (Invitrogen®), 0.1% BSA, and 2.5 mM Probenecid 160877.doc • 352-201247704 (Sigma) was prepared. A disposable dye-free stock solution was prepared and vortexed by adding 10 mL of water to a disposable dye powder (Molecular Devices). The stock solution of the disposable dye was diluted 1:10 in FLIPR buffer. The medium was removed from the plate and 80 μL of lx dye was added to each well. The samples were incubated on a slow shaker for 1.5 hours at room temperature. 200 standard strength ethanol containing PGE2 was diluted in FLIPR buffer from a 5 mM stock solution concentration. The antibody (or TVD-Ig) was diluted to 1000 ng/ml in FLIPR buffer. The antibody (or TVD-Ig) was serially diluted 1:3. PGE2 and antibody (or TVD-Ig) were combined and diluted in FLIPR buffer. 20 μM PGE2/antibody was added to each well and the sample was read on FLIPR. 8.6.2 KG-1 bioassay The potency of TVD-Ig semi-Ig binding protein against rhIL-18 was measured by KG-1 assay. The KG-1 cell line is a human acute myeloid leukemia cell line (ATCC catalog number CCL-246). Serial dilutions of mAb 2.5 or TVD-Ig were prepared in complete RPMI 1640 (10% FBS, 2 mM L-glutamic acid, 50 units/ml penicillin, 50 mg/ml clematis, and 0.075% sodium arhydride). Antibody dilutions were pre-incubated with recombinant human IL-18 (2 pg/mL) in 100 μL at 37 °C for 1 hour in 96-well tissue culture dishes (Costar®). KG-1 cells (100 μΐ) were seeded at a density of 1·〇·3.〇χ105 cells per well in the presence of 20 ng/mL TNF-α, and cultured at 37 ° C, 5% C02 16-20 hour. After incubation, cell-free supernatants were collected and the amount of human IFN-γ was measured by standard ELISA (R&D Systems). The percent inhibition was plotted against antibody concentration relative to the 2 ng/ml rIL-18 control. Analysis of the self-inhibition curve was determined by sigmoidal curve fitting IC5 〇 value. 160877.doc • 353 - 201247704 8.6.3 Human IL-12 bioassay The human IFN-γ line is released from PHA blast cells in a concentration-dependent manner in response to human IL-12 stimulation. To determine the neutralization potency, TVD-Ig was tested in the assay at a final concentration range of 10-7 Μ to 10 _14 在 in the presence of 200 pg/mL rhIL-12. 50 g of TVD semi-Ig binding protein was preincubated with 50 μM of human IL-12 in a 96-well flat-bottom microtiter plate for 1 hour at 37 °C in RPMI complete medium. The frozen blast cells were thawed and washed twice in the medium, followed by trypan blue counting. The cells were adjusted to a density of 2.5 χ 106 cells/ml in the medium. Subsequently, 100 μί PHA embryo cells were added to the TVD-Ig + IL-12 mixture. The final concentration of human IL-12 in the assay was 200 pg/mL. The mixture was incubated at 37 ° C, 5% CO 2 for 18 hours, after which the IFN-γ content in the supernatant was measured by human IFN-γ ELISA. The concentration of IFN-γ was plotted against the concentration of Ig (TVD-Ig 003 or monoclonal antibody) using the GraphPad Prism software to generate an IC5 〇 value. Each measurement was performed in quadruplicate and each experiment was performed at least twice. Table 43: IL-12 and IL-18 Neutralization Methods for TVD-Ig and TVD Semi-Ig Characterization

Ig neutralizing IL-12IC5〇(nM) neutralizing IL-18IC50(nM) control monoclonal antibody 6.105 0.1795 IL12/IL18/PGE2 TVD 4.565 0.5794 Semi-IL12/IL18/PGE2 TVD 12.52 0.9182 In cell-based bioassays, It was demonstrated that IL12/IL18/PGE2 TVD half Ig was able to neutralize IL-12 and IL-18 with a similar effect as the parental antibody alone (Table 43). 160877.doc -354· 201247704 Example 8.7: Affinity determination using BIAcore® technology Table 44: Reagent antigens used in BIAcore® analysis Supplier Nomenclature Supplier Catalog No. TOFa Recombinant Human TNF-a/TNFSF 1A R&D systems 210- TA IL-Ιβ Recombinant Human IL-Ιβ R&D systems 201-LB Example 8.7.1 BIAcore® Method BIAcore® Analytical Method (BIAcore®, Inc, Piscataway, NJ) Using Kinetic Measurements of Association Rate and Dissociation Rate Constant The affinity of the antibody is determined. Flow HBS-EP (10 mM HEPES [pH 7.4] ' 150 mM NaCl was used at 25 ° C on a BIAcore® 1000 or 3000 instrument (BIAcore® AB, Uppsala, Sweden) based on surface plasmon resonance measurements. '3 mM EDTA and 0.005% Surfactant P20) Determination of binding of an antibody to a target antigen (eg, purified recombinant target antigen) All chemicals were obtained from BIAcore® AB (Uppsala, Sweden) or otherwise obtained from the differences described herein source. For example, a standard amine coupling kit using a standard amine coupling kit according to the manufacturer's instructions and procedures will be diluted to 1 〇 1111 sodium acetate (1) 114.5) to approximately 5 〇〇〇 111 of 25 0§/1111; goat anti-mouse An IgG (FcY) fragment-specific polyclonal antibody (pierce Biotechnology Inc, Rockford, IL) was directly immobilized on a CM5 research grade biosensor wafer. The unreacted portion of the surface of the biosensor is blocked with ethanolamine. The surface of the modified carboxymethyl dextran in the flow cells 2 and 4 was used as a reaction surface. Flow trough! And unmodified carboxymethyl dextran having no goat anti-mouse IgG in 3 was used as a reference surface. For kinetic analysis, the rate equation derived from the 1:1 Langmuir binding model was simultaneously fitted to the association phase and dissociation phase of all 8 injections using Biae valuation 4.0.1 software. 160877.doc •355· 201247704 (using global fit analysis). The purified antibody was diluted in HEpES buffered saline to capture on the entire goat anti-mouse IgG specific reaction surface. The antibody (25 pg/ml) to be captured as a ligand was injected onto the reaction matrix at a flow rate of 5 μΐ/min. The association and dissociation rate constants kc > n (M-ls, and the rate constants were determined by kinetic binding measurements at different antigen concentrations ranging from 10 to 200 nM at a continuous flow rate of 25 μl / ηιίη. The equilibrium equilibrium dissociation constant between the antibody and the target antigen is then calculated by the automatic mechanical rate constant of the following formula ()^1): Kd = kt >ff/k〇n. Record the binding with time and calculate the kinetic rate constant In this analysis, the association rate of up to 1 〇6 M·1 s·1 can be measured and the dissociation rate as slow as 1 〇·6 s·1 is shown. Table 45. BIACORE analysis structure of semi-Ig Antigen IL-12 k〇n(MV) IWs·1) k〇(M) */2 IL12/IL18/PGE2 TVD 4.〇xl〇5 5.1X10'5 1.3xlO·10 '/2IL12/IL18/PGE2 TVD AB352 IL-18 1.3x10s 5.9xl0'5 4.4xlO'10 WhTNFaRFc TNF 1.9xl06 1.3xl〇·5 6.9xlO'10 ILlb DAI Fc IL-lb 1.5x10s 4.〇xl O'6* 2.7X10·11 '/2 ILlb DAI Fc IL-lb 1.2xl05 6.4x1 O'6* 5.3x10·" ILlb DA2 Fc IL-lb 3.4xl〇5 3.9XHT6* 1.2X10·11 */2 ILlb DA2 Fc IL-lb 1.7xl05 4.9x1 O '6* 2.8χ1〇·" Characterized by BIAcore® technology There are binding construct are maintained and comparable to full-length body build. For those structures with * after the dissociation rate, the dissociation rate reaches the dissociation limit after 1 hour of dissociation time. Example 8.8: Neutralization of huTNFcx L929 cells were grown to semi-confluent density and harvested using 0.05% trypsin (Gibco). The cells were washed with PBS, counted and resuspended in an assay medium containing 4 gg/mL actinomycin D in 6 160877.doc • 356· 201247704 cells per ml. The cells were seeded in 96-well plates (Costar®) at a volume of 50 pL and 5 x 1 Ο 4 cells per well. Samples were diluted to 4x concentration in assay medium and 1:3 fold serial dilutions were prepared. huTNFa was diluted to 400 pg/mL in assay medium. Antibody samples (200 μM were added to huTNFa (200 pL) in a 1:2 dilution procedure and incubated for 0.5 hours at room temperature. Sample/huTNFa solution was added to 100 μί of inoculated cells until a final concentration of 100 pg /mL huTNFa and 150 nM - 0.00014 η Μ samples. Each plate was incubated for 20 hours at 37 ° C, 5% CO 2 . To quantify the viability, 100 μιη was removed from each well and 10 pL of WST-1 reagent (Roche) was added. Each plate was incubated for 3.5 hours under assay conditions, centrifuged at 50 〇xg and transferred to 75 μL of supernatant to a ELISA plate (Costar). Read on a Spectromax® 190 ELISA disk reader at 〇D420-600 nm Each plate was taken. Example 8.9: IL-Ια/β bioassay and neutralization assay] \411€5 cells were inoculated at 1.5-2 per well<1〇4 cells in a volume of 1〇〇41^ and Incubate overnight at 37 ° C, 5% CO 2 . Prepare 20 pg/mL antibody working stock (4x concentration) in complete MEM medium. 8 point serial dilution in complete MEM in Marsh dilution plate (5 pg/mL -0.0003 pg/mL) Add 65 μM of each antibody dilution to each well in a 96-well v-type chassis (Costar®). Add 65 pL of 200 pg/mL IL-la or IL-Ιβ solution or 65 μί of a mixed solution containing 50 pg/mL IL-la solution and IL-Ιβ solution. Control wells accept 65 μί 200 pg/ml IL-la Or IL-Ιβ or 50 pg/mL mixed] Χ-1α/β (4-fold concentration) plus 65 μί MEM medium, and medium control well 160877.doc •357- 201247704 received 130 μί medium. After 1 hour of incubation, to MRC5 Add 100 μί Ab/Ag mixture to the cells. The volume of all wells is equal to 200 μί. Then all plate reagents are concentrated 1 time. After 16-20 hours of incubation, the contents of the wells (150 μΙ〇 are transferred to a 96-well round chassis (Costar) And placed in a -20 ° C freezer. The supernatant was tested for hIL by using a human IL-8 ELISA kit (R&D Systems, Minneapolis, MN) or hIL-8 chemiluminescence kit (MDS). -8 content. Neutralization potency was determined by calculating the percent inhibition relative to IL-la, IL-Ιβ or IL-Ια/β control values alone. The results are shown in Table 46. Table 46: Use of IL-Ιβ parent Antibody and semi-Ig binding protein construction of IL-ip neutralization assay construct IL-lp neutralization assay EC50 nM control antibody 0.0059 */ 2 ILlb DAI Fc 0.1546 ILlb DA2 Fc 0.0013 '/2 ILlb DA2 Fc 1.174 All constructs showed neutralization in the MRC5 IL-ΙΙα/β neutralization assay. Example 9: Characterization of cMet semi-Ig binding protein with FcRn binding mutation Example 9.1: Molecular colonization of cMet semi-Ig binding protein with FcRn binding mutations Mutant combinations T250Q/M428L and T3 07A/N434A/E3 80A were introduced into the construct pHyBE-cMetHC-Mutl8. The Fc mutations of the final construct are shown in Table 47. 160877.doc -358 - 201247704 Table 47: FcRn binding mutations in the cjet half Ig binding protein jjc sequence

HC Fc mutation hinge mutation cMetHC-Mutl8-QL T250Q > F405R > M428L C226S 'C229S cMetHC-Mutl 8-GTv2 T307A, E380A, F405R, N434A C226S, C229S Table 48: cMet half Ig binding protein VH with Fc binding mutation And constant region cDNA sequences

Protein region / domain sequence identifier cDNA sequence 12345678901234567890 cMet vH SEQ ID NO: 88 CAGGTCCAACTGCAGCAGTC TGGGCCTGAGCTGGTGAGGC CTGGGGCTTCAGTGAAGATG TCCTGCAGGGCTTCGGGCTA TACCTTCACCAGCTACTGGT TGCACTGGGTTAAACAGAGG CCTGGACAAGGCCTTGAGTG GATTGGCATGATTGATCCTT CCAATAGTGACACTAGGTTT AATCCGAACTTCAAGGACAA GGCCACATTGAATGTAGACA GATCTTCCAACACAGCCTAC ATGCTGCTCAGCAGCCTGAC ATCTGCTGACTCTGCAGTCT ATTACTGTGCCACATATGGT AGCTACGTTTCCCCTCTGGA CTACTGGGGTCAAGGAACCT CAGTCACCGTCTCCTCA CMetHC-Mutl8-QL CH SEQ ID NO: 89 GCGTCGACCAAGGGCCCATC GGTCTTCCCCCTGGCACCCT CCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTG CCTGGTCAAGGACTACTTCC CCGAACCGGTGACGGTGTCG TGGAACTCAGGCGCCCTGAC CAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCA GGACTCTACTCCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCA CAAGCCCAGCAACACCAAGG TGGACAAGAAAGTTGAGCCC AAATCTTGTGACAAAACTCA CACATCACCACCGTCTCCAG CACCTGAACTCCTGGGGGGG CCGTCAGTCTTCCTCTTCCC CCCAAAACCCAAGGACCAGC TCATGATCTCCCGGACCCCT GAGGTCACATGCGTGGTGGT GGACGTGAGCCACGAAGACC CTGAGGTCAAGTTCAACTG G TACGTGGACGGCGTGGAGGT -359· 160877.doc 201247704

GCATAATGCCAAGACAAAGC CGCGGGAGGAGCAGTACAAC AGCACGTACCGTGTGGTCAG CGTCCTCACCGTCCTGCACC AGGACTGGCTGAATGGCAAG GAGTACAAGTGCAAGGTCTC CAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCG AGAACCACAGGTGTACACCC TGCCCCCATCCCGCGAGGAG ATGACCAAGAACCAGGTCAG CCTGACCTGCCTGGTCAAAG GCTTCTATCCCAGCGACATC GCCGTGGAGTGGGAGAGCAA TGGGCAGCCGGAGAACAACT ACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTT CCGGCTCTACAGCAAGCTCA CCGTGGACAAGAGCAGGTGG CAGCAGGGGAACGTCTTCTC ATGCTCCGTGCTGCATGAGG CTCTGCACAACCACTACACG CAGAAGAGCCTCTCCCTGTC TCCGGGTAAATGA cMetHC-Mutl8-GTv2 Ch SEQ ID NO: 90 GCGTCGACCAAGGGCCCATC GGTCTTCCCCCTGGCACCCT CCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTG CCTGGTCAAGGACTACTTCC CCGAACCGGTGACGGTGTCG TGGAACTCAGGCGCCCTGAC CAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCA GGACTCTACTCCCTCAGCAG CGTGGTGACCGTGCCCTCCA GCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCA CAAGCCCAGCAACACCAAGG TGGACAAGAAAGTTGAGCCC AAATCTTGTGACAAAACTCA CACATCACCACCGTCTCCAG CACCTGAACTCCTGGGGGGG CCGTCAGTCTTCCTCTTCCC CCCAAAACCCAAGGACACCC TCATGATCTCCCGGACCCCT GAGGTC ACATGCGTGGTGGT GGACGTGAGCCACGAAGACC CTGAGGTCAAGTTCAACTGG TACGTGGACGGCGTGGAGGT GCATAATGCCAAGACAAAGC CGCGGGAGGAGCAGTACAAC AGCACGTACCGTGTGGTCAG CGTCCTCGCCGTCCTGCACC AGGACTGGCTGAATGGCAAG GAGTACAAGTGCAAGGTCTC CAACAAAGCCCTCCCAGCCC CCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCG AGAACCACAGGTGTACACCC

160877.doc -360- 201247704

TGCCCCCATCCCGCGAGGAG ATGACCAAGAACCAGGTCAG CCTGACCTGCCTGGTCAAAG GCTTCTATCCCAGCGACATC GCCGTGGCGTGGGAGAGCAA TGGGCAGCCGGAGAACAACT ACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTT CCGGCTCTACAGCAAGCTCA CCGTGGACAAGAGCAGGTGG CAGCAGGGGAACGTCTTCTC ATGCTCCGTGATGCATGAGG CTCTGCACGCCCACTACACG CAGAAGAGCCTCTCCCTGTC TCCGGGTAAATGA Table 49: amino acid sequence having mutations half cMet-Fc binding protein binding VH and Ig constant region

Protein region / domain of the amino acid sequence a sequence identifier 12345678901234567890 cMet VH SEQ ID MO: 91 QVQLQQSGPELVRPGASVKM SCRASGYTFTSYWLHWVKQR PGQGLEWIGMIDPSNSDTRF NPNFKDKATLNVDRSSNTAY MLLSSLTSADSAVYYCATYG SYVSPLDYWGQGTSVTVSS cMetHC-Mutl8-QL CH SEQ ID NO: 92 ASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEP KSCDKTHTSPPSPAPELLGG PSVFLFPPKPKDQLMISRTP EVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPV LDSDGSE ^ LYSKLTVDKSRW QQGNVFSCSVLHEALHNHYT qkslslspgk " cMetHC-Mut18-GTv2 Ch SEQ ID NO: 93 ASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEP KSCDKTHTSPPSPAPELLGG PSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYN STYRWSVLAVLHQDWLNGK EYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDI • 361 - 160877.doc 201247704

AVAWESNGQPENNYKTTPPV

LDSDGSFRLYSKLTVDKSRW

QQGNVFSCSVMHEALHAHYT _______I QKSLSLSPGK_ a table 50: structural protein molecule sequence of cMet semi-Ig binding protein with FcRn binding mutation cMet-Mutl8-QL heavy chain: SEQ ID NO: 91 + SEQ ID NO: 92 light chain: SEQ ID NO: 6 cMet-Mutl 8-GTv2 heavy chain: SEQ ID NO: 91 + SEQ ID NO: 93 Light chain: SEQ ID NO: 6 Example 9.2: Characterization and purification of cMet semi-Ig binding protein with FcRn binding mutation according to Example 2.1 Protocol, HEK293 cells were transfected with plastids of the cMet half with FcRn binding mutations. The amount of such molecules is comparable to that of the parental full antibody, indicating that these half molecules can be effectively expressed in mammalian cells. Table 51: cMet semi-Ig binding protein expression in 293 cells with FcRn binding mutation (Mg/ml) cMet-Mutl8-QL 27 cMet-Mutl 8-GTv2 35 Example 9.3: cMet semi-Ig binding with FcRn binding mutation FcRn binding assay of the protein cMet-Mutl8-QL and cMet were determined by binding assay using CHO-FcRnGPI cells (stabilized FcRn receptor expression) and CHO-pBudl 1 cells (non-FcRn representation) as described in Example 6. -Mutl8-GTv2 FcRn binding ability. The data indicate that both the T250Q/M428L and T307A/N434A/E380A mutant combinations significantly improved the FcRn binding capacity of the semi-Ig binding protein cMet-Mutl8 160877.doc -362- 201247704 (Table 52). Stronger FcRn semi-Ig binding protein interactions may result in prolonged in vivo half-life. Table 52: FcRn binding analysis of cMet semi-Ig binding protein EC50 (nM) at molecular pH 6_4 cMet-Mutl8-QL 35.02 cMet-Mutl 8-GTv2 14.99 cMet-Mutl8 N/A* cMet mAb 51.76 References The contents of all of the cited references (including references, patents, patent applications, and websites), which are hereby incorporated by reference in their entirety in their entireties in the entireties in The practice of the present invention will employ, unless otherwise indicated, conventional techniques of immunology, molecular biology, and cell biology well known in the art. Equivalents The present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing examples are therefore illustrative in all aspects and not restrictive of the invention. The scope of the invention is therefore intended to be limited by the scope of the claims and the scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1A is a schematic illustration of a semi-Ig construct containing variable numbers and types of variable domains. Figure 1B is a schematic representation of a semi-Ig construct and shows the strategy for generating a half Ig from a parent antibody. 160877.doc •363 - 201247704 Sequence Listing <110>

<120> Semi-immunoglobulin binding protein and use thereof <130> DOCKET NUMBER <14〇> 100148091 <141> 2011-12-22 <150> 61/426,207 <151> 2010-12- 22 <150> 61/539,130 <151> 2011-09-26 <160> 208 <170> Patcntln version 3.5 <210> 1 <211> 357 <212> DNA <213>

≪ 220 > < 223 > Description of Artificial Sequence of: synthetic polynucleotide < 400 > 1 caggtccaac tg ^ cagcagtc tgggcctgag ctggtgaggc ctggggcttc agtgaagatg 60 tcctgcaggg cttcgggcta taccttcacc agctactggt tgcactgggt taaacagagg 120 cctggacaag gccttgaitg gattggcaig attgatcctt ccaatagtga cactaggttt 180 aatccgaact tcaaggacaa ggccacattg aatgtagaca Gatcttccaa cacagcctac 240 atgctgctca gcagcctgac atctgctgac tctgcagtct attactgtgc cacatatggt 300 agctacgttt cccctctgga ciactggggt caaggaacct cagtcaccgt ctcctca 357 <210> 2 <211> 342 <212> DNA <2i3> artificial sequence <220><223> Description of artificial sequence : synthesis of polynucleotides gactttatga tgtcacagtc tccatcctcc ctaactgtgt cagttggaga gaaggttact 60 gtgagctgca agtccagtca gtccctttta tataclagca gtcagaagaa ctacttggcc 120 tggtaccagc agaaaccagg tcagtctcct aaactgctga tttactgggc atccactagg 180 gaatctgggg tccctgatcg cttcacaggc agtggatctg ggacagattt cactctcacc 240 atcaccagtg tgaaggctga cgacctggca gtttattact gtcagcaata ttatgcctat 300 ccfitggacgt tcggtg Gagg caccaagttg gagctcaaac gg 342 <210> 3 <211> 319 <212> PRT <213> artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide 160877 - Sequence Listing.doc 201247704 <400> 3

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Vai Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Tlir Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser AJa Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser 115 <210> 4 <211> 114 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Phe Met Met Ser Gin Ser Pro Ser Ser Leu Thr Val Ser Val Gly 15 10 15

Glu Lys Val Thr Val Ser Cys Lys Ser Ser Gin Ser Leu Leu Tyr Thr 20 25 30

Ser Ser Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45

Ser Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 SO

He Thr Ser Val Lys Ala Asp Asp Leu Ala Val Tyr Tyr Cys Gin Gin 85 90 95

Tyr Tyr Ala Tyr Pro Ding rp Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu 100 105 110

Lys Arg 160877 · Sequence Listing. doc 201247704 <210> 5 <211> 449 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <4〇〇 > 5

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Sex Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Scr Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255 160877 - Sequence Listing.doc 201247704

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Va] Asp Gly Val Glu Val His Asn 275 2S0 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro He G]u Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415

Scr Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pio Gly 435 440 445

Lys <210> 6 <211> 220 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Phe Met Met Ser Gin Ser Pro Ser Ser Leu Thr Val Ser Val Gly 15 10 15

Glu Lys Val Thr Val Ser Cys Lys Ser Scr Gin Ser Leu Leu Tyr Thr 20 25 30

Ser Ser Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45

Ser Pro Lys Leu Leu lie Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 160877 - Sequence Listing.doc 201247704 50 55 60

Pro Asp Arg Phe Thr Gly Ser G]y Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80

He Thr Ser Val Lys Ala Asp Asp Leu Ala Val Tyr Tyr Cys Gin Gin 85 90 95

Tyr Tyr Ala Tyr Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu 100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp 115 120 125

Glu Gin Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu 145 150 155 160

Gin Scr Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp 165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thx Leu Ser Lys Ala Asp Tyr 180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His G1n Gly Leu Ser 195 200 205

Scr Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 7 <21.1> 449 <212> PRT <213> Artificial Sequence <m><223> Description of Artificial Sequence: Synthetic peptide <400> 7

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met He Asp Pro Scr Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95 160877_ Sequence Listing.doc 201247704

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Aja Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Tlir Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Va] His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 160877 - Sequence Listing.doc 201247704 3S5 390 395 400

Asp Scr Asp Gly Ser Phe Phe Leu Tyr Scr Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 8 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide

<400> 8

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 1 5 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Tht Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala llir Leu Asn Val Asp Arg Sex Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Scr Scr Ala Scr Thr Lys Gly Pro Scr Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 160877 - Sequence Listing.doc 201247704

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val GIu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Cys Pro Pro Cys Fro Ala Pro GIu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met He Set 245 250 255

Arg Thr Pro GIu Val Tlir Cys Val Val Val Asp Val Ser His GIu Asp 260 265 270

Pro GIu Val Lys Phe Asn Trp Tyr Val Asp Gly Val GIu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg GIu GIu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Scr Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys GIu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro He GIu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg GIu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg GIu GIu Met Thr Lys Asn Gin Val Ser Leu Phe 355 360 365

Cys Phe Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val GIu Trp GIu 370 375 380

Scr Asn Gly Gin Pro GIu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Asp Leu Ήιγ Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His GIu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 9 <2I1> 449 <212> PRT <2]3> artificial sequence 160877 - Sequence Listing.doc 201247704 <220><223> Description of artificial sequence: synthetic polypeptide <400> 9

Gly Pro Glu Leu Val Arg Pro Gly Ala 10 15

Gin Val Gin Leu Gin Gin Scr 1 5

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met He Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 1)5 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Sei Gly Ala Leu Thr Scr Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Scr Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Giu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met He Scr 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 •9· 160877-Sequence List.doc 201247704

Ala Lys Tbr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr He Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Phe 355 360 365

Cys Phe Val Lys Gly Phe Tyr Pro Ser Asp He Ala Val Glu Trp Glu 370 375 380

Set Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 10 <211> 449 <232> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Tlir Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Tbr A]a Tyr 65 70 75 80 -10-

160877 - Sequence Listing.doc 201247704

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr vSer Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Scr Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Aig Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Scr Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Tlir 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380 -11 - 160877 - Sequence Listing.doc 201247704

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr ΓΙίγ Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210>11 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Scr Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 ]50 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 -12· 160877· Sequence Listing.doc 201247704 GJn Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205 ..

Ser Asn Thr Lys Val Asp Lys Lys Val GIu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Plie Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Scr Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyi Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Scr Asp Leu Thr Val Λερ Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 12 <211> 449 • 13-160877 - Sequence Listing.doc 201247704 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>; 12

Gin Val Gin Leu Gin Gin Scr Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Ttp He 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Ί\\τ Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Ήιγ Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Va] Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser VaJ Phe 115 〗 20 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Ser Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Cys Pro Ala Pto Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thi Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 • 14-

160877 - Sequence Listing.doc 201247704

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Scr Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Vai Phe Ser Cys Scr Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 13 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Sar Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60 -15- 160877 - Sequence Listing.doc 201247704

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser'Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365 •16·

160877· Sequence Listing.doc 201247704

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val «et His Glu

Ala Leu His Asn His Tyr Thr Gin Lys Scr Leu Ser Leu Ser Pro Gly 435 440 445

Lys

<210> 14 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Giy Pro Glu Leu Val Arg Pro Gly Ala i 5 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Sei Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp He 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Scr Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Scr Trp 145 150 155 160 • 17- 160877 · Sequence Listing.doc 201247704

Asn Ser Gly Ala Leu Thr 165

Ser Gly Val His Thr Phe 170

Pro Ala Val Leu 175

Gin Ser Ser Gly Leu Tyr 180

Ser Leu Ser Ser Val Val 185

Thr Val Pro Ser 190

Ser Ser Leu Gly Thr Gin 195

Thr Tyr lie Cys Asn Val 200

Asn His Lys Pro 205

Ser Asn Thr Lys Val Asp 210

Lys Lys Val Glu Pro Lys 215 220

Ser Cys Asp Lys

Thr His Thr Ser Pro Pro 225 230

Ser Pro Ala Pro Glu Leu 235

Leu Gly Gly Pro 240

Ser Val Phe Leu Phe Pro 245

Pro Lys Pro Lys Asp Thr 250

Leu Met lie Ser 255

Arg Thr Pro Glu Val Thr 260

Cys Val Val Val Asp Val 265

Ser His Glu Asp 270

Pro Glu Val Lys Phe Asn 275

Trp Tyr Val Asp Gly Val 280

Glu Val His Asn 285

Ala Lys Thr Lys Pro Arg 290

Glu Glu Gin Tyr Asn Ser 295 300

Thr Tyr Arg Val

Va) Ser Val Leu Thr Val 305 310

Leu His Gin Asp Trp Leu 315

Asn Gly Lys Glu 320

Tyr Lys Cys Lys Val Ser 325

Asn Lys Ala Leu Pro Ala 330

Pto lie Glu Lys 335

Thr lie Ser Lys Ala Lys 340

Gly Gin Pro Arg Glu Pro 345

Gin Val Tyr Thr 350

Leu Pro Pro Ser Aig Glu 355

Glu Met Thr Lys Asn Gin 360

Val Ser Leu Thr 365

Cys Leu Val Lys Gly Phe 370

Tyr Pro Ser Asp lie Ala 375 380

Val Glu Trp Glu

Ser Asn Gly Gin Pro Glu 385 390

Asn Asn Tyr Lys Thr Thr 395

Pro Pro Val Leu 400

Asp Ser Asp Gly Ser Phe 405

Arg Leu Arg Ser Asp Leu 410

Thr Val Asp Lys 415

Ser Arg Trp Gin Gin Gly 420

Asn Val Fhe Ser Cys Ser 425

Val Met His Giu 430

Ala Leu His Asn His Tyr 435

Thr Gin Lys Ser Leu Ser 440

Leu Ser Pro Gly 445

Lys 160877 - Sequence Listing. doc • 18. 201247704 <210> 15 <21 ]> 449 <212> PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <;400> 15

Gin Val Gin Leu Gin Gin Scr Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Scr Ser Leu Thr Ser Ala Asp Scr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pto Uu Asp Dyr yr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Scr Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Fhe Pro Ala Val Leu 165 170 175

Gin Scr Ser Gly Leu Tyr Ser Leu Ser Sei Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255 • 19· 160877 - Sequence Listing.doc 201247704

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Giy Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Arg Ser Asp Leu Ήιγ Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 16 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Va丨 Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp He 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Tlir Arg Phe Asn Pro Asn Phe • 20· 160877 - Sequence Listing.doc 201247704 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Tlir Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Scr Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phc Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195, 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 230 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe L«u Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350 -21 · 160877 - Sequence Listing.doc 201247704

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Va] Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Tyr Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Va) Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210>.17 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Va) Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp -22-

160877· Sequence Listing.doc 201247704 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Tlir Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Scr Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Va! Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr Lie Scr Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Tip Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Hie Arg Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Scr Leu Set Leu Scr Pro Gly 435 440 445 -23- 160877 - Sequence Listing, doc 201247704

Lys <210> 18 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Are Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phc Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thx Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Va] His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val ΊΊιγ Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser • 24·

160877 - Sequence Listing.doc 201247704 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Va] Asp Val Ser His GIu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Va] Glu Va] His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Scr Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp He Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Scr Asp Gly Ser Phe Phe Leu Arg Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Mel His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Scr Leu Ser Leu Ser Pro Gly 435 440 445

Lys

<210> 19 <21]> 449 <212> PRT β13" artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45 -25- 160877 - Sequence Listing.doc 201247704

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Gla Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Scr Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr He Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr -26-

160877 - Sequence Listing.doc 201247704 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 20 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 1 5 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe ITir Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp He 35 40 45

Gly Met He Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ma Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 -27- 160877 - Sequence Listing.doc 201247704

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Tip 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Fhe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met TTir Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Scr Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly •28·

160877 - Sequence Listing.doc 201247704 435 440 445

Lys <210> 21 <211> 449 <212> PRT c213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Tlir Ser Tyr 20 25 30

Txp Leu His Ttp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met He Asp Pro Ser Asn Ser Asp Thr Arg Phc Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pto Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Ser Giy Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Lea Tyr Scr Leu Ser Ser Val Val Thr Val Pro Set 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr He Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Pro Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 • 29· 160877 · Sequence Listing.doc 201247704

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Fhe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Tyr Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 22 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 •30·

160877 - Sequence Listing.doc 201247704

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Sei Val Thr Val Ser Ser Ala Ser Thi Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Set Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160

Asn Scr Gly Ala Leu Tbr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn. His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Lea Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 T\r Lys Cys Lys Val Sex Asn Lys Ala Leu Pro Ala Pto He Glu Lys 325 330 335 -31 - 160877 - Sequence Listing.doc 201247704

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp He Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Arg Ser Asp Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 23 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 1 5 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly G)n Gly Leu Glu Trp He 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr TyT Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 •32·

160877· Sequence Listing.doc 201247704

Pro Leu Ala Pro Ser Ser Lys Ser Thr Scr Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Va by Ser Trp 145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 G]n Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Va Kenting hr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300 .

