CN101094869A - Hk1-binding proteins - Google Patents

Abstract

The invention features hKl binding polypeptides as well as compositions comprising such polypeptides and methods of making and using such polypeptides.

Description

HK1 is conjugated protein

The application's case is advocated the U.S. patent application case the 60/598th of application on August 3rd, 2004, the U.S. patent application case the 60/615th of No. 506 and on October 4th, 2004 application, No. 721 right of priority, therefore the content of described application case is incorporated herein by reference.

Technical field

Do not have

Background technology

The human tissue kallidinogenase comprises hK1, a kind of protein by the KLK1 genes encoding, and be sometimes referred to as pancreas/kidney kallidinogenase, hPRK or GenBank  M25629 or M33105.Usually referring to people such as Yousef, (2001) Endocrine Reviews 22:184-204.

Summary of the invention

The present invention especially provides hK1 conjugated protein." hK1 is conjugated protein " refer to can with hK1 or its fragment, especially its protease activity fragment interacting proteins.HK1 is conjugated protein to be comprised with high-affinity and hK1 bonded antibody and the antibody that can suppress the hK1 enzymic activity.Can with the conjugated protein throwing of HK1 with to person under inspection's (for example) with the treatment illness, described illness such as asthma (for example, allergic asthma and non-allergic asthma), chronic obstructive pulmonary disease (COPD), multiple sclerosis, psoriasis, rheumatoid arthritis, osteoarthritis, rhinitis, sinusitis, inflammatory bowel (such as Crohn disease (Crohn ' s disease) and ulcerative colitis), immune-mediated diabetes, acute pancreatitis, interstitial cystitis and superfluous natural disposition illness (for example, transitivity carcinoma of the pancreas or tumor-blood-vessel growth) or other hK1 associated conditions.

On the one hand, the invention is characterized in the protein that combines and comprise heavy chain immunoglobulin variable domain sequence and light chain immunoglobulin variable territory sequence with hK1.Described protein can be incorporated into or the space on the avtive spot of interlock hK1.In one embodiment, described protein contacts one or more hK1 avtive spot residues 65,120 or 214, or the residue in 5 amino acid of described residue.In one embodiment, described protein suppresses the hK1 enzymic activity.For example, described protein can suppress hK1, wherein IC ₅₀Less than 200nM, 80nM, 50nM, 40nM, 30nM, 20nM, 10nM, 5nM, 4nM, 3nM, 2.5nM or 1nM, for example IC ₅₀Between 1nM and the 20nM with at other in scope wherein.Described protein can be incorporated into hK1, wherein K _dLess than 10 ^-7M, 10 ^-8M, 5 * 10 ^-9M, 10 ^-9M, 10 ^-10M, 10 ^-11M or 10 ^-12M, for example Kd is 10 ^-8M and 10 ^-11Between the M with at other in scope wherein.

In one embodiment, described protein stops hK1 and interacts greater than 12 amino acid whose protein substrates, described protein substrate for example, high molecular weight kininogen, low molecular weight kininogen, Urogastron, proinsulin, low-density lipoprotein, renninogen, vasoactive intestinal peptide, procollagenase and/or proangiotensin.In another embodiment, described protein stops hK1 and interacts less than 12 amino acid whose peptide substrates.

In one embodiment, described protein can not be in conjunction with by protease inhibitor or such as the hK1 molecule of other protein protease inhibitor deactivation of alpha1-antitrypsin or kallidinogenase conjugated protein (kallistatin).

In one embodiment, described protein does not cause human intravital immunogenic response.For example, described protein is humanized antibody, human antibodies or effective human antibodies.In one embodiment, described protein comprises one or more human CDR, for example at least 3,4,5 or 6 human CDR.In one embodiment, described protein comprises one or more CDR as herein described, for example the CDR of one or more SEQ ID NO:1-1020 and 1380-1385.In one embodiment, described protein comprises one or more, for example 3,4,5,6 SEQ ID NO:7,8,9,10,11 and 12 CDR.In another embodiment, described protein comprises one or more, for example 3,4,5 or 6 SEQ ID NO:1380,1381,1382,1383,1384 and 1385 CDR.In another embodiment, described protein comprises one or more, for example 3,4,5,6 SEQ ID NO:109,110,111,112,113 and 114 CDR.In another embodiment, described protein comprises one or more, for example 3,4,5,6 SEQ ID NO:151,152,153,154,155 and 156 CDR.In one embodiment, described protein comprises one or more human framework regions or human substantially framework region, for example comprises at least 3,4,5 or 6 human framework regions.In one embodiment, described protein comprises one or more framework region and/or one or more framework regions from the variable light chain of the aminoacid sequence with SEQ ID NO:1022-1198 and 1376 from the variable heavy chain with SEQ ID NO:1199-1369 and 1377 aminoacid sequences.In one embodiment, described protein comprises one or more framework region and/or one or more framework regions from the variable sequence of light chain of SEQ ID NO:1070 from SEQ ID NO:1245 variable heavy chain sequence.In another embodiment, described protein comprises one or more framework region and/or one or more framework regions from the variable sequence of light chain of SEQ ID NO:1029 from the variable heavy chain sequence of SEQ ID NO:1206.In another embodiment, described protein comprises one or more framework region and/or one or more framework regions from the variable sequence of light chain of SEQ ID NO:1183 from the variable heavy chain sequence of SEQ ID NO:1354.

HC CDR1 can comprise the aminoacid sequence with at least 5 amino acid lengths, wherein the HC CDR1 sequence of at least 3,4 or 5 amino acid and antibody as herein described (for example, SEQ ID NO:10,112,154 or 1383 HC CDR1) unanimity.

HC CDR2 can comprise the aminoacid sequence with at least 15,16 or 17 amino acid lengths, the HC CDR2 sequence of at least 10,12,14,15,16 or 17 amino acid and antibody as herein described (for example, SEQ ID NO:11,113,155 or 1384 HC CDR2) unanimity wherein.HC CDR2 can comprise the aminoacid sequence with at least 17 amino acid lengths, wherein the HC CDR2 sequence of at least 14,15,16 or 17 amino acid and antibody as herein described (for example, SEQ ID NO:11,113,155 or 1384 HC CDR2) unanimity.

HC CDR3 can comprise the aminoacid sequence with at least 7 or 8 amino acid lengths, wherein the HC CDR3 sequence of at least 5,6,7 or 8 amino acid and antibody as herein described (for example, SEQ ID NO:12,114,156 or 1385 HC CDR3) unanimity.

LC CDR1, CDR2 and/or CDR3 can comprise some aminoacid sequence, and it is different that consistent or per 10 amino acid lengths of the corresponding LC CDR sequence of described aminoacid sequence and antibody as herein described (for example SEQ ID NO:7,109,151 or 1380 LC CDR1, SEQID NO:8,110,152 or 1381 LC CDR2, SEQ ID NO:9,111,153 or 1382 LC CDR3) are less than two amino acid.

In one embodiment, H1 has the canonical structure identical with antibody as herein described with H2 hypermutation ring.In one embodiment, L1 has the canonical structure identical with antibody as herein described with L2 hypermutation ring.

In one embodiment, the aminoacid sequence of the aminoacid sequence of HC variable domain sequence and the HC variable domain of antibody as herein described (for example, SEQ ID NO:1206,1245 or 1354 HC variable domain) at least 70%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or 100% unanimity.In one embodiment, the aminoacid sequence of the aminoacid sequence of LC variable domain sequence and the LC variable domain of antibody as herein described (for example, SEQ ID NO:1029,1070 or 1183 LC variable domain) at least 70%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or 100% unanimity.For example, the aminoacid sequence of HC and LC variable domain sequence is consistent with the HC of antibody as herein described and the aminoacid sequence of LC variable domain (for example light chain variable territory of the HC variable domain of the light chain variable territory of the HC variable domain of the light chain variable territory of the HC variable domain of SEQ ID NO:1245 and SEQ ID NO:1070, SEQ ID NO:1206 and SEQ ID NO:1029, SEQ ID NO:1354 and SEQ ID NO:1183) at least 70%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or 100%.

The aminoacid sequence of HC and LC variable domain sequence can be by a sequence encoding, described sequence under the height stringent condition with nucleotide sequence as herein described or the coding variable domain nucleic acid array hybridizing or with the coding aminoacid sequence as herein described (for example aminoacid sequence of SEQ ID NO:1245 and/or SEQ ID NO:1070, SEQ ID NO:1206 and/or SEQ ID NO:1029, SEQ ID NO:1354 and/or SEQ ID NO:1183) nucleic acid hybridization.In one embodiment, the aminoacid sequence of one or more framework regions of HC and/or LC variable domain (for example, FR1, FR2, FR3 and/or FR4) is consistent with the HC of antibody as herein described and the corresponding framework region of LC variable domain (for example aminoacid sequence of SEQ ID NO:1245 and/or SEQ ID NO:1070, SEQ ID NO:1206 and/or SEQ ID NO:1029, SEQ ID NO:1354 and/or SEQ ID NO:1183) at least 70%, 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or 100%.In one embodiment, one or more heavy chain framework regions (for example, HC FR1, FR2 and FR3) with from the human reproduction be the corresponding framework region of antibody sequence (for example, VHIII reproductive tract antibody or with H1 and H2 hypermutation ring in the reproductive tract antibody sequence of 1-3 canonical structure compatibility) at least 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 100% consistent.

In one embodiment, one or more light chain framework regions and human reproduction are the sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 100% consistent of the corresponding framework region of antibody.

In one embodiment, the heavy chain variable domain sequence forms the variable domain of the 1-3 Chothia canonical structure with H1 and H2 hypermutation ring.

HC CDR3 can comprise R-(RV)-G-X-(WY)-(YG)-(AGS)-(FM)-D-(YIV)-W.HC CDR3 can comprise Y-(YP)-Y-(YG)-(AG)-(MF)-D-(VI).LC CDR1 can comprise (RG)-A-S-(QS)-S-(IV)-(SG)-(SG)-Y-(LY)-(NA).LC CDR1 can comprise (RGV)-A-S-(QS)-S-(IV)-(SG)-(ST)-(YN)-L-(NA).LC CDR1 can comprise R-A-S-Q-X-I-(SG)-(SLG)-X-(LY).LC CDR2 can comprise I-Y-(AG)-(AV)-S-(NS)-(RL)-(PA)-S-G-(VI).LC CDR2 can comprise I-Y-(AG)-(AV)-S-S-(RL)-(PAQ)-(ST)-G-(VI).LC CDR3 can comprise Q-Q-(YS)-(TANGY)-S-(SPT)-(PS)-X-T-F.LC CDR3 can comprise (CA)-Q-(QW)-(YD)-D-S-L-P-X-T-F or (CA)-Q-(QW)-(YD)-D-S-L-P-G-T-F.Be provided in amino acid indication the substituting in the parenthesis in giving position.

In one embodiment, described protein bound is by all or part of the epitope of the antibodies of for example DX-2300 as herein described, M093-F09, M137-E01 and M0097-B12 antibody.Described protein can suppress (for example competitive inhibition) for example DX-2300 as herein described, M093-F09, M137-E01 and the antibody of M0097-B12 antibody and combining of hK1.Protein can be in conjunction with for example epitope of configuration epitope or linear epitope, described epitope when in conjunction with the time stop for example DX-2300 as herein described, M093-F09, M137-E01 and the antibody of M0097-B12 antibody and combining of hK1.But on the described epitope proximity space or (for example overlapping or in abutting connection with) epitope that is associated on the function with linear order or configuration spacing near epitope by the antibody recognition of for example DX-2300 as herein described, M093-F09, M137-E01 and M0097-B12 antibody.

Described protein can be full length antibody (for example, IgG (for example, IgG1, IgG2, IgG3, IgG4), IgM, IgA (for example, IgA1, IgA2), IgD and IgE, but be preferably IgG) or can only comprise Fab (for example, Fab, F (ab ') ₂Or scFv fragment, or one or more CDR).Antibody or its Fab can comprise two heavy chains and two light chains or can be single-chain antibody.Antibody optionally can comprise the constant region that is selected from κ, λ, α, γ, δ, ε or μ constant region gene.Preferred antibody comprises substantially weight and the constant region of light chain from human antibodies, for example IgG 1 constant region, its part or concensus sequence.

In one embodiment, the conjugated protein hK1 catalytic activity of hK1 in conjunction with hK1 and inhibition or reduction hK1.In one embodiment, for hK1, the conjugated protein Ki that has less than 50nM, 40nM, 30nM, 20nM, 5nM, 1nM, 500pM, 100pM, 50pM, 30pM of hK1.Conjugated protein comparable another proteolytic enzyme of hK1 is at least 100,200,500 or 1000 times of inhibition hK1 enzymic activitys preferentially.

Feature of the present invention each the nucleic acid in the polypeptide described herein that also is to encode.Described nucleic acid can comprise homology codon or can be through translating to produce any codon group of polypeptide separately.For example, described nucleic acid can comprise one or more codons, described codon for can through translate with common password that produces polypeptide separately and for its cell type of expressing therein for common.In one embodiment, for the cell that encoded polypeptide is expressed therein, described nucleic acid comprises at least 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% or more common password.In one embodiment, the sequence of light chain of the sequence of heavy chain of the sequence of light chain of the sequence of heavy chain of the sequence of light chain of the sequence of heavy chain of described nucleic acid encoding SEQ ID NO:1245 and/or SEQ ID NO:1070, SEQ ID NO:1206 and/or SEQ ID NO:1029, SEQID NO:1354 and/or SEQ ID NO:1183.In one embodiment, the sequence of heavy chain of described nucleic acid encoding SEQ ID NO:1245 and (for example) are with regard to regard to expressing in Chinese hamster ovary (CHO) cell, one or more codons are common password, and for example, described nucleic acid comprises the nucleotide sequence of SEQ ID NO:1379.In one embodiment, the sequence of light chain of described nucleic acid encoding SEQ ID NO:1070 and (for example) are with regard to regard to expressing in Chinese hamster ovary (CHO) cell, one or more codons are common password, and for example, described nucleic acid comprises the nucleotide sequence of SEQ ID NO:1378.In addition, the invention is characterized in a kind of host cell that comprises nucleic acid as herein described.

On the other hand, the invention is characterized in the method for a kind of hK1 of generation binding antibody or its Fab.Described method comprises: provide and containing the host cell of first nucleotide sequence of polypeptide that coding comprises the variable region of heavy chain of variable region of heavy chain for example as described herein; Second nucleotide sequence of the polypeptide of the variable region of light chain that providing encodes comprises variable region of light chain for example as described herein; With under the condition that allows described light and variable region of heavy chain of assembling, in host cell, express described first and second nucleotide sequence to form and the interactional antigen-binding proteins of hK1.Described first nucleotide sequence can link to each other with described second nucleotide sequence or not link to each other, for example respectively at expressing on the identical or different carrier.The component that described first nucleotide sequence and described second nucleotide sequence can be with a part maybe can reside on the differing molecular (for example, coloured differently body or plasmid).

Host cell can be the eukaryotic cell of for example mammalian cell, insect cell, yeast cell or the prokaryotic cell prokaryocyte of intestinal bacteria (E.coli) for example.For example, mammalian cell can be culturing cell or cell strain.Exemplary mammalian cell comprises lymphocyte strain (for example, NSO), CHO, COS cell, ovocyte and from the cells (for example mammary epithelial cell) of transgenic animal.For example, the encode nucleic acid of antibody as herein described can be expressed in the transgenic animal.In one embodiment, antibody is descended and produces in the control that described nucleic acid is placed in tissue-specific promoter's (for example, mammary gland-specific promotor) in transgenic animal.For example, antibody molecule is secreted in the emulsion such as immunocow, pig, horse, sheep, goat or rodentine transgenic animal.

Feature of the present invention also is a kind of method of the hK1 of improvement associated conditions.Described method comprises: the hK1 conjugated protein (for example, as described herein) that will effectively improve the amount of described illness or its at least a symptom throw with to the person under inspection.Described illness can be the inflammatory illness.Described illness can be (for example) COPB, asthma, rheumatoid arthritis, osteoarthritis, multiple sclerosis.Described illness can be rhinitis, sinusitis, inflammatory bowel (such as Crohn disease and ulcerative colitis), immune-mediated diabetes, acute pancreatitis, interstitial cystitis or superfluous natural disposition illness (for example, transitivity carcinoma of the pancreas or tumor-blood-vessel growth).Described protein can comprise further feature as herein described.Described protein comprises the antigen binding site that comprises light chain (LC) variable domain sequence and heavy chain (HC) variable domain sequence usually.Described protein can be IgG or Fab form.Usually, described protein does not have immunogenicity in person under inspection's body, and for example described protein comprises the mankind or effective people's class framework and constant domain, for example, and modified human constant domain.In one embodiment, described protein suppresses hK1.Described method can comprise further feature as herein described.

Feature of the present invention also is the method for a kind of prevention or treatment hK1 associated conditions.Described method comprises: will effectively prevent or the hK1 conjugated protein (for example, as described herein) that treats described illness, postpone its at least a paresthesia epilepsy or improve the amount of its at least a symptom throw with to the person under inspection.For example, described illness is: asthma (for example, allergic asthma and non-allergic asthma), chronic obstructive pulmonary disease (COPD), multiple sclerosis, psoriasis, rheumatoid arthritis, osteoarthritis, rhinitis, sinusitis, inflammatory bowel (such as Crohn disease and ulcerative colitis), immune-mediated diabetes, acute pancreatitis, interstitial cystitis or superfluous natural disposition illness (for example, transitivity carcinoma of the pancreas or tumor-blood-vessel growth).Described protein can comprise further feature as herein described.Described protein comprises the antigen binding site that comprises light chain (LC) variable domain sequence and heavy chain (HC) variable domain sequence usually.Described protein can be IgG or Fab form.Usually, described protein does not have immunogenicity in person under inspection's body, and for example, described protein comprises the mankind or effective people's class framework and constant domain, for example, and modified human constant domain.In one embodiment, described protein suppresses hK1.Described method can comprise further feature as herein described.

Feature of the present invention also is tracheae inflammation or the over-reactive method of tracheae among a kind of adjusting person under inspection.Described method comprises: effectively (i) reduces bronchial tissue's kallidinogenase activity, the hK1 conjugated protein (for example, as described herein) that (ii) reduces tracheae inflammation among the person under inspection and/or (iii) reduce the over-reactive amount of tracheae among the person under inspection throw with to the person under inspection.In one embodiment, described protein be (for example) use metered dose inhaler through suck throw with.In another embodiment, described protein be through subcutaneous injection throw with.

On the other hand, the invention is characterized in the method for a kind of prevention or treatment vasculogenesis associated conditions.Described method comprises: will effectively reduce vasculogenesis and/or for example improve the hK1 conjugated protein (for example, as described herein) of the amount of the vasculogenesis associated conditions of superfluous natural disposition illness throw with to the person under inspection.For example, described illness is for being the superfluous natural disposition illness of feature with the malignant growth.Described protein can by several different methods throw with, for example, for example subcutaneous, intramuscular or intravenous injection.In one embodiment, with the local throwing of described protein and to tumor site.

HK1 conjugated protein (first medicament) can with second medicament combination of the superfluous natural disposition illness of effective treatment throw with.For example, second medicament is regulated the activity of VEGF class somatomedin, and for example, second medicament is regulated the activity of VEGF or vegf receptor.In one embodiment, second medicament is and VEGF bonded antibody.This paper provides other example of second medicament.

As used herein, " combination throw with " mean with two or more medicament simultaneously or in certain intervals, throw with to the person under inspection so that there be each medicament overlapping to patient's effect.Preferably, the throwing of described first medicament and described second medicament is consequently reached combined effect with the interval close enough.Described interval can be a minute interval, hour interval, day interval or weekly interval.Usually, medicament has biological usability simultaneously in person under inspection's body, for example can detect.Described first medicament and described second medicament can be arbitrary order throw with.In a preferred embodiment, with one in the described medicament (for example first medicament) throw with several minutes, 1 hour, 2 hours, 3 hours of another medicament (for example, second medicament) or 4 hours or even one or two day in throw with at least once.

In one embodiment, described first medicament and described second medicament are thrown simultaneously with.For example, described first medicament and described second medicament are allocated jointly.In another embodiment, with described first medicament and described second medicament different time throw with.