Val Ser Vai Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr He Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Ήιγ Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Pbe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp GIti Gin Gly Asn Val Hie Scr Cys Scr Val Met His Glu 420 425 430 -33- 160877 - Sequence Listing.doc 201247704

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <21Q> 24 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 )05 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly C]y Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Tr 145 150 155 16i

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Scr Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 -34-

160877 - Sequence Listing.doc 201247704

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pto Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Vai Leu Thr Val Leu His Gin Asp Ding rp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Arg Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Lea Ser Pro Gly 435 440 445

Lys <210> 25 <213> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15 • 35- 160877 · Sequence Listing.doc 201247704

Ser Val Lys Met Ser Cys Arg Ala Ser Cly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Ding rp Gly G]n Gly 100 105 110

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly rrhr Ala Ala Leu 130 135 140

Cly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 ]55 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser L80 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr He Cys Asn Va] Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Va] His Asn 275 280 285

Ala Lys Thr Lys Pro Arg G!u Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

VaJ Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Ly £ G]u 305 310 315 320 •36-

160877 - Sequence Listing.doc 201247704

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr He Scr Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Scr Asp Gly Ser Phe Phe Leu Arg Ser Lys Leu Thr Val Asp Lys 405 410 415

Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Scr Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 26 <211> 449 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Leu His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110 -37- 160877 · Sequence Listing _doc 201247704

Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 】60

Asn Set Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 G】n Ser Ser G】y Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220

Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met lie Ser 245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val 290 295 300

Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie Glu Lys 325 330 335

Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr 340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr 355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu Trp Glu 370 375 380

Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Asp Leu Thr Val Asp Lys 405 410 415 -38-

160877· Sequence Listing.doc 201247704

Scr Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430

Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445

Lys <210> 27 <211> 12] <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Lys Leu Gin Gin Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 15 10 15

Scr Val Arg Leu Scr Cys Lys Ala Scr Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 lie lie His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Trp Phe Tyr Pro Gly Ser Asn Asp lie Gin Tyr Asn Ala Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 28 <211> 108 <212> PRT <213>Artificial Sequence<220><223> Description of Artificial Sequence··Synthetic Polypeptide<400> 28

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Val Scr Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Thr Asn Glu Asn lie Tyr Ser Asn 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu lie 35 40 45 • 39- 160877 · Sequence Listing.doc 201247704

Tyr Ala Ala Thr His Leu Val Glu Gly Val Pro Scr Arg Phc Scr Gly 50 55 60

Ser Gly Scr Gly Thr Gin Tyr Ser Leu Lys lie Thr Ser Leu Gin Ser 65 70 75 80

Glu Asp Phe Gly Asn Tyr Tyr Cys Gin His Phe Trp Gly Thr Pro Cys 85 90 95

Thr Fhc Gly Gly Gly Thr Lys Leu Glu He Lys Arg 100 105 <210> 29 <211> 363 <212> DNA <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Poly nucleotide < 400 > 29 caggtgaagc tgcagcagag tggcgctgag ctcgtgaagc ctsgcgcctc agtccggctg 60 agttgtaagg catcaggtta cacgtttacc gagtatatta tccattggat taaactcasg 120 tctgggcaag gattggaatg gataggctgg ttctatcctg gatcaaatga catccagtac 180 aatgctaagt tcaaggggaa ggccacactg accgcagaca agtcctcctc tacagtgtat 240 atggaactca ctgggclgac cagcgaagac agtgcagtgt atttctgcgc gaggagggac 300 gatttctccg gttatgacgc tctgccatat tgggggcagg gcacgatggt taccgtgtct 360 Age 363

<210> 30 <211> 324 <212> im <213> artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide <400> 30 gacatacaga tgacccaaag cccggcctcc ctctccgtct cagtagggga gactgtaaca 60 atcacatgta ggactaatga gaatatctac tctaatctgg cgtggtacca gcaaaagcag 120 ggcaaatccc cccagctgct catctatgct gccacccatc ttgtagaagg agtcccctct 180 cgcttcagcg gctccgggLc cgggacacaa tattctctga aaattaccag cctccaatca 240 gaagaetteg ggaactacta ttgccagcac ttttggggaa ccccctgtac ctttggaggc 300 eecacaaagc tegagataaa gegg 324

<210> 31 <211> 451 <212> PRT <2丨3> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide 160877 - Sequence Listing. doc -40 - 201247704 <400> 31

Gin Val Lys Leu Gin Gin Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15

Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 lie lie His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Trp Phe Tyr Pto Gly Ser Asn Asp lie Gin Tyr Asn Ala Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Ήιγ Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Asp Asp Phe Scr Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val ThT Val Set Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pto Ala Pro Glu Leu Leu Gly 225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 lie Ser Arg ThT Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 290 295 300 •41 · 160877-Sequence List.doc 201247704

Are Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie 325 330 335

Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser 355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu 370 375 380

Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415

Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser 435 440 445

Pro Gly Lys 450 <210> 32 <211> 214 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp He Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Val Ser Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Thr Asn Glu Asn lie Tyr Ser Asn 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu lie 35 40 45

Tyr Ala Ala Thr His Leu Val Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Tyr Ser Leu Lys Tie Thr Ser Leu Gin Ser 65 70 75 80

Glu Asp Phe Gly Asn Tyr Tyr Cys Gin His Phe Trp Gly Thr Pro Cys 85 90 95 • 42·

160877 - Sequence Listing.doc 201247704

Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 145 150 155 160

Glu Ser Val Thr Glu GLn Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205

Phe Asn Arg Gly Glu Cys 210 <210> 33 <211> 451 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Lys Leu Gin Gin Sec Gly Ala Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Arg Leu Ser Cys Lys Ala Scr Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 lie He His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp He 35 40 45

Gly Trp Phe Tyr Pro Gly Ser Asn Asp lie Gin Tyt Asn Ala Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 •43· 160877·Sequence List.doc 201247704

Val Phe Pro Leu 130

Ala Pro Ser Ser Lys Ser 135

Thr Ser Gly Gly Thr Ala 140

Ala Leu Gly Cys 145

Leu Val Lys Asp Tyr Phe 150

Pro Glu Pro Val Thr Val 155 160

Set Trp Asn Ser

Gly Ala Leu Thr Ser Gly 165 170

Val His Thr Phe Pro Ala 175

Val Leu Gin Ser 180

Ser Gly Leu Tyr Ser Leu 185

Ser Ser Val Val Thr Val 190

Pro Ser Ser Ser 195

Leu Gly Thr Gin Thr Tyr 200

He Cys Asn Val Asn His 205

Lys Pro Ser Asn 210

Thr Lys Val Asp Lys Lys 215

Val Glu Pro Lys Ser Ser 220

Asp Lys Thr His 225

Thr Ser Pro Pro Cys Pro 230

Ala Pro Glu Leu Leu Gly 235 240

Gly Pro Ser Val

Phe Leu Phe Pro Pro Lys 245 250

Pro Lys Asp Thr Leu Met 255 lie Ser Arg Thr 260

Pro Glu Va丨 Thr Cys Val 265

Val Val Asp Val Ser His 270

Glu Asp Pro Glu 275

Val Lys Phe Asn Trp Tyr 280

Val Asp Gly Val Glu Val 285

His Asn Ala Lys 290

Thr Lys Pro Arg Glu Glu 295

Gin Tyr Asn Ser Thr Tyr 300

Arg Val Val Ser 305

Val Leu Thr Val Leu His 310

Gin Asp Trp Leu Asn Gly 315 320

Lys Glu Tyr Lys

Cys Lys Val Ser Asn Lys 325 330

Ala Leu Pro Ala Pro lie 335

Glu Lys Thr lie 340

Ser Lys Ala Lys Gly Gin 345

Pro Arg Glu Pro Gin Val 350

Tyr Thr Leu Pro 355

Pro Ser Arg Glu Glu Met 360

Thr Lys Asn Gin Val Ser 365

Leu Thr Cys Leu 370

Val Lys Gly Phe Tyr Pro 375

Ser Asp lie Ala Val Glu 380

Trp Glu Ser Asn 385

Gly Gin Pro Glu Asn Asn 390

Tyr Lys Thr Thr Pro Pro 395 400

Val Leu Asp Ser

Asp Gly Ser Phe Phe Leu 405 410

Tyr Ser Lys Leu Thr Val 415

Asp Lys Ser Arg 420

Trp Gin Gin Gly Asn Val 425

Phe Ser Cys Ser Val Met 430 160877 · Sequence Listing. doc -44- 201247704

His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser 435 440 445

Pro Gly Lys 450 <210> 34 <211> 450 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <4〇〇> 34

Gin Val Lys Leu Gin Gin Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30

Lie lie His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Trp Fhe Tyr Pro Gly Ser Asn Asp He Gin Tyr Asn Ala Lys Phe 50 55 60 L^s Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 - 95

Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val Thr Val Ser Scr Ala Ser Thr Lys Gly Pro Ser 115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Scr Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Ser 210 215 220 -45- 160877 · Sequence Listing.doc 201247704

Asp Lys Thr His Ώιγ Ser Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie 325 330 335

Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser 355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu 370 375 380

Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Ala Val 385 390 395 400

Leu Asp Scr Asp Gly Ser Phe Ala Leu Ala Ser Asp Leu Thr Val Asp 405 410 415

Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Scr Val Met His 420 425 430

Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro 435 440 445

Gly Lys 450

<210> 35 <211> 450 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400> 35 Gin Val Lys Leu Gin Gin Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 160877 · Sequence Listing _doc -46- 201247704 20 25 30 lie lie His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Trp Phe Tyr Pro Giy Ser Asn Asp lie Gin Tyr Asn Ala Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125

Val Phe Pro Leu Ala Pro Scr Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gla Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thi Val 180 185 190

Pro Ser Ser Ser Uu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205

Lys Pro Scr Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Scr Ser 210 215 220

Asp Lys Thr His Thr Scr Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240

Gly Pro Scr Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Va] Asp Gly Val Glu Val 275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 290 295 300

Arg Val Val Scr Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 305 310 315 320 -47· 160877-Sequence List.doc 201247704

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro He 325 330 335

Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Va] Ser 355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu 370 375 3S0

Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Ala Val 385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Arg Leu ATg Ser Asp Leu Thr Val Asp 405 410 415

Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Scr Cys Ser Val Met His 420 425 430

Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro 435 440 445

Gly Lys 450 <210> 36 <211> 450 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Lys Leu Gin Gin Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 lie lie His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp He 35 40 45

Gly Trp Phe Tyr Pro Gly Ser Asn Asp lie Gin Tyr Asn Ala Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser _ 48 ·

160877 - Sequence Listing.doc 201247704 115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Giy Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Ser Leu Giy Thr Gin Thr Tyr lie Cys Asn Val Asn His 】95 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Ser 210 215 220

Asp Lys Thr His Thr Ser Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 lie Ser Arg Thr Pro Giu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 290 295 300

Arg Val Val Scr Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie 325 330 335

Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser 355 360 365

Leu Phe Cys Phe Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu 370' 375 380

Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Ala Val 385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Ala Leu Ala Ser Asp Leu Thr Val Asp 405 410 415 • 49. 160877 · Sequence Listing.doc 201247704

Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430

Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro 435 440 445

Gly Lys 450 <210> 37 <211> 450 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Lys Leu Gin Gin Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr 20 25 30 lie lie His Trp lie Lys Leu Arg Ser Gly Gin Gly Leu Glu Trp He 35 40 45

Gly Trp Phe Tyr Pro Gly Ser Asn Asp lie Gin Tyr Asn Ala Lys Phe 50 55 60

Lys Giy Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80

Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Scr Gly Val His Thr Phe Pro Ala 165 170 175

Va] Leu Gin Scr Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Ser Leu Gly Ήιγ Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Scr -50·

160877 - Sequence Listing.doc 201247704 210 215 220

Asp Lys Thr His Thr Ser Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240

LY Ser Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 305 310 315 320

Lys Glu Tyi Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro He 325 330 335

Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Tin Lys Asn Gin Val Ser 355 360 365

Leu Phe Cys Phc Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Glu 370 375 380

Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Ala Val 385 390 395 400

Leu Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Asp Leu Thr Val Asp 405 410 415

Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430

Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro 435 440 445

Gly Lys 450 <210> 38 <211> 119 <2]2> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 1. 5 10 15 -51 · 160877 - Sequence Listing.doc 201247704

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asa Fro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser 115 <210> 39 <211> 106 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Leu Thr Gin Ser Pro Ala lie Met Ser Ala Ser Pro Gly 15 10 15

Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30

Asn Trp Tyr Gin Gin Lys Ser Gly Thr Ser Pro Lys Arg Trp lie Tyr 35 40 45

Asp Thr Ser Lys Val Ala Ser Gly Val Pro Tyr Arg Phe Set Gly Ser 50 55 60

Gly Set Gly Tlir Ser Tyr Ser Leu Thr He Ser Ser Met Glu Ala Glu 65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Trp Ser Ser Asn Pro Leu Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 40 <211> 124 <212> PRT <2I3>Artificial Sequence<220><223> Description of Artificial Sequence··Synthetic Polypeptide- 52-

160877-SEQ ID NO.doc 201247704 <400> 40

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Val Arg Pro Gly Ser 1 5 10 15

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr 20 25 30

Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp 100 105 310

TyrTrpGlyG, nG, y^ThrVarrVal Ser Ser <210> 41 <211> 111 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Leu Thr Gin Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15

Gin Arg Ala Tht He Ser Cys Lys Ala Ser Gin Ser Val Asp Tyr Asp 20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin He Pro Gly Gin Pro Pro 35 40 45

Lys Leu Leu lie Tyr Asp Ala Ser Asn Leu Val Ser Gly He Pro Pro 50 55 60

Arg Phe Ser Gly Scr Gly Ser Gly Thr Asp Phe Thr Leu Asn lie His 65 70 75 80

Pto Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gin Gin Ser Thr 85 90 95

Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110 <210> 42 <211> 357 •53·160877-SEQ ID NO.doc 201247704 <212> DNA <213>Artificial Sequence<; 220 > < 223 > description of artificial sequence of: synthetic polynucleotide < 400 > 42 gatatcaaac tgcagcagtc aggggctgaa ctggcaagac ctggggcctc agtgaagat £ 60 tcctgcaaga cttctggcta cacctttact aggtacacga tgcactgggt aaaacagagg 120 cctggacagg gtctggaatg gattggatac attaatccta gccgtggtta tactaattac 180 aatcagaagt tcaaggacaa ggccacattg actacagaca aatcctccag cacagcctac 240 atgcaactga gcagcctgac atctgaggac tctgcagtct attactglgc aagatattat 300 gaigatcatt actgccttga ctactggggc caaggcacca ctctcacagt ctcctca 357 <210> 43 <211> 318 <2]2> DNA <2]3> artificial sequence <220><223> Description: Synthetic Polynucleotide <400> 43 gacattcagc tgacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc 60 atgacctgca gagccasttc aagtgtaagt tacatgaact ggiaccagca gaagtcaggc 120 acctccccca aaagatggat ttatgac aca tccaaagtgg cttctggagt cccttatcgc 180 ttcagtggca gtgggtctgg gacctcatac tctctcacaa tcagcagcat ggaggctgaa 240 gatgctgcca cttattactg ccaacagtgg agtagtaacc cgctcacgtt cggtgctggg 300 accaagctgg agctgaaa 318 < 210 > 44 < 211 > 372 < 212 > DNA < 213 > artificial sequence < 220 > < 223 > description of artificial sequences: synthetic polynucleotide < 400 > 44 caggtgcagc tgcagcagtc tggggctgag ctggtgaggc ctgggtcctc agtgaagatt 60 tcctgcaagg cttctggcta tgcattcagt agctactgga tgaactgggt gaagcagagg 120 cctggacagg gtcttgagtg gattggacag atttggcctg gagatggtga tactaactac 180 aatggaaagt tcaagggtaa agccactctg actgcagacg aatcctccag cacagcctac 240 atgcaactca gcagcctagc atctgaggac tctgcggtct atttctgtgc aagacgggag 300 actacgacgg taggccgtta ttactatgct atggactact ggggccaagg gaccacggtc 360 accgtctcct cc 372 <210> 45 <2]1> 333 <212> DNA <213>Artificial sequence<220> -54-160877-sequence table.doc 201247704 < 223 > Description of Artificial Sequence: Synthetic Polynucleotide <400> 45 gatatccagc tgaccca gtc tccagcttct ttggctgtgt ctctagggca gagggccacc atctcctgca aggccagcca aagtsttgat tatgatggtg atagttattt gaactggtac caacagattc caggacagcc acccaaactc ctcatctatg atgcatccaa tctagtttct gggatcccac ccaggtttag tggcagtggg tctgggacag acttcaccct caacatccat cctgtggaga aggtggatgc tgcaacctat cactgtcagc aaagtaclga ggatccgtgg acgttcggtg gagggaccaa gctcgagatc aaa < 210 > 46 < 211 > 13 < 212 > PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 46

Ala vSer Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 1 5 10 <210> 47 <211> 39 <212> DNA <213> Artificial Sequence <220><?23> Description of Artificial Sequence : synthetic oligonucleotide <400> 47 gcgtcgacca agggcccatc ggtcttcccc ctggcaccc <210> 48 <2U> 6 <212> PRT <213>Artificial sequence<220><223> Description of artificial sequence: synthesis Peptide <400> 48

Ala Ser Thr Lys Gly Pro 1 s <210> 49 <211> 18 <212> DNA <213>Artificial Sequence <220><223> Description of Artificial Sequence··Synthetic Oligonucleotide<400> 49 gcgtcgacca agggccca <210> 50 <211> 12 <212> PRT <213> Artificial sequence 55-160877 - Sequence Listing doc 201247704 <220><223> Description of artificial sequence: synthetic peptide <;400> 50

Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro 1 5 10 <210> 51 <211> 36 <212> DNA <233>Artificial Sequence <220><223> Description of Artificial Sequence: Synthesis Nucleotide <400> 51 acggtggctg caccatctgt cttcatcttc ccgcca 36 <210> 52 <211> 5 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400> 52

Thr Val Ala Ala Pro 1 5 <210> 53 <211> 15 <212> DNA <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Oligonucleotide <400> 53 acggtggctg caeca 15 <210> 54 <211> 586 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Atg Pro Gly Ala 15 10 15

Ser Val Lys Met Scr Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60 56-

160877 - Sequence Listing.doc 201247704

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Scr Ser Tht Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Τντ Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Set Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Val 130 135 ]40

Arg Pro Gly Ser Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Ala 145 150 155 160

Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp lie Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr 180 185 190

Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser 195 200 205

Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr 225 230 235 240

Tyr Ala Met Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Scr Scr 245 250 255

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 260 265 270

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 275 280 285

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 290 295 300

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 305 310 315 320

Leu Ser Set Val Val Thr Val Pro Ser Ser Ser Leu Gly Ήιγ Gin Thr 325 330 335

Tyr lie Cys Asn Va! Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 340 345 350

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 355 360 365 • 57· 160877· Sequence Listing.doc 201247704

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phc Pro Pro 370 375 380

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 385 390 395 400

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 405 410 415

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 420 425 430

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 435 440 445

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 450 455 460

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 465 470 475 480

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 485 490 495

Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 500 505 510

Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 515 520 525

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 530 535 540

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 545 550 555 560

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 565 570 575

Gin Lys Ser Leu Ser Leu Scr Pro Gly Lys 580 585 <210> 55 <211> 335 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide<400> 55

Asp lie Gin Leu Thr Gin Ser Pro Ala He Met Ser Ala Ser Pro Gly L 5 10 15

Glu Lys Val Thr 5<et Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met 20 25 30 -58-

160877 - Sequence Listing.doc 201247704

Asn Trp Tyr Gin Gin Lys Ser Gly Thr Ser Pro Lys Arg Trp lie Tyr 35 40 45

Asp Ser Lys Val Ala Ser Gly Va! Pro Tyr Arg Phe Ser Gly Ser

Gly Ser Gly Thr Ser Tyr Ser Leu Thr ile Ser Ser Met Glu Ala Glu 65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Trp Ser Ser Asn Pro Leu Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Thr Val Ala Ala Pro Ser 100 105 110

Val Phe Ile Phe Pro Pro Asp lie Gin Leu Thr Gin Ser Pro Ala Ser 115 120 125

Leu Ala Val Ser Leu Gly Gin Arg Ala Thr Ile Ser Cys Lys Ala Ser 130 135 140

Gin Ser Val Asp Tyr Asp Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin 145 150 155 160

Ile Pro Gly Gin Pro Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu 165 170 175

Val Ser Gly Ile Pro Pro Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 180 185 190

Phe Thr Leu Asn Ile His Pro Val Glu Lys Val Asp Ala Ala Tht Tyr 195 200 205

His Cys Gin Gin Ser Thr Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr 210 215 220

Lys Leu Glu He Lys Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro 225 230 235 240

Pro Scr Asp Glu Gin Leu Lys Ser Gly Thr Ala Scr Val Val Cys Leu 245 250 255

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp 260 265 270

Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp 275 280 285

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Scr Thr Leu Thr Leu Ser Lys 290 295 300

Ala Asp Tyr Gl\i Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gin 305 310 315 320

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 325 330 335 • 59· 160877 - Sequence Listing.doc 201247704 <210> 56 <211> 579 <212> PRT <213>Artificial Sequence<;220><223> Description of artificial sequence: synthetic polypeptide <400> 56

Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Gin Val Gin 115 120 125

Leu Gin Gin Ser Gly Ala Glu Leu Val Arg Pro Gly Ser Ser Val Lys 130 135 140

He Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met Asn 145 150 355 160

Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie Gly Gin lie 165 170 175

Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly Lys 180 185 190

Ala Thr Leu Thr Ala Asp Glu Ser Ser Sct Thr Ala Tyr Met Gin Leu 195 200 205

Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala ktg Arg 210 215 220

Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly 225 230 235 240

Gin Gly TT,r Thr Val Thr Vai Ser Ser Ala Ser TT,r Lys Gly Pro Ser -60-

160877 - Sequence Listing.doc 201247704

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 260 265 270

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phc Pro Glu Pro Val Thr Val 275 280 285

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 290 295 300

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 305 310 315 320

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 325 330 335

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 340 345 350

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 355 360 365

Gly Pro Ser Val Phc Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 370 375 380 lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 385 390 395 400

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 405 410 415

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 420 425 430

Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 435 440 445

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie 450 455 460

Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 465 470 475 480

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser 485 490 495

Leu Thr Cys Leu Val Lys Gly Phc Tyr Pro Ser Asp lie Ala Val Glu 500 505 510

Trp Glu Ser Asn Gly Gin Fro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 515 520 525

Val Leu Asp Ser Asp Gly Ser Phc Phe Leu Tyr Ser Lys Leu Thr Val 530 535 540

Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phc Ser Cys Ser Val Met 545 550 555 560 -61 · 160877 - Sequence Listing.doc 201247704

His Glu Ala Leu His Asn His Tyr Thr Gin Lys Scr Leu Ser Leu Ser 565 570 575

Pro Gly Lys <210> 57 <211> 328 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Leu Thr Gin Ser Pro Ala lie Met Ser Ala Ser Pro Gly 15 10 15

Glu Lys Val Thr Met Thr Cys Arg Ala Scr Ser Ser Val Ser Tyr Met 20 25 30

Asn Trp Tyr Gin Gin Lys Ser Gly Thr Ser Pro Lys Arg Trp lie Tyr 35 40 45

Asp Thr Ser Lys Val Ala Ser Gly Val Pro Tyr Arg Phe Ser Gly Ser 50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr lie Ser Ser Met Glu Ala. Glu 65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Trp Ser Ser Asn Pro Leu Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Thr Val Ala Ala Pro Asp 100 105 110 lie Gin Leu Thr Gin Ser Pro Ala Ser Leu Ala Val Ser Leu Gly Gin 115 120 125

Arg Ala Thr lie Ser Cys Lys Ala Ser Gin Ser Val Asp Tyr Asp Gly 130 135 240

Asp Ser Tyr Leu Asn Trp Tyr Gin Gin lie Pro Gly Gin Pro Pro Lys 145 150 155 160

Leu Leu lie Tyr Asp Ala Ser Asn Leu Val Ser Gly lie Pro Pro Arg 165 170 175

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn lie His Pro 180 185 190

Val Glu Lys Val Asp Ala Ala Thr Tyr His Cys Gin Gin Ser Thr Glu 195 200 205

Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu He Lys Thr Val 210 215 220 • 62- 160877 - Sequence Listing.doc 201247704

Ala Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Giu Gin Leu Lys 225 230 235 240

Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg 245 250 255

Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn 260 265 270

Ser Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser 275 280 285

Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys 290 295 300

Val Tyr Ala Cys Glu Val Thr His Gin G!y Leu Ser Ser Val Val 305 310 315 320

Lys Ser Phe Asn Arg Gly Glu Cys 325

<210> 58 <211> 586 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Lys Leu G3n Gin Ser G!y Ala Glu Leu Ala Arg Pro Gly Ala 1 5 10 15

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30 rrhr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Scr Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Sex Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Ήιγ Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 1.25

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Val 130 135 140 • 63- 160877 · Sequence Listing.doc 201247704

Arg Pro Gly Ser Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Ala 145 150 155 160

Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp lie Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr 180 185 190

Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser 195 200 205

Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr 225 230 235 240

Tyr Ala Met Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser 245 250 255

Ala Ser Thr Lys Gly Pro Ser Val Phe Fro Leu Ala Pro Ser Ser Lys 260 265 270

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tvr 275 2S0 285

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 290 295 300

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 305 310 315 320

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 325 330 335

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 340 345 350

Lys Val Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Cys 355 360 365

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 370 375 380

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 385 390 395 400

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 405 410 415

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 420 425 430

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu ΊΤιγ Val Leu 435 440 445 •64-

160877· Sequence Listing.doc 201247704

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Scr Asn 450 455 460

Lys Ala Leu Pro Ala Pro He Glu Lys Thr lie Ser Lys Ala Lys Gly 465 470 475 480

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 485 490 495

Met Thr Lys Asn Gin Val Ser Lea Thr Cys Leu Val Lys Gly Phe Tyr 500 505 510

Pro Scr Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 515 520 525

Asn Tyr Lys Thr Thr Pro Ala Val Leu Asp Ser Asp Gly Ser Phe Arg 530 535 540

Leu Arg Ser Asp Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 545 550 555 560

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 565 570 575

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 580 585 <210> 59 <211> 579 <2\2> PRT <213>Artificial Sequence<220><223> Description of Artificial Sequence: Synthetic Peptide <400> 59

Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg Fro Gly Ala 1 5 10 15

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110 -65- 160877 · Sequence Listing.doc 201247704

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Gin Val Gin 115 120 125

Leu Gin Gin Ser Gly Ala Glu Leu Va] Arg Pro Gly Ser Ser Val Lys 130 135 140 lie Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Tyr Trp Met Asn 145 150 155 160

Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie Gly Gin lie 165 170 175

Trp Pro Gly Asp Gly Asp Thr Asn Tyr Asn Gly Lys Phe Lys Gly Lys 180 185 190

Ala Thr Leu Thr Ala Asp Glu Ser Ser Ser Thr Ala Tyr Met Gin Leu 195 200 205

Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Arg 210 215 220

Glu Thr Thr Thr Val Gly Arg Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly 225 230 235 240

Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 245 250 255

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 260 265 270

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 275 280 285

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 290 295 300

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 305 310 315 320

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 325 330 335

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Scr 340 345 350

Asp Lys Thr His Thr Ser Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 355 360 365

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 370 375 380 lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 385 390 395 400

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 405 410 415 _66·

160877· Sequence Listing.doc 201247704

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 420 425 430

Arg Val Val Ser Val Leu Ίϊγ Val Leu His Gin Asp Trp Leu Asn Gly 435 440 445

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro He 450 455 460

Glu Lys Thr lie Scr Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 465 470 475 480

Tyr Thr Leu Pro Pro Scr Arg Glu Glu Met Thr Lys Asn Gin Val Ser 485 490 495

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val Giu 500 S05 510

Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Ala 515 520 525

Val Leu Asp Ser Asp Gly Ser Phe Arg Leu Arg Ser Asp Leu Thr Val 530 535 540

Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Scr Cys Ser Val Met 545 550 555 560

His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser 565 570 575

Pro Gly Lys <210> 60 <211> 586 <212> PRT <2i3> Artificial sequence <220><2 is > Description of artificial sequence: synthetic polypeptide <400> 60

Asp He Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Ser Arg Giy Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 -67- 160877 - Sequence Listing.doc 201247704

Met Gin Leu Ser Ser Leu rrhr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Val 130 135 140

Arg Pro Gly Ser Ser Val Lys lie Scr Cys Lys Ala Ser Gly Tyr Ala 145 150 155 160

Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp lie Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr 180 185 190

Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser 195 200 205

Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr 225 230 235 240

Tyr Ala Met Asp Tyi Trp Gly GJn Gly Thr Thr Val Thr Val Ser Ser 245 250 255

Ala Ser Thr Lys Gly Pro Ser Val F*he Pro Leu Ala Pro Ser Ser Lys 260 265 270

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 275 280 285

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 290 295 300

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 305 310 315 320

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr G!n Thr 325 330 335

Tyr He Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 340 345 350

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro 355 360 365

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 370 375 380 -68 -

160877 - Sequence Listing.doc 201247704

Lys Pro Lys Asp Thr Leu Met He Ser Arg Thr Pro Glu Val Thr Cys 385 390 395 400

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 405 410 415

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 420 425 430

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 435 440 445

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 450 455 460

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 465 470 475 480

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 485 490 495

Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phc Tyr 500 505 510

Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 515 520 525

Asn Tyr Lys Thr Tlir Pro Ala Val Leu Asp Ser Asp Gly Ser Phe Arg 530 . 535 540

Leu Arg Ser Asp Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 545 550 555 560

Val Phe Scr Cys Scr Val Met His Glu Ala Leu His Asn His Tyr Thr 565 570 575

Gin Lys Ser Leu Scr Leu Ser Pro Gly Lys 580 585 <210> 61 <211> 586 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide<400> 61

Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala L 5 10 15

Ser Val Lys Met Ser Cys Lys rrhr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45 -69- 160877 - Sequence Listing.doc 201247704

Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 1)0

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Val 130 135 140

Arg Pro Gly Ser Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Ala 145 150 155 160

Phe Ser Ser tyr Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp He Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr 180 185 190

Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser 195 200 205

Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr 225 230 235 240

Tyr Ala Met Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser 245 250 255

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 260 265 270

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 275 280 285

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 290 295 300

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 305 310 315 320

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 325 330 335

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 340 345 350 • 70· 160877 - Sequence Listing.doc 201247704

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro Scr 355 360 365

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 370 375 380

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thi Cys 3S5 390 395 400

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 405 410 415

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 420 425 430

Glu Gin Tyr Asn Ser Thr Ding yr Arg Val Val Ser Val Leu Thr Val Leu 435 440 445

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 450 455 460

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr He Ser Lys Ala Lys Gly 465 470 475 480

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 485 490 495

Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Plie Tyr 500 505 510

Pro Ser Asp He Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 515 520 525

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Arg 530 535 540

Leu Arg Ser Asp Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 545 550 555 560

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 565 570 575

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 580 585 <210> 62 <211> 586 <212> PRT <2]3>Artificial Sequence<220><223> Description of Artificial Sequence: Synthetic Peptide <400> 62

Asp He Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala 15 10 15 -71 - 160877 - Sequence Listing.doc 201247704

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Val 130 135 140

Arg Pro Gly Ser Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Ala 145 150 155 160

Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp lie Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr 180 185 190

Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser 195 200 205

Ser Thr Ala Tyr Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Ar^ Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr 225 230 235 240

Tyr Ala Met Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser 245 250 255

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 260 265 270

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 275 280 285

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 290 295 300

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 305 310 315 320 -72-

160877 - Sequence Listing.doc 201247704

Leu Scr Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 325 330 335

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 340 345 350

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro 355 360 365

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 370 375 3S0

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 385 390 395 400

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 405 410 415

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 420 425 430

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Va丨 Leu Thr Val Leu 435 440 445

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 450 455 460

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 465 470 475 480

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 485 490 495

Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 500 505 510

Pro Scr Asp lie Ala Val Glu Trp Glu Scr Asn Gly Gin Pro Glu Asn 515 520 525

Asn Tyr Lys Thr Thr Pro Ala Val Leu Asp Ser Asp Gly Ser Phe Phe 530 535 540

Leu Tyr Ser Asp Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 545 550 555 560