Described protein can comprise further feature as herein described.Described protein comprises the antigen binding site that comprises light chain (LC) variable domain sequence and heavy chain (HC) variable domain sequence usually.Described protein can be the form of IgG or Fab.Usually, described protein does not have immunogenicity in person under inspection's body, and for example, described protein comprises the mankind or effective people's class framework and constant domain, for example, and modified human constant domain.In one embodiment, described protein suppresses hK1.Described method can comprise further feature as herein described.

Definition

As used herein, term " antibody " refers to comprise the protein of at least one immunoglobulin variable territory or immunoglobulin variable territory sequence.For example, antibody can comprise a weight (H) chain variable region (this paper is abbreviated as VH) and light (L) chain variable region (this paper is abbreviated as VL).In another example, antibody comprises two weight (H) chain variable regions and two light (L) chain variable regions.Antigen-binding fragments of antibodies (for example, single-chain antibody, Fab fragment, F (ab ') contained in term " antibody " ₂, Fd fragment, Fv fragment and dAb fragment) and complete antibody.

VH and VL district can further be further divided into the hypervariable region that is called " complementary judgement district " (" CDR "), wherein scatter more conservative being called " framework region " zone (FR).The scope of explication framework region and CDR (referring to, Kabat, E.A., Deng the people, (1991) Sequences of Proteins ofImmunological Interest, the 5th edition, U.S.Departmentof Health and Human Services, open case of NIH 91-3242 number and Chothia, people such as C., (1987) J.Mol.Biol.196:901-917).This paper just defines with Kabat.Each VH and VL generally include three CDR and four FR, arrange in the following order from N-terminal to C-terminal: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

" immunoglobulin (Ig) territory " refers to from the variable domain of immunoglobulin molecules or the territory of constant domain.The immunoglobulin (Ig) territory contain usually two βZhe Dies that form by about seven β chains and conservative disulfide linkage (referring to, for example A.F.Williams and A.N.Barclay 1988 Ami.Rev Immunol.6:381-405).The canonical structure of the hypermutation ring of immune globulin variable region can be inferred by its sequence, as people such as Chothia, and (1992) J.Mol.Biol.227:799-817; People such as Tomlinson, (1992) J.Mol.Biol.227:776-798) and people such as Tomlinson, (1995) EMBO is (18) J.14: 4628-38 is described.

As used herein, " immunoglobulin variable territory sequence " refers to form the aminoacid sequence of the structure in immunoglobulin variable territory.For example, described sequence can comprise naturally occurring variable domain aminoacid sequence in whole or in part.For example, described sequence can be saved one, two or more N-terminal or C-terminal amino acid, internal amino acid, can comprise one or more insertions or other not terminal amino acid maybe can comprise other variation.In one embodiment, comprising immunoglobulin variable territory polypeptide of sequence can associate to form target integrated structure (or " antigen binding site ") with another immunoglobulin variable territory sequence, the structure of for example interact with hK1 (for example be incorporated into or suppress hK1).

Thereby the VH chain of antibody or VL chain can further comprise forming respectively in whole or in part of heavy chain or constant region of light chain weighs immunoglobulin chain or light immunoglobulin chain.In one embodiment, antibody is the tetramer of two heavy immunoglobulin chains and two light immunoglobulin chains, and wherein said heavy immunoglobulin chain is connected with light immunoglobulin (Ig) chain warp (for example) disulfide linkage is interior.The heavy chain constant domain comprises three territories, CHI, CH2 and CH3.The light chain constant domain comprises the CL territory.The territory that combines with AI is contained in the variable region of heavy chain and light chain.The common mediate antibody of the constant region of antibody combines with the host tissue or the factor, and the described host tissue or the factor comprise first component (Clq) of immune various kinds of cell (for example, effector cell) and classical complement system.Term " antibody " comprises the complete immunoglobulin (Ig) of IgA, IgG, IgE, IgD, IgM (with its hypotype) type.The light chain of immunoglobulin (Ig) can be κ type or λ type.In one embodiment, antibody is through glycosylation.But the cytotoxicity of antibody antagonist dependent cellular cytotoxicity and/or complement-mediated works.

The one or more zone of antibody can be the mankind's or effective mankind's.For example, one or more variable regions can be the mankind's or effective mankind's.For example, one or more CDR can be human, for example HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and LC CDR3.Among the light chain CDR each all can be human.HC CDR3 can be human.One or more framework regions can be human, for example the FR1 of HC or LC, FR2, FR3 and FR4.In one embodiment, all framework regions are human, for example are derived from human somatocyte, for example produce the hematopoietic cell or the non-hematopoietic cell of immunoglobulin (Ig).In one embodiment, the human sequence is the reproductive tract sequence, for example, and by the reproductive tract sequence of reproductive tract nucleic acid encoding.One or more constant regions can be the mankind's or effective mankind's.In another embodiment, at least 70%, 75%, 80%, 85%, 90%, 92%, 95% or 98% framework region (for example, FR1, FR2 and FR3 are all or FR1, FR2, FR3 and FR4 all) or whole antibody can be human or effectively human.For example, FR1, FR2 and FR3 are all can at least 70%, 75%, 80%, 85%, 90%, 92%, 95%, 98% or 99% consistent with the human sequence who by the human reproduction is VH section coding.

Can encoding by immunoglobulin gene or its section in whole or in part of antibody.Exemplary human immunoglobulin gene comprises κ, λ, α (IgA1 and IgA2), γ (IgG1, IgG2, IgG3, IgG4), δ, ε and μ constant region gene and countless immune globulin variable region gene.Total length immunoglobulin (Ig) " light chain " (about 25Kd or 214 amino acid) is by the variable region gene (about 110 amino acid) of NH2 end and the κ or the λ constant region genes encoding of COOH end.Total length immunoglobulin (Ig) " heavy chain " (about 50Kd or 446 amino acid) is similarly by variable region gene (about 116 amino acid) and one other above-mentioned constant region genes encoding of γ (about 330 amino acid of encoding) for example.

As used herein, " Fab " of term full length antibody (or be called for short " antibody moiety " or " fragment ") refers to keep one or more fragments of full length antibody that specificity is incorporated into the ability of the target of being paid close attention to.The example that is covered by the binding fragment in " Fab " of term full length antibody comprises (i) Fab fragment, the unit price fragment of being made up of VL, VH, CL and CH1 territory; (ii) F (ab ') 2 fragments, comprise two at hinge area by the segmental divalence fragment of the Fab of disulfide bridge bond binding; (iii) the Fd fragment is made up of VH and CH1 territory; (iv) Fv fragment is made up of the VL and the VH territory of the single armed of antibody; (v) dAb fragment (people such as Ward, (1989) Nature 341:544-546), it is made up of the VH territory; (vi) (CDR) distinguished in the isolating complementary judgement of reservation function.In addition, though segmental two territories of Fv (VL and VH) are by independent genes encoding, but can use recombination method that it is engaged by synthetic connexon, described synthetic connexon makes described two territories can become the simple protein chain, and the pairing of VL and VH district forms the monovalent molecule that is called strand Fv (scFv) in described simple protein chain.Referring to, people such as Bird for example, people such as (1988) Science 242:423-426 and Huston, (1988) Proc.Natl.Acad.Sci.USA 85:5879-5883.

Antibody fragment can use any proper technology to obtain, and described technology comprises the known known techniques of those skilled in the art.Term " monospecific antibody " refers to the special target of for example epitope is shown the antibody of single binding specificity and avidity.Described term is included in " monoclonal antibody " used herein or " monoclonal antibody combination ", and it is meant antibody or its segmental preparation of single molecular composition.As used herein, " homotype " refers to the antibody classification (for example, IgM or IgG1) by the weight chain constant area gene coding.

" effectively human " immune globulin variable region is to comprise the human framework amino acid position of enough numbers so that immune globulin variable region does not cause immunogenic response in normal human subject immune globulin variable region." effectively human " antibody is to comprise the human amino acid position of enough numbers so that antibody does not cause immunogenic response in normal human subject antibody.

" humanization " immune globulin variable region is the modified immune globulin variable region that does not cause immunogenic response with the human framework amino acid position that comprises enough numbers so that described immune globulin variable region in normal human subject.The description of " humanization " immunoglobulin (Ig) is including (for example) US 6,407, and 213 and US 5,693,762.

As used herein, " binding affinity " refers to apparent association constant or K _aK _aBe dissociation constant (K _d) inverse.Conjugated protein (for example) can have less than 10 special target molecule ^-5, 10 ^-6, 10 ^-7Or 10 ^-8The K of M _dBinding partner and first target combine than with second target have higher avidity can be by than K in conjunction with second target _a(or numerical value K _d) the higher K in conjunction with first target _a(or littler numerical value K _d) indication.Under described situation, with respect to second target (for example, the protein except that hK1, for example serum albumin, ln or sphaeroprotein), conjugated protein (for example, hK1) have a specificity to first target.The difference of binding affinity (for example, specificity or other comparison) can be at least 1.5 times, 2 times, 3 times, 4 times, 5 times, 10 times, 15 times, 20 times, 50 times, 70 times, 80 times, 100 times, 500 times, 1000 times or 10 ⁵Doubly.

Binding affinity can be measured by several different methods, and described method comprises equilibrium dialysis, balance combination, gel-filtration, ELISA, surface plasma resonance or spectroscopy (for example, using fluorescent assay).The assessment binding affinity exemplary condition be in PBS (phosphate buffered saline (PBS)) under the pH7.2 under 30 ℃.Described technology can be used for measuring combination and the free protein-bonded concentration as the function of conjugated protein (or target) concentration.The concentration of protein-bonded binding site is relevant on the protein-bonded concentration of bonded ([Bound]) and protein-bonded concentration ([Free]) and the target freely, as shown in the formula, wherein (N) is the number of the binding site of every target molecule;

[Bound]＝N·[Free]/((1/Ka)+[Free])。

Always must not carry out the accurate mensuration of Ka, although because (for example) used such as the method for ELISA or facs analysis and measured the quantitative measurment that is enough to obtain with the proportional avidity of Ka sometimes, and therefore can be used for comparison (whether (for example) is high 2 times such as determining higher affinity) is obtained avidity by the activity in the function calibrating (for example in vitro or in vivo calibrating) with the observational measurement that obtains avidity or (for example) deduction." composition for separating " refers at least 90% composition that shifts out from least a component of the natural sample that can obtain composition for separating.Composition artificial or natural generation can be " composition of purity at least to a certain degree ", and is at least 5%, 10%, 25%, 50%, 75%, 80%, 90%, 92%, 95%, 98% or 99% pure if the colony of species of being paid close attention to or species counts with weight-weight.

" epitope " refers to by the site on conjugated protein the antibody of Fab or full length antibody (for example, such as) bonded target compound.At target compound is under the proteinic situation, and described site can be made of amino acid composition fully, constitutes by proteinic chemistry of amino acids modification (for example glycosyl part) formation or by it fully.Eclipsed antigen decision base comprises at least one common amino acid residue.

" homology " or " sequence identity " (described term exchanges use in this article) between two sequences be calculated as follows execution.For the best purpose relatively, arrange described sequence (for example, be that the best is aimed at, can in one or two of first and second amino acid or nucleotide sequence, introduce the gap, and for comparison purposes, can ignore non-homogeneous sequence).Use that GAP program in the GCG software package is 12 in gap punishment, gap expansion punishment be 4 and frame to shift gap punishment be with Blossum62 score matrix the best to be aimed under 5 the situation to be defined as best score.Subsequently, the amino-acid residue or the Nucleotide at more corresponding amino acid position or nucleotide position place.When the position in first sequence by with second sequence in the same amino acid residue of corresponding position or Nucleotide when occupying, then described molecule is described position consistency (as used herein amino acid or nucleic acid " consistence " are equivalent to amino acid or nucleic acid " homology ").Consistence percentage between two sequences is the function of the number of the described sequence consistent position of sharing.

In a preferred embodiment, the length of the reference sequences of arranging for purpose relatively be reference sequences length at least 30%, preferably at least 40%, more preferably at least 50%, even more preferably at least 60% and even more preferably at least 70%, 80%, 90%, 92%, 95%, 97%, 98% or 100%.For example, reference sequences can have the length of immunoglobulin variable domain sequence.

As used herein, term as used herein " consistent substantially " (or " homology substantially ") refers to first amino acid or nucleotide sequence, its contain enough numbers with second amino acid or consensus nucleic acid sequence or equivalence (for example, have the similar side chain that conserved amino acid for example replaces) amino-acid residue or Nucleotide so that described first and second amino acid or nucleotide sequence have similar activity (or coding has similar active protein), for example in conjunction with active, in conjunction with priority or biological activity.Under the situation of antibody, with respect to same antigen, second antibody has identical specificity and has at least 50% avidity.

Similar or homologous sequence (for example, at least about 85% sequence identity) also is the part of the application's case with the disclosed sequence of this paper.In certain embodiments, sequence identity can be about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher.In addition, during down with the complementary sequence hybridization of described chain, there is property unanimous on the whole in selective cross condition (for example, highly Yan Ge hybridization conditions) when nucleic acid segment.Nucleic acid can be present in the intact cell, is present in the cell lysates or pure or pure substantially form exists with part.

As used herein, term " low strict, moderate is strict, hybridize under the highly strict or high stringent condition " condition of hybridizing and washing of being used to described.The guide of carrying out hybridization is found in Current Protocols in MolecularBiology, John Wiley﹠amp; Sons, N.Y. (1989), 6.3.1-6.3.6, it is incorporated herein by reference.Described reference is described moisture and anhydrous process, and can use any one.Specific cross condition as referred to herein is as follows: (1) low stringent hybridization condition, under about 45 ℃, 6X sodium chloride/sodium citrate (SSC) is then under 50 ℃, with 0.2XSSC, 0.1%SDS washed twice (with regard to low stringent condition, wash temperature can increase to 55 ℃); (2) moderate stringent hybridization condition, under about 45 ℃, 6X SSC is then under 60 ℃, with 0.2X SSC, 0.1%SDS washing once or once; (3) height stringent hybridization condition, under about 45 ℃, 6X SSC is then under 65 ℃, with 0.2X SSC, 0.1%SDS washing once or once; (4) high stringent hybridization condition is under 65 ℃, and 0.5M sodium phosphate, 7%SDS are then under 65 ℃, with 0.2X SSC, 1%SDS washing once or once.High stringent condition (4) is an optimum condition and except as otherwise noted, otherwise should use described high stringent condition.The present invention is included under minuent, moderate, height or the high strict situation nucleic acid with nucleic acid as herein described or its complementary sequence hybridization, the protein-bonded nucleic acid as herein described of for example encoding.Described nucleic acid can have equal length or be 30%, 20% or 10% of reference nucleic acid length with reference nucleic acid.Described nucleic acid can be corresponding to the district of coding immunoglobulin variable territory sequence.

Conjugated protein with respect to as herein described, hK1 is conjugated protein to have sudden change (for example, conservative or non-essential amino acid replaces), and described sudden change does not exert an influence to protein function substantially.No matter whether special replacement can allow, its equal (for example) should not influence biological property unfriendly, and described biological property waits the people such as can (for example) using Bowie, and the combination of the method prediction of (1990) Science 247:1306-1310 is active.

" conserved amino acid replacement " is the replacement of the radical amino acid replacement of amino-acid residue through having similar side chain.Amino-acid residue family with similar side chain defines in affiliated field.Described family comprise have basic side chain amino acid (for example, Methionin, arginine, Histidine), amino acid with acid side-chain (for example, aspartic acid, L-glutamic acid), amino acid with uncharged polar side chain (for example, glycine, l-asparagine, glutamine, Serine, Threonine, tyrosine, halfcystine), amino acid with non-polar sidechain (for example, L-Ala, Xie Ansuan, leucine, Isoleucine, proline(Pro), phenylalanine, methionine(Met), tryptophane), amino acid with β-branched building block (for example, Threonine, Xie Ansuan, Isoleucine) and amino acid (for example, tyrosine with aromatic side chains, phenylalanine, tryptophane, Histidine).Numerous frameworks and cdr amino acid residue might comprise one or more conservative replacement.

The concensus sequence of biological polymer can comprise the position that can change between multiple amino acids.For example, in described context, symbol " X " is made a general reference any amino acid (for example, any in any in 20 kinds of natural amino acids or the 19 kinds of non-cysteine amino acids).Other allows amino acid also can (for example) use parenthesis and oblique line indication.For example, " (A/W/F/N/Q) " means at described specific position and allows L-Ala, tryptophane, phenylalanine, l-asparagine and glutamine.

" nonessential " amino-acid residue is not eliminated or is more preferably changed bioactive residue substantially for changing from the wild-type sequence of the wedding agent of for example antibody; And " essential " amino-acid residue produces described variation.

Term " polypeptide " or " peptide " (it is used interchangeably) refer to three or three above polymer of amino acid through the peptide bond binding, and for example length or between 30 and 300 amino acid, or surpasses 300 amino acid between 3 and 60 amino acid.Polypeptide can comprise one or more alpha-non-natural amino acids.Usually, polypeptide only comprises natural amino acid." protein " can comprise one or more polypeptide chains.Therefore, polypeptide contained in term " protein ".Protein or polypeptide also can comprise one or more modifications, for example, and glycosylation, amidation, phosphorylation etc.

Term " homology substrate " refers to naturally occurring hK1 substrate, and it comprises naturally occurring its varient (for example, splicing variants, naturally occurring mutant and with the merit iso series).

As used herein, term " common password " refers to be used for common password of sequence expression special acid of the cell category of marking protein." inferior common password " is for being present in usually in the specialized species but be not the codon of common password.All codons except that common password and time common password are " non-common codon ".

" hK1 be correlated with inflammatory illness " refers to so that small part is the illness of feature by the inflammatory reaction of hK1 (for example, the protease activity of hK1) mediation.With regard to described illness, the active reduction of hK1 makes the inflammatory reaction reduce.The relevant inflammatory illness of exemplary hK1 comprises asthma (for example, allergic asthma and non-allergic asthma), chronic obstructive pulmonary disease (COPD), multiple sclerosis, psoriasis, rheumatoid arthritis, osteopathy sacroiliitis, osteoarthritis, rhinitis, sinusitis, inflammatory bowel (such as Crohn disease and ulcerative colitis), immune-mediated diabetes, acute pancreatitis and interstitial cystitis.

Statistical significance can be measured by the known method in field under any.Exemplary statistical test comprises: Student T check, Mann Whitney U nonparameter test and Wilcoxon nonparametric statistics check.Some statistically evident relations have the P value less than 0.05 or 0.02.Special combination albumen can be showed the difference of (for example) specificity, combination or biological activity aspect, and described difference is statistically evident (for example, P value＜0.05 or 0.02).Diacritic qualitative or quantitative difference between two states of (for example) expression such as term " is induced ", " inhibition ", " enhancing ", " raising ", " increase ", " reduction " and can refer to the difference of for example statistically evident difference between two states.

Unless otherwise defined, otherwise employed all technology of this paper and scientific terminology all have with the present invention under the identical implication of implication of those skilled in the art's common sense in field.Though in practice of the present invention or test, can use with method as herein described and material type like or the method and the material of equivalence, suitable method and substance description are as follows.Mentioned all open cases, patent application case, patent and other reference of this paper all is incorporated herein by reference with its whole content.

Description of drawings

Fig. 1. the SPR grouping of epitope.Biotin labeled hK1 being trapped on the chip of streptavidin coating until final ligand density is 88 RU.Use Biacore 3000 instruments that X-2300 is injected in fixedly on the hK1 with the concentration of 50nM, the flow velocity of following under 25 ℃ with the concentration 30 μ L/min of 50nM is injected in the 2nd Fab among the PBST.Injection DX-2300, the signal when then injecting the 2nd Fab end is R1.Signal when injection Fab finishes separately is R2.Figure A shows the influence chart in conjunction with the Fab (M0112-D07) of the epitope identical with DX-2300.Figure B shows the influence chart in conjunction with the Fab (M0139-A09) of the epitope different with DX-2300.