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 565 570 575

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 580 585 <210> 63 <211> 586 <212> PRT <213>Artificial Sequence<220> •73·160877-Sequence List.doc 201247704 <223&gt Description of artificial sequence: synthetic peptide <400> 63

Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Lea Ala Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Ihi Arg Tyr 20 25 30

Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala Thr Leu Thr llir Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Va] Tyr Tyr Cys 85 90 95

Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 325

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Giy Ala Glu Leu Val 130 135 140

Arg Pro Gly Ser Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Ala 145 150 155 160

Phe Ser Ser Tyr Trp Met Asn Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp lie Gly Gin lie Trp Pro Gly Asp Gly Asp Thr Asn Tyr 180 185 190

Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Glu Ser Ser 195 200 205

Scr Thr Ala Tyr Met Gin Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Arg Arg Glu Thr Thr Thr Val Gly Arg Tyr Tyr 225 230 235 240

Tyr Ala Met Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser 245 250 255

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 260 265 270

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 275 280 285 •74·

160877· Sequence Listing.doc 201247704

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 290 295 300

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 305 310 315 320

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 325 330 335

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 340 345 350

Lys Val Glu Pro Lys Scr Cys Asp Lys Thr His Thr Ser Pro Pro 355 360 365

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 370 375 380

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 385 390 395 400

Val Val Val Asp Va! Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 405 410 415

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 420 425 430

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 435 440 445

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 450 455 460

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 465 470 475 480

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 485 490 495

Met Thr Lys Asn Gin Val Ser Leu rihr Cys Leu Val Lys Gly Phe Tyr 500 505 510

Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 515 520 525

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Arg 530 535 540

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 545 550 555 560

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 565 570 575

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 580 585 • 75· 160877· Sequence Listing.doc 201247704 <210> 64 <211> 1104 <212> DNA <213>Artificial Sequence<220><223>; description of artificial sequences: synthetic polynucleotide < 400 > 64 gaggtcacct tgagggagtc tggtcctgcg ctggtgaaac ccacacagac cctcacactg 60 acctgcacct tctctgggtt ctcactcagc aaatctgtta tgggtgtgag ctggatccgt 120 cagcccccag ggaaggccct ggagtggctt gcacacattt actgggatga tgacaagtac 180 tataatccat ccctaaagag caggctcacc atctccaagg acacctccaa aaaccaggtg 240 gtccttacaa tgaccaacat ggaccctgts gacacagcca cgtattattg tgcacggaga 300 gggatacgaa gtgctatgga ctattggggg caagggacca cggtcaccgt ctcctcagcg 360 tcgaccaagg gcccagaggt gcagctggtg cagtctggaa cagaggtgaa aaaacccggg 420 sagtctctea agatctccts taagggttct ggatacactg ttaccagtta ctggatcggc 480 tgggtgcgcc agatgcccgg gaaaggcctg gagtggatg £ gattcatcta tcctggtgac 540 tctgaaacca gatacagtcc gaccttccaa ggccaggtca ccatctcagc cgacaagtcc 600 ttcaataccg ccttcctgca giggagcagt ctaaaggcct cggacaccgc catg tattac 660 tgtgcgcgag tcggcagtgg ctggtaccct tatacttttg atatctgggg ccaagggaca 720 atggicaccg tctcttcagc gtcgaccaag ggcccagagg tgcagctggt gcagagcggc 780 gccgaggtga agaagcccgg cgccagcgtg aaggtgagct gcaaggccag cggctacacc 840 ttcaccaagt actggctggg ctgggtgcgg caggcccccg gccagggcct ggagtggatg 900 ggcgacatct accccggcta cgactacacc cactacaacg agaagttcaa ggaccgggtg 960 accctgacca ccgacaccag caccagcacc gcctacatgg agctgcggag cctgcggagc 1020 gacgacaccg ccgtgtacta ctgcgcccgg agcgacggca gcagcaccta ctggggccag 1080 ggcaccctgg tgaccgtgag cage 1104 <210> 65 <211> 993 <212> DNA <213>Artificial sequence<220>c223> Description of artificial sequence: synthetic polynucleotide <400> 65 gcgtcgacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 60 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 120 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 180 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 240 tacatctgca aegtgaatea caagcccagc aacac caagg tggacaagaa agttgagccc 300 aaatcttgtg acaaaactca cacatcacca ccgtctccag cacctgaact cctggmgg 360 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 420 -76- 160877- Sequence Listing .doc 201247704 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gicctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc igcccccatc ccgcgaggag atgaccaaga accaggtcag cctgacctgc ctgglcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaacl acaagaccac gcctcccgtg ctggactccg acggctcctt ccggcictac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc tccgggtaaa tga < 210 > 66 < 211 > 993 < 21.2 > DNA <213>Artificial Sequence<220>

≪ 223 > Description of Artificial Sequence of: synthetic polynucleotide < 400 > 66 gcgtcgacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca Sgactctact ccctcagcag cgtgstgacc gtgccctcca gcagcttggg cacccagacc tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgg £ agga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgcgaggag atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt Gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggac Tccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc iccgggtaaa tga <210> 67 <211> 1020 <212> DNA <213> artificial sequence <220><223> Description of artificial sequence: synthesis of polynucleotide sequences of table -77-160877- .doc 201247704 < 400 > 67 gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc atcaactgca aggccagtca gagtgteagt aatgatgtag cttggtacca gcagaaacca ggacagcctc ctaagctgct catttactat gcatccaatc gctacactgg egtccctgac cgattcagtg gcagcgggtc tgggacagat ttcactctca ccatcagcag cctscaggct gaagatgtgg cagtttatta ctgtcagcag gattataact ctccgtggac gttcggcgga gggaccaagg tggagatcaa acgtacggtg gctgcaccag aaatagtgat gac ^ cagtct ccagccaccc tgtctgtgtc tccaggggaa agagccaccc tctcctgcag ggccagtgag agtauagca gcaacttagc ctggtaccag cagaaacctg gccaggctcc caggctcttc atctatactg catccaccag ggccactgat atcccagcca ggttcagtgg cagtgggtct gggacagagt tcactctcac catcagcagc ctgcagtctg aagattttgc agtttattac tgtcagcagt ataataactg gccttcgatc accttcggcc aagggacacg actggagatt aaacgaacgg tggctgcacc agacgtgctg atgacccaga cccccctgag cctgcccgtg acccccggcg agcccgccas catcagctgc accagcagcc agaacatcgi gcacagcaac ggcaacacct acctggaglg gtacctgcag aagcccggcc agagccccca gctgctgatc tacaaggtga gcaaccggtt cagcggcgtg cccgaccggt tcagcggcag cggcagcggc accgacttca ccctgaagat cagccgggtg gaggccgagg acgtgggcst gtactactgc ttccaggtga gccacgtgcc ctacaccttc ggcggcggca ccaaggtgga gatcaagcgg 60 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020

<210> 68 <211> 321 <212> DNA <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide <400> 68 acggtggctg caccatctgt cttcatcttc actgcctctg ttgtgtgcct gctgaataac aaggtggata acgccctcca atcgggtaac aaggacagca cctacaecct cagcagcacc cacaaagtct acgcctgcga agtcacccat ttcaacaggg gagagtgttg a ccgccatctg atgagcagtt gaaatctgga ttctatccca gagaggccaa agtacagtgg tcccaggaga gtgtcacaga gcaggacagc ctgacgctga gcaaagcaga ctacgagaaa cagggcctga gctcgcccgt cacaaagagc 60 120 180 240 300 321

<210> 69 <211> 729 <212> DNA <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide <400> 69 gatatcaaac tgcagcagtc aggggctgaa ctggcaagac ctggggcctc aglgaagalg tcctgcaaga cttctggcta cacctttact aggtacacga tgcactgggt aaaacagagg cctggacagg gtctggaatg gattggatac attaatccta gccgtggtta tactaattac 60 120 180 160877- sequence Listing .doc • 78 - 201247704 aatcagaagt tcaaggacaa ggccacattg actacagaca aatcctccag cacagcctac 240 atgcaactga gcagcctgac atctgaggac tctgcagtct attactgtgc aagatattar 300 gatgatcatt actgccttga ctactggggc caaggcacca ctctcacagt ctcctcagtc 360 gaaggtggaa stggaggttc tggtggaagt ggaggttcag gtggagtcga cgacattcag 420 ctgacccagt ctccagcaat catgtctgca tctccagggg agaaggtcac catgacctgc 480 agagccagtt caagtgtaag ttacatgaac tggtaccagc agaagtcagg cacctccccc 540 aaaagatgga tttatgacac atccaaagtg gcttctggag tcccttatcg cttcagtg £ c 600 agtgggtctg ggacctcata ctctctcaca atcagcagca tggaggctga agatgctgcc 660 acttattact gccaacagtg gagtagtaac ccgct Cacgt tcggtgctgg gaccaagctg 720 gagctgaaa 729

<210> 70 <211> 366 <212> m < 213 > artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide <400> 70 gaagttcagc tgttggaaag cggcggaggt ttggtgcag ctigagggtc tctccggctc tcttgtgccg catcagggtt taccttcgct gatgagggaa tgatgtgggt tcggcaggcc ccaggaaagg gactggagtg ggtgtcacga atcacctata gcggcaagaa tacctactat gccgactccg tgaaagggcg gtttaccatt tcacgcgaca acagtaagaa caccctgtac ctgcaaatga attcactccg cgcggaagac actgcggtgt actactgcgc gaaatataca ggtcggattc tgggacacca tctgttcgac tactggggac aaggcacctt ggicacagtc tcttca 60 120 180 240 300 360 366

<210> 71 <211> 366 <212> DNA <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide <400> 71 gaggtgcaac tgctcgaatc tggcggggga ctggtacaac ctgggggtag ccttcgactc agctgcgccg cctccggatt taccttcgcc gaggaaagct ggaigtgggt gagacaggcg cctggaaaag ggctggagtg egtgagtcgg attggtcagg atggaaaaaa cacctattac cgagaggatg tgaagggacg attcaccata tccagggata atagtaaaaa cactctgtat ctccagatga acagccttcg cgcggaagac accgccgtct actattgtgc taagtacact ggacggatca tgggccatca tctgtttgat tactggggcc aggggacatt ggtgaccgtt tcctca 60 120 180 240 300 360 366 < 210 > 72 < 211 > 771 <212> DNA <213> artificial sequence 160877. Sequence Listing. doc • 79-201247704 <220><223> Description of artificial sequence: synthetic polynucleotide <400> 72 atggcccctg ttgccstctg ggctgcgttg gctglggggc tggaactgtg ggccgcagcg 60 cacgctctgc Ctgctcaggt agcatttact ccttacgcac ccgagcccgg atccacgtgc 120 agactcasge agtattatga tcaaaccgca cagatgtgtt gtagcaaatg tagcccaggc 180 cagcatgcca aggtgtt ctg caccaagacc tccgatacag tctgtgatag ctgtgaggat 240 tctacctaca cacagctcts gaactgggtt ccggagtgct tgtcctgtgg ttctcggtgc 300 agcagcgatc aagtcgaaac tcaggcctgc acccgagagc agaatcgcat ctgtacctgc 360 agaccgggtt ggtactgcgc actgtcaaaa caggaaggtt gccgactgtg cgccccactc 420 agaaagtgca ggcccggatt cggagttgct agacccggaa cagaaaccag tgatgtggtt 480 tgtaaacctt gtgctccggg gacctttagt aacacaacta gcagcacgga catctgcaga 540 ccccaccaga tctgcaatgt tgtggcaatt ccaggaaacg cctccatgga tgccgtdgc 600 acttcaacct ctcccacgag Aagtatggct cccggcgccg tgcatctccc gcagccggtg 660 tcaactcggt ccca^catac ccagcccaca ccagagccaa gcaccgcccc ttcaacttca 720 tttttgctgc caatgggacc ctccccacca gccgaaggaa gcactggcga c 771 <210> 73 <211> 450 <212> DNA <213> artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide <400> 73 atgggcgttc tgctgacaca gaggacgctg ctttcactcg tgttggctct cctcttcccg 60 tctatggcgt ctatggcaat gcacgtggcc cagccggcgg tcgttctggc ttcctcaaga 120 ggaatcgcat cttttgtttg cgaatatgcc tctc caggca aggcaacaga agtccgagta 180 acggtcctca gacaggccga ttcccaggtg acagaggtct gtgccgctac ttacatgatg 240 ggcaatgaac tgacatttct ggatgattca atctgcaccg gcacctccag cggtaatcaa 300 gtgaatctta ccatccaggg ccttcgcgcc atggatacag gactgtatat ctgcaaagtg 360 gaactgatgt atccgcctcc ctactatctg ggaatcggaa acgggacaca aatatatgtg 420 atcgatcccg aaccgtgtcc cgatagcgac 450 < 230 > 74 < 211 > 699 < 212 > DNA <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polynucleotide <400> 74 gagcccaaat ctagcgacaa aactcacaca tcaccaccgt ctccagcacc tgaactcctg 60

Egggggccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 120 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 180 • 80 - 160877- Sequence Listing .doc 201247704 aactggtacg tggacegcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 240 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 300 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 360 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgc 420 gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 480 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 540 cccgtgctgg actccgacgg ctccttccgg ctctacagca agctcaccgt ggacaagagc 600 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 660 tacacgcaga agagcctctc cctgtctccg ggiaaatga 699 < 210 > 75 < 21i > 699 < 212 > DNA <213>Artificial sequence <220><223> Description of artificial sequence: synthetic polynucleotide

≪ 400 > 75 60 120 180 240 300 360 420 480 540 600 660 699 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgc gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatga < 210 > 76 < 211 > 368 < 212 > PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 76

Glu Val Thr Leu Arg Glu Scr Gly Pro Ala Leu Val Lys Pro Thr Gin 15 10 15

Thr Leu Thr Leu Thr Cys Thr Phc Ser Gly Phe Ser Leu Ser Lys Ser 20 25 30

Va, Met Gly Val Ser Trp He Arg Gin Pro Pro Gly Lys Ala Leu Glu • 81 · 160877 · Sequence Listing.doc 201247704

Trp Leu Ala His lie Tyr Trp Asp Asp Asp Lys Tyr Tyr Asn Pro Ser 50 55 60

Leu Lys Ser Arg Leu Thr lie Ser Lys Asp Thr Ser Lys Asn Gin Val 65 70 75 80

Va]Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95

Cys Ala Arg Arg Gly lie Arg Ser Ala Met Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Giu Val Gin 115 120 125

Leu Val Gin Ser Gly Thr Glu Val Lys Lys Pro Gly Glu Ser Leu Lys 130 135 140 lie Ser Cys Lys Gly Ser Gly Tyr Thr Val Thr Ser Tyr Trp lie G1 145 150 155 16

Trp Val Arg Gin Met Pro Gly Lys Gly Leu Glu Trp Met Gly Phe lie 165 170 175

Tyr Pro Gly Asp Ser Glu Thr Arg Tyr Ser Pro Thr Phe Gin Gly Gin 180 185 190

Vai Thr He Ser Ala Asp Lys Ser Phe Asn Thr Ala Phe Leu Gin Trp 195 200 205

Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys Ala Arg Val 210 215 220

Gly Ser Gly Trp Tyr Pro Tyr Thr Phe Asp lie Trp Gly Gin Gly Thr 225 230 235 240

Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Glu Val Gin Leu 245 250 255

Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val 260 265 270

Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Lys Tyr Trp Leu Gly Trp 275 280 285

Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met Gly Asp lie Tyr 290 295 300

Pro Gly Tyr Asp Tyi Thr His Tyr Asn Glu Lys Phe Lys Asp Aig Val 305 310 315 320

Thr Leu Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr Met Glu Leu Arg 325 330 335

Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Asp -82 -

160877 - Sequence Listing.doc 201247704 340 345 350

Gly Ser Ser Thr Tyr Trp Gly Gin Gly Thx Leu Val Thr Val Ser Ser 355 360 365 <210> 77 <211> 330 <212> PRT <213> Artificial Sequence <220><223> Artificial Sequence Description: Synthetic Peptide <400> 77

Ala Ser Tlir Lys Gly Pro Scr Val Phc Pro Leu Ala Pro Ser Ser Lys 15 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 65 70 75 80

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thx Lys Val Asp Lys 85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro 100 100 110 110

Pro Ala Pro Glu Leu Leu Gly Giy Pro Ser Val Phe Leu Phe Pro Pro 115 120 125

Lys Pro Lys Asp Thr Leu Met He Ser Arg Thr Pro Glu Val Thr Cys 130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205

Lys Ala Leu Pro Ala Pro He Glu Lys Thr lie Ser Lys Ala Lys Gly 210 215 220

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 225 230 235 240 -83- 160877 · Sequence Listing.doc 201247704

Met Thr Lys Asn Gin Val Scr Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255

Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phc Arg 275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 78 <211> 330 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide<400> 78

Ala Scr Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Scr Lys 15 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Scr Leu Gly Thr Gin Thr 65 70 75 80

Tyr He Cys Asn VaJ Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 -84-

160877 - Sequence Listing.doc 201247704

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 210 215 220

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 225 230 235 240

Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255

Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phc Phe 275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 79 <211> 340 <212> PRT <213>Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide<400> 79

Asp lie Val Met Thr Gin Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 15 10 15

Glu Arg Ala Thr lie Asn Cys Lys Ala Ser Gin Ser Val Ser Asn Asp 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie 35 40 45

Tyr Tyr Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phc Thr Leu Thr lie Ser J5cr Leu Gin Ala 65 70 75 80 -85 - 160877 - Sequence Listing.doc 201247704

Glu Asp Val Ala Val Tyr Tyr Cys Gin Gin Asp Tyr Asn Ser Pro Trp 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu He Lys Arg Thr Val Ala Ala 100 105 HO

Pro Glu lie Val Met Thr Gin Ser Pro Ala Thr Leu Ser Val Ser Pro 115 120 125

Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser lie Ser Ser 130 135 140

Asn Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Phe 145 150 155 160 lie Tyr Thr Ala Ser Thr Arg Ala Thr Asp lie Pro Ala Arg Phe Ser 】 65 170 175

Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin 180 185 190

Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Tyr Asn Asn Trp Pro 195 200 205

Ser lie Thr Phe Gly Gin Gly Thr Arg Leu Glu lie Lys Arg Thr Val 210 215 220

Ala Ala Pro Asp Val Leu Met Thr Gin Thr Pro Leu Ser Leu Pro Val 225 230 235 240

Thr Pro Gly Glu Pro Ala Ser lie Ser Cys Thr Ser Ser Gin Asn lie 245 250 255

Val His Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gin Lys Pro 260 265 270

Gly Gin Ser Pro Gin Leu Leu lie Tyr Lys Val Ser Asn Arg Phe Ser 275 280 285

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 290 295 300

Leu Lys lie Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys 305 310 3]5 320

Phe Gin Val Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val 325 330 335

Glu lie Lys Arg 340 <210> 80 <211> 106 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide -86-

160877-SEQ ID NO.doc 201247704 <400> 80

Thr Val Ala Ala Pro Ser Val Phc lie Phe Pro Pro Ser Asp Glu Gla 15 10 15

Leu Lys Ser Cly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phc Tyr 20 25 30

Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser 35 40 45

Gly Asn Ser Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr 50 55 60

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 65 70 75 80

His Lys Val Tyr Ala Cys Glu Val Thr Ris Gin Gly Lea Ser Ser Pro 85 90 95

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 81 <211> 262 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide<>81

Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala lie Leu Lys Gly 15 10 15

Val Gin Cys Asp lie Lys Leu Gin Gin Ser Gly Ala Glu Leu Ala Arg 20 25 30

Pro Gly Ala Ser Val Lys Met Ser Cys Lys Iht Ser Gly Tyr Thr Phe 35 40 45

Thr Arg Tyr Thr Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu 50 55 60

Glu Trp lie Gly Tyr lie Asn Pro Ser Arg Gly Tyr Thr Asn Tyr Asn 65 70 75 80

Gin Lys Phe Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser 85 90 95

Thr Ala Tyr Met Gin Leu Sex Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105 110

Tyr Tyr Cys Ala Arg Tyr Tyr Asp Asp His Tyr Cys Leu Asp Tyr Trp 115 120 125

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser Val Glu Gly Gly Ser Gly 130 135 140 • 87· 160877 · Sequence Listing.doc 201247704

Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Val Asp Asp lie Gin Leu 145 150 155 160

Thr Gin Ser Pro Ala lie Met Ser Ala Ser Pro Gly Glu Lys Val Thr 165 170 175

Met Thr Cys Arg Ala Ser Ser Ser Ser Ser Serr Met Asn Trp Tyr Gin 180 185 190

Gin Lys Ser Gly Thr Ser Pro Lys Arg Trp lie Tyr Asp Thr Ser Lys 195 200 205

Val Ala Ser Gly Val Pro Tyr Arg Phe Ser Gly Ser Gly Ser Gly Thr 210 215 220

Ser Tyr Ser Leu Thr lie Ser Ser Met Glu Ala Glu Asp Ala Ala Thr 225 230 235 240

Tyr Tyr Cys Gin Gin Trp Ser Ser Asn Pro Leu Thr Phe Gly Ala Gly 245 250 255

Thr Lys Leu Glu Leu Lys 260 <210> 82 <21l> 141 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala lie Leu Lys Gly 15 10 15

Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin 20 25 30

Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45

Ala Asp Glu Gly Met Met Trp Val Arg Gin Ala Pro Gly Lys Gly Leu 50 55 60

Glu Trp Val Ser Arg lie Thr Tyr Ser Gly Lys Asn Thr Tyr Tyr Ala 65 70 75 80

Asp Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn 85 90 95

Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110

Tyr Tyr Cys Ala Lys Tyr Thr Gly Arg lie Leu Gly His His Leu Phe 115 120 125 -88 -

160877 - Sequence Listing.doc 201247704

Asp Tyr Trp Gly Gin Gly TTir Leu Val Thr Val Ser Ser 130 135 140 <210> 83 <211> 141 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic polypeptide <400> 83

Met Glu Phc Gly Leu Scr Trp Leu Phc Leu Val Ala He Leu Lys Gly 15 10 15

Val Gin Cys Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin 20 25 30

Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45

Ala Glu Ciu Ser Trp Met Trp Val Arg Gin Ala Pro Gly Lys Gly Leu

50 55 60

Glu Trp Val Ser Arg lie Gly Gin Asp Gly Lys Asn Thr Tyr Tyr Arg 65 70 75 80

Glu Asp Val Lys Gly Arg Phe Thr lie Scr Arg Asp Asn Scr Lys Asn 85 90 95

Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110

Tyr Tyr Cys Ala Lys Tyr Thr Gly Arg lie Met Gly His His Leu Phe 115 120 125

Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 130 135 140 <210> 84 <211> 257 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic polypeptide <400> 84

Met Ala Pro Val Ala Val Trp Ala Ala Leu Ala Val Gly Leu Glu Leu 10 15

Trp Ala Ala Ala His Ala Leu Pro Ala Gin Val Ala Phe Thr Pro Tyr 20 25 30

Ala Pro Glu Pro Gly Ser Thr Cys Arg Leu Arg Glu Tyr Tyr Asp Gin 35 40 45

Thr Ala Gin Met Cys Cys Ser Lys Cys Ser Pro Gly Gin His Ala Lys 50 55 60 -89- 160877 · Sequence Listing.doc 201247704

Val Phe Cys Thr Lys Thr Ser Asp Thr Val Cys Asp Ser Cys Glu Asp 65 70 75 80

Ser Thr Tyr Thr Gin Leu Trp Asn Trp Val Pro Glu Cys Leu Ser Cys 85 90 95

Gly Ser Arg Cys Ser Ser Asp Gin Val Glu Thr Gin Ala Cys Thr Arg 100 105 110

Glu Gin Asn Arg lie Cys Thr Cys Arg Pro Gly Trp Tyr Cys Ala Leu 115 120 125

Ser Lys Gin Glu Gly Cys Arg Leu Cys Ala Pro Leu Arg Lys Cys Arg 130 135 140

Pro Gly Phe Gly Val Ala Arg Pro Gly Thr Glu Thr Ser Asp Vai Val 145 丨50 155 160

Cys Lys Pro Cys Ala Pro Gly Thr Phe Ser Asn Thr Thr Ser Ser Thr 165 170 175

Asp lie Cys Arg Pro His Gin lie Cys Asn Val Val Ala lie Pro Gly 180 185 190

Asn Ala Ser Met Asp Ala Val Cys Thr Ser Thr Ser Pro Thi1 Arg Ser 195 200 205

Met Ala Pro Gly Ala Val His Leu Pro Gin Pro Val Ser Thr Arg Ser 210 215 220

Gin His Thr Gin Pro Thr Pro Glu Pro Ser Thr Ala Pro Ser Thr Ser 225 230 235 240

Phe Leu Leu Pro Met Gly Pro Ser Pro Pro Ala Glu Gly Ser ITir Gly 245 250 255

Asp <210> 85 <211> 150 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Met Gly Val Leu Leu Thr Gin Arg Thr Leu Leu Ser Leu Val Leu Ala 15 10 15

Leu Leu Phe Pro Ser Met Ala Ser Met Ala Met His Val Ala Gin Pro 20 25 30

Ala Val Val Leu Ala Ser Ser Arg Gly lie Ala Ser Phe Val Cys Glu 35 40 45 -90·

160877 - Sequence Listing.doc 201247704

Tyr Ala Ser Pro Gly Lys Ala Thr Glu Val Arg Val Thr Val Leu Arg 50 55 60

Gin Ala Asp Ser Gin Val T.r Glu Val Cys Ala A]a Lr Tyr Met Met

Gly Asn Glu Leu thr Phe Leu Asp Asp Ser lie Cys Thr Gly Thr Ser 85 90 95

Ser Gly Asn Gin Val Asn Leu Thr lie Gin Gly Leu Arg Ala Met Asp 100 105 110

Thr Gly Leu Tyr lie Cys Lys Val Glu Leu Met Tyr Pro Pro Pro Tyr 115 120 125

Tyr Leu Gly lie Gly Asn Gly Thr Gin lie Tyr Vai lie Asp Pro Glu 130 135 140

Pro Cys Pro Asp Ser Asp 145 150

<210> 86 <211> 232 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala 15 10 15

Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 20 25 30

Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 50 55 60

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin 65 70 75 80

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin 85 90 95

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 100 105 110

Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro 115 120 125

Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Scr Arg Glu Glu Met Thr 130 135 140 -91 - 160877 - Sequence Listing.doc 201247704

Lys Asn Gin Val Scr Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser ]45 150 155 160

Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr 165 170 175

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Arg Leu Tyr 180 185 190

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe 195 200 205

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gin Lys 210 215 220

Ser Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 87 <211> 232 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 87

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 15 10 15

Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 20 25 30

Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val 35 40 45

Val Asp Val Ser His Glu Asp Pro Glu Va丨 Lys Phe Asn Trp Tyr Val 50 55 60

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin 65 70 75 80

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin 85 90 95

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Scr Asn Lys Ala 100 105 110

Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro 115 120 125

Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr 130 135 140

Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 145 150 155 160

Asp lie Ala Val Glu Trp Glu Scr Asn Gly Gin Pro Glu Asn Asn Tyr -92- 160877 - Sequence Listing.doc 201247704 165 170 175

Lys Thr Thr Ρπ> Pro Va丨 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 180 185 190 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe 195 200 205

Ser Cys Ser Val Met His GIu Ala Leu His Asn His Tyr Ήιτ Gin Lys 210 215 220

Ser Leu Ser Leu Scr Pro Gly Lys 225 230 <210> 88 <211> 357 <212> m <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polynucleotide

≪ 400 > 88 caggtccaac tgcagcagtc tgggcctgag ctggtgaggc ctggggcttc agtgaagatg tcctgcaggg cttcgggcta taccttcacc agctactggt tgcactgggt taaacagagg cctggacaag gccttgagtg gattggcatg attgatcctt ccaatagtga cactaggttt aatccgaact tcaaggacaa ggccacattg aatgtagaca gatcttccaa cacagcctac atgctgctca gcagcctgac atctgctgac tctgcagtct attactgtgc cacatatggt agctacgttt cccctctgga ctactggggt caaggaacct cagtcaccgt ctcctca 60 120 180 240 300 357 < 210 & gt 89 <211> 993 <212> DNA <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polynucleotide

≪ 400 > 89 gcgtcgacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc aaatcttgtg acaaaactca cacatcacca ccgtctccag cacctgaact cctggggggg ccgtcagtct tcctcttccc cccaaaaccc aaggaccagc tcatgatctc ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgcgaggag 60 120 180 240 300 360 420 480 540 600 660 160877 · sequence Listing _doc -93- 720 201247704 atgaccaaga Accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 780 gccgtggagt gggagagcaa tgggcagccg gagaa caact acaagaccac gcctcccgtg 840 ctggactccg acggctcctt ccggctctac agcaagctca ccgtggacaa gagcaggtgg 900 cagcagggga acgtcttctc atgctccgtg ctgcatgagg ctctgcacaa ccactacacg 960 cagaagagcc tctccctgtc tccgggtaaa tga 993 < 210 > 90 < 211 > 993 < 212 > DNA < 2] 3 > artificial sequence < 220 > < 223 > description of artificial sequence of: synthetic polynucleotide < 400 > 90 gcgtcgacca agg £ cccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 60 ggcacagcgg ccctgggctg cctsgtcaag gactacttcc ccgaaccggt gacggtgtcg 120 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 180 ggactctact ccctcascag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 240 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 300 aaatcttgtg acaaaactca cacatcacca ccgtctccag cacctgaact cctsggggge 360 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 420 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 480 tacgtggac ^ scgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 540 agc acgtacc gtgtggtcag cgtcctcgcc gtcctgcacc aggactggct gaatg ^ caag 600 gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 660 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgcgaggag 720 atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 780 gccgtggcgt magagcaa tgsscagccg gagaacaact acaagaccac gcctcccgtg S40 ctggactccg acggctcctt ccggctctac agcaagctca ccgtggacaa gagcaggtgg 900 cagca ^ gga acgtcttctc atgctccgtg atgcatgagg Ctctgcacgc ccactacacg 960 cagaasagcc tctccctgtc tccgggtaaa tga 993 <210> 91 <211> 119 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Arg Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 -94-

160877· Sequence Listing.doc 201247704

TrpLcu His TrpValLys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie

Gly Met lie Asp Pro Ser Asn Ser Asp Thr Arg Phe Asn Pro Asn Phe 50 55 60

Lys Asp Lys Ala Thr Leu Asn Val Asp Arg Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Lea Leu Ser Ser Leu Thr Ser Ala Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Tyr Gly Ser Tyr Val Ser Pro Leu Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Ser Val Thr Val Ser Ser 115

<210> 92 <211> 330 <212> PRT <213> Artificial sequence <220><2:23> Description of artificial sequence: synthetic polypeptide <400>

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 15 10 15

Scr Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 65 70 75 80

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Scr Pro Pro Scr 100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125

Lys Pro Lys Asp Gin Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu -95- 160877 - Sequence Listing.doc 201247704 165 170 175

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190

His Gin Asp Trp Leu Asn GJy Lys Glu Tyr Lys Cys Lys VaJ Scr Asn 195 200 205

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 210 215 220

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 225 230 235 240

Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255

Pro Ser Asp He Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn 260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Arg 275 280 2S5

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 93 <211> 330 <212> PRT <213>Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide<400> 93

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 15 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 50 55 60

Leu Scr Ser Val Val Thr Val Pto Ser Ser Ser Leu Gly Thr Gin Thr 65 70 75 80

Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 • 96- 160877 - Sequence Listing.doc 201247704

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Ser Pro Pro 100 100 110 110

Pro AJa Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125

Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys 130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175

Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Ala Val Leu 1.80 185 190

His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205

Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly 210 215 220

Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu 225 230 235 240

Met Thr Lys Asn Gin Val Ser Lea Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255

Pro Ser Asp lie Ala Val Ala Trp Glu Ser Asn Gly Gin Pro Glu Asn 260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Arg 275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn 290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr 305 31.0 315 320

Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 94 <211> 16 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide<400> 94

Ala Lys Thr Thr Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Arg 1 5 10 15 •97· 160877-sequence table.doc 201247704 <210> 95 <211> 17 <212> PRT <213> Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 95

Ala Lys Thr Thr Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Arg 15 10 15

Val <210> 96 <211> 9 <212> PRT <213> Artificial sequence Description of artificial sequence: synthetic peptide <400> 96