The inhibition mechanism of Fig. 2 .DX-2300 (this paper is also referred to as " M0131-F07 ").Displaying as the inhibition mechanism of the DX-2300 of Fab (figure A) for emulative and have the K of 60 ± 2pM _iDisplaying as the inhibition mechanism of the DX-2300 of IgG (figure B) for emulative and have the K of 39 ± 4pM _i

Fig. 3. the inhibition mechanism of other hK1 inhibitor.Displaying as the inhibition mechanism of the M0093-F09 of Fab (figure A) for noncompetitive and have the K of 1.9nM _iDisplaying as the inhibition mechanism of the M0137-E01 of Fab (figure B) for emulative and have the K of 0.9nM _i

The march line analysis of Fig. 4 .DX-2300.The curve (figure A) that dynamically carries out in the presence of the DX-2300 of cumulative amount inhibitor is fit to following equation acquisition kobs value: F=v _s+ (v ₀-v _s) (1-exp (k _Obs* t))/k _Obs+ C, F=fluorescence wherein, v _s=final speed of steady state, v ₀=initial speed of steady state, k _Obs=index suppresses constant, and t=is the time of unit with the second, and C is the constant of explanation t=0 initial background fluorescence during second.In figure B, kobs value mapped to inhibitor concentration and be fit to the equation of the agent of combining closely at a slow speed of following isomerization steps; k _Obs=k ₆+ (k ₅* [I]/(K _{I, app}+ [I])), k wherein ₆Be the reversed reaction rate constant of enzyme inhibitors isomerization steps, k ₅Be the positive reaction rate constant of described step, K _{I, app}For the apparent equilibrium that forms initial enzyme-inhibitor complex suppresses constant, and [I] is for being present in the concentration of the inhibitor in the calibrating.

Fig. 5. the active DX-2300 of class kallidinogenase suppresses in the animal urine.

Class kallidinogenase activity is measured as previously discussed in the animal.Suppressing percentage is viewed initial rate (R in the presence of DX-2300 _Suppress) with its not in the presence of viewed initial rate (R _o) ratio, therefore suppress percentage=100-(R _o-R _Suppress)/R _o* 100.

Fig. 6. the active DX-2300 IC of class kallidinogenase in the mankind or the sheep urine ₅₀Mensuration.The DX-2300 of varied concentration exist or non-existent situation under, carry out the plain active measurement of releiving of activation urine medium vessels as mentioned above.The measured active DX-2300 IC of class kallidinogenase during sheep urine is urinated with the people ₅₀Value is respectively 2.9 ± 1.6nM and 3.0 ± 0.6nM for suitable.

Fig. 7. from the active inhibition of class kallidinogenase among the patient's of rhinovirus infection future trouble mild asthma the human BAL.0.5 μ M DX-2300 exist or non-existent situation under, use the Pro-Phe-Arg-AMC substrate (table 4) of 100 μ M in the KAL damping fluid in 80 μ LBAL, to measure kallidinogenase activity among the BAL.Before adding substrate, under 30 ℃, BAL was cultivated 30 minutes with DX-2300.

Fig. 8 .DX-2300 is to the effect of the tracheae reaction of antigen induction in the sheep.Throw and 10mgDX-2300 through sucking, attack poison with ascaris suum (Ascaris suum) anaphylactogen after 12 hours 30 minutes to sheep (n=2).After anaphylactogen was attacked poison, figure A was by measuring the animal (square) and control animal (rhombus) that lung resistance is relatively handled through DX-2300 through 8 hours.Figure B compares the tracheae overreaction of throwing after viewed attacking against each other malicious 24 hours with carbachol in animal that DX-2300 handles and control animal.Hollow strips indication surpasses the amount of the required carbachol of baseline 400% in order to induce bronchoconstriction before attacking poison at anaphylactogen.Packing indication induces bronchoconstriction to surpass the amount of the required carbachol of baseline 400% in order to attack poison at anaphylactogen after 24 hours.

Fig. 9 .DX-2300 is to inducing the effect of the high molecular weight kininogen of bronchoconstriction in asthma sheep model.Diamond symbols is illustrated under the situation that does not have medicine, the bronchoconstriction of the HMWK (100 μ g) that response is sucked.Square symbol is illustrated in the DX-2300 that 1mg sucks and has down the bronchoconstriction of the HMWK (100 μ g) that response is sucked.Trilateral is illustrated in 5mg DX-2300 and has down the bronchoconstriction of the HMWK (100 μ g) that response is sucked.

Embodiment

hK1

Exemplary hK1 albumen comprises following sequence; Human kallidinogenase 1 precursor (EC3.4.21.35) (tissue kallikrein) (kidney/pancreas/sialisterium kallidinogenase) of＞sp|P06870|KLKl_-homo sapiens (mankind).

MWFLVLCLALSLGGTGAAPPIQSRIVGGWECEQHSQPWQAALYHFSTFQCGGILVHRQ

WVLTAAHCISDNYQLWLGRHNLFDDENTAQFVHVSESFPHPGFNMSLLENHTRQADED

YSHDLMLLRLTEPADTITDAVKVVELPTQEPEVGSTCLASGWGSIEPENFSFPDDLQC

VDLKILPNDECEKAHVQKVTDFMLCVGHLEGGKDTCVGDSGGPLMCDGVLQGVTSWGY

VPCGTPNKPSVAVRVLSYVKWIEDTIAENS (SEQ ID NO：1021)

Described protein is generally the proteinic protein through maturation processing that for example comprises about amino acid 25 to 262, or for example it has the segmental fragment of proteolytic activity.

HK1 albumen can comprise one or more following features.

Signal properties chain action site action site action site carbohydrate carbohydrate carbohydrate carbohydrate carbohydrate carbohydrate disulfide bond disulfide bond disulfide bond disulfide bond disulfide bond

1 19 25 65 120 214 93 102 104 108 165 167 31 50 153 185 210

18 24 262 65 120 214 93 102 104 108 165 167 174 66 220 199 235

18 6 238

Possible activated peptide (possibility) the callicrein 1 charge transfer System Charges transmission system charge transfer O-of system binding N-binding (GlcNAc...) O-binding N-binding (GlcNAc...) N-binding (GlcNAc...); Part O-binding similitude similitude similitude similitude similitude

HK1 albumen also can comprise following exemplary varient;

Varient 77 R-＞H (in dbSNP:5515) [NCBl/Ensembl]. VAR_014567

Varient 145 Q-＞E (in dbSNP:5516) [NCBl/Ensembl]. VAR_006625

Varient 186 E-＞K (in dbSNP:5517) [NCBl/Ensembl]. VAR_006626

Varient 193 V-＞E (in dbSNP:5518) [NCBl/Ensembl]. VAR_014568

The exemplary substrate of hK1 comprises precursor, renninogen, vasoactive intestinal peptide, procollagenase and the proangiotensin of proinsulin, low-density lipoprotein, atrium natriuretic factor.HK1 as herein described is conjugated protein to can be used for regulating described substrate, for example reduces the proteolytic cleavage of described substrate.

Can be used to make the antigen of animal immune or obtain by using with hK1 bonded antibody as (for example) as the hK1 that (for example) is used to screen the target in recombinant antibodies storehouse.Also can use peptide and the fragment of hK1.

Present the storehouse

In one embodiment, use presents storehouse discriminating and hK1 bonded protein.Present the set that the storehouse is an entity, each entity comprises the recyclable component (for example, nucleic acid) that can reach protein component (for example, Fab or scFv) and coding or differentiate protein component.Described protein component can have any length, and for example 3 amino acid are to surpassing 300 amino acid.When selecting, make each member's in storehouse protein component contact with hK1 and combine with hK1 as if protein component, then (for example) discriminating presents the library member on the upholder by being stranded in.Described protein component can comprise the varient in one or more immunoglobulin variable territories or another territory.The method in the immunoglobulin (Ig) territory that use is used to present be described below (referring to, for example " antibody presents storehouse (Antibody Display Libraries) ").

The self-supporting thing reclaim be detained present library member and analysis.Analysis can comprise amplification and the selection under similar or different condition subsequently.For example, positive and negative selection can replace.Analyze and also can comprise the aminoacid sequence of measuring protein component and be detailed featuresization protein purification component.

Can use multiple form for presenting the storehouse.Example comprises following each person.

Phage presents.A kind of form utilizes virus, especially phage.Described form is called " phage presents ".The common covalent bond of protein component is in the phage sheath protein.Described binding result forms the translating of nucleic acid that coding is blended in the protein component of sheath protein.Described binding can comprise flexible peptide connexon, protease site or because of restraining the amino acid that terminator codon is incorporated into.Phage presents and is described in following each document: for example United States Patent (USP) the 5th, 223, No. 409; Smith (1985) Science 228:1315-1317; WO 92/18619; WO 91/17271; WO 92/20791; WO 92/15679; WO 93/01288; WO 92/01047; WO 92/09690; WO 90/02809; People such as de Haard, (1999) J.Biol.Chem 21A:18218-30; People such as Hoogenboom, (1998) Immunotechnology 4:1-20; People such as Hoogenboom, (2000) Immunol Today 2:371-8; People such as Fuchs, (1991) Bio/Technology 9:1370-1372; People such as Hay, (1992) Hum Antibod Hybridomas 3:81-85; People such as Huse, (1989) Science 246:1275-1281; People such as Griffiths, (1993) EMBO J 12:725-734; People such as Hawkins, (1992) JMol Biol 226:889-896; People such as Clackson, (1991) Nature 352:624-628; People such as Gram, (1992) PNAS 89:3576-3580; People such as Garrard, (1991) Bio/Technology 9:1373-1377; People such as Rebar, (1996) Methods Enzymol.267:129-49; People such as Hoogenboom, people such as (1991) Nuc Acid Res 19:4133-4137 and Barbas, (1991) PNAS 88:7978-7982.

Developed and be used for filobactivirus (phage f1, fd and M13) and other phage (for example T7 phage and lambdoid phage; Referring to, Santini (1998) J.Mol.Biol.282:125-135 for example; People such as Rosenberg, (1996) Innovations 6:1-6; People such as Houshm, (1999) Anal Biochem 268:363-370) phage presents system.Filobactivirus presents system and usually uses and fusion such as proteic less important sheath protein of gene III and gene VIII albumen (main sheath protein), but also can use with such as the fusion in gene VI albumen, gene VII albumen, proteic other sheath protein of gene IX or its territory (referring to, for example WO 00/71694).In one embodiment, described fusion is to be blended in the proteic territory of gene III, for example, grappling territory or " undesirable root " (stump) (referring to, the localized United States Patent (USP) of gene III albumen anchor is for example described the 5th, 658, No. 727).Also might use the non-peptide bond of non covalent bond for example or non-peptide covalent linkage that the protein physics that is presented is associated in described sheath.For example, can use disulfide linkage and/or c-fos and c-jun coiled coil with physics associate (referring to, people such as Crameri for example, (1993) Gene 137:69 and WO 01/05950).