97. A < 211 > 10 <

Ser Ala Lys Thr Thr Pro Lys Leu Gly Gly 1 5 10 <210> 98 <211> 6 <212> PRT <2]3>Artificial Sequence <220><223> Description of Artificial Sequence: Synthesis Peptide <400> 98

Ser Ala Lys Thr Thr Pro 1 5 <210> 99 <211> 6 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Arg Ala Asp Ala Ala Pro 160877 · Sequence Listing.doc 98-

201247704 <210> 100 <211> 9 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400>

Arg Ala Asp Ala Ala Pro Thr Val Ser 1 5 <210> 101 <211> 12 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>; 101

Arg Ala Asp Ala Ala Ala Ala Gly Gly Pro Gly Ser 1 5 10

<2I0> 102 <211> 27 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400>

Arg Ala Asp Ala Ala Ala Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly I 5 10 15

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 20 25 <210> 103 <211> 18 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <;400> 103

Ser Ala Lys Thr Thr Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala 15 10 15

Arg Val <210> 104 <211> 5 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400>

Ala Asp Ala Ala Pro 99·160877 - Sequence Listing.doc 201247704 1 5 <210> 105 <211> 12 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic peptide <400> 105

Ala Asp Ala Ala Pro Thr Val Ser lie Phe Pro Pro ] 5 10 <210> 106 <211> 6 <212> PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthesis Peptide <400> 106

Gin Pro Lys Ala Ala Pro <210> 107 <211> 13 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400>

Gin Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro 1 5 10 <210> 108 <211> 6 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic peptide <400> 108

Ala Lys Thr Thr Pro Pro <210> 109 <211> 13 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Ala Lys Thr Thr Pro Pro Ser Val Thr Pro Leu Ala Pro 1 5 10 <210> 110 <211> 6 100-

160877· Sequence Listing.doc 201247704 <2I2> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Ala Lys Thr Thr Ala Pro <210> 111 <211> 13 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400>

Ala Lys Thr Thr Ala Pro Ser Val Tyt Pro Leu Ala Pro 1 5 10

<210> 112 <211> 15 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic peptide <400> 112

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Giy Ser 15 10 15 <210> 113 <211> 15 <212> PRT <213>Artificial Sequence <220><223> Artificial Sequence Description: Synthetic peptide <400> 113 G3y Glu Asn Lys Val GIu Tyr Ala Pro Ala Leu Met Ala Leu Ser 15 10 15 <210> 114 <211> 15 <212> PRT <213> Artificial sequence<220><223> Description of artificial sequence: synthetic peptide <400> 114

Gly Pro Ala Lys Glu Leu Thr Pro Leu Lys Glu Ala Lys Val Ser 15 10 15 <210> 115 <211> 15 <212> PRT <213> Artificial Sequence <220><223> Artificial Sequence Description: Synthetic peptide-101 - 160877 · Sequence Listing.doc 201247704 <400> 115

Gly His Glu Ala Ala Ala Val Met Gin Val Gin Tyr Pro Ala Ser 15 10 15 <210> 116 <211> 24 <212> PRT <2]3> Artificial Sequence <220><223> Description of the sequence: synthetic peptide <400> 116

Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala 15 10 15

Pro Ser Va丨Phe lie Phe Pro Pro 20 <210> 117 <211> 26 <2I2> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 117

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ala Ser Thr 15 10 15

Lys Gly Pro Ser Val Phe Pro Leu Ala Fro 20 25 <210> 118 <211> 119 <212> PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide <4〇0> 118

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp lie Glu Trp lie Lys Gin krg Pro Gly His Gly Leu Glu Trp lie 35 40 45

Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe 50 55 60

Ar£ Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Scr Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 -102-

160877 - Sequence Listing.doc 201247704

Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly 100 105 110

119 <211> 108 &lt

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 15 30 15

Glu Thr Val Thr lie Thr Cys Arg Thr Ser Glu Asn lie Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro His Leu Leu Val 35 40 45

Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pto Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro Leu 85 90 95

Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys Arg 100 105 <210> 120 <211> 119 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 120

Glu Val Gin I-eu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Lys Leu Scr Cys Ala Ala Ser Giy Phe Thr Phe Asn Asti Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45

Ala Tyr He Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Scr Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr •103· 160877_ Sequence Listing.doc 201247704 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Fhe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser 115 <210> 121 <211> 108 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys He Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 100 105 <210> 122 <2]1> 118 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic polypeptide <400> 122

Lys lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 15 10 15

Thr Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30

Gly Met Asn Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 •104·

160877 - Sequence Listing.doc 201247704

Gly Trp He Asn lie Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Leu Cys 85 90 95

Ala Arg Asp Ser Tyr Ser Gly Gly Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 lie Val Thr Val Ser Ser 115 <210> 123 <211> 114 <212> PRT <213>Artificial Sequence<220>;<223> Artificial sequence description: synthetic peptide

<400> 123

Asp lie Val Niet Thr Gin Ser Pro Scr Ser Leu Ser Val Ser Ala Gly 15 10 15

Glu Lys Val Thr Leu Ser Cys Lys Ser Ser Gin Ser Leu Leu lie Ser 20 25 30

Gly Asp Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45

Pro Pro Lys Leu Leu lie Tyr Gly Ala Ser Thr Arg Asp Ser Gly Val 50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr 65 70 75 80 lie Scr Ser Va) Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn 85 90 95

Asp His Ser Phe Pro Pro Thi Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110

Lys Arg <210> 124 <211> 122 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400> 124

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 15 10 15 -105- 160877 - Sequence Listing.doc 201247704

Val Lys lie Ser Cys Lys Ala Ser G.y Tyr Thr Phe ?〇r Asp Tyr

Ser Met His Trp Val Lys Gin Ala Pro Gly Lys G]y Leu Lys Trp Met 35 40 45

Gly Trp lie His Thr Glu Thr Gly GIu Pro Arg Tyr Val Asp Asp Fhe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95

Ala Arg Asp Ser Tyr Tyr Phe Gly Ser Ser Tyr Tyr Phe Asp Tyr Trp 100 105 110

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 125 <211> 108 <212> PRT <213> Artificial Sequence <220><M3> Description of Artificial Sequence: Synthetic Polypeptide<400> 125

Asp Thr Val Met Thr Gin Ser His Lys Phe Met Scr Thr Ser Val Gly 15 10 15

Asp Arg Val Scr Jle Thr Cys Lys Ala Ser Gin Asp Val Ser Ser Ala 20 25 30

Vai Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Ser Ala Scr Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Ser Gly Ser Gly Met Asp Phe Thr Phe Thr lie Ser Ser Val Gin Ala 65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gin Gin His Tyr Ser Thr Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Glu Arg 100 105 <210> 126 <2tl> 123 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide -106-

160877·Sequence list.doc 201247704 <400> 126

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Met Lys He Ser Cys Lys Ala Ser Asp Tyr Ser Phe Thr Ala Tyr 20 25 30

Thr lie His Trp Met Lys Gin Scr His Gly Lys Asn Leu Glu Trp lie 35 40 45

Gly Leu lie Asn Pro Tyr Asn Gly Gly Ser Tyr Asn Gin Lys Phe

Gin Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Ser Ser lie Ala Tyr 65 70 75 80

Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Arg Gly Tyr Asp Arg Glu Gly His Tyr Tyr Ala Met Asp Tyr 100 105 110

Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 127 <211> 108 <2\2> PRT <213>Artificial Sequence <220><223> Description of Artificial Sequence: Synthesis Peptide <400> 127

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ala Ala Ser Val Gly 15 10 15

Glu Thr Val Thr He Thr Cys Arg Ala Ser Glu Asn lie Tyr Thr Phe 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Thr Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Scr Gly 50 55 60

Ser Gly Ser Gly Hir Gin Phe Ser Leu Lys lie Lys Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Gly Leu Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 100 105

<210> 128 <211> 122 <212> PRT • 107·160877· Sequence Listing.doc 201247704 <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 128

Glu Val Gin Leu Gin Gin Ser Gly Pro Giu Leu Val Gin Pro Gly Ala 15 10 15

Ser Met Lys lie Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr 20 25 30

Thr Met Asn Trp Val Lys Gin Ser His Gly Lys Asn Leu Glu Trp lie 35 40 45

Gly Leu lie Asn Pro Tyr Asn Gly Gly Ser Arg Tyr Asn Gin Lys Phe 50 55 60

Met Ala Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Glu Leu Leu Ser Val Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Asp Ala Gly Tyr Phe Gly Ser Gly Phe Tyr Phe Asp Tyr Trp 100 105 110

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 129 <231> 107 <212> FRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide <400> 129

Asp lie Val Met Thr Gin Ser His Lys Phe Met Ser Thr Ser Val Gly 35 10 15

Asp Arg Val Ser lie Thr Cys Lys Ala Ser Gin Asp Vai Ser Thr Ala 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Ser Ala Ser Tyr Arg Ser Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr 11c Ser Ser Val Gin Ala 65 70 75 80

Giu Asp Leu Ala Val Tyr Tyr Cys Gin Gin His Tyr Ser Thr Pro Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 100 105 -108-

160877 - Sequence Listing.doc 201247704 <210> 130 <211> 118 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide <400>

Gin Val Gin Leu Gin Gin Pro Gly Ser Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Glv Asn lie Tyr Pto Gly Thr Val Asn Tht Asn Tyr Asp Glu Lys Phe 50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyt Cys 85 90 95

Thr Arg Asp Tyr Tyr Gly Gly Gly Leu Asn Tyr Trp Gly Gin Gly Thr 100 105 110

Thr Leu Thr Val Ser Ser 115 <210> 131 <211> 108 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Ser lie Val Met Thr Gin llir Pro Lys Phe Leu Leu Val Ser Ala Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Ser Val Ser Asn Asp 20 25 30

Val Ala Trp Phe Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Tyr Ala Ser Asn Arg Tyr Ala Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Ser Gly Phe Gly Thr Asp Phe Thr Phe Thr lie Ser Thr Val Gin Ala 65 70 75 80

Glu Asp Leu Ala Val Tyr Phe Cys His Gin Asp Tyr Ser Ser Pro Arg -109- 160877 - Sequence Listing.doc 201247704 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu GIu lie Lys Arg 100 105 <210> 132 <211> 117 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 132

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Arg Lys Pro Gly Glu 15 10 15

Thr Val Lys lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr 20 25 30

Gly lie Asn Trp Va] Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met 35 40 45

Gly Trp lie Asn Thr His Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Asn Tlir Ala Tyr 65 70 75 80

Leu Gin He Asn Asn Leu Asn Asn Gly Asp Met Gly Thr Tyr Phe Cys 85 90 95

Thr Arg Ser His Arg Phe Gly Leu Asp Tyr Trp Gly Gin Gly Thr Ser 100 105 110

Val Thr Vai Ser Ser 115 <210> 133 <211> 112 <212> FRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Asn Val Leu Thr Gin Ser Pro Pro Ser Leu Ala Val Ser Leu Gly 15 10 15

Gin Arg Ala Thr He Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asn 20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45

Lys Phe Leu He Tyr Ala Ala Ser Asn Leu Glu Scr Gly lie Pro Ala 50 55 60 •110-

160877· Sequence Listing.doc 201247704

Arg Phc Scr Gly Sex Gly Ser Gly Thr Asp Phe Asn Leu Asn He His 65 70 75 80

Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn 85 90 95

Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu He Lys Arg 100 105 110 <210> 134 <211> 112 <212> PRT <213> Artificial Sequence <220><223> Artificial Sequence Description: Synthetic Peptide <400> 134

Gin Val Gin Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met Asn Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp IJe 35 40 45

Gly Arg 11c Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phc 50 55 60

Lys Asp Lys Ala !le Leu Tlir Val Asp Lys Ser Ser Ser Thr Ala Phe 65 70 75 80

Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Val Ser Asp Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 100 105 110 <210> 135 <211> 113 <212> m <213> Artificial Sequence <220><223> Artificial Sequence Description: Synthetic Peptide <400> 135

Asp Va] Val Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly 15 10 15

Gin Pro Ala Ser He Ser Cys Lys Ser Ser Gin Ser Leu Leu Asp Ser 20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Giy Glu Set 35 40 45

Pro Lys Leu Leu lie Tyr Val Val Ser Lys Leu Glu Ser Gly Val Pro 50 55 60 -111- 160877 - Sequence Listing.doc 201247704

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 80

Ser Are Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala 85 90 95

Thr His Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110

Arg <210> 136 <211> 112 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Vai Gin B eu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met Asn Trp Va) Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp lie 35 40 45

Gly Arg lie Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe 65 70 75 80

Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Val Ser Asp G]y Tyf Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 100 105 110 <210> 137 <211> 113 <212> PRT <213> Manual Sequence <220><223> Description of the artificial sequence: synthetic peptide <400> 137

Asp Val Val Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly 15 10 15

Gin Pro Ala Ser lie Ser Cys Lys Ser Ser Gin Ser Leu Leu Asp Ser 20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser 35 40 45 •112·

160877 - Sequence Listing.doc 201247704

Pro Lys Leu Leu lie Tyr Va! Thr Asp lie Leu Glu Ser Gly Val Pro 50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 SO

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala 85 90 95

Thr His Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110

Arg <210> 138 <2ll> 119 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide

<400> 138

Glu Val Gin Leu Gin Gin Ser Gly Pro Asp Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Arg He Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Asn Leu His Trp Val Lys Gin Ser His Gly Lys Ser Leu Glu Trp lie 35 40 45

Gly Tyr lie Tyr Pro Tyr Asn Gly lie Thr Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Ser Lys Ala Thr Leu Thr Val Asp Ser Scr Ser Asn Thr Ala Tyr 65 70 75 80

Met Asp Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Asp Ala Tyr Asp Tyr Asp Tyr Leu Thr Asp Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala 115 <210> 139 <211> 108 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Leu Leu Thr Gin Ser Pro Val lie Leu Ser Val Scr Pro Gly -113- 160877 - Sequence Listing.doc 201247704 5 10 15

Glu Arg Val Ser Phe Ser Cys Arg Thr Ser Lys Asn Val Gly Thr Asn 20 25 30 lie His Trp Tyr Gin Gin Arg Thr Asn Gly Ser Pro Arg Leu Leu lie 35 40 45

Lys Tyr Ala Ser Glu Arg Leu Pro Gly lie Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser lie Asn Ser Val Glu Ser 65 70 75 80

Glu Asp lie Ala Asp Tyr Tyr Cys Gin Gin Ser Asn Asn Trp Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg 100 105 <210> 140 <211> 244 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 140

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Tlir Ser Tyr 20 25 30

Trp lie Glu Trp He Lys Gin Arg Pro Gly His Gly Leu Glu Trp lie 35 40 45

Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe 50 55 60

Arg Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly loo 105 no

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Gin Val Gin 115 120 125

Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala Ser Val Lys 130 135 140 lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp lie Glu 145 150 155 160 •114·

160877 - Sequence Listing.doc 201247704

Trp lie Lys Gin Arg Pro Gly His Gly Leu Glu Trp lie Gly Glu lie 165 170 175

Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe Arg Asp Lys 180 185 190

Ala Thr Phe Tlir Ala Asp Thr Ser Ser Asn Thr Ala Tyr Met Gin Leu 195 200 205

Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Gly 210 215 220

Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly Thr Leu Val 225 230 235 240

Thr Val Scr Ala

<210> 141 <211> 221 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Scr Leu Ser Ala Ser Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Thr Ser Glu Asn lie Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro His Leu Leu Val 35 40 45

Tyr Asn Thr Lys Thr Leu Ala C3u Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro Leu 85 90 95

Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys ATg Thr Val Ala Ala 100 105 110

Pro Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val 115 120 125

Gly Glu Thr Val Thr lie Thr Cys Arg Thr Ser Glu Asn lie Tyr Ser 130 135 140

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro His Leu Leu 145 150 155 160 •115· 160877-Sequence List.doc 201247704

Val Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser 165 170 175

Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin 180 185 190

Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro 195 200 205

Leu Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys Arg 210 215 220 <210> 142 <211> 251 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic polypeptide <400> 142

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Scr Tyr 20 25 30

Trp lie Glu Trp lie Lys Gin Arg Pro Gly His Gly Leu Glu Trp lie 35 40 45

Gly Glu He Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe 50 55 60

Arg Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met 130 135 140

Lys Pro Gly Ala Ser Val Lys lie Ser Cys Lys Ala Scr Gly Tyr Thr 145 150 155 160

Phe Thr Ser Tyr Trp lie Glu Trp lie Lys Gin Arg Pro Gly His Gly 165 170 175

Leu Glu Trp lie Gly Glu ile Leu Pro Gly Thr Gly Ser Leu Asn Asn 180 185 390 •116-

160877· Sequence Listing.doc 201247704

Asn G\w Lys Phe Arg Asp Lys Ala Thi Phe Thr Ala Asp Thr Ser Ser 195 200 205

Asn Thr Ala Tyr Met Gin Leu Ser Sei Leu Thr Ser Glu Asp Set Ala 210 215 220

Val Tyr Tyr Cys Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr 225 230 235 240

Trp Gly Gin Gty Thr Leu Val Thr Val Ser Ala 245 250 <210> 143 <211> 228 <212> PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <;4〇〇> 143

Asp lie Gin Met Thr Gin Ser Pro Ala Scr Leu Ser Ala Ser Val Gly

15 10 15

Glu Thr Val Thr lie Thr Cys Arg Thr Scr Glu Asn lie Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro His Leu Leu Val 35 40 45

Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro Leu 85 90 95

Thr Phe Gly Ser G]y Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Asp lie Gin Met Thr Gin Sei Pro 115 120 125

Ala Ser Leu Ser Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg 130 135 140

Thr Ser Glu Asn lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro 145 150 155 160

Gly Lys Ser Pro His Leu Leu Val Tyr Asn Thr Lys Thr Leu Ala Glu 165 170 175

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser ISO 185 190

Leu Lys lie Asn Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys • 117- 160877 - Sequence Listing.doc 201247704 195 200 205

Gin His His Tyr Asp Ser Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu 210 215 220

GL <211&gt

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 15 10 35

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp lie Glu Trp lie Lys Gin Arg Pro Gly His Gly Leu Glu Trp lie 35 40 45

Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe 50 55 60

Arg Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser A]a Val Tyr Tyr Cys 85 90 95

Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140

Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly 145 150 155 160

Ala Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser 165 170 175

Tyr Trp He Glu Trp lie Lys Gin Arg Pro Gly His Gly Leu Glu Trp 180 185 190 lie Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys 195 200 205

Phe Arg Asp Lys Ala Thr Phe Thr Ata Asp Thr Ser As As Thr Thr Ala 210 215 220 -118·

160877 - Sequence Listing.doc 201247704

Tyr Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 225 230 235 240

Cys Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin 245 250 255

Gly Thr Leu Val Thr Val Ser Ala 260 <2I0> 145 <211> 240 <2I2> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 15 10 15

Glu Thr Val Thr He Thr Cys Arg llir Ser Glu Asn lie Tyr Ser Tyr

20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro His Leu Leu Val 35 40 45

Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Giu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro Leu 85 90 95

Thr Phe Gly Ser Gly Thr lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala Pro Ser Val Phe 115 120 125 lie Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Scr 130 135 140

ZA Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro 165 170 175

His Leu Leu Val Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser 180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn 195 200 205 •119- 160877·Sequence List.doc 201247704

Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr 210 215 220

Asp Ser Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys Arg 225 230 235 240 <210> 146 <211> 255 <212> PRT <213> Manual Sequence <220><223> Description of the sequence: synthetic peptide <400> 146

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp lie Glu Trp lie Lys Gin Arg Pro Gly His Gly Leu Glu Trp lie 35 40 45

Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe 50 55 60

Arg Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val 130 135 140

Lys Pro Gly Aia Ser Met Lys I】e Ser Cys Lys A]a Ser Asp Tyr Ser 145 150 155 160

Phe Thr Ala Tyr Thr lie His Trp Met Lys Gin Ser His Gly Lys Asn 165 170 175

Leu Glu Trp lie Gly Leu lie Asn Pro Tyr Asn Gly Gly Thr Ser Tyr 180 185 190

Asn Gin Lys Phe Gin Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Ser 195 200 205

Ser lie Ala Tyr Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala 210 215 220 -120-

160877 - Sequence Listing.doc 201247704

Val Tyr Tyr Cys Ala Arg Arg Gly Tyr Asp Arg Glu G3y His Tyr Tyr 225 230 235 240

Ala Met Asp Tyr Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser 245 250 255 <210> 147 <211> 228 <212> PRT <213> Artificial Sequence <220><223> Artificial Sequence Description: Synthetic Peptide <400> 147

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly L 5 10 15

Glu Thr Val Thr lie Thr Cys Arg Thr Ser Glu Asn lie Tyr Ser Tyr 20 25 30

Leu Ala rp Tyr Gin Gin Lys Pro Gly Lys Ser Pro His Leu Leu Val 35 40 45

Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Tlir Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro Leu 85 90 95

Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe He Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro 115 120 125

Ala Ser Leu Ala Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg 130 135 140

Ala Ser Glu Asn lie Tyr Thr Phe Leu Ala Trp Tyr Gin Gin Lys Gin 145 150 155 160

Gly Lys Ser Pro Gin Leu Leu Val Tyr Thr Thr Lys Thr Leu Ala Glu 165 170 175

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser 180 185 190

Leu Lys He Lys Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys 195 200 205

Gin His His Tyr Gly Leu Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu 210 215 220

Glu Leu Lys Arg 121 - 160877 · Sequence Listing.doc 201247704 225 <210> 148 <231> 268 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 148

Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp He Glu Trp He Lys Gin Arg Pro Gly His Gly Leu Glu Trp lie 35 40 45

Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn Asn Asn Glu Lys Phe 50 55 60

Arg Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala Tyr Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140

Pro Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Lys Pro Gly 145 150 155 160

Ala Ser Met Lys lie Ser Cys Lys Ala Ser Asp Tyr Ser Phe Thr Ala 165 170 175

Tyr Thr lie His Trp Met Lys Gin Ser His Gly Lys Asn Leu Glu Trp 180 185 190 lie Gly Leu lie Asn Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gin Lys 195 200 205

Phe Gin Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Ser Ser lie Ala 210 215 220

Tyr Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 225 230 235 240

Cys Ala Aig Arg Gly Tyr Asp Arg Glu Gly His Tyr Tyr Ala Met Asp 245 250 255 -122-

160877 - Sequence Listing.doc 201247704

Tyr Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser 260 265 <210> 149 <211> 240 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400> 149

Asp He Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Thr Ser Glu Asn lie Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gla Gin Lys Pro Gly Lys Ser Pro His Leu Leu Val 35 40 45

Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Asp Ser Pro Leu 85 90 95.

Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala Pro Ser Val Phe 115 120 125 lie Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ala 130 135 140

Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn 145 150 155 160 lie Tyr Thr Phe Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Sex Pro 165 170 175

Gin Leu Leu Val Tyr Tlir Thr Lys Thr Leu Ala Glu Gly Val Pro Ser 180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys He Lys 195 200 205

Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr 210 215 220

Ply <213&gt Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Met Lys He Ser Cys Lys Ala Ser Asp Tyr Ser Phe Thr Ala Tyr 20 25 30

Thr lie His T卬 Met Lys Gin Ser His Gly Lys Asn Uu Glu Trp lie 35 40 45

Gly Leu lie Asn Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gin Lys Phe 50 55 60

Gin Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Ser Ser lie Ala Tyr 65 70 75 80

Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Arg Gly Tyr Asp Arg Glu Gly His Tyr Tyr Ala Met Asp Tyr 100 105 110

Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly US 120 125

Pro Ser Val Phe Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly 130 135 140

Ala Glu Leu Met Lys Pro Gly Ala Ser Val Lys lie Ser Cys Lys A]a 145 150 155 160

Ser Gly Tyr Thr Phe Thr Scr Tyr Trp lie Glu Trp lie Lys Gin Arg 165 170 175

Pro Gly His Gly Leu Glu Trp lie Gly Glu lie Leu Pro Gly Thr Gly 180 185 190

Ser Leu Asn Asn Asn Glu Lys Phe Arg Asp Lys Ala Thr Phe Thr Ala 195 200 205

Asp Thr Ser Ser Asn Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser 210 215 220

Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Gly Tyr Arg Tyr Asp Gly 225 230 235 240

Trp Phe Ala Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ala 245 250 255 -124-

160877 - Sequence Listing.doc 201247704 <210> 151 <211> 228 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Set Leu Ala Ala Ser Val Gly 1 5 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala SeT Glu Asn lie Tyr Thr Phc 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Thr Thr Lys TTir Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Lys Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Gly Leu Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phc lie Phe Pro Pro Asp ilc Gin Met Thr Gla Ser Pro 115 120 125

Ala Ser Leu Ser Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg 130 135 140

Thr Ser Glu Asn lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro 145 150 155 160

Gly Lys Ser Pro His Leu Leu Val Tyr Asn Thr Lys Thr Leu Ala Glu 165 170 175

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Fhe Ser 180 185 190

Leu Lys lie Asn Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys 195 200 205

Gin His His Tyr Asp Ser Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu 210 215 220

Glu Leu Lys Arg 225 <210> 152 <211> 268 <212> PRT <213> Artificial sequence -125·160877 - Sequence Listing.doc 201247704 <220><223> Description of artificial sequence: synthesis Peptide <400> 152

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Met Lys lie Ser Cys Lys Ala Ser Asp Tyr Ser Phc Thr Ala Tyr 20 25 30

Thr lie His Trp Met Lys Gin Ser His Gly Lys Asn Leu Glu Trp He 35 40 45

Gly Leu lie Asn Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gin Lys Phe 50 55 60 GJn G]y Arg A]a Thr Leu Thr Va]Asp Lys Ser Ser Ser lie Ala Tyr 65 70 75 80

Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Arg Gly Tyr Asp Arg Glu Gly His Tyr Tyr Ala Met Asp Tyr 100 105 110

Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125

Pro Ser Val Phe Pro Leu Ala Pro Ala Ser Thr Lys Gly Pro Ser Val 130 135 140

Phe Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Ser Gly Ala Glu Leu 145 150 155 160

Met Lys Pro Gly Ala Ser Val Lys lie Ser Cys Lys Ala Ser Gly Tyr 165 170 175

Thr Phe Thr Ser Tyr Trp lie Glu Trp lie Lys Gin Arg Pro Gly His 180 185 190

Gly Leu Glu Trp He Gly Glu lie Leu Pro Gly Thr Gly Ser Leu Asn 195 200 205

Asn Asn Glu Lys Phe Arg Asp Lys Ala Thr Phe Thr Ala Asp Thr Ser 210 215 220

Ser Asn Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser 225 230 235 240

Ala Val Tyr Tyr Cys Ala Arg Gly Tyr Arg Tyr Asp Gly Trp Phe Ala 245 250 255

Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser A1a 260 265 <2'i0> 153 <211> 240 -126-

160877 - Sequence Listing.doc 201247704 <212> PRT <2]3> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ala Ala Ser Val Gly 15 10 15

Glu ThT Val Thr lie Thr Cys Arg Ala Ser Glu Asn lie Tyr Thr Phe 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Thr Thr Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Lys Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr Gly Leu Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala Pro Set Val Phe 115 120 125 lie Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser 130 135 140

Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg Thr Ser Glu Asn L45 ISO 155 160

Lie Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ser Pro 165 170 175

His Leu Leu Val Tyr Asn Thr Lys Thr Leu Ala Glu Gly Val Pro Ser 180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys He Asn 195 200 205

Ser Leu Gin Pro Glu Asp Phe Gly Ser Tyr Tyr Cys Gin His His Tyr 210 215 220

Asp Ser Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys Arg 225 230 235 240 <210> 154 <211> 257 <212> PRT <213> Manual Sequence <220><223> Description of the sequence: synthetic peptide -127- 160877 · Sequence Listing.doc 201247704 <400> 154

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 15 10 15

Thr Vai Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Ser Met His Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45

Gly Trp lie His Thr Glu Thr Gly Glu Pro Arg Tyr Val Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95

Ala Arg Asp Ser Tyr Tyr Phe Gly Ser Ser Tyr Tyr Phe Asp Tyr Trp 100 105 110

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125

Ser Val Phe Pro Leu Ala Pro Gin lie Gin Leu Val Gin Ser Gly Pro 130 135 140

Glu Leu Lys Lys Pro Gly Glu Thr Va] Lys I]e Ser Cys Lys Ala Ser 145 150 155 160

Gly Tyr Thr Phe Thr Asp Tyr Ser Met His Trp Val Lys Gin Ala Pro 165 170 175

Gly Lys Gly Leu Lys Trp Met Gly Trp lie His Thr Glu Thr Gly Glu 180 185 190

Pro Arg Tyr Val Asp Asp Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu 195 200 205

Thr Ser Ala Ser Thr Ala Tyr Leu Gin 11c Asn Asn Leu Lys Asn Glu 210 215 220

Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp Ser Tyr Tyr Phe Gly Ser 225 230 235 240

Ser Tyr Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Thr Leu Thr Val Ser 245 250 255

Ser <210> 155 <211> 228 <212> PRT <213> artificial sequence -128-

160877-SEQ ID NO.doc 201247704 <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Thr Val Met Thr Gin Ser His Lys Phe Met Ser Thr Ser Val Gly 15 10 15

Asp Arg Val Ser He Thr Cys Lys Ala Ser Gin Asp Val Ser Scr Ala 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Scr Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Ser Gly Ser Gly Met Asp Phe Thr Phe Thr lie Ser Ser Val Gin Ala 65 70 75 80

Glu Asp Leu Ala Va! Tyr Tyr Cys Gin Gin His Tyr Ser Thr Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Glu Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Asp Thr Val Met Thr Gin Ser His 115 120 125

Lys Phe Met Scr Thr Ser Val Gly Asp Arg Val Ser lie Thr Cys Lys 130 135 140

Ala Ser Gin Asp Val Scr Ser Ala Val Ala Trp Tyr Gin Gin Lys Pro 145 150 155 ]60

Gly Gin Ser Pro Lys Leu Leu lie Tyr Ser Ala Ser Tyr Arg Tyr Thr 165 170 175

Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Met Asp Phe Thr 180 185 190

Phe Tlir lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys 195 200 205

Gin Gin His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu 210 215 220

Glu Leu Glu Arg 225 <210> 156 <211> 250 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide-129-160877· Sequence Listing.doc 201247704 <400> 156

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 】 5 10 15

Thr Val Lys Tie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Ser Met His Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Mel 35 40 45

Gly Trp lie His Thr Glu Thr Gly Glu Pro Arg Tyr Val Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95

Ala Arg Asp Ser Tyr Tyr Phe Gly Ser Ser Tyr Tyr Phe Asp Tyr Trp 100 105 110

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125

Gin He Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 130 135 140

Thr Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 145 150 155 160

Ser Met His Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met 165 370 175

Gly Trp lie His Thr Glu Thr Gly Glu Pro Arg Tyr Val Asp Asp Phe 180 185 190

Lys Gly Arg Phe Ala Phe Ser Leu Glu ΊΤίγ Ser Ala Ser Thr Ala Tyr 195 200 205

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 210 215 220

Ala Arg Asp Ser Tyr Tyr Phe Gly Ser Ser Tyr Tyr Phe Asp Tyr Trp 225 230 235 240

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 245 250 <210> 157 <211> 221 <212> m < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide <400> 157 -130·

160877 - Sequence Listing.doc 201247704

Asp Thr Val Met Thr Gin Ser His Lys Phe Met Ser Thr Ser Val Gly 15 10 15

Asp Arg Val Ser He Thr Cys Lys Ala Ser Gin Asp Val Ser Scr Ala 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Ser Gly Ser Gly Met Asp Phe Thr Phe Thr lie Ser Ser Val Gin Ala 65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gin Gin His Tyr Ser Thr Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Glu Arg Thr Val Ala Ala 100 105 110

Pro Asp Thr Val Met Thr Gin Ser His Lys Phe Met Ser Thr Ser Val 115 120 125

Gly Asp Arg Val Ser lie Thr Cys Lys Ala Ser Gin Asp Val Ser Ser 130 135 140

Ala Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ser Pro Lys I^u Leu 145 150 155 160 lie Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr 165 170 175

Gly Ser Gly Ser Gly Met Asp Phe Thr Phe Thr lie Scr Ser Va! Gin 180 185 190

Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Gin His Tyr Ser Thr Pro 195 200 205 I^eu Thr Phe Gly Ala Gly Thr Lys Leu Glu I^eu Glu Arg 210 215 220 <210> 158 <211> 257 &lt ;212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Glu Val Gin Leu Gin Gin Ser Gly Pro G!u Leu Val Gin Pro Gly Ala 15 10 15