Can make the phage growth that presents protein component and use the pre-Preparation Method of the sedimentary standard phage of substratum PEG of for example growing certainly to collect.Select to present individually after the phage, the described nucleic acid self-infection of the selected protein component of can will encoding after amplification is selected the cellular segregation of phage or is separated from phage itself.Can select indivedual groups or bacteriolyze spot, with separate nucleic acid and carry out sequential analysis.

Other presents form.Other present form comprise based on the presenting of cell (referring to, for example WO 03/029456), the protein-nucleic acid fusion (referring to, for example US 6,207,446) and rrna present (referring to, people such as Mattheakis for example, (1994) people such as Proc.Natl.Acad.Sci.USA 91:9022 and Hanes, (2000) Nat Biotechnol18:1287-92; People such as Hanes. people such as (2000) Methods Enzymol.328:404-30 and Schaffitzel, (1999) JImmunol Methods.231 (1-2): 119-35).

The epitope binding proteins specific.It is conjugated protein that the technology that presents also can be used for the special epitope bonded that obtains with target, for example antibody.Epitope can be divided into " configuration type " or " sequence type ".The configuration epitope comprises amino-acid residue, though described amino acid in sequence may for substantially isolating (for example, separate by at least 1,2,4,6,8 or 10 amino acid), but it has definite relative orientation on suitably folding target.The sequence epitope comprises short polypeptide chain part, regardless of Protein Folding state (for example, primary or unfolded), its equal binding antibody.Can (for example) the non-target molecule that lacks special epitope or suddenly change in described epitope be competed with (for example) L-Ala and differentiated the conjugated protein of configuration epitope by utilization.Described non-target molecule can be used in the following negative select procedure, when making when presenting the storehouse and being incorporated into target as the competition molecule, or catches in washing soln as pre-scrub solution (for example) and target not to be had specific dissociating presents the library member.In another embodiment, the epitope binding proteins specific by use with target molecule on the epitope bonded paid close attention to compete conjugated protein elution and present the library member and differentiate.Can (for example) use small peptide to select the conjugated protein of binding sequence epitope with the aminoacid sequence in the target protein of seeing.Usually, conjugated protein being incorporated into a little less than also that is incorporated into the configuration epitope contained some and is contained in amino acid whose peptide or other peptide in the configuration epitope.Therefore, can be chosen under the utmost point low stringency condition and combine, and select subsequently to combine with folding target protein with peptide.

Affinity maturation.In one embodiment, conjugated protein by (for example) mutagenesis modification and target bonded to provide modification conjugated protein pond.It is conjugated protein conjugated protein to differentiate one or more transformations with functional performance of change (for example, the in vivo stability of the stability of the associativity of raising, raising, prolongation) to assess described modification subsequently.In one embodiment, use presents the storehouse choice of technology or screens the conjugated protein pond of described modification.Subsequently, (for example) used higher stringent condition or had more emulative combination and wash conditions differentiates to have the conjugated protein of high-affinity more from second storehouse.Also can use other triage techniques.

In certain embodiments, mutagenesis is the zone that the target known region maybe may be positioned at bonding interface.If what (for example) differentiated is conjugated protein for antibody, then mutagenesis can be at the CDR district of heavy chain as herein described or light chain.In addition, mutagenesis can near or in abutting connection with the framework region of CDR, framework regions in 10 of the CDR land, 5 or 3 amino acid especially for example.Under the situation of antibody, mutagenesis also can be confined among the CDR one or several (for example) progressively to improve.

In one embodiment, use mutagenesis so that antibody more is similar to one or more reproductive tract sequence.A kind of exemplary reproductive tract method can comprise: the reproductive tract sequence of differentiating the sequence similar (for example, similar in the particular database) of one or more and separation antibody.Subsequently, can be in separation antibody with increment type, combination or both suddenly change (on amino acid levels).For example, produce the nucleic acid library of the sequence comprise the some or all of possible reproductive tracts sudden changes of coding.Assess the antibody that described mutant antibody (for example) has one or more other reproductive tract residues with discriminating with respect to separation antibody and still is suitable for (for example, having functionally active) subsequently.In one embodiment, reproductive tract residue as much as possible is introduced in the separation antibody.

In one embodiment, use mutagenesis replaces one or more reproductive tract residues or is inserted in the CDR district.For example, reproductive tract CDR residue can from the reproductive tract sequence of the variable region of being modified similar (for example, similar).After the mutagenesis, can assess the activity (for example, in conjunction with or other functionally active) of antibody and whether be allowed to measure described reproductive tract residue.Can in framework region, carry out similar mutagenesis.

Can different methods carry out the selection of reproductive tract sequence.For example, if the predetermined criterion of reproductive tract sequence coincidence selectivity or similarity, for example reach a certain at least consistence percentage (for example at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% consistence), then can select described reproductive tract sequence.Can use at least 2,3,5 or 10 reproductive tract sequences execution selections.Under the situation of CDR1 and CDR2, differentiate that similar reproductive tract sequence can comprise described sequence of selection.Under the situation of CDR3, differentiate that similar reproductive tract sequence can comprise described sequence of selection, but can comprise the reproductive tract sequence of using two to help N-terminal part and C-terminal part respectively.In other embodiments, use more than one or two reproductive tract sequence (for example) to form concensus sequence.

In one embodiment, special reference variable domain sequence with respect to for example sequence described herein, relevant variable domain sequence have with reference to the residue in the CDR sequence, with the human reproduction be residue at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, the 95% or 100% inconsistent cdr amino acid position of the residue unanimity of corresponding position in the sequence (that is, be the aminoacid sequence of nucleic acid encoding) by the human reproduction.

In one embodiment, special reference variable domain sequence with respect to for example sequence described herein, relevant variable domain sequence has and from the human reproduction is the FR sequence at least 30%, 50%, 60%, 70%, 80%, 90% of sequence (for example, reproductive tract sequence) relevant with reference variable domain sequence or the FR district of 100% unanimity.

Therefore, might separate the given antibody similar activity that has and paid close attention to, but more be similar to one or more reproductive tract sequences, especially one or more human reproductions are the antibody of sequence.For example, antibody and reproductive tract sequence can have at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% consistence in the district of CDR outside (for example, framework region).In addition, antibody can comprise at least 1,2,3,4 or 5 reproductive tract residues in the CDR district, described reproductive tract residue be from the reproductive tract sequence of the variable region of being modified similar (for example, similar).The reproductive tract sequence of mainly being paid close attention to is human reproductive tract sequence.The activity of antibody (for example, in conjunction with active) can be 1 times or 100 times, 10 times, 5 times, 2 times, 0.5 times, 0.1 times and 0.001 times of original antibody.Exemplary reproductive tract sequence comprises VKI-O2, VL2-1, VKIH-L2::JK2, vg3-23, V3-23::JH4 and V3-23::JK6.

Some exemplary induced-mutation techniques comprise: fallibility PCR (error-prone PCR) (people such as Leung, reorganization (1989) Technique 1:11-15), (referring to, for example USSN 10/279,633), use at random cracked DNA reorganization (Stemmer (1994) Nature 389-391; Be called " nucleic acid reorganization "), RACHITT ^TM(people such as Coco, (2001) site-directed mutagenesis (people such as Zoller Nature Biotech.19:354),, Nucl Acids Res 10:6487-6504), sequence box mutagenesis (Reidhaar-Olson (1991) Methods Enzymol.208:564-586) and incorporate degeneracy oligonucleotide (people such as Griffiths, (1994) EMBO J 13:3245) into.In an example of affinity maturation, use method as herein described at first to differentiate with minimum at least binding specificity or lowest activity (for example, the bonded equilibrium dissociation constant is less than 1nM, 10nM or 100nM) conjugated protein in conjunction with hK1 to target from presenting the storehouse.The initial protein-bonded nucleotide sequence of differentiating of coding had the second conjugated protein of enhanced propertied (for example, binding affinity, kinetics or stability) as the template nucleic acid (for example) of introducing variation to differentiate with respect to initial conjugated protein.Perhaps, the aminoacid sequence of one or more CDR can be used as the guide of the nucleic acid library that designs the nucleic acid that comprise coding separation sequence and numerous flanking sequences.Described diversified nucleic acid can be introduced and contain being in the expression vector of the initial separation that is selected from described storehouse and improved varient.

The dissociation rate back-and-forth method.Because slow dissociation rate can indicate high-affinity, especially about the interaction between polypeptide and its target spot, therefore can use method as herein described to have the conjugated protein of the dynamic dissociation rate of being wanted (that is reduction) to separate for binding interactions with target spot.

Select slowly to dissociate conjugated protein for presenting the storehouse certainly, make described storehouse and contact through the fixed target.Subsequently, with fixed target first solution washing that removes non-specific binding or weak bonded biomolecules.Subsequently, with the second solution elution of fixed target with the free target of the duplicate of the target that for example is not attached to particle that comprises saturation capacity.Free target is incorporated into from the dissociated biomolecules of described target.With respect to the fixed target of much lower concentration, effectively stop in conjunction with free target once more by saturation capacity.

Second solution can have solution condition or the strict solution condition that is substantially physiological conditions.Usually, the solution condition of second solution is consistent with the solution condition of first solution.Elution part of second solution is collected to distinguish early stage elution part and elution in late period part with time sequence.Elution in late period part comprise with than the slower speed of the biomolecules in early stage elution part from the dissociated biomolecules of target.

In addition, also might reclaim even after prolonging cultivation, still keep being incorporated into the library member that presents of described target.Can use from the liquid condition and it be dissociated or can when being connected in target, it be increased.For example, the phage that is incorporated into target is contacted with bacterial cell.

Specific selection and screening.Presenting under the situation in storehouse, " selection " refers to allow many library members of presenting to contact the process that target and recovery and propagation bonded present the library member.Selection can be from the storehouse with a large amount of members, for example more than 10 ¹⁰Individual member.Presenting under the situation in storehouse, " screening " refers to be incorporated into one by one the separation member's of target test library process.By automatic operation, can be in a highly parallel process the thousands of material standed fors of screening.The storehouse back-and-forth method that presents as herein described can comprise and gives up the chosen process that presents the library member that is incorporated into non-target molecule.

(for example) be used for the example of non-target molecule of hK1 binding antibody including (for example) the proteolytic enzyme except that hK1, for example, the kallidinogenase except that hK1 (for example hK2, hK3, hK4, hK5, hK6, hK7, hK8, hK9, hK10, hK11, hK12, hK13, hK14 or hK15).Being suitable for the hK1 binding antibody may have specificity to the kallidinogenase subclass, and for example, it can interact with a plurality of kallidinogenases, and wherein at least one is hK1.