Ser Met Lys lie Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr 20 25 30

Thr Met Asn Trp Val Lys Gin Ser His Gly Lys Asn Leu Glu Trp lie -131 - 160877 · Sequence Listing.doc 201247704 35 40 45

Gly Leu lie Asn Pro Tyr Asn Gly Gly Ser Arg Tyr Asn Gin Lys Phe 50 55 60

Met Ala Lys Ala Thr Leu Thr Val Asp Lys Scr Ser Asn Thr Ala Tyr 65 70 75 80

Met Glu Leu Leu Ser Val Thr Ser Glu Asp Ser Ala Vai Tyr Tyr Cys 85 90 95

Ala Arg Asp Ala Gly Tyr Phe Gly Ser Gly Phe Tyr Phe Asp Tyr Trp 100 105 110

Gly Gin Gly Thr Thr Leu Tlu Val Scr Ser Ala Ser Thr Lys Gly Pro 115 120 125

Ser Val Phe Pro Leu Ala Pro Glu Val Gin Leu Gin Gin Ser Gly Pro 130 135 140

Glu Leu Val Gin Pro Gly Ala Ser Met Lys lie Ser Cys Lys Ala Ser 145 150 155 160

Gly Tyr Ser Phe Thr Asp Tyr Thr Met Asn Trp Val Lys Gin Ser His 165 170 175

Gly Lys Asn Leu Glu Trp lie Gly Leu lie Asn Pro Tyr Asn Gly Gly 180 185 190

Ser Arg Tyr Asn Gin Lys Phe Met Ala Lys Ala Thr Leu Thr Val Asp 195 200 205

Lys Ser Ser Asn Thr Ala Tyr Met Glu Leu Leu Ser Val Thr Ser Glu 210 215 220

Asp Ser Ala Val Tyr Tyr Cys Ala Arg Asp Ala Gly Tyr Phe Gly Ser 225 230 235 240

Gly Phe Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Thr Leu Thr Val Ser 245 250 255

Ser <210> 159 <211> 226 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Val Met Thr Gin Ser His Lys Phe Met Ser Thr Ser Val Gly 15 10 15

Asp Arg Val Scr lie Thr Cys Lys Ala Ser Gin Asp Val Ser Thr Ala 20 25 30 -132- 160877 - Sequence Listing.doc 201247704

Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu He 35 40 45

Tyr Sei Ala Ser Tyr Arg Ser Thr Gly Val Pro Asp Arg Phe Ήιγ Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe TTir Phe Thr lie Ser Ser Val Gin Ala 65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gin Gin His Tyr Ser Thr Pro Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro 100 105 110

Scr Val Phe lie Phe Pro Pro Asp lie Val Met Thr Gin Ser His Lys 115 120 125

Phe Met Ser Thr Ser Val Gly Asp Arg Val Ser lie Thr Cys Lys Ala 130 135 140

Ser Gin Asp Val Ser Thr Ala Val Ala Trp Tyr Gin Gin Lys Pro Gly 145 150 155 160

Gin Ser Pro Lys Leu Leu lie Tyr Ser Ala Scr Tyr Arg Ser Thr Gly 165 170 175

Val Pro Asp Arg Phe Thr Gly Ser Gly Scr Gly Thr Asp Phe Thr Phe 180 185 190

Thr lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin 195 200 205

Gin His Tyr Ser Thr Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 210 215 220

Lys Arg 225 <210> 160 <211> 251 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Ixu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45 •133- 160877·Sequence List.doc 201247704

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser GIu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val 130 135 140

Gin Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr 145 150 155 160

Phe Asn Asn Tyr Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg 165 170 175

Leu Glu Trp Val Ala Tyr lie Ser Ser Ser Gly Giy Ser Thr Dyr Tyr 180 1S5 190

Ser Asp Ser Val Arg Gly Arg Phe Thr lie Ser Arg Asp ThT Ala Arg 195 200 205

Asn Thr Leu Tyr Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala 210 215 220

Met Tyr Tyr Cys Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr 225 230 235 240

Trp Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 245 250 <210> 161 <211> 228 <212> PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <;400> 161

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45 -134-

160877 - Sequence Listing.doc 201247704

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe- Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro 115 120 125

Ala Ser Leu Ser Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg 130 135 140

Ala Ser Glu Asn Phe Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Gin 145 150 155 160

Gly Lys Ser Pro Gin Leu Leu Val Tyr Asn Ala Lys Thr Leu Ala Glu 165 170 175

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser 180 185 190

Leu Lys lie Asn Ser Leu Gin Pro Glu Asp Phe Gly Thr Tyr Tyr Cys 195 200 205

Gin His His Tyr Asp lie Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu 210 215 220

Glu Leu Lys Arg 225 <210> 162 <211> 244 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <4〇0> 162

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr -135- 160877 - Sequence Listing.doc 201247704 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Glu Val Gin 115 120 125

Leu Val Glu Ser Gly Gly Gly Leu Va] Gin Pro Gly Gly Ser Leu Lys 130 135 140

Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr Tyr Met Ser 145 150 155 160

Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val Ala Tyr lie 165 170 175

Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val Arg Gly Arg 180 185 190

Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr Leu Gin Met 195 200 205

Thr Ser Leu Lys Scr Glu Asp Thr Ala Met Tyr Tyr Cys Ala Arg His 210 215 220

Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Thr Leu 225 230 235 240

Thr Val Ser Ser <210> 163 <2Π> 221 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 15 10 15

Glu Thr Val Thr He Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser G]y Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80 136·

160877· Sequence Listing.doc 201247704

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pto Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Scr Ala Ser Val 115 120 125

Gly Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser 130 135 140

Tyr Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu 145 150 155 160

Val Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser 165 170 175

Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin 180 185 190

Pro Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro 195 200 205

Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 210 215 220 <210> 164 <211> 249 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic peptide <400> 164

Lys lie Gin Leu Vai Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 15 10 15

Thr Val Lys He Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30

Gly Met Asn Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45

Gly Trp lie ^sn He Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Leu Cys 85 90 95

Ala Arg Asp Ser Tyr Sei Gly Gly Phe Asp Tyr Trp Gly Gin Gly Thi 100 105 110 -137- 160877 - Sequence Listing.doc 201247704 lie Va] Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125

Leu Ala Pro Lys lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys 130 135 140

Pro Gly Glu Thr Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe 145 150 155 160

Thr Asn Tyr Gly Met Asn Trp Val Lys Gin Ala Pro Gly Lys Gly Leu 165 170 175

Lys Trp Met Gly Trp lie Asn lie Asn Thr Gly Glu Pro Thr Ding yr Ala 180 185 190

Glu Glu Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr 195 200 205

Thr Ala Phe Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr 210 215 220

Tyr Leu Cys Ala Arg Asp Ser Tyr Ser Gly Gly Phe Asp Tyr Trp Gly 225 230 235 240

Gin Gly Thr I]e Val Thr Vai Ser Ser 245 <210> 165 <211> 240 <212> PRT' <2]3>Artificial Sequence<220><223> Description of Artificial Sequence: Synthesis Peptide <400> 165

Asp lie Val Met Thr Gin Ser Pro Ser Ser Leu Ser Val Ser Ala Gly 15 10 15

Glu Lys Val Thr Leu Ser Cys Lys Ser Ser Gin Ser Leu Leu lie Ser 20 25 30

Gly Asp Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45

Pro Pro Lys Leu Leu lie Tyr Gly Ala Ser Thr Arg Asp Ser Gly Val 50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr 65 70 75 80 lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn 85 90 95

Asp His Ser Phe Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 -138-

160877· Sequence Listing.doc 201247704

Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp lie 115 120 125

Val Met Thr Gin Ser Pro Ser Ser Leu Ser Val Ser Ala Gly Glu Lys 130 135 140

Val Thr f^eu Ser Cys Lys Ser Ser Gin Ser Leu Leu lie Ser Gly Asp 145 150 155 160

Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 165 170 175

Lys Leu Leu He Tyr Gly Ala Ser Thr Arg Asp Scr Gly Val Pro Asp 180 185 190

Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr lie Ser 195 200 205

Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn Asp His 210 215 220

Ser Phe Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 225 230 235 240 <210> 166 <211> 242 <212> PRT <213> Manual Sequence <220><223> Description of the sequence: synthetic peptide <400> 166

Lys He Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Giu 15 10 15

Thr Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30

Gly Met Asn Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 G]y Trp lie Asn lie Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Leu Cys 85 90 95

Ala Arg Asp Ser Tyr Ser Gly Gly Phe Asp Tyr Trp Gly Gin Giy Thr 100 105 110 lie Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Lys lie Gin Leu 115 120 125

Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu Thr Val Lys He • 139· 160877· Sequence Listing.doc 201247704 130 135 140

Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Gly Met Asn Trp 145 150 155 160

Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met Gly Trp lie Asn 165 170 175 lie Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe Lys Gly Arg Phe 180 185 190

Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe Leu Gin lie Asn 195 200 205

Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Leu Cys Ala Arg Asp Ser 210 215 220

Tyr Ser Gly Gly Phe Asp Tyr Trp Gly Gin Gly Thr He Val Thr Val 225 230 235 240

Ser Ser <210> 167 <211> 233 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Val Met Thr Gin Ser Pro Ser Ser Leu Scr Val Ser Ala Gly 1 5 10 15

Glu Lys Val Thr Leu Ser Cys Lys Ser Ser Gin Ser Leu Leu lie Ser 20 25 30

Gly Asp Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45

Pro Pro Lys Leu Leu lie Tyr Gly Ala Ser Thr Arg Asp Ser Gly Val 50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr 65 70 75 80 lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn 85 90 95

Asp His Ser Phe Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110

Lys Arg Thr Val Ala Ala Pro Asp lie Val Met Thr Gin Ser Pro Ser 115 120 125

Ser Leu Ser Val Ser Ala Gly Glu Lys Val Thr Leu Ser Cys Lys Ser 130 135 140 -140- 160877 - Sequence Listing.doc 201247704

Scr Gin Scr Leu Leu lie Scr Gly Asp Gin Lys Asn Tyr Leu Ala Trp 145 150 155 160

Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Gly Ala 165 170 175

Scr Thr Arg Asp Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser 180 185 190

Gly Ala Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp Leu 195 200 205

Ala Val Tyr Tyr Cys Gin Asn Asp His Ser Phe Pro Pro Thr Phe Gly 210 215 220

Ala Gly Thr Lys Leu Glu Leu Lys Arg 225 230

<210> 168 <211> 250 <212> FRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45

Ala Tyr He Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Ήιγ Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Set Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Lys lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys 130 135 140

Lys Pro Gly Glu Thr Val Lys lie Scr Cys Lys Ala Scr Gly Tyr Thr 145 150 155 160 -141 - 160877 · Sequence Listing.doc 201247704

Phe Thr Asa Tyr Gly Met Asn Trp Val Lys Gin Ala Pro Giy Lys Gly 165 170 175

Leu Lys Trp Met Gly Trp lie Asn I]e Asn Thr Gly Glu Pro Ding hr Tyr 180 185 190

Ala Glu Glu Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala 195 200 205

Thr Thr Ala Phe Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala 210 215 220

Thr Tyr Leu Cys Ala Arg Asp Ser Tyr Ser Gly Gly Phe Asp Tyr Trp 225 230 235 240

Gly Gin Gly Thr He Val Thr Val Ser Ser 245 250 <210> 169 <211> 234 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide<400> 169

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 1 5 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Ding yr 20 25 . 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Giy Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Asp lie Val Met Thr Gin Ser Pro 115 120 125

Scr Ser Leu Ser Val Ser Ala Gly Glu Lys Val Thr Leu Ser Cys Lys 130 135 140

Ser Ser Gin Scr Leu Leu ]le Ser Gly Asp Gin Lys Asn Tyr Leu Ala 145 150 155 160 •142·

160877 - Sequence Listing.doc 201247704

Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Leu Leu lie Tyr Gly 165 170 175

Ala Ser Thr Arg Asp Ser Gly Val Pro Asp Arg Phe Thr Gly Sex Gly 180 185 190

Ser Gly Ala Asp Phe Thr Leu Thr lie Ser Ser Val Gin Ala Glu Asp 195 200 205 l^u Ala Val Tyr Tyr Cys Gin Asn Asp His Ser Phe Pro Pro Thr Phe 210 215 220

Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 225 230 <210> 170 <211> 243 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide

<400> 170

Glu Val Gin Leu Val Glu Set Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Lys lie Gin 115 120 125

Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Giy Glu Thr Val Lys 130 135 140 lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Gly Met Asn 145 150 〗 55 160

Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met Gly Trp lie 165 170 175

Asn 11c Asn Thr Gly Glu Pro ITir Tyr Ala Glu Glu Phe Lys Gly Arg -143- 160877 · Sequence Listing.doc 201247704 180 185 190

Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe Leu Gin lie 195 200 205

Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Leu Cys Ala Arg Asp 210 215 220

Ser Tyr Ser Gly Gly Phe Asp Tyr Trp Giy Gin Gly Thr lie Val Thr 225 230 235 240

Val Ser Ser <210> 171 <211> 227 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Scr Ala Ser Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Asp lie Val Met Thr Gin Ser Pro Ser Ser Leu Ser Val Ser Ala 115 120 125

Gly Glu Lys Val Thr Leu Ser Cys Lys Ser Ser Gin Ser Leu Leu lie 130 135 140

Ser Gly Asp Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly 145 150 155 160

Gin Pro Pro Lys Leu Leu lie Tyr Gly Ala Ser Thr Arg Asp Ser Gly 165 170 175

Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu 180 185 190 •144·

160877 - Sequence Listing.doc 201247704

Thr lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin 195 200 205

Asn Asp His Ser Phe Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu 210 215 220

Leu Lys Arg 225 <210> 172 <211> 250 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Lys lie Gin Leu Val Gin Scr Gly Pro Glu Leu Lys Lys Pro Gly Glu 15 10 15

Thr Val Lys He Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30

Gly Met Asn Trp Val Lys Gin Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45

Gly Trp lie Asn lie Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Leu Cys 85 90 95

Ala Arg Asp Ser Tyr Ser Gly Gly Phe Asp Tyr Trp Gly Gin Gly Thr 100 105 110 lie Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125

Leu Ala Pro Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin 130 135 140

Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 145 150 155 160

Asn Asn Tyr Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu 165 170 175

Glu Trp Val Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser 180 185 190

Asp Ser Val Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asa 195 200 205 -145· 160877 - Sequence Listing.doc 201247704

Thr Leu Tyr Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met 210 215 220

Tyr Tyr Cys Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp 225 230 235 240

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 245 250 <210> 173 <211> 234 <212> PRT <2]3>Artificial Sequence<220><223> Description of Artificial Sequence: Synthetic Peptide <400> 173

Asp lie Val Met Hir Gin Ser Pro Ser Ser Leu Ser Val Ser Ala Gly 1 5 10 15

Glu Lys Val Thr Leu Sex Cys Lys Ser Ser Gin Ser Leu Leu lie Ser 20 25 30

Gly Asp Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin 35 40 45

Pro Pro Lys Leu Leu lie Tyr Gly Ala Ser Thr Arg Asp Ser Gly Val 50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr 65 70 75 80

He Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn 85 90 95

Asp His Ser Phe Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp lie 115 120 125

Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly Glu Thr 130 135 140

Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr Leu Ala 145 150 155 160

Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val Tyr Asn 165 170 175

Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly Ser Gly 180 185 190

Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro Glu Asp 195 200 205 -146-

160877· Sequence Listing.doc 201247704

Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu Thr Phe 210 215 220

Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 225 230 <210> 174 <211> 243 <212> PRT <213> Artificial Sequence <220><2Ϊ3> Description of Artificial Sequence: Synthetic Peptide<400> 174

Lys lie Gin Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 15 10 15

Thr Val Lys lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30

Gly Met Asn Trp Val Lys Gin Ala Pro Gly Lys Gly Lea Lys Trp Met 35 40 45

Gly Trp lie Asn lie Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Thr Ala Phe 65 70 75 80

Leu Gin lie Asn Asn Leu Lys Asn Glu Asp Oir A】a Thr Tyr Leu Cys 85 90 95

Ala Arg Asp Ser Tyr Scr Gly Gly Phe Asp Tyr Trp Gly GJn Gly Thr 100 105 110 lie Val Thr Va] Ser Ser Ala Ser TTu Lys Gly Pro Glu Val Gin Leu 115 120 125

Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser Leu Lys Leu 130 135 140

Scr Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr Tyr N5et Ser Trp 145 150 155 160

Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val Ala Tyr lie Ser 165 170 175

Ser Ser Gly Gly Ser Thr Tyr Tyr -Ser Asp Ser Val Arg Gly Arg Phe 180 185 190

Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr Leu Gin Met Thr 195 200 205

Ser Leu Lys Ser Glu Asp Thr Aia Met Dyr Tyr Cys Ala Arg His Phe 210 215 220

Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Thr Leu Thr -147- 160877 · Sequence Listing.doc 201247704 225 230 235 240

Val Ser Ser <210> 175 <2ll> 227 <2I2> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp lie Val Met Thr Gin Ser Pro Ser Ser Leu Ser Val Ser Ala Gly 15 10 15

Glu Lys Val Thr Leu Ser Cys Lys Ser Ser Gin Ser Leu Leu lie Ser 20 25 30

Gly Asp Gin Lys Asn Tyr Leu Ala Trp Tyr Gin Gin Lys Pro G]y Gin 35 40 45

Pro Pro Lys Leu Leu lie Tyr Gly Ala Ser Thr Arg Asp Ser Gly Val 50 55 60

Pro Asp Arg Phe Thr Gly Ser Oly Ser Gly Ala Asp Phe Thr Leu Thr 65 70 75 80 lie Ser Ser Val Gin Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gin Asn 85 90 95

Asp His Ser Phe Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110

Lys Arg Thr Val Ala Ala Pro Asp lie Gin Met Thr Gin Ser Pro Ala 115 120 125

Ser Leu Ser Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg Ala 130 135 140

Ser Glu Asn Phe Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Gin Gly 145 150 155 160

Lys Ser Pro CJn Leu Leu VaJ Tyr Asn Ala Lys Thr Leu Ala Glu Gly 165 170 175

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu 180 185 190

Lys He Asn Ser Leu Gin Pro Glu Asp Phe Gly Thr Tyr Tyr Cys Gin 195 200 205

His His Tyr Asp lie Fro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu 210 215 220

Leu Lys Arg 225 -148-

160877 - Sequence Listing.doc 201247704 <210> 176 <211> 243 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Polypeptide <400>

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Scr Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Gin Val Gin 115 120 125

Leu Gin Gin Pro Gly Ser Glu Leu Val Arg Pro Gly Ala Ser Val Lys 130 135 140

Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His 145 150 155 160

Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie Gly Asn lie 165 170 175

Tyr Pro Gly Thr Val Asn Thr Asn Tyr Asp Glu Lys Phe Lys Asn Lys 180 185 190 A]a Thr Leu Thr Vat Asp THir Ser Ser Ser Thr Ala Tyr Met Leu Leu 195 200 205

Ser Ser Leu Thr Scr Glu Asp Ser Ala Val Tyr Tyr Cys Thr Arg Asp 210 215 220

Tyr Tyr Gly Gly Gly Leu Asn Tyr Trp Gly Gin Gly Thr Thr Leu Thr 225 230 235 240

Val Ser Ser -149-160877 - Sequence Listing.doc 201247704 <210> 177 <211> 221 <212> PRT <213>Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <;400> 177

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Giu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser lie Val Met Thr Gin Thr Pro Lys Phe Leu Leu Val Ser Ala 115 120 125

Gly Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Ser Val Ser Asn 130 135 140

Asp Val Ala Trp Phe Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu 145 150 ).55 】60

He Tyr Tyr Ala Ser Asn Arg Tyr Ala Gly Val Pro Asp Arg Phe Thr 165 170 175

Gly Ser Gly Phe Gly Thr Asp Phe Thr Phe llir lie Ser Thr Val Gin 180 185 190

Ala Glu Asp Leu Ala Val Tyr Phe Cys His Gin Asp Tyr Ser Ser Pro 195 200 205

Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg 210 215 220 <210> 178 <211> 250 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic peptide 150-

160877·Sequence list.doc 201247704 <400> 178

Glu Val Gin Leu Vai Glu Ser Gly Gly Gly Leu Vai Gin Pro Gly Gly 15 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 35 40 45

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Gin Val Gin Leu Gin Gin Pro Gly Ser Glu Leu Val 130 135 140

Arg Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr 145 150 155 160

Phe Thr Ser Tyr Trp Met His Trp Val Lys Gin Arg Pro Gly Gin Gly 165 170 175

Leu Glu Trp lie Gly Asn lie Tyr Pro Gly Thr Val Asn Thr Asn Tyr 180 185 190

Asp Glu Lys Phe Lys Asn Lys Ala Thr Leu Thr Val Asp Thr Ser Ser 195 200 205

Ser Thr Ala Tyr Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Sex Ala 210 215 220

Val Tyr Tyr Cys Thr Arg Asp Tyr Tyr Gly Gly Gly Leu Asn Tyr Trp 225 230 235 240

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 245 250 <210> 179 <211> 228 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide-151 - 160877 - Sequence Listing.doc 201247704 <400> 179

Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Sex Va] Gly 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Va! 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser lie Val Met Thr Gin Thr Pro 115 120 125

Lys Phe Leu Leu Val Ser Ala Gly Asp Arg Val Thr lie Thr Cys Lys 130 135 140

Ala Ser Gin Ser Val Ser Asn Asp Val Ala Trp Phe Gin Gin Lys Pro 145 150 155 160

Gly Gin Ser Pro Lys Leu Leu lie Tyr Tyr Ala Ser Asn Arg Tyr Ala 165 170 175

Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Phe Gly Thr Asp Phe Thr 180 185 190

Phe Thr lie Ser Thr Val Gin Ala Glu Asp Leu Ala Val Tyr Phe Cys 195 200 205

His Gin Asp Tyr Ser Ser Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu 210 215 220

Glu lie Lys Arg 225 <210> 180 <211> 263 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15 .152- 160877 · Sequence Listing.doc 201247704

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 65 W 75 80

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130 135 140

Pro Gin Val Gin Leu Gin Gin Pro Gly Ser Glu Leu Val Arg Pro Gly 145 150 155 160

Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser 165 170 175

Tyr Trp Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp 180 185 190 lie Gly Asn lie Tyr Pro Gly Thr Val Asn Thr Asn Tyr Asp Glu Lys 195 200 205

Phe Lys Asn Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala 210 215 220

Tyr Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyt Tyr 225 230 235 240

Cys Thr Arg Asp Tyr Tyr Gly Gly Gly Leu Asn Tyr Trp Gly Gin Gly 245 250 255

Thr Thr Leu Thr Val Ser Ser 260 <210> 181 <211> 240 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <400>

Asp lie Gin Met Thr Gin Ser Pro Ala Scr Leu Ser Ala Ser Val Gly -153- 160877 - Sequence Listing.doc 201247704 15 10 15

Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu Val 35 40 45

Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyt Asp lie Pro Leu 85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala Pro Ser Val Phe 115 120 125

He Phe Pro Pro Ser lie Val Met Thr Gin Thr Pro Lys Phe Leu Leu 130 135 140

Val Ser Ala Gly Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Ser 145 150 155 160

Val Sex Asn Asp Val Ala Trp Phe Gin Gin Lys Pro Gly Gin Ser Pro 165 170 175

Lys Leu Leu lie Tyr Tyr Ala Ser Asn Arg Tyr Ala Gly Val Pro Asp 180 185 190

Arg Phe Thr Gly Ser Gly Phe Gly Thr Asp Phe Thr Phe Thr lie Ser 195 200 205

Thr Val Gin Ala Glu Asp Leu Ala Val Tyr Phe Cys His Gin Asp Tyr 210 215 220

Ser Ser Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg 225 230 235 240 <210> 182 <211> 243 <212> PRT <213> Manual Sequence <220><223> Description of the sequence: synthetic peptide <400> 182

Gin Val Gin. Leu Gin Gin Pro Gly Ser Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 -154-

160877 - Sequence Listing.doc 201247704

Trp Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Tip lie 35 40 45

Gly Asn lie Tyr Pro Gly Thr Val Asn Thr Asn Tyr Asp Glu Lys Phe 50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Thr Arg Asp Tyr Tyr Gly Gly Gly Leu Asn Tyr Trp Gly Gin Gly Thr 100 105 110

Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Glu Val Gin Leu 115 120 125

Val Glu Ser Gly Gly Gly Leu Val Gin Fro Gly Gly Ser Leu Lys Leu 130 135 140

Ser Cys Ala Ala Ser Gly Phe Thr Phc Asn Asn Tyr Tyr Met Ser Trp 145 150 155 160

Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val Ala Tyr lie Ser 165 170 175

Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val Arg Gly Arg Phe 180 185 190

Thr lie SeT Arg Asp Thr Ala Arg Asn Thr Leu Tyr Leu Gin Met Thr 195 200 205

Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys Ala Arg His Phe 210 215 220

Gly Asp Tyi Ser Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Thr Leu Thr 225 230 235 240

Val Ser Ser <210> 183 <211> 221 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Ser lie Val Met Thr Gin Tlir Pro Lys Phc Leu Leu Val Scr Ala Gly 15 10 15

Asp Arg Val Thr He Thr Cys Lys Ala Ser Gin Ser Val Ser Asn Asp 20 25 30 -155- 160877 - Sequence Listing.doc 201247704

Val Ala Trp Phe Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Tyr Ala Ser Asn Arg Tyr Ala Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Scr Gly Phe Gly Thr Asp Phe Thr Phe Thr lie Ser Thr Val Gin Ala 65 70 75 80

Glu Asp Leu Ala Val Tyr Phe Cys His Gin Asp Tyr Ser Ser Pro Arg 85 90 95

Thr Phe Gly Gly Gly Thf Lys Leu Glu He Lys Arg Thr Val Ala Ala 100 105 110

Pro Asp He Gin Met Thr Gin Ser Pro Ala Ser Leu Ser Ala Ser Val 115 120 125

Gly Glu Thr Val Thr lie Thr Cys Arg Ala Ser Glu Asn Phe Tyr Ser 130 135 140

Tyr Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro Gin Leu Leu 145 150 155 160

Val Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser 165 170 175

Gly Scr Gly Ser Gly Thr Gin Phe Ser Leu Lys lie Asn Ser Leu Gin 180 185 190

Pro Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr Asp lie Pro 195 200 205

Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 210 215 220 <210> 184 <211> 250 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic polypeptide <400> 184

Gin Val Gin Lea Gin Gin Pro Gly Ser Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Asn lie Tyr Pro Gly Thr Val Asn Thr Asn Tyr Asp Glu Lys Phe 50 55 60 •156-

160877 - Sequence Listing.doc 201247704

Lys Asn Lys Ala Thr Leu Thr Val Asp Thr Ser 65 70 75

Ser Ser Ala l¥

Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Ser 85 90

Ala Val Tyr Tyr Cys 95 llir Arg Asp Tyr Tyr Gly Gly Gly Leu Asn Tyr 100 105

Trp Gly Gin Gly Thr 110

Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120

Pro Ser Val Phe Pro 125

Leu Ala Pro Glu Val Gin Leu Val Glu Ser Gly 130 135

Gly Gly Leu Val Gin 140

Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala 145 150 155

Ser Gly Phe Thr Phe 160

Asn Asn Tyr Tyr Met Ser Trp Val Arg Gin Thr 165 170

Pro Glu Arg Arg Leu 175

Glu Trp Val Ala Tyr He Ser Ser Ser Gly Gly 180 185

Ser Thr Tyr Tyi Ser 190

Asp Ser Val Arg Gly Arg Phe Thr He Ser Arg 195 200

Asp Thr Ala Arg Asn 205

Thr Leu Tyr Leu Gin Met Thr Ser Leu Lys Ser 210 215

Glu Asp Thr Ala Met 220

Tyr Tyr Cys Ala Arg His Phe Gly Asp Tyr Ser 225 230 235 Dyr yr Phe Asp Tyr Trp 240

Gly Gin Gly Thr Thr Leu Thr Val Ser Ser 245 250

<210> 185 <211> 228 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400> 185 Ser He Val Met Thr Gin Thr Pro Lys Phe Leu 1 5 10

Leu Val Ser Ala Gly 15

Asp Arg Val Thr He Thr Cys Lys Ala Ser Gin 20 25

Ser Val Ser Asn Asp 30

Val Ala Trp Phe Gin Gin Lys Pro Gly Gin Ser 35 40

Pro Lys Leu Leu He 45

Tyr Tyr Ala Ser Asn Arg Tyr Ala Gly Val Pro 50 55

Asp Arg Phe Thr Gly 60

Ser Gly Phe Gly Thr Asp Phe Thr Phe Thr lie

Ser Thr Val Gin Ala 160877 - Sequence Listing.doc -157- 201247704 65 70 75 80

Glu Asp Leu Ala Val Tyr Phe Cys His Gin Asp Tyr Ser Ser Pro Arg 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro 115 120 125

Ala Ser Leu Ser Ala Ser Val Gly Glu Thr Val Thr lie Thr Cys Arg 130 135 140

Ala Ser Glu Asn Phe Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Gin 145 150 155 160

Gly Lys Ser Pro Gin Leu Leu Val Tyr Asn Ala Lys Thr Leu Ala Glu 165 170 175

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser 180 185 190

Leu Lys lie Asn Ser Leu Gin Pro Glu Asp Phe Gly Thr Tyr Tyr Cys 195 200 205

Gin His His Tyr Asp lie Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu 21.0 215 220

Glu Leu Lys Arg 225 <210> 186 <211> 263 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Pro Gly Ser Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met His Trp Val Lys Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45

Gly Asn lie Tyr Pro Gly Thr Val Asn Thr Asn Tyr Asp Glu Lys Phe 50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Leu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 -158- 160877 - Sequence Listing.doc 201247704

Thr Arg Asp Tyr Tyr Gly Gly Gly Leu Asn Tyr Trp Gly Gin Gly Thr 100 105 110

Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125

Leu Ala Pro Ala Ser Thr Lys Gly Pro Set Val Phc Pro Leu Ala Pro 130 135 140

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 145 150 155 160

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Asn Tyr 165 170 175

Tyr Met Ser Trp Val Arg Gin Thr Pro Glu Arg Arg Leu Glu Trp Val 180 185 190

Ala Tyr lie Ser Ser Ser Gly Gly Ser Thr Tyr Tyr Ser Asp Ser Val 195 200 205

Arg Gly Arg Phe Thr lie Ser Arg Asp Thr Ala Arg Asn Thr Leu Tyr 210 215 220

Leu Gin Met Thr Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 225 230 235 240

Ala Arg His Phe Gly Asp Tyr Ser Tyr Phe Asp Tyr Trp Gly Gin Gly 245 250 255

Thr Thr Leu Thr Val Ser Ser 260 <210> 187 <211> 240 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Ser lie Val Met Thr Gin Thr Pro Lys Phe Leu Leu Val Ser Ala Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Ser Val Ser Asn Asp 20 25 30

Val Ala Trp Phe Gin Gin Lys Pro Gly Gin Ser Pro Lys Leu Leu lie 35 40 45

Tyr Tyr Ala Sex Asn Arg Tyr Ala Gly Val Pro Asp Arg Phe Thr Gly 50 55 60

Ser Gly Phe Gly Thr Asp Phe Thr Phe Thr lie Ser Tlir Val Gin Ala 65 70 75 80 -159- 160877 - Sequence Listing.doc 201247704

Glu Asp Leu Ala Val Tyr Phe Cys His Gin Asp Tyr Ser Ser Pro Arg 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu He Lys Arg TTir Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala Pro Ser Val Phe 115 120 125 lie Phe Pro Pro Asp lie Gin Met Thr Gin Ser Pro Ala Ser Leu Ser 130 135 140

Ala Ser Val Gly Glu Thr Val Thr He Thr Cys Arg Ala Ser Glu Asn 145 150 155 160

Phe Tyr Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Gin Gly Lys Ser Pro 165 170 175

Gin Leu Leu Val Tyr Asn Ala Lys Thr Leu Ala Glu Gly Val Pro Ser 180 185 190

Arg Phe Ser Gly Ser Gly Ser Gly Thr Gin Phe Ser Leu Lys He Asn 195 200 205

Ser Leu Gin Pro Glu Asp Phe Gly Thr Tyr Tyr Cys Gin His His Tyr 210 215 220

Asp Jle Pro Leu Thr Phe G]y A]a Gly Thr Lys Leu Glu Leu Lys Arg 225 230 235 240 <210> 188 <211> 247 <212> PRT <213>Artificial Sequence<220>;223> Description of artificial sequence: synthetic peptide <400> 188

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Arg Lys Pro Gly Glu 15 10 15

Thr Val Lys lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Hu His Tyr 20 25 30

Gly lie Asn Trp Val Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met 35 40 45

Gly Trp lie Asn Thr His Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Asn Thr Ala Tyr 65 70 75 80

Leu GJn He Asn Asn Leu Asn Asn Gly Asp Met Gly Thr Tyr Phe Cys 85 90 95 • 160- 160877 · Sequence Listing.doc 201247704

Thr Arg.Ser His Arg Phe Gly Leu Asp Tyr-Trp Gly Gin Gly Thr Ser 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Arg Lys Pro 130 135 140

Gly Glu Thr Val Lys lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr 145 150 155 160

His Tyr Gly Lie Asn Trp Val Lys Gin Thr Pro Arg Lys Asp Leu Lys 165 170 175

Trp Met Gly Trp lie Asti Thr His Thr Gly Glu Ala Tyr Tyr Ala Asp 180 185 190

Asp Phe Lys Gly Arg Phe Ala Phe Scr Leu Glu Thr Ser Ala Asn Thr 195 200 205

Ala Tyr Leu Gin lie Asn Asn Leu Asn Asn Gly Asp Met Gly Thr Tyr 210 215 220

Phe Cys Thr Arg Scr His Arg Phe Gly Leu Asp Tyr Trp Gly Gin Gly 225 230 235 240

Thr Ser Val Thr Val Ser Ser 245 <210> 189 <211> 236 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Asn Val Leu Thr Gin Ser Pro Pro Ser Leu Ala Val Ser Leu Gly 15 10 15

Gin Arg Ala ΊΤιγ I]e Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asn 20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Fro 35 40 45

Lys Phe Leu lie Tyr Ala Ala Ser Asn Leu Glu Ser Gly lie Pro Ala 50 55 60

Arg Phe Ser G1y Ser Gly Ser Gly Thr Asp Phe Asn Leu Asn lie His 65 70 75 SO

Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn 85 90 95

Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu lie Lys Arg -161 - 160877 · Sequence Listing.doc 201247704 loo 105 no

Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp Asn Val Leu 115 120 125

Thr Gin Ser Pro Pro Ser Leu Ala Val Ser Leu Gly Gin Arg Ala Thr 130 135 140 lie Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asn Cly Asp Ser Tyr 145 150 155 160

Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Phe Leu lie 165 170 175

Tyr Ala Ala Ser Asn Leu Glu Ser Gly lie Pro Ala Arg Phe Ser Gly 180 〗 85 190

Ser Gly Ser Gly Thr Asp Phe Asn Leu Asn lie His Pro Val Glu Glu 195 200 205

Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn Glu Asp Pro Phe 210 215 220

Thr Phe Gly Ser Cly Thr Lys Leu Glu He Lys Arg 225 230 235 <210> 190 <211> 240 <212> PRT <213> Artificial Sequence <220><2 Magic > Description of Artificial Sequence : Synthetic Peptide <400> 190

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Arg Lys Pro Gly Glu 15 10 15

Thr Val Lys lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr 20 25 30

Gly lie Asn Trp Val Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met 35 40 45

Gly Trp lie Asn Thr His Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Asn Thr Ala Tyr 65 70 75 80

Leu Gin lie Asn Asn Leu Asn Asn Gly Asp Met Gly Thr Tyr Phe Cys 85 90 95

Thr Arg Ser His Arg Phe Gly Leu Asp Tyr Trp Gly Gin Gly Thr Ser 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Fro Gin lie Gin Leu Val 115 120 125 -162-

160877 - Sequence Listing.doc 201247704

Gin Ser Gly Pro Glu Leu Arg Lys Pro Gly Glu Thr Val Lys lie Ser 130 135 140

Cys Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr Gly lie Asn Trp Val 145 150 155 160

Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met Gly Trp lie Asn Thr 165 170 175

His Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe Lys Gly Arg Phe Ala 180 185 190

Phe Ser Leu Glu Thr Ser Ala Asn Thr Ala Tyr Leu Gin lie Asn Asn 195 200 205

Leu Asn Asn Gly Asp Met Giy Tlir Tyr Phe Cys Thr Arg Ser His Arg 210 215 220

Phe Gly Leu Asp Tyr Trp Gly Gin Gly Thr Ser Val Thr Val Ser Ser 225 230 235 240

<210> 191 <211> 229 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Asn Val Leu Thr Gin Ser Pro Pro Ser Leu Ala Val Ser Leu Gly 15 10 15

Gin Arg Ala Thr lie Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asn 20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45

Lys Phe Leu lie Tyr Ala Ala Ser Asn Leu Glu Ser Gly lie Pro Ala 50 55 60 ATg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Asn Leu Asn lie His 65 70 75 80

Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn 85 90 95

Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu lie Lys Arg 100 105 110

Thr Val Ala Ala Pto Asp Asn Val Leu Thr Gin Ser Pro Pro Ser Leu 115 120 125

Ala Val Ser Leu Gly Gin Arg Ala Thr lie Scr Cys Lys Ala Asn Trp 130 135 140 -163- 160877-Sequence List.doc 201247704

Pro Val Asp Tyr Asn Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin Lys 145 150 155 160

Pro Gly Gin Pro Pro Lys Phe Leu lie Tyr Ala Ala Ser Asn Leu Glu 165 170 175

Ser Gly lie Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 180 185 190

Asn Leu Asn lie His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr 195 200 205

Cys Gin Gin Ser Asn Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys 210 215 220

Leu Glu lie Lys Arg 225 <210> 192 <211> 237 <212> PRT <213>Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Gin Val Gin Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met Asn Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp lie 35 40 45

Gly Arg lie Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe 65 70 75 80

Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Val Ser Asp Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Scr Ser 100 105 110

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Gin Val Gin 115 120 125

Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala Ser Val Lys 130 135 140

Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met Asn 145 150 155 160 -164-

160877 - Sequence Listing.doc 201247704

Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp He Gly Arg He 165 170 175

Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phc Lys Asp Lys 180 185 190

Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe Val Gin Leu 195 200 205

Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Val Ser Asp 210 215 220

Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 225 230 235 <210> 193 <211> 238 <212> PRT <213>Artificial Sequence <220>

<223> Description of artificial sequence: synthetic polypeptide <400> 193

Asp Val Val Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly 15 10 15

Gin Pro Ala Ser lie Scr Cys Lys Ser Scr Gin Ser Leu Leu Asp Ser 20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser 35 40 45

Pro Lys Leu Leu lie Tyr Val Val Ser Lys Leu Glu Ser Gly Val Pro 50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 SO

Ser Arg Va) Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala 85 90 95

Thr His Phc Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp Val Val 115 120 125

Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly Gin Pro Ala 130 135 140

Ser lie Ser Cys Lys Ser Scr Gin Scr Leu Leu Asp Ser Asp Gly Lys 145 150 155 160

Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser Pro Lys Leu 165 170 175

Leu lie Tyr Val Val Scr Lys Leu Glu Ser Gly. Val Pro Asp Arg Phe -165- 160877 - Sequence Listing.doc 201247704

ISO 185 190

Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie Ser Arg Val 195 200 205

Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala Thr His Phe 210 215 220

Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg 225 230 235 <210> 194 <211> 242 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence : synthetic peptide <400> 194

Gin Val Gin Leu Gin Gin Pro Gly Ala Glu Leu Val Are Pro Gly Ala 1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met Asn Trp Val Lys Gin Arg Pro Glu Gin Giy Leu Glu Trp lie 35 40 45

Gly Arg lie Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala He Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe 65 70 75 80

Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Val Ser Asp Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 100 105 110

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Gin lie Gin 115 120 125

Leu Val Gin Ser Gly Pro Glu Leu Arg Lys Pro Gly Glu Thr Val Lys 130 135 140 lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr Gly He Asn 145 150 155 160

Trp Val Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met Gly Trp He 165 170 175

Asn Thr His Thr G]y Glu Ala Tyr Tyr Ala Asp Asp Phe Lys Gly Arg 180 185 190

Phe Ala Phe Ser Leu Glu Thr Ser Ala Asn Thr Ala Tyr Leu Gin lie 195 200 205 160877 - Sequence Listing.doc -166-

201247704

Asn Asn Leu Asn Asn Gly Asp Met Gly Thr T^r Phe Cys Thi Kxg Ser 210 215 220

His Arg Phe Gly Leu Asp Tyr Trp Gly Gin Gly Thr Ser Val Thr Val 225 230 235 240

Ser Ser <210> 195 <211> 237 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Val Val Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly 15 10 15

Gin Pro Ala Ser lie Ser Cys Lys Ser Ser Gin Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser 35 40 45

Pro Lys Leu Leu lie Tyr Val Val Ser Lys Leu Glu Ser Gly Vai Pro 50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala 85 90 95

Thr His Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110

Arg llir Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp Asn Val 115 120 125

Leu Thr Gin Ser Pro Pro Ser Leu Ala Val Ser Leu Gly Gin Arg Ala 130 135 140

Thr He Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asn Gly Asp Ser 145 150 155 160

Tyr Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Phe Leu 165 170 175 lie Tyr Ala Ala Ser Asn Leu Glu Ser Gly lie Pro Ala Arg Phe Ser 180 185 190

Gly Ser Gly Ser Gly Thr Asp Phe Asn Leu Asn lie His Pro Val Glu 195 200 205 -167- 160877 · Sequence Listing.doc 201247704

Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn Glu Asp Pro 210 215 220

Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu lie Lys Arg 225 230 235 <210> 196 <211> 255 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic peptide <400> 196

Gin Val Gin Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30

Trp Met Asn Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp lie 35 40 45

Gly Arg lie Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe 65 70 75 80

Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Val Ser Asp Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 100 105 110

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ala Ser Thr 115 120 125

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Gin lie Gin Leu Val Gin 130 135 140

Ser G]y Pro Glu Leu Arg Lys Pro Gly Glu Thr Val Lys lie Ser Cys 145 150 155 160

Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr Gly lie Asn Trp Val Lys 165 170 175

Gin Thr Pro Arg Lys Asp Leu Lys Trp Met Gly Trp lie Asn Thr His 180 185 190

Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe Lys Gly Arg Phe Ala Phe 195 200 205

Ser Leu Glu Thr Ser Ala Asn Tlir Ala Tyr Leu Gin lie Asn Asn Leu 210 215 220 •168-

160877· Sequence Listing.doc 201247704

Asn Asn Gly Asp Met Gly Thr Tyr Phe Cys Thr Arg Ser His Arg Phe 225 230 235 240

Gly Leu Asp Tyr Trp Gly Gin Gly Thr Sei Val Thr Val Scr Ser 245 250 255 <210> 197 <2il> 249 <212> PRT <213> Artificial Sequence <220><223> Artificial Sequence Description: Synthetic Peptide <400> 197

Asp Val Val Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly 15 10 15

Gin Pro Ala Ser lie Ser Cys Lys Ser Ser Gin Ser Leu Leu Asp Ser 20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser 35 40 45

Pro Lys Leu Leu lie Tyr Val Val Ser Lys Leu Glu Ser Giy Val Pro 50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 SO

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala 85 90 95

Thr His Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110

Arg Thr Val Ala Ala Fro Ser Val Phe lie Phe Pro Pro Thr Val Ala 115 120 125

Ala Pro Ser Val Phe lie Phe Pro Pro Asp Asn Val Leu Thr Gin Ser 130 135 140

Pro Pro Ser Leu Ala Val Ser Leu Gly Gin Arg Ala Tlir lie Ser Cys 145 150 155 160

Lys Ala Asn Trp Pro Val Asp Tyr Asn Gly Asp Ser Tyr Leu Asn Trp 165 170 175

Tyr Gin Gin Lys Pro Gly Gin Pro Pro Lys Phe Leu lie Tyr Ala Ala 180 185 190

Ser Asn Leu Glu Ser Gly lie Pro Ala Arg Phe Ser Gly Ser Gly Ser 195 200 205

Gly Thr Asp Phe Asn Leu Asn lie His Pro Val Glu Glu Glu Asp Ala 210 215 220

Ala Thr Tyr Tyr Cys Gin Gin Ser Asn Glu Asp Pro Phe Thr Phe Gly • 169- 160877 · Sequence Listing.doc 201247704 225 230 235 240

Ser Gly Thr Lys Leu Glu lie Lys Arg 245 <210> 198 <211> 242 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <40Q> 198

Gin lie Gin Leu Val Gin Ser Gly Pro Glu Leu Arg Lys Pro Gly Glu 15 10 15

Thr Val Lys lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr 20 25 30

Gly lie Asn Trp Val Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met 35 40 45

Gly Trp lie Asn Thr His Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Asn Thr Ala Tyr 65 70 75 80

Leu Gin He Asn Asn Leu Asn Asn Gly Asp Met Gly Thr Tyr Phe Cys 85 90 95

Thr Arg Ser His Arg Phe Gly Leu Asp Tyr Trp Gly Gin Gly Thr Ser 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Gin Val GJn Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro 130 135 140

Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 145 150 155 160

Ser Tyr Trp Met Asn Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu 165 170 175

Trp lie Gly Arg lie Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin 180 185 190

Lys Phe Lys Asp Lys Ala He Leu Thr Val Asp Lys Ser Ser Ser Thr 195 200 205

Ala Phe Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr 210 215 220

Tyr Cys Val Ser Asp Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val 225 230 235 240 • 170-

160877 - Sequence Listing.doc 201247704

Scr Ser <210> 199 <21]> 237 <212> PRT <213> Artificial sequence <220><223> Description of artificial sequence: synthetic polypeptide <400>

Asp Asn Val Leu Tlir Gin Ser Pro Pro Scr Leu Ala Val Ser Leu Gly 15 10 15

Gin Arg Ala Thr lie Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asa 20 25 30

Gly Asp Ser Tyr Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45

Lys Phe Leu lie Tyr Ala Ala Ser Asn Leu Glu Ser Gly lie Pro Ala 50 55 60

Arg Phc Ser Gly Ser Gly Ser Gly Thr Asp Phe Asn Leu Asn lie His 65 70 75 80

Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn 85 90 95

Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu lie Lys Arg 100 105 110

Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp Val Val Met 115 120 125

Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly Gin Pro Ala Ser 130 135 140 lie Ser Cys Lys Ser Ser Gin Ser Leu Leu Asp Ser Asp Gly Lys rrhr 145 150 155 160

Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser Pro Lys Leu Leu 165 170 175 lie Tyr Val Val Ser Lys Leu Glu Ser Gly Val Pro Asp Arg Phe Thr 180 185 190

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie Ser Arg Val Glu 195 200 205

Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala Thr His Phe Pro 210 215 220

Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg 225 230 235 -171 - 160877 · Sequence Listing.doc 201247704 <210> 200 <211> 255 <212> PRT <213>Artificial Sequence<220>;<223> Description of artificial sequence: synthetic polypeptide <400> 200

Gin lie ύΐη Leu Val G!n Ser Gly Pro Glu Leu Arg Lys Pro Gly G】u 1 5 10 15

Thr Val Lys lie Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr His Tyr 20 25 30

Gly lie Asn Trp Val Lys Gin Thr Pro Arg Lys Asp Leu Lys Trp Met 35 40 45

Gly Trp lie Asn Thr His Thr Gly Glu Ala Tyr Tyr Ala Asp Asp Phe 50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Asn Thr Ala Tyr 65 70 75 80

Leu Gin lie Asn Asn Leu Asn Asn Gly Asp Met Gly Thr Tyr Phe Cys 85 90 95

Thr Arg Ser His Arg Phe Gly Leu Asp Tyr Trp Gly Gin Gly Thr Ser 100 105 110

Val Ήιγ Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Gin 130 135 140

Val Gin Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala Ser 145 150 155 160

Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp 165 170 175

Met Asn Trp Val Lys Gin Arg Pro Glu Gin Gly I^eu Glu Trp He Gly 180 185 190

Arg He Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe Lys 195 200 205

Asp Lys Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe Val 210 215 220

Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Val 225 230 235 240

Ser Asp Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 245 250 255 -172- 160877 · Sequence Listing.doc 201247704 <210> 201 <211> 249 <212> PRT <213> Artificial Sequence <;220><223> Description of artificial sequence: synthetic polypeptide <400> 201

Asp Asn Val Leu Thr Gin Ser Pro Pro Ser Leu Ala Val Ser Leu Gly 1 5 10 15

Gin Arg Ala Thr lie Ser Cys Lys Ala Asn Trp Pro Val Asp Tyr Asn 20 25 30

Giy Asp Ser Tyr Leu Asn Trp Tyr Gin Gin Lys Pro Gly Gin Pro Pro 35 40 45

Lys Phe Leu He Tyr Ala Ala Ser Asn Leu GIu Ser Gly lie Pro Ala 50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Tlir Asp Phe Asn Leu Asn 31e His 65 70 75 80

Pro Val Glu GIu Glu Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Ser Asn 85 90 95

Glu Asp Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu lie Lys Arg 100 105 110

Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Thr Val Ala Ala 115 120 125

Pro Ser Val Phe lie Phe Pro Pro Asp Val Val Met Thr Gin Thr Pro 130 135 140

Leu Thr Leu Scr Val Thr Thr Gly Gin Pro Ala Scr He Ser Cys Lys 145 150 155 160

Ser Ser Gin Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn Trp 165 170 175

Leu Phe Gin Arg Pro Giy Glu Ser Pro Lys Leu Leu lie Tyr Val Val 180 185 190

Ser Lys Leu Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser 195 200 205

Gly Thr Asp Phe Thr Leu Lys lie Ser Arg Val Glu Ala Glu Asp Leu 210 215 220

Gly Val Tyr Tyr Cys Leu Gin Ala Thr His Phe Pro Trp γΠιγ Phe Gly 225 230 235 240

Gly Gly Thr Lys Leu Glu He Lys Arg 245 <210> 202 •173·160877·Sequence List.doc 201247704 <211> 237 <212> PRT <213> Artificial Sequence <220><223> Description of the sequence: synthetic peptide <400> 202

Gin Val Gin Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 15 10 15

Ser Val Lys Leu Scr Cys Lys Ala Ser Gly Tyr Thr Phe Ίϊιγ Ser Tyr 20 25 30

Trp Met Asn Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp lie 35 40 45

Gly Arg lie Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe 50 55 60

Lys Asp Lys Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe 65 70 75 80

Val Gin Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Val Ser Asp Gly Tyi Ding rp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 100 105 110

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Gin Val Gin 115 120 125

Leu Gin Gin Pro Gly Ala Glu Leu Val Arg Pro Gly Ala Ser Val Lys 130 135 140

Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met Asn 145 150 155 160

Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp lie Gly Arg lie 165 170 175

Asp Pro Tyr Asp Ser Glu Thr His Tyr Asn Gin Lys Phe Lys Asp Lys 180 185 190

Ala lie Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Phe Val Gin Leu 195 200 205

Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Val Ser Asp 210 215 220

Gly Tyr Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 225 230 235 <210> 203 <211> 238 <212> PRT <213> Artificial Sequence <220> -174·

160877 - Sequence Listing.doc 201247704 <223> Description of Artificial Sequence: Synthetic Peptide <400> 203

Asp Val Val Met Hir Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly 15 10 15

Gin Pro Ala Ser lie Ser Cys Lys Ser Ser Gin Ser Leu Leu Asp Ser 20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser 35 40 45

Pro Lys Leu Leu lie Tyr Val Thr Asp lie Leu Glu Scr Gly Val Pro 50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 80

Ser Are Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala 85 90 95

Thr His Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys 100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe lie Phe Pro Pro Asp Val Val 115 120 125

Met Thr Gin Thr Pro Leu Thr Leu Ser Val Thr Thr Gly Gin Pro Ala 130 135 140

Ser lie Ser Cys Lys Ser SeT Gin Ser Leu Leu Asp Ser Asp Gly Lys 145 150 155 160

Thr Tyr Leu Asn Trp Leu Phe Gin Arg Pro Gly Glu Ser Pro Lys Leu 165 170 175

Leu lie Tyr Val Thr Asp lie Leu Glu Ser Gly Val Pro Asp Arg Phe 180 185 190

Thr Gly Ser Gly Ser Gly Hir Asp Phe Th Leu Lys lie Ser Arg Val 195 200 205

Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Leu Gin Ala Thr His Phe 210 215 220

Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg 225 230 235 <210> 204 <211> 251 <212> PRT <213> Artificial Sequence <220><223> Description of Artificial Sequence : Synthetic Peptide <400> 204

Glu Val Gin Leu Gin Gin Ser Gly Pro Asp Leu Val Lys Pro Gly Ala -175- 160877 - Sequence Listing.doc 201247704 15 10 15

Ser Val Arg lie Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Asn Leu His Trp Val Lys Gin Ser His Gly Lys Ser Leu Glu Trp lie 35 40 45

Gly Tyr lie Tyr Pro Tyr Asn Gly He Thr Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Ser Lys Ala Thr Leu Thr Val Asp Ser Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Asp Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95

Ala Arg Asp Ala Tyr Asp Tyr Asp Tyr Leu Thr Asp Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125

Pro Leu Ala Pro Glu Val Gin Leu Gin Gin Ser Gly Pro Asp Leu Val 130 135 140

Lys Pro Gly Ala Ser Val Arg lie Ser Cys Lys Ala Ser Gly Tyr Thr 145 150 155 160

Phe Thr Asp Tyr Asn Leu His Trp Val Lys Gin Ser His Gly Lys Ser 165 170 175

Leu Glu Trp lie Gly Tyr lie Tyr Pro Tyr Asn Gly lie Thr Gly Tyr 180 185 190

Asn Gin Lys Phe Lys Ser Lys Ala Thr Leu Thr Val Asp Ser Ser Ser 195 200 205

Asn Thr Ala Tyr Met Asp Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala 210 215 220

Val Tyr Phe Cys Ala Arg Asp Ala Tyr Asp Tyr Asp Tyr Leu Thr As 225 230 235 24

Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ala 245 250 <210> 205 <211> 228 <212> PRT < 213 > Artificial Sequence <220><223> Description of Artificial Sequence: Synthetic Peptide <;400> 205

Asp lie Leu Leu Thr Gin Ser Pro Val lie Leu Scr Val Ser Pro Gly 15 10 15 •176·

160877 - Sequence Listing.doc 201247704

Glu Arg Val Ser Phe Ser Cys Arg Thr Ser Lys Asn Val Gly Thr Asn 20 25 30 lie His Trp Tyr Gin Gin Arg Thr Asn Gly Ser Pro Arg Leu Leu lie 35 40 45

Lys Tyr Ala Ser Glu Arg Leu Pro Gly lie Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser lie Asn Ser Vai Glu Ser 65 70 75 80

Glu Asp lie Ala Asp Tyr Tyr Cys Gln'Gln Ser Asn Asn Trp Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu lie Lys Arg Hir Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Asp lie Leu Leu Thr Gin Sex Pro 115 120 125

Val lie Leu Ser Val Ser Pro Gly Glu Arg Val Ser Phe Ser Cys Arg 130 135 140

Thr Ser Lys Asn Val Gly Thr Asn lie His Trp Tyr Gin Gin Arg Thr 145 150 155 160

Asn Gly Ser Pro Arg Leu Leu lie Lys Tyr Ala Ser Glu Arg Leu Pro 165 170 175

Gly lie Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 180 185 190

Leu Ser lie Asn Ser Val Glu Ser Glu Asp lie Ala Asp Tyr Tyr Cys 195 200 205

Gin Gin Ser Asn Asn Trp Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu 210 215 220

Glu lie Lys Arg 225 <210> 206 <211> 125 <212> PRT <213> Homo sapiens <400> 206

Ala Met His Val Ala Gin Pro Ala Val Val Leu Ala Ser Ser Arg Gly 15 10 15 lie Ala Ser Phe Val Cys Glu Tyr Ala Ser Pro Gly Lys Ala Thr Glu 20 25 30

Val Arg Val Thr Val Leu Arg Gin Ala Asp Ser Gin Val Thr Glu Val 35 40 45 -177· 160877-Sequence List.doc 201247704

Cys Ala Ala Thr Tyr Met Met Gly Asn Glu Leu Thr Phe Leu Asp Asp 50 55 60

Ser He Cys Thr Gly Thr Ser Ser Gly Asn Gin Val Asn Leu Thr lie 65 70 75 80

Gin Gly Leu Arg Ala Met Asp Thr Gly Leu Tyr lie Cys Lys Val Glu 85 90 95

Leu Met Tyr Pro Pro Pro Tyr Tyr Leu Gly lie Gly Asn Gly Thr Gin 100 105 110 lie Tyr Val He Asp Pro Glu Pro Cys Pro Asp Ser Asp 115 120 125 <210> 207 <211> 161 <212> PRT <213> Homo sapiens <400> 207

Ala Gin Val Ala Phe Thr Pro Tyr Ala Pro Glu Pro Gly Ser Thr Cys 15 10 15

Arg Leu Arg Glu Tyr Tyr Asp Gin Thr Ala Gin Met Cys Cys Ser Lys 20 25 30

Cys Ser Pro Gly Gin His Ala Lys Val Phe Cys Thr Lys Thr Ser Asp 35 40 45

Thr Val Cys Asp Sei Cys Glu Asp Ser Thr Tyr Thr Gin Leu Trp Asn 50 55 60

Trp Val Pro Glu Cys Leu Ser Cys Gly Ser Arg Cys Ser Ser Asp Gin 65 70 75 80

Val Glu Thr Gin Ala Cys Thr Arg Glu Gin Asn Arg He Cys Thr Cys 85 90 95

Arg Pro Gly Trp Tyr Cys Ala Leu Ser Lys Gin Glu Gly Cys Arg Leu 100 105 Π0

Cys Ala Pro Leu Arg Lys Cys Arg Pro Gly Phe Gly Val Ala Arg Pro IIS 120 125

Gly Thr Glu Thr Ser Asp Val Val Cys Lys Pro Cys Ala Pro Gly Thr 130 135 140

Phe Ser Asn Thr ΤΗτ Ser Ser Thr Asp lie Cys Arg Pro His Gin lie 145 150 155 160

Cys <210> 208 <211> 5 <2I2> PRT <213> artificial sequence •178-

160877·SEQ ID NO.doc 201247704 <220><223> Description of artificial sequence: synthetic peptide <400> 208

Gly Gly Gly Gly Ser 1 5

160877 - Sequence Listing. doc • 179-

Claims (1)