In one embodiment, use so-called " the negative selection " step debate other target and relevant non-target molecule and be correlated with but the non-target molecule of uniqueness.Make and present the storehouse or its pond contacts with non-target molecule.Collect and the sample member of the non-target of debond and be used for subsequently select with the target molecule bonded or even the feminine gender that is used for subsequently select.The negative step of selecting can be before or after selection and target molecule bonded library member.

In another embodiment, use the screening step.To present the library member and separate, respectively separate the library member with regard to its aptitude tests that are incorporated into non-target molecule (for example, above listed non-target) to be incorporated into after the target molecule.For example, can use high-throughput ELISA screening to obtain described data.Also can use the ELISA screening to obtain the quantitative data that each library member is incorporated into target.More non-target and target binding data (for example, using computer and software) are incorporated into the library member of hK1 to differentiate specificity.

Storehouse back-and-forth method and the sieve method of presenting as herein described can comprise selection that presents the library member and the screening process of selecting to be incorporated into specificity site on the target molecule.For example, select to be incorporated into phage with the antibody elution as herein described of high density by the epitope of described antibodies.Can be by in damping fluid, under the situation that has or do not have the competition antibody of the described epitope of identification to exist, carrying out the phage that the ELISA screening is incorporated into the special epitope of hK1.

Antibody presents the storehouse

In one embodiment, present the stock in proteinic diversity pond, wherein each all comprises at least one and common two immunoglobulin variable territories.Present the storehouse and be particularly useful for the antibody that (for example) differentiates the human antigenic mankind of identification or effective mankind.Because the constant region of antibody and framework region are human, thus described treatment antibody can make himself avoid being used as antigen recognition for or target.Also with the effector function of constant region optimization with recruitment human immunity system.In vitro present chosen process and overcome the ability that the normal human subject immunity system can not produce antibody to autoantigen.

Typical antibody presents the storehouse and presents the protein that comprises VH territory and VL territory.Present the antibody that the storehouse can present the Fab pieces (for example, using two polypeptide chains) or be the antibody (for example, using single polypeptide chain) of strand Fv form.Also can use other form.

Under the situation of Fab and other form, the antibody that is presented can comprise the constant region as the part of light chain or heavy chain.In one embodiment, each chain comprises a constant region, for example under the situation of Fab.In other is implemented, present other constant region.

Antibody library can construct by several different methods (referring to, people such as de Haard for example, (1999) J.Biol.Chem274:18218-30; People such as Hoogenboom, (1998) Immunotechnology 4:1-20; People such as Hoogenboom, (2000) Immunol Today 21:371-8; US 2003-0232333 and US 2004-0029113).In addition, the key element of each method can with the factor combination of other method.Can use described method to introduce in the single immunoglobulin (Ig) territory (for example, VH or VL) or in a plurality of immunoglobulin (Ig)s territory (for example, VH and VL) making a variation.Variation can be introduced in the immunoglobulin variable territory, for example in the one or more described district among CDR1, CDR2, CDR3, FR1, FR2, FR3 and the FR4, it refers to any one of heavy chain and light chain variable territory or both described districts.In one embodiment, variation is introduced among all three CDR of given variable domain.In another preferred embodiment, among will make a variation introducing CDR1 and the CDR2, for example among the CDR1 and CDR2 of heavy chain variable domain.Any combination is feasible.In a method, the multiple oligonucleotide of antibody library by the CDR that will encode inserts in the respective area of nucleic acid that coding presents protein or its part and constructs.Described oligonucleotide can use the subunit of multiple for example monomer Nucleotide or trinucleotide synthetic.For example, people such as Knappik, (2000) J.Mol.Biol.296:57-86 the structure of transvers plate that describe to use the trinucleotide synthesis method and be used to the accept oligonucleotide method of CDR of described oligonucleotide of encoding with through engineering approaches restriction site.

In other method, make for example rodentine animal immune with hK1.Optionally, use antigen stimulation with further irritant reaction described animal.Subsequently, splenocyte is separated from animal, and the nucleic acid amplification in will encode VH and/or VL territory and clone in presenting the storehouse, to express.

In another method, antibody library is by the nucleic acid structure from natural reproductive tract immunoglobulin gene (for example, Human genome) amplification.Described amplification of nucleic acid comprises the nucleic acid in coding VH and/or VL territory.The source of the nucleic acid of coding immunoglobulin (Ig) is in following description.Amplification can be including (for example) PCR or another amplification method of use with conservative constant region annealed primer.

Coding immunoglobulin (Ig) territory or its segmental nucleic acid can be available from (for example) mankind, primate, mouse, rabbit, camel or rodentine immunocytes.Can select cell with regard to special property.For example, can be chosen in the B cell in various stage of maturity, comprise the natural B cell.

Can use fluorescent activation cell sorter (FACS) sorting to express the B cell of surface bonding IgM, IgD or IgG molecule.In addition, the different isostructural B cell of separable expression IgG.Can be with B and T cell (for example) by cultivating with feeder cell or by mitogen or other conditioning agent such as antibody being added in CD40, CD40 part or CD20, phorbol tetradecanoate acetic ester, bacteria lipopolysaccharide, con A, phytohemagglutinin or the Phytolacca acinosa mitogen and in vitro cultivated and stimulate.

The person under inspection that cell also can have the immunology illness certainly separates, described illness for example, systemic lupus erythematosus (SLE), rheumatoid arthritis, vasculitis, Xiu Gelian syndrome (Sjogren syndrome), Sjogren's syndrome disease or anti-phosphatide syndromes.The person under inspection can be the mankind or animal, for example, and the animal model of human diseases or have the animal of similar conditions.Cell can self-contained human immunoglobulin gene's seat the transgenic nonhuman animal separate.

Described cell may activate the program of somatic hypermutation.Can (for example) by coming irritation cell experience immunoglobulin gene with anti-immunoglobulin, anti-CD40 and anti-CD38 antibody treatment somatocyte mutagenesis (referring to, people such as Bergthorsdottir for example, (2001) JImmunol.166:2228).In another embodiment, cell is natural.

Can in natural storehouse, separate by will the encode nucleic acid in immunoglobulin variable territory of following exemplary methods.At first, RNA is separated from immunocyte.MRNA separates (for example, by making the cap cleaved rna dephosphorylation with the calf intestinal phosphatase enzyme) with total length (for example, band cap).Subsequently, cap is removed and use reverse transcription produce cDNA with tobacco acid pyrophosphatase.

The reverse transcription of first (antisense) chain can be finished by any way with any suitable primer.Referring to, people such as Haard for example, (1999) J.Biol.Chem 274:18218-30.PBR can be the different homotypes of constant (for example) with the reverse transcription immunoglobulin (Ig) between different immunoglobulin (Ig)s.PBR also can have specificity to the special homotype of immunoglobulin (Ig).Usually, primer to for the coding at least one CDR sequence 3 ' the zone have specificity.In another embodiment, can use poly-dT primer (and for heavy chain gene may for preferred).

Composition sequence can be engaged in 3 of reverse transcription chain ' end.During the pcr amplification, composition sequence can be used as primer binding site after the reverse transcription in conjunction with forward primer.The use of composition sequence can be exempted the different forward primers of use pond to catch available multifarious needs fully.

Subsequently, (for example) uses one or more circulations to make the gene amplification of coding variable domain.If use a plurality of circulations, then can use nested primer (nested primer) for the fidelity of reproduction that increases.Subsequently, with the amplification nucleic acid clone in presenting the storehouse carrier.

Antibody produces

Some for example the antibody of Fab can in the bacterial cell of for example intestinal bacteria (E.coli) cell, produce.For example, if Fab comprises the phage that can restrain terminator codon and is sequence encoding in the expression vector by presenting between entity and the phage protein (or its fragment), then vector nucleic acid can be shuttled back and forth and to restrain in the bacterial cell of terminator codon.In the case, Fab does not merge with gene III albumen and secretes in substratum.

Antibody also can produce in eukaryotic cell.In one embodiment, antibody (for example, scFv) such as pichia spp (Pichia) (referring to, people such as Powers for example, (2001) JImmunol Methods.251:123-35), express in the yeast cell of Hansenula anomala (Hanseula) or yeast (Saccharomyces).

In one embodiment, antibody, especially for example the full length antibody of IgG produces in mammalian cell.Be used for lymphocyte strain that recombinant expressed exemplary mammalian host cell comprises Chinese hamster ovary cell (Chinese hamster ovary celI) (comprise the dhfr-CHO cell that is described in Urlaub and Chasin (1980) Proc.Natl.Acad.Sci.USA 77:4216-4220, use with the DHFR selective marker that (for example) is described among Kaufman and Sharp (1982) Mol.Biol.159:601-621), for example NSO myeloma cell and SP2 cell, COS cell, K562 and from the cells of the transgenic animal of for example transgene mammal.For example, described cell is a mammary epithelial cell.

Except that the nucleotide sequence in coding immunoglobulin (Ig) territory, recombinant expression vector can have other such as the sequence of duplicating (for example, replication orgin) of regulating carrier in the host cell and the sequence of selectable marker gene.Selectable marker gene be beneficial to the host cell of introducing carrier selection (referring to, for example United States Patent (USP) the 4th, 399, No. 216, the 4th, 634, No. 665 and the 5th, 179, No. 017).Exemplary selectable marker gene comprises Tetrahydrofolate dehydrogenase (DHFR) gene (for using) and neo gene (selecting to use for G418) in the dhff host cell with the selection/amplification of methylamine petrin.

Being used for antibody one (for example, full length antibody or its antigen-binding portion thereof in) the recombinant expressed illustrative system, introduces the recombinant expression vector of encoding antibody heavy chain and light chain of antibody in the dhfr-CHO cell by the transfection of calcium phosphate mediation.In recombinant expression vector, each operability of heavy chain of antibody and light chain gene (for example is connected in the enhancers/promoters regulatory element, be derived from SV40, CMV, adenovirus etc., such as cmv enhancer/AdMLP modulator promoter element or SV40 enhanser/AdMLP modulator promoter element) transcribe with the high level that drives gene.Recombinant expression vector also has the DHFR gene, and described gene allows to use the selection/amplification of methylamine petrin to select to have used the Chinese hamster ovary celI of carrier transfection.Cultivate selected transformed host cell to allow expressing antibodies heavy chain and light chain and to reclaim complete antibody from substratum.Use standard molecular biological technique to prepare recombinant expression vector, transfection host cell is selected transformant, cultivates host cell and reclaims antibody from substratum.For example, some antibody can separate with a-protein or protein G by the affinity chromatograph method.