201247704 VII. Patent Application Range: 1. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-X2, wherein: VD1 comprises a heavy chain antigen binding domain; XI comprises a group selected from the group consisting of Domain: polypeptide, CH1 domain, CH2 domain, CH1 domain and CH2 domain, and linker; N is 0 or 1; and X2 comprises a polypeptide having at least a portion of the CH3 domain, φ wherein the binding protein comprises at least A mutation to inhibit dimerization, wherein the binding protein forms a functional antigen binding site. 2. The binding protein of claim 1, wherein the VD1 is selected from the group consisting of a heavy chain variable domain, a double heavy chain variable domain, a triple heavy chain variable domain, a light chain variable domain, and a double light chain. Variable domain, triple light chain variable domain, combination of heavy chain variable domain and light chain variable domain, combination of two heavy chain variable domains and light chain variable domains, heavy chain variable domain and two light chains Combination of variable domains, domain_antibody, camelid antibody, scFv, receptor and backbone antigen binding protein. 3. The binding protein of claim 1 or 2, wherein the binding protein further comprises a hinge (H) region between VD1 and X2. The binding protein of any one of claims 1 to 3, wherein the at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. The binding protein according to any one of claims 1 to 4, wherein the at least one mutation is at a residue selected from the group consisting of C220, C226, C229, T366, L368, P395 according to Kabat nomenclature, F405, Y407 160877.doc 201247704 and K409. The binding protein of any one of claims 1 to 5, further comprising a second polypeptide chain, wherein the second polypeptide chain comprises vd1_(x1)n, wherein VD1 comprises a light chain antigen binding domain; The domains of the following composition are selected: polypeptide, cL domain, CL-CH2 domain, CH1 domain, CH2 domain, CH1 domain, and CH2 domain, and a linker; and N is 0 or 1. 7. The binding protein of claim 6, wherein the vm is selected from the group consisting of a light chain variable domain, a double light chain variable domain, a triple light chain variable domain, a heavy chain variable domain, and a double heavy chain. Variable domain, triple heavy chain variable domain, heavy chain variable domain and light chain variable domain combination, two heavy chain variable domain and light chain variable domain group 〇, heavy bond variable domain and two light A combination of key variable domains, a camelid antibody, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. 8. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1_X1-X2, wherein: VD1 comprises a first heavy chain variable domain; XI comprises a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain, and a linker; and X2 comprises at least a portion of a ch3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit CH3-CH3 dimerization, wherein the binding protein forms a functional antigen Binding site. 9. The binding protein of claim 8, wherein the binding protein further comprises a chining region between VD1 and 乂2 of 160877.doc 201247704. 10. The binding protein of any one of clauses 8 or 9, wherein the at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. The binding protein according to any one of claims 8 to 10, wherein the at least one mutation that inhibits CH3-CH3 dimerization is at a residue selected from the group consisting of C220, C226 according to Kabat nomenclature, C229, T366, L368, P395, F405, Y407 and K409. 12. A binding protein comprising a first polypeptide chain and a second polypeptide chain, wherein: the first polypeptide chain comprises VD1-X1-X2, wherein: VD1 comprises a first heavy chain variable domain; a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain, and a linker; and X2 comprises at least a portion of the CH3 domain; and wherein the second polypeptide chain comprises VD1-XI, wherein VD1 comprises a light chain variable domain; and XI comprises a light chain constant domain, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain; wherein the binding protein comprises at least one mutation at the residue to inhibit CH3-CH3 dimerization, And the binding protein forms a functional antigen binding site. 13. The binding protein of claim 12, wherein the binding protein further comprises a Syrian bond region between VD1 and X2. The binding protein of any one of claims 12 or 13, wherein the at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. The binding protein according to any one of claims 12 to 14, wherein the at least one mutation that inhibits CH3-CH3 dimerization is at a residue selected from the group consisting of: according to Kabat nomenclature C220, C226, C229, T366, L368, P395, F405, Y407 and K409. 16. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-X1-X2, wherein: VD1 comprises a heavy chain variable domain; XI comprises a CH1 domain and a hinge region, wherein the hinge region is located in the CH1 domain C-terminal; and φ X2 comprises at least a portion of the CH3 domain; wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, C229, T366, L368, P395, F405, Y407 and K409, lend! T匕 inhibits CH3-CH3 dimerization; and the binding protein forms a functional antigen binding site. 17. A binding protein comprising a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises VD1-X1-X2, wherein; LuD VD1 comprises a heavy chain variable domain; XI comprises a CH1 domain And a hinge region, wherein the hinge region is located at the C-terminus of the CH1 domain; and X2 comprises at least a portion of the CH3 domain; and wherein the second polypeptide chain comprises VD1-XI, wherein VD1 comprises a light chain variable domain; and X1 comprises light a chain constant domain; 160877.doc 201247704 wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226, C229, T366, L368, P395, F405, Y407 and K409, thereby inhibiting CH3-CH3 dimerization; and the binding protein forms a functional antigen binding site. 18. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-X3, wherein: VD1 comprises a first heavy chain antigen binding domain; φ XI is a linker VD2 comprises a second heavy chain antigen binding domain; X2 comprises a domain selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain and a CH2 domain, a light chain constant region, and a linker; each N independently Is selected from 0 and 1; and X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit • CH3-CH3 dimerization, wherein the binding protein forms a functional antigen binding site . 19. The binding protein of claim 18, wherein VD1 and VD2 are each independently selected from the group consisting of a heavy chain variable domain, a light chain variable domain, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. 20. The binding protein of claim 18 or 19, wherein the binding protein further comprises a Qinling region between VD2 and X3. 21. The binding protein of claim 18 or 19, wherein the at least one mutation that inhibits CH3-CH3 dimerization 160877.doc 201247704 is in the CH3/CH3 dimerization contact zone or in the hinge region. The binding protein according to any one of claims 18 to 21, wherein the at least one mutation that inhibits CH3-CH3 dimerization is at a residue selected from the group consisting of C220, C226 according to Kabat nomenclature, C229, T366, L368, P395, F405, Y407 and K409. The binding protein of any one of claims 18 to 32, further comprising a second polypeptide chain, wherein the second polypeptide chain comprises VD1-(X1)n-VD2-(X2)n, wherein VD1 comprises a first light chain antigen binding domain; XI is a linker; VD2 comprises a second light chain antigen binding domain; X2 comprises a domain selected from the group consisting of: a polypeptide, a light chain constant domain, a CH1 domain, a CH2 domain, a CH1 domain And a CH2 domain; and each N is independently selected from 0 and 1. The binding protein according to any one of claims 18 to 23, wherein the VD1 and the VD2 are selected from the group consisting of a light chain variable domain, a heavy chain variable domain, a domain antibody, a scFv, a receptor, and Skeletal antigen binding protein. 25. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-X3, wherein: VD1 comprises a first heavy chain variable domain; XI is a linker; Each N is independently selected from 0 and 1; VD2 comprises a second heavy chain variable domain; 160877.doc _6· 201247704 X2 comprises a heavy chain constant 1 (CH1) domain, and X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein The binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: T366, L368, P395, F405, Y407 and K409; wherein the binding protein forms a functional antigen binding site. 26. The binding protein of claim 25, wherein the binding protein further comprises a hinge region between VD2 and X3. Φ 27. The binding protein of claim 26, wherein the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226 and C229 » 28-binding protein A first polypeptide chain comprising a VD1-(X1)N-VD2-(X2)N-X3, wherein: VD1 comprises a first heavy bond variable domain; a linker; each N is independently selected from 0 and 1; Φ VD2 comprises a second heavy chain variable domain; X2 comprises a heavy chain constant l (CH1) domain; and X3 comprises a polypeptide having at least a portion of the CH3 domain, wherein the The dipeptide chain comprises VD1-(X1)N-VD2-(X2)N, wherein VD1 comprises a first light chain variable domain; XI is a linker; VD2 comprises a second light chain variable domain; X2 comprises a light chain constant And 160877.doc 201247704 each N is independently selected from 0 and 1; wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: Τ366, L368, Ρ395 , F405, Υ407 and Κ409; and wherein the binding protein forms a functional antigen binding site. 29. The binding protein of claim 28, wherein the binding protein further comprises a hinge region between VD2 and Χ3. The binding protein of claim 29, wherein the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226 and C229. 31. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)n-VD2-(X2)n-VD3-(X3)n-X4, wherein: VD1 comprises a first heavy chain antigen binding Domain; XI is the first linker; VD2 comprises the second heavy chain antigen binding domain; X2 is the second linker; VD3 comprises the third heavy chain antigen binding domain; X3 comprises a domain selected from the group consisting of: polypeptide, a CH1 domain, a CH2 domain, a CH1 domain and a CH2 domain, a light chain constant domain, and a linker; each N is independently selected from 0 and 1; and X4 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein is at a residue At least one mutation is included to inhibit emeus dimerization, and wherein the binding protein forms a functional antigen junction 160877.doc 201247704. 32. The binding protein of claim 31, wherein the binding protein further comprises a more bond region between VD3 and X4. The binding protein according to any one of claims 25 to 32, wherein each of VD1, VD2 and VD3 is selected from the group consisting of a heavy chain variable domain, a light chain variable domain, a domain antibody, a scFv, a receptor and Skeletal antigen binding protein. The binding protein of any one of claims 25 to 33, wherein the at least one mutation that inhibits CH3-CH3 dimerization is in the CH3/CH3 dimerization contact zone φ or in the hinge region. The binding protein according to any one of claims 25 to 34, wherein the at least one mutation is at a residue selected from the group consisting of C220, C226, C229, T366, L368, P395 according to Kabat nomenclature The binding protein according to any one of claims 18 to 35, further comprising a second polypeptide chain, wherein the second polypeptide chain comprises VD1-(X1)n-VD2-( X2)n-VD3-(X3)n, wherein #VD1 comprises a first light chain antigen binding domain; XI is a first linker; VD2 comprises a second light chain antigen binding domain; X2 is a second linker; VD3 comprises a triple light chain antigen binding domain; X3 comprises a domain selected from the group consisting of a polypeptide, a light chain constant domain, a CH1 domain, a CH2 domain, and a CH1 domain and a CH2 domain; and each N is independently selected from 0 and 1. The binding protein of claim 36, wherein each of VD1, VD2 and VD3 is selected from the group consisting of a light chain variable domain, a heavy chain variable domain, a domain antibody, a scFv, And Skeletal Antigen Binding Proteins A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-VD3-(X3)N-X4, wherein: VD1 comprises the first a heavy chain variable domain; XI is a first linker; VD2 comprises a second heavy chain variable domain; X2 is a second linker; VD3 comprises a third heavy bond variable domain; each N is independently selected from 〇 and 1; X3 comprises a heavy chain constant 1 (CH1) domain; and X4 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: T366, L368, P395, F405, Y407 and K409; and wherein the binding protein forms a functional antigen binding site. A binding protein comprising a first and a second polypeptide chain, wherein the first polypeptide chain comprises VD1-(X1)N-VD2-(X2)N-VD3-(X3)N-X4, wherein; VD1 comprises a first heavy chain variable domain; XI is a first linker; VD2 comprises a second heavy chain variable domain; X2 is a second linker; 160877.doc • 10. 201247704 VD3 comprises a third heavy chain variable domain; N is independently selected from the group consisting of 〇 and 1; X3 comprises a heavy chain constant i(CH1) domain; and X4 comprises a polypeptide having at least a portion of the CH3 domain; and wherein the second polypeptide chain comprises VD1-(X1)n-VD2-( X2)n-VD3-(X3)n, wherein VD1 comprises a first light chain variable domain; XI is a first linker; φ VD2 comprises a second light chain variable domain; X2 is a second linker; VD3 comprises a three light chain variable domain; X3 comprises a light chain constant domain; and each N is independently selected from the group consisting of 〇 and 1; wherein the binding protein comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 Polymerization: T366, L368, P395, F405, Y407, and K409, and wherein the binding protein forms a successful Φ-energy antigen binding site. 40. The binding protein of claim 38 or 39, wherein the binding protein further comprises a hinge region between VD3 and X4. 41. The binding protein of claim 40, wherein the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226 and C229. 42. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises a format selected from the group consisting of: R-(X1)N-(VD1)N-(X2)N-X3, or 160877.doc 201247704 (VD1)n-(X1)nR-(X2)n-X3, or (VD1)N-(X2)N-X3-(X1)NR, wherein: R comprises a receptor; XI is a linker; VD1 comprises heavy a strand antigen binding domain; X2 comprises one or more domains selected from the group consisting of: a polypeptide, a CH1 domain, a CH2 domain, a CH1 domain and a CH2 domain, a hinge region, and a linker; each N is independently selected from 0 and And X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein the binding protein comprises at least one mutation at the residue to inhibit dimerization, and wherein the binding protein forms a functional antigen binding site. 43. The binding protein of claim 24 or 25, wherein the VD2 is selected from the group consisting of a heavy chain variable domain, a light chain variable domain, a domain antibody, a scFv, a receptor and a backbone antigen binding protein. The binding protein of any one of claims 39 to 43, wherein the at least one mutation is in the CH3/CH3 dimerization contact zone or in the hinge region. The binding protein according to any one of claims 39 to 44, wherein the at least one mutation is at a residue selected from the group consisting of C220, C226, C229, T366, L368, P395 according to Kabat nomenclature, The conjugated protein according to any one of claims 39 to 45, further comprising a second polypeptide chain, wherein the second polypeptide chain comprises a group selected from the group consisting of 160877.doc -12 · Format of 201247704: R-(X1)n-VD1-(X2)n, or VD1-(X1)nR-(X2)n, or VD1-(X2)n-(X1)nR, where R contains a receptor XI is a linker; VD1 comprises a light chain antigen binding domain; X2 comprises a domain selected from the group consisting of: a polypeptide, a light chain constant domain, a CH1 domain, a CH2 domain, a CH1 domain, and a CH2 domain; and each N independently Selected from 0 and 1. Φ 47. The binding protein of claim 46, wherein the VD2 is selected from the group consisting of a light chain variable domain, a heavy chain variable domain, a domain antibody, a scFv, a receptor, and a backbone antigen binding protein. 48. A binding protein comprising a polypeptide chain, wherein the polypeptide chain comprises R-(X1)n-VD1-(X2)n-X3, wherein; R comprises a receptor; XI is a linker; each N is independently selected From 0 and 1; #VD1 comprises a heavy chain variable domain; X2 comprises a heavy chain constant 1 (CH1) domain; and X3 comprises a polypeptide having at least a portion of a CH3 domain, wherein X3 comprises at a residue selected from the group consisting of At least one mutation to inhibit CH3-CH3 dimerization: T366, L368, P395, F405, Y407, and K409; wherein the binding protein forms a functional antigen binding site. 49. A binding protein comprising a first and a second polypeptide chain, wherein the first 160877.doc-13·201247704 polypeptide chain comprises R-(X1)N-VD2-(X2)N-X3, wherein; R Include a receptor; XI is a linker; each N is independently selected from 〇 and 1; VD2 comprises a heavy chain variable domain; X2 comprises a heavy chain constant 1 (CH1) domain; and X3 comprises a polypeptide having at least a portion of the CH3 domain, wherein X3 comprises at least one mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: T366, L368, P395, F405, Y407, and K409; and wherein the second polypeptide chain comprises R-(X1) N-VD1-(X2)N, wherein R is a receptor; XI is a linker; VD1 is a light bond variable domain; X2 is a light chain constant domain; and each N is independently selected from 0 and 1; wherein the binding The protein forms a functional antigen binding site. 50. The binding protein of claim 48 or 49, wherein the binding protein further comprises a hinge region between VD3 and X4. 5. A binding protein according to claim 50, wherein the binding protein comprises a mutation at a residue selected from the group consisting of: inhibiting CH3-CH3 dimerization: C220, C226 and C229 » 52. A binding protein according to any one, wherein the at least one mutation is at a residue selected from the group consisting of C220, C226 and 160877.doc 201247704 C229. 53. The binding protein of claim 52, wherein the at least one mutation is two mutations selected from the group consisting of C220S, C226S and C229S. 54. The binding protein of claim 52, wherein the at least one mutation is all three mutations C220S, C226S and C229S. 55. The binding protein of any of the preceding claims, wherein at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75% of the population of binding proteins At least 80%, at least • 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% Dimerization via this CH3 domain. The binding protein according to any one of the preceding claims, wherein the binding protein comprising at least one mutation that inhibits CH3-CH3 dimerization corresponds to at least one mutation that does not comprise inhibition of CH3-CH3 dimerization. Binding proteins have altered biological activity. The binding protein according to any of the preceding claims, wherein the binding protein is a 9 antagonist. 58. The binding protein of any of the preceding claims, wherein the binding protein is an agonist. The binding protein according to any one of the preceding claims, wherein at least one Fc function of the binding protein that is not dimerized via the CH3 domain is altered compared to the corresponding binding protein that it will dimerize. The binding protein according to any one of the preceding claims, wherein the binding activity of the FcRn in the binding protein that is not dimerized via the CH3 domain is compared to the corresponding binding of its fusion of 160877.doc -15-201247704 The protein changes. 61. The binding protein of claim 60, wherein the binding protein in the binding protein that is not dimerized via the CH3 domain has an increased FcRn binding efficiency compared to its corresponding binding protein that will dimerize. 62. The binding protein of claim 60, wherein the binding protein in the binding protein that is not dimerized via the CH3 domain has a reduced FcRn binding efficiency compared to its corresponding binding protein that will dimerize. 63. The binding protein of any of the preceding claims, wherein the FcyR binding potency of the binding protein that is not dimerized via the CH3 domain is altered compared to the corresponding binding protein that it will dimerize. 64. The binding protein of claim 63, wherein the FcyR binding efficiency of the binding protein that is not dimerized via the CH3 domain is increased compared to the corresponding binding protein that will dimerize. 65. The binding protein of claim 63, wherein the FcyR binding efficiency of the binding protein that is not dimerized via the CH3 domain is reduced compared to the corresponding binding protein that will dimerize. 66. The binding protein of any of the preceding claims, wherein the binding activity of the binding protein in the binding protein that is not dimerized via the CH3 domain is altered compared to the corresponding binding protein that it will dimerize. 67. The binding protein of claim 66, wherein the binding activity of the C1 qR in the binding protein that is not dimerized via the CH3 domain is increased compared to the corresponding binding protein that it dimerizes. 68. The binding protein of claim 66, wherein the binding activity of the C1 qR in the binding protein that is not dimerized via the CH3 domain is comparable to its corresponding dimerization. 160877.doc • 16- 201247704 69. 70 71. 72. 73. • 74. 75. 76. Binding protein is reduced. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, T366F, T368F, P395A, F405R, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, T366F 'T368F, P395A, F405A, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, P395A, F405R, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S 'C226S, C229S, P395A, F405A, Y407A and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, P395A, F405R and Y407A. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, F405R, Y407A and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, P395A, Y407A and K409D. The binding protein of any one of claims 1 to 68 wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S, 160877.doc 201247704 P395A, F405R and K409D » 77. as in claims 1 to 68 The binding protein of any of the following has a mutation at the following residues: C220S, P395A and F405R » 78. The binding protein of any one of claims 1 to 68 has a mutation at the following residues: C220S, P395A and Y407R. 79. The binding protein of any one of claims 1 to 68 having a mutation at the following residues: C220S, P395A and K409D. 80. The binding protein of any one of claims 1 to 68 having a mutation at the following residues: C220S, F405R and F407R. 81. The binding protein of any one of claims 1 to 68 having a mutation at the following residues: C220S, F405R and K409D. 82. The binding protein of any one of claims 1 to 68 having a mutation at the following residues: C220S, F407R and K409D. 83. The binding protein of any one of claims 1 to 68 having a mutation at the following residues: C220S, P395A. The binding protein according to any one of claims 1 to 68 having a mutation at the following residue: C220S, wherein the binding protein C226S, C229S, wherein the binding protein C226S, C229S, , wherein the binding protein C226S, C229S, wherein the binding proteins C226S, C229S, wherein the binding proteins C226S, C229S, wherein the binding proteins C226S, C229S, wherein the binding proteins C226S, C229S and, wherein the binding proteins C226S, C229S and 160877.doc • 18 · 201247704 K405R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S and F407R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, C229S and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein φ has a mutation at the following residues: T366F, T368F, P395A, F405R, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: T366F, T368F, P395A, F405A, Y407R, and K409D ° 89. As claimed in claims 1, 7 to 9 The binding protein of any one of 15 to 17, 23 to 25 or 3, wherein the binding protein has a mutation at the following residues: P395A, F405R, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: P395A, F405A, Y407A and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: P395A, F405R and Y407A. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: F4〇5R, Y407A and K4〇9D. The binding protein of any one of claims 1 to 68, wherein the binding protein 160877.doc -19-201247704 has a mutation at the following residues: P395A, Y407A and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: P395A, F405R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: P395A and F405R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: P395A and Y407R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: P395A and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: F405R and F407R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: F405R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: F407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the residue: Ρ395Α. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the residue: K405R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the residue: F407R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the residue: K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein 160877.doc -20 2012044704 comprises a wild type hinge region sequence. The binding protein according to any one of claims 1 to 68, wherein the binding protein comprises a wild type amino acid at at least one of C220, C226 and C229. The binding protein of any one of claims 1 to 68, wherein the binding protein comprises a wild-type amino acid at at least two of C220, C226 and C229. The binding protein of any one of claims 1 to 68, wherein the binding protein comprises a wild type amino acid at C220, C226 and C229. The binding protein of any one of claims 1 to 108, wherein the CH3 domain is at least 50% identical to the wild type CH3 domain, at least 55%, at least 60%, at least 65%, At least 70%, at least 75%, at least 80%, at least 85%, at least 87.5%, at least 90%, at least 92%, at least 94%, at least 96 % - or at least 98%. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, T366F, T368F, P395A, F405R, Y407R and K409D. The binding protein of any one of claims 11 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, T366F, T368F, P395A, F405R, Y407R and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, T366F, T368F, P395A, F405A, Y407A and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, T366F, 160877.doc -21 - 201247704 T368F, P395A, F405A, Y407A and The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, P395A, F405R, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, P395A, F405R, Y407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, P395A, F405A, Y407A and K409D. 117. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C220S, C226S, P395A, F405A, Y407A and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, P395A, F405R and Y407A. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, F405R, Y407A, and K409D » 120. As claimed in any one of claims 1 to 68 The binding protein, wherein the binding protein has the following residues: C226S, C229S, P395A, Y407A, and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein is at the following residue There are mutations: C226S, C229S, P395A, 160877.doc •22· 201247704 F405R and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, P395A and F405R ° 123. The combination according to any one of claims 1 to 68 Protein, wherein the binding protein has mutations at the following residues: C226S, C229S, P395A and Y407R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, P395A and K409D. The binding protein according to any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, F405R and F407R ° 126. The combination according to any one of claims 1 to 68 Protein, wherein the binding protein has mutations at the following residues: C226S, C229S, F405R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S, F407R and K409D. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S and P3 95A. 129. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S and K405R. The binding protein according to any one of claims 1 to 68, wherein the binding protein 160877.doc • 23· 201247704 has mutations at the following residues: C226S, C229S and F407R. The binding protein of any one of claims 1 to 68, wherein the binding protein has a mutation at the following residues: C226S, C229S and K409D. The binding protein of any one of claims 1 to 131, wherein the binding protein forms a functional antigen binding site against an antigen selected from the group consisting of: a cell surface binding molecule, a soluble molecule, a cytokine, a trend Factors, enzymes, haptens, lipids and receptors. 133. The binding protein of any one of claims 1 to 132, wherein the binding protein forms a functional antigen binding site against an antigen selected from the group consisting of c-Met; Muc-1; CD28; CD40; CD19 , CD3 , TWEAK , TNFR , TREM - 1 , ABCF1 , ACVR1 , ACVR1B , ACVR2 , ACVR2B , ACVRL1 , ADORA2A , Aggrecan , AGR2 , AICDA , AIF1 , AIG1 , AKAP1 , AKAP2 , AMH , AMHR2 ; ANGPT1 , ANGPT2 , ANGPTL3 , ANGPTL4 , ANPEP , APC , APOC1 , AR , AZGP1 ( zinc - a - glycoprotein ) , B7.1 , B7.2 , BAD , BAFF , BAG1 , BAI1 , BCL2 , BCL6 , BDNF , BLNK ; BLR1 (MDR15); BlyS; BMP1; BMP2; BMP3B (GDF10); BMP4; BMP6; BMP8; BMPR1A; BMPR1B; BMPR2; BPAG1 (plectin); BRCA1; C19orflO (IL27w); C3; C4A; C5; C5R1; CANT1; CASP1; CASP4; CAV1; CCBP2(D6/JAB61); CCL1(1-309); CCL11 (eotaxin); CCL13(MCP-4); CCL15(MIP-ld ); CCL16(HCC-4); 160877.doc -24· 20124770 4 CCL17 (TARC); CCL18 (PARC); CCL19 (MIP-3b); CCL2 (MCP-1); MCAF; CCL20 (MIP-3a); CCL21 (MIP-2); SLC; non-lymphoid tissue chemokine (exodus) CCL22 (MDC/STC-1); CCL23 (MPIF-1); CCL24 (MPIF-2/eosinophil chemoattractant-2); CCL25 (TECK); CCL26 (eosinophil chemotaxis) Factor-3); CCL27 (CTACK/ILC); CCL28; CCL3 (MIP-la); CCL4 (MIP-lb); CCL5 (RANTES); CCL7 (MCP-3); CCL8 (mcp-2); CCNA1; CCNA2 CCND1 ; CCNE1 ; CCNE2 ; CCR1 (CKR1/HM145) ; CCR2 (mcp-1RB/RA) ; CCR3 (CKR3/CMKBR3) ; CCR4 ; CCR5 (CMKBR5/ ChemR13) ; CCR6 (CMKBR6/CKR-L3/STRL22/DRY6 CCR7 (CKR7/EBI1); CCR8 (CMKBR8/TER1/CKR-L1); CCR9 (GPR-9-6); CCRL1 (VSHKl); CCRL2 (L-CCR); CD164; CD19; CD1C; CD20; CD200 CD-22 ; CD24 ; CD28 ; CD3 ; CD37 ; CD38 ; CD3E ; CD3G ; CD3Z ; CD4 ; CD40 ; CD40 ; CD44 ; CD72 ; CD74 ; CD79 ; CD72 ; ; CD86; CDH1 (E-cadherin); C CDH13; CDH13; CDH18; CDH19; CDH20; CDH5; CDH7; CDH8; CDH9; CDK2; CDK3; CDK4; CDK5; CDK6; CDK7; CDK9; CDKNlA (p21Wapl/Cipl); CDKNlB (p27Kipl); CDKN1C; CDKN2A CDKN2B; CDKN2C; CDKN3; CEBPB; CER1; CHGA; CHGB; chitinase 160877.doc -25-201247704 (Chitinase); CHST10; CKLFSF2; CKLFSF3; CKLFSF4; CKLFSF5; CKLFSF6; CKLFSF7; CKLFSF8; CLDN3; CLDN7 (Clindin-7); CLN3; CLU (aggregin (. 11^61^11)); (: fox 1^1; €] ^ 0 ft 1 (11 〇 (: 1); € grab 1; COL18A1; COL1A1; COL4A3; COL6A1; CR2; CRP; CSFl (M-CSF CSF2 (GM-CSF); CSF3 (GCSF); CTLA4; CTNNBl (b-catenin); CTSB (cathepsin B); CX3CL1 (SCYD1); CX3CR1 (V28) CXCL1(GROl) ; CXCLIO(IP-IO) ; CXCL11 (I-TAC/IP- φ 9) ; CXCL12(SDF1) ; CXCL13 ; CXCL14 ; CXCL16 ; CXCL2(GR02); CXCL3(GR03); CXCL5(ENA- 78/LIX); CXCL6(GCP-2); CXCL9(MIG); CXCR3(GPR9/CKR-L2); CXCR4; CXCR6(TYMSTR/STRL33/Bonzo); CYB5; CYC1; CYSLTR1; DAB2IP; DES; DKFZp451 JO 118 DNCL1 , DPP4 , E2F1 , ECGF1 , EDG1 , EFNA1 , EFNA3 , EFNB2 , EGF , EGFR , ELAC2 , ENG , ENOl , EN02 , EN03 , EPHB4 , EPO , ERBB2 ( Her-2 ) , EREG , ERK8 , ESR1 , ESR2 F3 (TF); FADD; FasL; FASN; FCER1A; FCER2; FCGR3A; FGF; FGF1 (aFGF); FGF10; FGF11; FGF12; FGF12B; FGF13; FGF14; FGF16; FGF17; FGF18 FGF19; FGF2 (bFGF); FGF20; FGF21; FGF22; FGF23; FGF3(int-2); FGF4(HST); FGF5; FGF6(HST-2); FGF7(KGF); FGF8; FGF9; FGFR3; FIGF(F VEGFD); I60877.doc -26- 201247704 FIL1 (EPSILON); FIL1 (ZETA); FLJ12584; FLJ25530 FLRT1 (fibronectin); FLT1; FOS FOSL1 (FRA-1); FY (DARC); GABRP (GABAa) GAGEB1; GAGEC1; GALNAC4S-6ST; GATA3 GDF5; GFI1; GGT1; GM-CSF; GNAS1; GNRH1 GPR2 (CCR10); GPR31; GPR44; GPR81 (FKSG80) GRCCIO(CIO); GRP; GSN (Gelsolin); GSTP1 HAVCR2; HDAC4; HDAC5 HDAC7A; HDAC9 HGF; HIF1A; HIP1; histamine and histamine receptor; HLA-A HLA-DRA; HM74; HMOX1; HUMCYT2A; ICEBERG ICOSL; ID2; IFN-a; IFNA1; IFNA2; IFNA4 IFNA5; IFNA6; IFNA7 IFNB1 , IFNy , IFNW1 IGBP1 , IGF1 , IGF1R , IGF2 , IGFBP2 , IGFBP3 , IGFBP6 , IL - 1 , IL10 , IL10RA , IL10RB , IL11 IL11RA , IL-12 , IL12A , IL12B , IL12RB1 , IL12RB2 IL13 , IL13RA1 , IL13RA2 , IL14 IL15 , IL15RA , IL16 , IL17 , IL17B , IL17C , IL17R , IL18 , IL18BP , IL18R1 , IL18RAP , IL19 , ILIA , IL1B , IL1F10 , IL1F5 , IL1F6 , IL1F7 , IL1F8 , IL1F9 , IL 1HY1 IL1R1 , IL1R2 , IL1RAP , IL1RAPL1 , IL1RAPL2 IL1RL1 , IL1RL2 , IL1RN , IL2 , IL20 , IL20RA , IL21R , IL22 , IL22R , IL22RA2 , IL23 , IL24 , IL25 , IL26 , IL27 , IL28A , IL28B , IL29 , IL2RA , IL2RB , IL2RG , IL3 IL30 , IL3 , IL4 , IL4 , IL4 , IL4 , IL4 , IL4 , IL6 , IL6 , IL6 , IL6 , IL6 , IL6 , IL6 , IL7 , IL7 , IL8 , IL8 , IL8 , IL8 , IL8 , IL8 , IL8 , IL8 , IL8 , IL8 INHBA; INSL3; INSL4; IRAKI; IRAK2; ITGA1; ITGA2; ITGA3; ITGA6 (a6 integrin); ITGAV; ITGB3; ITGB4 (b4 integrin); JAG1; JAK1; JAK3; JUN; K6HF; KAI1; KDR; KITLG; KLF5 (GC box BP); KLFK; KLK10; KLK12; KLK13; KLK14; KLK15; KLK3; KLK4; KLK5; KLK6; KLK9; KRT1; KRT19 (Keratin 19); KRT2A; KRTHB6 (hair _ specificity Type II keratin); LAMA5; LEP (leptin); Lingo-p75; Lingo-Troy; LPS; LTA (TNF-b); LTB; LTB4R (GPR16); LTB4R2; LTB R; MACMARCKS; MAG or Omgp; MAP2K7 (c-Jun); MDK; MIB1; midkine; MIF; MIP-2; MKI67 (Ki-67); MMP2; MMP9; MS4A1; MSMB; Protein-III (metallothionectin-III); MTSS1; MUC1 (mucin); MYC; MYD88; NCK2; neurocan (neurocan); φ NFKB1; NFKB2; NGFB (NGF); NGFR; NgR-Lingo NgR-Nogo66 (Nogo); NgR-p75; NgR-Troy; NME1 (NM23A); NOX5; NPPB; NR0B1; NR0B2; NR1D1; NR1D2; NR1H2; NR1H3; NR1H4; NRII2; NRII3; NR2C1; NR2C2; NR2E1; NR2E3 NR2F1 , NR2F2 , NR2F6 , NR3C1 , NR3C2 , NR4A1 , NR4A2 , NR4A3 , NR5A1 , NR5A2 , NR6A1 , NRP1 , NRP2 , NT5E , 160877.doc • 28 · 201247704 NTN4 , ODZ1 , OPRD1 , P2RX7 , PAP , PARTI , PATE , PAWR , PCA3 , PCNA , PDGFA , PDGFB , PECAM1 , PF4 ( CXCL4 ) , PGF , PGR , phosphoglycan , PIAS2 , PIK3CG , PLAU PLG; PLXDC1; PPBP(CXCL7); PPID; PR1; PRKCQ; PRKD1; PRL; PROC; PROK2; PSAP; PSCA; PTAFR; PTEN; PTGS2(COX-2); PTN; RAC2(p21Rac2); RARB; RGS13; RGS3; RNF110(ZNF144); R0B02; SI00A2; SCGB1D2 (lipophilin B); SCGB2A1 (mammaglobin 2); SCGB2A2 (mammaglobin 1); SCYE1 (endothelial single) Nuclear cell activated cytokines; SDF2; SERPINA1; SERPINA3; SERPINB5 (maspin); SERPINE1 (PAI-1); SERPINF1; SHBG; SLA2; SLC2A2; SLC33A1; SLC43A1; SLIT2; SPP1; SPRRlB (Sprl ST6GAL1 , STAB1 , STAT6 , STEAP , STEAP2 , TB4R2 , TBX21 , TCP10 , TDGF1 , TEK , TGFA , TGFB1 , TGFB1 , TGFB2 , TGFB3 , TGFB1 , TGFBR1 , TGFBR3 , TGFBR3 TH1L; THBS1 (thrombospondin-l); THBS2; THBS4; ΤΗΡΟ; TIE(Tie-1); TIMP3; tissue factor; TLR10; TLR2; TLR3; TLR4; TLR5; TLR6; TLR7; TLR8; TLR9; TNF; TNF-a; TNFAIP2 (B94); TNFAIP3; TNFRSF11A; TNFRSF1A; TNFRSF1B; TNFRSF21; TNFRSF5; TNFRSF6 (Fas); TNFRSF7; TNFRSF8; 160877.doc -29- 201247704 TNFRSF9; TNFSFIO (TRAIL); TNFSF11 ( TRANCE); TNFSF12 (AP03L); TNFSF13 (April); TNFSF13B; TNFSF14 (HVEM-L); TNFSF15 (VEGI); TNFSF18; TNFSF4 (OX40 ligand); TNFSF5 (CD4〇g hexasome); TNFSF6 (FasL TNFSF7 (CD27 ligand); TNFSF8 (CD30 ligand); TNFSF9 (4-1BB ligand); TOLLIP; Toll-like receptor; TOP2A (topoisomerase Iia); TP53; TPM1; TPM2; TRADD; TRAF1; TRAF2; TRAF3; TRAF4; TRAF5; TRAF6; TREM1; TREM2; TRPC6; TSLP; TWEAK; VEGF; VEGFB; VEGFC; versican; VHL C5; VLA-4; XCL1 Chemokine Lymphotactin)); XCL2 (SCM-lb); XCR1 (GPR5/CCXCR1); YY1; and ZFPM2. 134. The binding protein of any one of claims 2, 7, 8, 12, 16, 19, 24, 25, 28, 33, 36, 38, 39, 43 and 47 to 49, wherein at least one of the heavy chains The variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 27, 38, 40, 76, 81-83, 85, 91, 118, 120, 122, 124, 126, 128, 130, 132, 138, 160, 162, 164, 166, 168, 170, 172 '174, 176, 178, 180, 182 > 184, 186, 188, 190, 192, 194 '196, 198, 200' 202 And a binding protein according to any one of claims 2, 7, 12, 17, 19, 24, 28, 33, 36, 39, 43, 47, and 49, wherein the light chain variable domain comprises The amino acid sequence of the group consisting of: SEQ ID NO: 4, 160877.doc -30- 201247704 28, 39, 41, 79, 81-83 '85, 119, 121, 123 '125, 127, 129, 131, 133 > 135, 137, 139, 161, 163, 165, 167, 169, 171, 173, 175 '177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199 , 201 and 203. 136. The binding protein of any one of claims 2, 7, 8, 12, 16, 19, 24, 25, 28, 33, 36, 38, 39, 43 and 47 to 49, wherein the heavy chain R Or the acceptor comprises an amino acid sequence φ 选自 selected from the group consisting of SEQ ID NOS: 84, 206 and 207. 137. The binding protein of claim 2, 7, 12, 17, 19, 24, 28, 33, 36, 39, 43, 47 and 49, wherein the R or receptor of the light chain comprises a group selected from the group consisting of Amino acid sequence: SEQ ID NOS: 84, 206 and 207. 138. The binding protein of any of the preceding claims, wherein the binding protein is capable of binding one or more targets. 139. The binding protein of claim 138, wherein the one or more target lines are selected from the group consisting of: c-Met; CD-28; CD-3; CD-19; ABCF1; ACVR1; ACVR1B; ACVR2 ACVR2B; ACVRL1; ADORA2A; aggrecan; AGR2; AICDA; AIF1; AIG1; AKAP1; AKAP2; AMH; AMHR2; ANGPT1; ANGPT2; ANGPTL3; ANGPTL4; ANPEP; APC; APOC1; AR; AZGP1 (zinc-a- Glycoprotein); Β7·1; Β7.2; BAD; BAFF; BAG1; BAI1; BCL2; BCL6; BDNF; BLNK; BLR1 (MDR15); BlyS; BMP1; BMP2; 160877.doc • 31· 201247704 BMP3B (GDF10) BMP4; BMP6; BMP8; BMPR1A; BMPR1B; BMPR2; BPAG1 (retinin); BRCA1; C19orflO(IL27w); C3; C4A; C5; C5R1; CANT1; CASP1; CASP4; CAV1; CCBP2(D6/JAB61); CCL1 (1-309); CCL11 (eosinophil chemotactic factor); (:(:1^13(]^€?-4); CCL15(MIP-ld); CCL16(HCC-4) ; CCL17(TARC CCL18 (PARC); CCL19 (MIP-3b); CCL2 (MCP-1); MCAF; CCL20 (MIP-3a); CCL21 (MIP-2) ; SLC; non-lymphoid tissue chemokine (exodus)-2; CCL22 (MDC/STC-1); CCL23 φ (MPIF-1); CCL24 (MPIF-2/eosinophil chemotactic factor-2); CCL25 (TECK); CCL26 (eosinophil chemoattractant-3); CCL27 (CTACK/ILC); CCL28; CCL3 (MIP-la); CCL4 (MIP-lb); CCL5 (RANTES); CCL7 (MCP- 3); CCL8(mcp-2); CCNA1; CCNA2; CCND1; CCNE1; CCNE2; CCR1(CKR1/HM145); CCR2(mcp-1RB/RA); CCR3(CKR3/ CMKBR3); CCR4; CCR5(CMKBR5/ChemRl3 CCR6 (CMKBR6/CKR-L3/STRL22/DRY6); CCR7 (CKR7/EBI 1); · CCR8 (CMKBR8/TER1/CKR-L1); CCR9 (GPR-9-6); CCRL1 (VSHK1); CCRL2 CDL ; CD21 ; CD22 ; CD24 ; CD28 ; CD3 ; CD3 ; CD3 ; CD3 ; CD3 ; CD4 ; CD72; CD74; CD79A; CD79B; CD8; CD80; CD81; CD83; CD86; CDH1 (E·about mucin); CDH10; CDH12; CDH13; CDH18; 160877.doc -32· 201247704 CDH19 ; CDH8 ; CDH5 ; CDH7 ; CDK3 ; CDK9 ; CDK2 ; CDK3 ; CDK7 ; CDK5 ; CDKN3; CEBPB; CER1; CHGA; CHGB; chitinase; CHST10; CKLFSF2; CKLFSF3; CKLFSF4; CKLFSF5; CKLFSF6; CKLFSF7; CKLFSF8; CLDN3; CLDN7 (Claudin-7); CLN3; CLU (clusters); CMKLR1 ; CMKORl (RDCl); CNR1; COL18A1; COL1A1; COL4A3; COL6A1; CR2; CRP; CSF1 (M-CSF); CSF2 (GM-CSF); CSF3 (GCSF); CTLA4; CTNNB1 (b_sodacin); CTSB (Cathepsin B); CX3CL1 (SCYD1); CX3CR1 (V28); CXCL1 (GRO1); CXCLIO (IP-IO); CXCL11 (I-TAC/IP-9); CXCL12 (SDF1); CXCL13; CXCL14; CXCL16 CXCL2(GR02) ; CXCL3(GR03) ; CXCL5(ENA-78/LIX) ; CXCL6(GCP-2) ; CXCL9(MIG) ; CXCR3(GPR9/ CKR-L2) ; CXCR4 ; CXCR6(TYMSTR/STRL33/Bonzo CYB5; CYC1; CYSLTR1; DAB2IP; DES; DKFZp451J0118; DNCL1; DPP4; E2F1 ECGF1 , EDG1 , EFNA1 , EFNA3 , EFNB2 , EGF , EGFR , ELAC2 , ENG , ENOl , EN02 , EN03 , EPHB4 , EPO , ERBB2 ( Her-2 ) , EREG , ERK8 , ESR1 , ESR2 , F3 (TF ) , FADD FasL , FASN , FCER1A , FCER2 , FCGR3A , FGF , FGF1 ( aFGF ) , FGF10 , FGF11 , FGF12 , 160877.doc -33 - 201247704 FGF12B , FGF13 , FGF14 , FGF16 , FGF17 , FGF18 , FGF19 , FGF2 ( bFGF ) ; FGF21; FGF21; FGF23; FGF3(int-2); FGF4(HST); FGF5; FGF6(HST-2); FGF7(KGF); FGF8; FGF9; FGFR3; FIGF (VEGFD); FIL1 (EPSILON); FIL1 (ZETA); FLJ12584 FLJ25530; FLRT1 (fibrous binding protein); FLT1; FOS FOSL1 (FRA-1); FY (DARC); GABRP (GABAa); GAGEB1 GAGEC1; GALNAC4S-6ST; GATA3; GDF5; GFI1 GGT1; GM-CSF; GNAS1; GNRH1; GPR2 (CCR10) GPR31; GPR44; GPR81 (FKSG80); GRCCIO(CIO) GRP; GSN (plycosin); GSTP1; HAVCR2; HDAC4 HDAC5; HDAC7A; HDAC9; HGF; HIF1A HIP1 ; amide and histamine receptors ; HLA - A , HLA - DRA , HM74 HMOX1 , HUMCYT2A , ICEBERG , ICOSL , ID2 , IFN - a , IFNA1 , IFNA2 , IFNA4 , IFNA5 , IFNA6 , IFNA7 , IFNB1 , IFNy ; IGW1 , IGF1 , IGF1R , IGF2 , IGFBP2 , IGFBP3 , IGFBP6 , IL - 1 , IL10 , IL10RA , IL10RB , IL11 , IL11B , IL-12 , IL12B , IL12B , IL12RB1 , IL12RB2 , IL13 , IL13RA1 , IL13RA2 , IL14 ; IL15 , IL15 , IL16 , IL17 , IL17B , IL17C , IL17R , IL18 , IL18BP , IL18R1 , IL18RAP , IL19 , ILIA , IL1B , IL1F10 , IL1F5 , IL1F6 , IL1F7 , IL1F8 , IL1F9 , IL1HY1 , IL1R1 , IL1R2 , IL1RAP , IL1R1 , IL1R2 , IL1RAP , IL1 IL1RAPL2; IL1RL1; IL1RL2; IL1RN; 160877.doc -34- 201247704 IL20 , IL20 , IL20RA , IL21R , IL22 , IL22R , IL22RA2 , IL23 , IL24 , IL25 , IL26 , IL27 , IL28 , IL28 , IL29 , IL2 , IL2 , IL2 , IL3 , IL3 , IL3 , IL2 , IL3 , IL3 , IL3 IL6; IL6R; IL6ST (glycoprotein 130); IL7; IL7R; IL8; IL8RA; IL8RB, IL8RB, IL9, IL9R, ILK, INHA, INHBA, INSL3, INSL4, IRAKI, IRAK2, ITGA1, ITGA2, ITGA3, ITGA6(a6 Integrin); ITGAV; ITGB3; ITGB4 (b4 integrin); JAG1; JAK1; JAK3; JUN; K6HF; KAI1; KDR; KITLG; KLF5 (GC box BP); KLF6; KLK10; KLK12; KLK13; KLK14; KLK15; KLK3; KLK4; KLK5; KLK6; KLK9; KRT1; KRT19 (keratin 19); KRT2A; KRTHB6 (hair-specific type II keratin); LAMA5; LEP (alkaline); Lingo-p75; Lingo-Troy; LPS LTA (TNF-b); LTB; LTB4R (GPR16); LTB4R2; LTBR; MACMARCKS; MAG or Omgp; MAP2K7 (c-Jun); MDK; MIB1; midkine; MIF; MIP-2; MKI67 (Ki-67) MMP2 ; MMP9 ; MS4A1 ; MSMB MT3 (metallothionein-III); MTSS1; MUC1 (mucin); MYC; MYD88; NCK2; neuroglycan; NFKB1; NFKB2; NGFB (NGF); NGFR; NgR-Lingo; NgR-Nogo66 (Nogo) NgR-p75; NgR-Troy; NME1 (NM23A); NOX5; NPPB; NR0B1; NR0B2; NR1D1; NR1D2; NR1H2; NR1H3; NR1H4; NRII2; NRII3; NR2C1; 160877.doc -35- 201247704 NR2C2; NR2E1; NR2E3 NR2F1 , NR2F2 , NR2F6 , NR3C1 , NR3C2 , NR4A1 , NR4A2 , NR4A3 , NR5A1 , NR5A2 , NR6A1 , NRP1 , NRP2 , NT5E , NTN4 , ODZ1 , OPRD1 , P2RX7 , PAP , PARTI , PATE , PAWR , PCA3 , PCNA , PDGFA PDGFB; PECAM1; PF4 (CXCL4); PGF; PGR; phosphoproteoglycan; PIAS2; PIK3CG; PLAU (uPA); PLG; PLXDC1; PPBP (CXCL7); PPID; PR1; PRKCQ; PRKD1; PRL; PROC; PROK2 PSAP; PSCA; PTAFR; PTEN; PTGS2 (COX-2); PTN; RAC2 (p21Rac2); RARB; RGS1; RGS13; RGS3; RNF110(ZNF144); ROB02; SI00A2; SCGB1D2 (lipophilic B); SCGB2A1 (L-globin 2); SCGB2A2 (|L-globin 1); SCYE1 (endothelial mononuclear cell cytokine); SDF2; SERPINA1; SERPINA3; SERPINB5 SERPINEl ( PAI-l) , SERPINF1 , SHBG , SLA2 , SLC2A2 , SLC33A1 , SLC43A1 , SLIT2 , SPP1 , SPRR1B ( Sprl ) , ST6GAL1 , STAB1 , STAT6 , STEAP , STEAP2 , TB4R2 , TBX21 , TCP10 , TDGF1 , TEK ; TGFA; TGFB1; TGFB111; TGFB2; TGFB3; TGFBI; TGFBR1; TGFBR2; TGFBR3; TH1L; THBS1 (platelet-reactive protein-1); THBS2; THBS4; ΤΗΡΟ; TIE(Tie-l); ΤΙΜΡ3; tissue factor; TLR10; TLR2 TLR3; TLR4; TLR5; TLR6; TLR7; TLR8; TLR9; TNF; TNF-a; TNFAIP2 (B94); TNFAIP3; TNFRSF11A; TNFRSF1A; 160877.doc-36-201247704 TNFRSF1B; TNFRSF21; TNFRSF5; TNFRSF6 (Fas); TNFRSF7; TNFRSF8; TNFSF10 (TRAIL); TNFSF11 (TRANCE); TNFSF12 (AP03L); TNFSF13 (April); TNFSF13B; TNFSF14 (HVEM-L); TNFSF 15 (VEGI); TNFSF18; TNFSF4 (OX40 ligand); TNFSF5 (CD40 ligand); TNFSF6 (FasL); TNFSF7 (CD27 ligand); TNFSF8 (CD30 ligand); TNFSF9 (4-1BB ligand); TOLLIP Toll-like receptor; TOP2A ( topoisomerase Iia ) , TP53 , TPM1 , TPM2 , TRADD , TRAF1 , TRAF2 , TRAF3 , TRAF4 , TRAF6 , TRAF6 , TREM1 , TREM2 , TRPC6 , TSLP , TWEAK , VEGF , VEGFB ; VEGFC; versican; VHL C5; VLA-4; XCL1 (lymphocyte chemotactic factor); XCL2 (SCM-lb); XCR1 (GPR5/CCXCR1); YY1; and ZFPM2. The binding protein of any one of claims 1 to 139, wherein the binding protein is capable of binding two targets, wherein the two target lines are selected from the group consisting of: c-Met and CD-28; c- Met and CD-3 ; c-Met and CD-19 ; CD-28 and CD-3 ; CD-28 and CD-19 ; CD-3 and CD-19 ; CD138 and CD20 ; CD138 and CD40 ; CD20 and CD3 ; CD38 and CD138; CD38 and CD20; CD38 and CD40; CD40 and CD20; CD19 and CD20; CD-8 and IL-6; PDL-1 and CTLA-4; CTLA-4 and BTN02; CSPGs and RGM A; IGF1 and IGF2 IGF1/2 and Erb2B; IL-12 and IL-18; IL-12 and TWEAK; IL-13 and ADAM8; IL-13 and CL25; IL-13 and IL-Ιβ; IL-13 and IL-25; .doc -37- 201247704 IL-13 and IL-4; IL-13 and IL-5; IL-13 and IL-9; IL-13 and LHR agonists; IL-13 and MDC; IL-13 and MIF IL-13 and PED2; IL_13 and SPRR2a; il-13 and SPRR2b; IL-13 and TARC; IL-13 and TGF-β; IL-1-α and IL-Ιβ.; MAG and RGM A; NgR and RGM A; NogoA and RGM A; OMGp and RGM A; RGM A and RGM B; Te38 and TNF-α; TNF-α and IL-12; TNF-α and IL-12p40; TNF-α And IL-13; TNF-α and IL-15; TNF-α and IL·17; TNF-α and IL-18; TNF-α and IL-Ιβ; TNF-α and IL-23; TNF-α and MIF TNF-α and PEG2; TNF-α and PGE4; TNF-α and VEGF; and VEGFR and EGFR; TNF-α and RANK ligand; TNF-α and Blys; TNF-α and GP130; TNF-α and CD -22; and TNFa and CTLA-4. 141. The binding protein of claim 140, wherein the binding protein is capable of modulating the biological function of one or more targets. 142. The binding protein of claim 140, wherein the binding protein is capable of neutralizing one or more targets. 143. The binding protein of any one of claims 139 and 141 to 142, wherein one or more are 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. 144. The binding protein of claim 109, wherein the cytokine is selected from the group consisting of lymphokines, mononuclear hormones, and polypeptide hormones. 145. The binding protein of claim 144, wherein the cytokines are IL-la and IL-Ιβ » 146. The binding protein of claim 144, wherein the cytokines are TNF-a and IL-13 » 160877. Doc • 38 · 201247704 147. The binding protein of claim 144, wherein the cytokines are IL-12 and IL-18. 148. The binding protein of claim 144, wherein the chemokine is selected from the group consisting of CCR2, CCR5, and CXCL-13. 149. The binding protein of claim 109, wherein the cell surface protein is an integrin. 150. The binding protein of claim 149, wherein the cell surface protein is selected from the group consisting of CD-20 and CD3. Φ 151. The binding protein of claim 109, wherein the enzyme is selected from the group consisting of a kinase and a protease. 152. The binding protein of claim 109, wherein the recipient system is selected from the group consisting of a lymphokine receptor, a mononuclear hormone receptor, and a polypeptide hormone receptor. 153. The binding protein of any one of claims 1 to 152, wherein the linker is selected from the group consisting of: ASTKGPSVFPLAP (SEQ ID NO: 46); ASTKGP (SEQ ID NO: 48); TVAAPSVFIFPP (SEQ • ID NO: 50); TVAAP (SEQ ID NO: 52); AKTTPKLEEG EFSEAR (SEQ ID NO: 94); AKTTPKLEEGEFSEARV (SEQ ID NO: 95); AKTTPKLGG (SEQ ID NO: 96); SAKTTPKLGG (SEQ ID NO: 97) SAKTTP (SEQ ID NO: 98); RADAAP (SEQ ID NO: 99); RADAAPTVS (SEQ ID NO: 100); RADAAAAGGPGS (SEQ ID NO: 101); RADAAAA(G4S)4 (SEQ ID NO: 102) SAKTTPKLEEG EFSEARV (SEQ ID NO: 103); ADAAP (SEQ ID NO: 160877.doc -39- 201247704 104); ADAAPTVSIFPP (SEQ ID NO: 105); QPKAAP (SEQ ID NO: 106); QPKAAPSVTLFPP (SEQ ID NO: SEQ ID NO: : AKTTPP (SEQ ID NO: 108); AKTTPPSVTPLAP (SEQ ID NO: 109); AKTTAP (SEQ ID NO: 110); AKTTAPSVYPLAP (SEQ ID NO: 111); GGGGSGGGGS GGGGS (SEQ ID NO: 112); GENKVEYAPALMALS (SEQ ID NO: 113); GPAKELTPLKEAKVS (SEQ ID NO: 114); GHEAAAVMQVQYPAS (SEQ ID NO: 115); TVAAPSVFIFPPTV AAPSVFIFPP (SEQ ID NO: 116); and ASTKGPSVFPLAPASTKGPSVFPLAP (SEQ ID NO: 117). 154. The binding protein of any one of claims 1 to 153, wherein the binding protein has a association rate constant selected from the group consisting of the one or more targets as measured by surface plasmon resonance ( Km): at least about 1〇2 Μ·,·1; at least about ΙΟ3 Μ···1; at least about Μ4 Μ,·1; at least about Μ5 Μ,·1; and at least about Μ6 Μ·, _1 » 155. The binding protein of any one of claims 1 to 154, wherein the binding protein has a dissociation rate constant (Koff) selected from the group consisting of: one or more targets, as measured by surface plasmon resonance: At most about ]3]^,·1; at most about; at most about Η^Μ, -1; and at most about. 156. The binding protein of any one of claims 1 to 155, wherein the binding protein has a dissociation constant (KD) selected from the group consisting of: at most about ΙΟ·6 对该 for the one or more targets; About 1〇_7 μ; at most about ΙΟ-8 Μ; at most about ΙΟ·9 Μ; at most about ΙΟ·10 Μ; at most about ΐ〇&&;;Μ; and at most about ΙΟ·12 Μ. 157. The conjugate of the protein of claim 1, wherein the binding protein conjugate further comprises an agent selected from the group consisting of: immunoadhesion Molecules, developers, therapeutic agents and cytotoxic agents. 158. The binding protein conjugate of claim 157, wherein the agent is a developer selected from the group consisting of: a radioactive label, an enzyme, a fluorescent label 'luminescent label, a bioluminescent label, a magnetic label, and biotin. 159. The binding protein conjugate of claim 57, wherein the developer is selected from the group consisting of The radioactive label of the group consisting of: "TC, UlIn, 丨25, 131l, 丨, 丨66 3H, Ho and 丨4C, 35S, 90γ 153Sm. 160. A binding protein conjugate according to 157, wherein the agent is a therapeutic or cytotoxic agent selected from the group consisting of an antimetabolite, an alkylating agent antibiotic, a growth factor, a cytokine, an anti-angiogenic agent , anti-mitosis, j:: i-cycline (anthracycHne), toxic t and cells> weekly death agent. The binding protein according to any one of the items (1), wherein the binding protein is a crystal binding protein. 162. The binding protein of claim 161 of May wherein the crystallized binding protein crystal is a drug-free controlled release crystal. 163. The binding protein of Item 161, wherein the crystal binding protein has a ratio of μ, ·'. The in vivo half-life of the soluble counterpart of the D protein. 164. The protein of claim 161, wherein the crystal binds to egg activity. 165. A binding protein according to any one of (1), wherein the protein is 160877.doc • 41 · 201247704 Manufactured according to a method comprising culturing a host cell in a medium under conditions sufficient to produce the binding protein The cell comprises a vector; the vector comprises a nucleic acid encoding the binding protein. S 166. A pharmaceutical composition comprising the gentleman human protein of any of the preceding claims and a pharmaceutically acceptable carrier. 167. The pharmaceutical composition of claim 166, further comprising at least one of the other agents. The pharmaceutical composition of claim 167, wherein the other agent is selected from the group consisting of: a therapeutic agent; a developer; a cytotoxic agent; an angiogenesis inhibitor; a kinase inhibitor; a costimulatory molecule blocker; Adhesion molecule blocker; anti-cytokine antibody or functional fragment thereof; methotrexate; cyclosporin (rapamycm); FK506; detectable marker or reporter Body; tnf antagonist; antirheumatic drug; muscle relaxant; anesthetic; non-steroidal anti-inflammatory drug (NSAID); analgesic; anesthetic; sedative; local anesthetic; neuromuscular blocker; antimicrobial; antipsoriatic; Corticosteroids; anabolic steroids; erythropoietin; immunization; immunoglobulins; immunosuppressants; growth hormone; hormone replacement drugs; radiopharmaceuticals; 3H; 14c; 35s; 9°Y; 99Tc; ηιιη; 125][ ; 177Lu, 166H〇; 153Sm; fluorescent label; luminescent label; bioluminescent label; magnetic label; biotin; antidepressant; antipsychotic; stimulant; Agonist; inhaled steroids; epinephrine or analog; cytokines and cytokine antagonists. 169. A pharmaceutical composition comprising a binding protein according to claim 157 in combination with 160877.doc • 42-201247704 and a pharmaceutically acceptable carrier. 170. A nucleic acid, which encodes a polypeptide according to any one of the claims 171. 171. A representation construct comprising a nucleic acid according to claim 17 172. A cell comprising the request Item 171. The use of a binding protein according to any one of claims 165, which is for the preparation of a drug for the treatment of a disease or condition selected from the group consisting of: arthritis , osteoarthritis, adolescent chronic arthritis, septic arthritis, Lyme arthritis, psoriatic arthritis, reactive arthritis, spondyloarthropathy, systemic lupus erythematosus, Crohn's disease ( Crohn's disease), ulcerative colitis, inflammatory bowel disease, insulin-dependent diabetes mellitus, verrucous, asthma, allergic disease, psoriasis, inflammatory scleroderma, graft versus host disease, organ transplantation, Related to organ transplantation "Acute or chronic immune disease, sarcoidosis, atherosclerosis, disseminated intravascular coagulation, Kawasaki, s disease, disease", renal syndrome Chronic fatigue syndrome, Wegener's granulomatosis, Henoch-Schoenlein purpurea, renal microscopic vasculitis, chronic active hepatitis, uveitis, septic shock, Toxic shock syndrome, sepsis syndrome, cachexia, infectious disease, parasitic disease, acquired immunodeficiency syndrome, acute transverse myelitis, Huntington's chorea, Parkins〇n, s disease Alzheimer's disease, Alzheimer's disease, primary biliary cirrhosis of the stroke, hemolytic anemia, malignant disease, heart failure 160877.doc -43- 201247704 Exhaustion, myocardial infarction, Addison's disease Addison's disease), sporadic polyglandular deficiency type I and sputum polygland secretion deficiency, Schmidt's syndrome, adult (acute) respiratory distress syndrome, hair loss , plaque alopecia, seronegative joint disease, joint disease, Reiter's disease, psoriasis Arthropathy, ulcerative colitis, arthritis, intestinal synovitis, joint disease associated with chlamydia, yersinia, and salmonella, spinal cord disease, atheroma Disease/arteriosclerosis, atopic hypersensitivity, autoimmune bullous disease, pemphigus vulgaris, phyllodes pemphigus, pemphigoid, linear IgA disease, autoimmune hemolytic anemia, Comm's positive hemolytic Anemia (Coombs positive haemolytic anaemia), Acquired Pernicious Anemia, Adolescent Pernicious Anemia, Myalgesic Encephalitis/R〇yal Free Disease, Chronic Skin Mucosal Candidiasis, Giant Cell Arteritis Primary Severe Hepatitis Unexplained autoimmune hepatitis, acquired immunodeficiency syndrome, acquired immunodeficiency-related diseases, hepatitis B, hepatitis C, general variant immunodeficiency (common variant hypoxemia globulinemia), dilated myocardium Disease, female infertility, ovarian failure, (four) premature aging, fibrotic lung disease, unexplained fibrotic alveolitis, post-inflammatory interstitial lung disease, interstitial lung Inflammation, connective tissue disease-related interstitial lung disease, mixed connective tissue disease-related lung disease, systemic sclerosis-related interstitial lung disease, rheumatoid arthritis-related diseases, systemic neonatal lupus erythematosus-related lung disease, Dermatomy/polymyositis-associated lung disease, Hugh's disease-associated lung disease (Sj6grenis 160877.doc 201247704 associated lung disease) 'Asthmatic spondylitis-associated lung disease, vasculitic diffuse lung disease, hemosiderin deposition (haem〇sider〇sis^g Guanzhi's lung disease, drug-induced interstitial lung disease, fibrosis, radiofibrosis, obstructive bronchiolitis, chronic eosinophilic pneumonia' lymphocytic infiltrating lung disease, post-infection interstitial Pulmonary disease, gouty arthritis, autoimmune hepatitis, type 1 autoimmune hepatitis (typical autoimmune or lupus-like hepatitis), type 2 autoimmune hepatitis (anti-LKM antibody hepatitis), autoimmune Mediated hypoglycemia, type B tamsin resistance with black • acanthosis, parathyroid gland dysfunction, acute immune disease associated with organ transplantation, and organ transplantation Related chronic immune diseases, osteoarthritis, primary sclerosing cholangitis, type 1 psoriasis, type 2 psoriasis, idiopathic leukopenia, autoimmune neutropenia, NOS type nephropathy, Mitochondrial nephritis, renal microscopic vasculitis, Lyme disease, discoid lupus erythematosus, idiopathic or N〇s male infertility, sperm autoimmune, multiple sclerosis (all subtypes) ), sympathetic ophthalmia, pulmonary hypertension secondary to connective tissue disease, GoodPasture's syndrome, pulmonary manifestations of nodular polyarteritis, acute rheumatic fever, rheumatoid spondylitis, Stirling Stm, s disease, systemic sclerosis, Hugh's syndrome, Takayasu's disease/arteritis, autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmune Sickle adenosis, hyperthyroidism, verrucous gland autoimmune verrucous hypoglycemia (Hashimoto's disease), atrophic autoimmune thyroid dysfunction, primary mucinous edema , Salivary uveitis, original 160877.doc -45· 201247704 vasculitis, leukoplakia acute liver disease, chronic liver disease, alcoholic cirrhosis, alcohol-induced liver injury, biliary stagnation, characteristic liver disease, drug induction Hepatitis, nonalcoholic steatosis hepatitis, allergies and asthma, B group of key bacteria (GBS) infections, mental disorders (such as depression and schizophrenia), Th2 and Th1 type mediated diseases, acute and chronic Pain (different forms of pain), and cancers such as lung cancer, breast cancer, stomach cancer, bladder cancer, colon cancer, pancreatic cancer, ovarian cancer, prostate cancer, and rectal cancer, and hematopoietic malignancies (leukemia and lymphoma), no beta Lipoproteinemia, sweetness of the hands and feet, acute and chronic parasitic or infection processes, acute leukemia, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute or chronic bacterial infection, acute pancreatitis, acute renal failure , adenocarcinoma, atrial ectopic beat, AIDS dementia complex, alcohol-induced hepatitis, allergic conjunctivitis, allergic contact dermatitis, allergies Rhinitis, allograft rejection, α_1_antitrypsin deficiency, amyotrophic lateral sclerosis, anemia, angina pectoris, anterior horn cell degeneration, anti-ccj3 therapy, antiphospholipid syndrome, anti-receptor allergic reaction, aorta and peripheral Aneurysm, aortic dissection, arterial hypertension, atherosclerosis, arteriovenous fistula, ataxia, atrial fibrillation (sustained or paroxysmal), atrial flutter, atrioventricular block, B-cell lymph Tumor, bone graft rejection, bone marrow transplantation (BMT) rejection, bundle branch block, Burkitt's lymphoma, burns, arrhythmia, cardiac stun syndrome, cardiac tumor, cardiomyopathy, cardiopulmonary bypass Inflammatory response, cartilage transplant rejection, cerebellar cortical degeneration, cerebellar disorders, turbulent or multifocal atrial tachycardia, chemotherapy-related disorders, 160877.doc -46· 201247704 Chronic myeloid leukemia (CML), chronic alcohol Poisoning, chronic inflammatory disease, chronic lymphocytic leukemia (CLL), chronic obstructive pulmonary disease (COPD), chronic poplar Acidosis, colorectal cancer, congestive heart failure, conjunctivitis, contact dermatitis, pulmonary heart disease, coronary artery disease, Creutzfeldt-Jakob disease, culture-negative sepsis, cystic fibrosis, cells Hormone therapy-related disorders, boxer dementia, demyelinating disease, dengue hemorrhagic fever, dermatitis, dermatological conditions, diabetes (diabete/diabetes mellitus), urinary arteriosclerosis, diffuse Diffuse Lewy body disease, dilated cardiomyopathy, basal ganglia disease, D〇wn, s Syndrome in middle age, induced by drugs that block CNS dopamine receptors Drug-induced dyskinesia, drug sensitivity, eczema, encephalomyelitis, endocarditis, endocrine disease, epiglottis, epstein-barr virus infection, acromegaly, extrapyramidal and cerebellar disorders , familial hemophagocytic histiocytosis, fetal thymic transplant rejection, Friedreich's ataxia (Friedreich's at Axia), functional peripheral arterial disease, fungal sepsis, gas gangrene, gastric ulcer, glomerulonephritis, graft rejection of any organ or tissue, gram negative sepsis, Gram-positive sepsis , granuloma caused by intracellular organisms, hairy cell leukemia, Haiierror (jen-Spatz disease), hashimoto's thyroiditis, hay fever, heart transplant rejection, Hemochromatosis, hemodialysis, 160877.doc •47· 201247704 Hemolytic uremic syndrome/thrombotic thrombocytopenic purpura, hemorrhage, hepatitis (A), Hisbundie arrythmias, HIV infection/HIV neuropathy , H〇dgkin, s disease, hyperactive motor disorder, allergic reaction, hypersensitivity pneumonitis, hypertension, hypokinesia, hypothalamic-pituitary-adrenal axis assessment, idiopathic Eddie Sjogren's disease, idiopathic pulmonary fibrosis, antibody-mediated cytotoxicity, debilitation, infant spinal muscular atrophy, aortic inflammation , influenza A, exposure to ionizing radiation, iridocyclitis / uveitis / optic neuritis, ischemia-reperfusion injury, ischemic stroke, juvenile rheumatoid arthritis, adolescent spinal muscular atrophy, Kaposi Sarcoma (Kap〇si, s sarcoma), renal transplant rejection, Legionnaires disease 〇eg〇neUa), leishmaniasis, leprosy, cortical vertebral system lesions, edema, liver transplant rejection Reaction, lymphedema, dysentery, malignant lymphoma, malignant histiocytosis, malignant melanoma, meningitis, meningococcalemia, metabolic/idiopathic disease, migraine, mitochondrial multisystemic disease, Mixed connective tissue disease, gamma globulin disease, multiple myeloma, multi-system degeneration (Mencel's Dejerine-Thomas Shi-Drager and Machado-Joseph ))' Myasthenia gravis, mycobacterium avium intracellulare, mycobacterium tuberculosis, myelodysplastic syndrome, myocardial infarction , myocardial ischemic disease, nasopharyngeal carcinoma, neonatal chronic lung disease, nephritis, nephropathy, neurodegenerative disease, type I neuromuscular atrophy, neutrophilic fever, non-Hodgkin's lymphoma (n〇n_ 160877 .doc •48- 201247704 hodgkins lymphoma), abdominal aorta and its branch blockage, obstructive arterial disease, okt3 therapy, testicular inflammation / epididymitis, testicular inflammation / vas deferens, organ enlargement, osteoporosis, pancreas Transplant rejection, pancreatic cancer, paraneoplastic syndrome/malignant hypercalcemia, paratypic gland transplant rejection, pelvic inflammatory disease, perennial rhinitis, pericardial disease, peripheral atherosclerotic disease, peripheral vascular disease, Abdominal inflammation, pernicious anemia, pneumocystis carinii pneumonia, pneumonia, POEMS syndrome (polyneuropathy, organ enlargement, internal division, disease, gamma globulin disease and skin variability syndrome), Post-perfusion syndrome, after-effect syndrome, post-surgical syndrome, pre-eclampsia, progressive supranuclear palsy, primary pulmonary hypertension, radioactivity Law, Raynaud's phenomenon and disease 'Rayn〇ud, s disease, Refsum's disease, regular stenosis QRS tachycardia, renal vascular hypertension, reperfusion injury , restricted cardiomyopathy, sarcoma, scleroderma, senile chorea, Senile Dementia of Lewy body type 'Serum-negative; disease, shock, sickle cell anemia, skin allograft Rejection, skin change syndrome, small bowel transplant rejection, solid tumor, specific arrhythmia, spinal ataxia, spinocerebellar degeneration, streptococcal myositis, cerebellar structural lesions, subacute sclerosing panencephalitis, fainting, heart Vascular system syphilis, systemic allergy, systemic inflammatory response syndrome, systemic adolescent rheumatoid arthritis, T-cell or FAB ALL, telangiectasia, thromboembolic vasculitis, thrombocytopenia, poisoning, transplantation, trauma/bleeding , sputum allergic reaction, 160877.doc •49· 201247704 ιν type allergy, unstable angina 'Urine, urinary septicemia, urticaria, cardiac disease, varicose veins, vasculitis, venous disease, venous thrombosis, ventricular fibrillation, viral and fungal infections, viral encephalitis/aseptic meningitis, Xenograft rejection of virus-associated hemophagocytic syndrome, Wemicke-Korsakoff syndrQme:, Wilson's disease, or any organ or tissue. 160877.doc 50·