With regard to the antibody that comprises the Fc territory, antibody producing system can synthesize wherein the Fc district by glycosylated antibody.For example, the l-asparagine 297 places glycosylation of the Fc territory of IgG molecule in the CH2 territory.The site of described l-asparagine for modifying with two feeler type oligosaccharides.Described glycosylation participates in effector function (Burton and Woof (1992) Adv.Immunol.51:1-84 by Fc γ acceptor and complement Clq mediation; People such as Jefferis, (1998) Immunol.Rev.163:59-76).The Fc territory can result from and make corresponding in the suitable glycosylated mammalian expression system of the residue of l-asparagine 297.The Fc territory also can comprise other eucaryon and translate the back modification.

Antibody also can be including (for example) the modification that changes the Fc function.For example, IgG 1 constant region can be at one or more residue places, for example one or more place's sudden changes in the residue 234 and 237 (for example according to US 5,648, the number in 260).Other exemplary modification comprises and is described in US 5,648, the modification in 260.

Antibody also can be produced by transgenic animal.For example, U.S 5,849, and 992 are described in the method for expressing antibodies in the mammary gland of transgene mammal.Transgenosis comprises the nucleic acid of emulsion specificity promoter and the antibody paid close attention to of coding and is used for the excretory signal sequence through structure.Emulsion by the female generation of described transgene mammal comprises the antibody of secreting in wherein of being paid close attention to.Can be with antibody from the emulsion purifying, or with regard to some application, can directly use.

Also might (for example) use (for example) to have the animal of the natural mankind or groups of people's immunoglobulin like protein locus by immunity generation and hK1 bonded antibody.In one embodiment, the non-human animal comprises the immunoglobulin gene to a minority.For example, might make mouse antibodies produce defective mouse species through engineering approaches with the big fragment of human Ig locus.Use hybridoma technology, can produce and select to be derived from antigen-specific monoclonal antibody with the specific gene of wanting.Referring to, XenoMouse for example ^TM, people such as Green, Nature Genetics 7:13-21 (1994); U.S.2003-0070185 and WO 96/34096.

HK1 can be used as immunogen in whole or in part.

The non-human antibody also can be modified to comprise the replacement of inserting human immunoglobulin sequence, for example, at for example more than one or one (preferably at least 5,10,12 or all) the consistent human amino acid residue at the specific position place of column position down: (in the FR of the variable domain of light chain) 4L, 35L, 36L, 38L, 43L, 44L, 58L, 46L, 62L, 63L, 64L, 65L, 66L, 67L, 68L, 69L, 70L, 71L, 73L, 85L, 87L, 98L and/or (in the FR of the variable domain of heavy chain) 2H, 4H, 24H, 36H, 37H, 39H, 43H, 45H, 49H, 58H, 60H, 67H, 68H, 69H, 70H, 73H, 74H, 75H, 78H, 91H, 92H, 93H and/or 103H (according to the Kabat numbering).Referring to, U.S.6 for example, 407,213.

Exemplary human antibodies

In one embodiment, hK1 binding antibody as herein described comprises one or more human frame sequences, for example human or effectively human FR1, FR2, FR3 and/or the FR4 in heavy chain and/or the light chain variable territory sequence.

In one embodiment, the heavy chain variable domain sequence has H1 and H2 hypermutation ring, described hypermutation ring have the 1-3 canonical structure (according to people such as Chothia, (1992) J.Mol.Biol.227:799-817; People such as Tomlinson, (1992) J.Mol.Biol.227:776-798).With regard to FR1, FR2 and FR3, the exemplary framework compatible with described structure comprises following sequence respectively;

EVQLVESGGGLVQPGGSLRLSCAASGFTF WVRQAPGKGLEWVA RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR

EVQLVESGGGLVQPGRSLRLSCAASGFTF WVRQAPGKGLEWVS RFTISRDNAKNSLYLQMNSLRAEDTALYYCAKD

QVQLVESGGGLVKPGGSLRLSCAASGFTF WIRQAPGKGLEWVS RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR

EVQLVESGGGVVRPGGSLRLSCAASGFTF WVRQAPGKGLEWVS RFTISRDNAKNSLYLQMNSLRAEDTALYHCAR

EVQLVESGGGLVKPGGSLRLSCAASGFTF WVRQAPGKGLEWVS RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR

EVQLLESGGGLVQPGGSLRLSCAASGFTF WVRQAPGKGLEWVS RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK

QVQLVESGGGVVQPGRSLRLSCAASGFTF WVRQAPGKGLEWVA RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK

QVQLVESGGGVVQPGRSLRLSCAASGFTF WVRQAPGKGLEWVA RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR

QVQLVESGGGVVQPGRSLRLSCAASGFTF WVRQAPGKGLEWVA RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK

QVQLVESGGGVVQPGRSLRLSCAASGFTF WVRQAPGKGLEWVA RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR

EVQLVESGGVVVQPGGSLRLSCAASGFTF WVRQAPGKGLEWVS RFTISRDNSKNSLYLQMNSLRTEDTALYYCAKD

Usually, compatible with the 1-3 canonical structure framework from any human reproductive tract antibody sequence can be used for heavy chain.Similarly, with regard to light chain CDR separately, the optional self-consistent human reproduction of light chain framework is an antibody sequence.The framework of VH or VL section can be used for FR1, FR2 and FR3.

The generation of target protein

HK1 can produce by the recombination and expression techniques of for example expressing in intestinal bacteria or pichia pastoris phaff (Pichia pastoris).Basically as people such as Irie, (1986) Biochem.Int.13,375-382 is described, and human kallidinogenase hK1 also can be purified to homogeneous in urine.

In conjunction with calibrating

ELISA。Use the ELISA calibrating can be just for example in conjunction with the interactive property assessment of character for example can with the protein of the interactional antibody of hK1.For example, make each protein contact bottom surface scribble the microtiter plates of target (target of for example limiting the quantity of).Described dish is washed to remove the polypeptide of non-specific binding with damping fluid.Subsequently, measure the proteinic amount that is incorporated into described dish by surveying described dish with the antibody that can discern polypeptide (for example mark part of polypeptide or constant portion).Described antibody is the enzyme that is connected in such as alkaline phosphatase, and when suitable substrate was provided, it produced the colorimetric product.Can be with protein from cell purification or to present library format, for example be form calibrating with the fusion of filobactivirus sheath.Perhaps, be applied in the microtiter plates and be used for testing being present in and present the storehouse or the avidity by peptide/antibody of selecting to obtain in presenting the storehouse expressing the cell of the target molecule of hK1 for example (for example, live or fixed).

In another pattern of ELISA calibrating, use the different holes of each the polypeptide coating microtiter plates in the diversity chain storehouse.Subsequently, use the constant target molecule to carry out ELISA to detect each hole.

Calibrating all combines.The binding interactions of candidate albumen matter and target can use the homogeneous phase calibrating to analyze, and for example after all components that adds calibrating, does not need other liquid-phase operation.For example, fluorescence resonance energy transmission (FRET) can be used as homogeneous phase calibrating (referring to, for example, people such as Lakowicz, United States Patent (USP) the 5th, 631, No. 169; Stavrianopoulos waits the people, United States Patent (USP) the 4th, 868, No. 103).If contiguous first molecule of second molecule, the fluorescent mark of then selecting fluorophore mark (for example, the molecule that identifies in elution part) on first molecule so that its emitted fluorescence energy to can be on second molecule (for example, target) absorbs.When the fluorescent mark on second molecule absorbed energy delivered, it sent fluorescence.Because the efficient of the transmission ofenergy between the mark and the distance dependent of separating molecule, so can assess spatial relation between the molecule.Taking place between the molecule under the bonded situation, the fluorescent emission of ' acceptor ' molecule marker should be maximum in the calibrating.Be configured to can measure easily by the standard fluorescence detection method of knowing in the affiliated field (for example, using photofluorometer) by the FRET monitoring in conjunction with the result.By the amount of the titration first or second binding molecule, can produce binding curve with the estimation equilibrium association constant.

Another example of homogeneous phase calibrating is α screening (Packard Bioscience, Meriden CT).Two beads through mark are used in the α screening.A bead produces creating singlet oxygen by using when Stimulated Light excites.When creating singlet oxygen by using contacted from the diffusion of first bead and with another bead, another bead produced optical signal.Only when two beads were contiguous, described signal just produced.A bead can be connected in and present the library member, another person can be connected in target.Measurement signal is to measure the bonded degree.

When candidate albumen matter be connected in phage for example present the storehouse carrier time or use candidate albumen matter as free molecule, can carry out the homogeneous phase calibrating.

Surface plasma resonance (SPR).Can use SPR to be analyzed from the binding interactions that presents isolating molecule in storehouse and target.SPR or biomolecular interaction analysis (BIA) detection of biological specificity in real time interact, and need not in the labeling reaction any.Quality change on the mating surface of BIA chip (indication is in conjunction with the result) causes the refraction index changing (optical phenomena of surface plasma resonance (SPR)) near the light on described surface.Change of refractive produces detectable signal, and described signal measurement is the indication of the real time reaction between the biomolecules.Use the method for SPR to be described in following each document, for example United States Patent (USP) the 5th, 641, No. 640; Raether (1988) Surface Plasmons SpringerVerlag; Sjolander and Urbaniczky (1991) Anal.Chem.63:2338-2345; People such as Szabo, (1995) Curr.Opin.Struct.Biol.5:699-705 and by BIAcore International AB (Uppsala, the online resource that Sweden) provides.

Bonded equilibrium dissociation constant (the K that biomolecules and target are provided providing from the information of SPR _d) and comprise K _OnAnd K _OffThe accurate and quantitative measurement of kinetic parameter.Described data can be used for the comparison different biological molecules.For example, can be relatively to differentiate target be had high-affinity or has slow K by the protein of the nucleic acid encoding in the storehouse that is selected from the diversity chain _OffIndividuality.Described information also can be used for illustrating structure-activity relation (SAR).For example, can be relatively with the parameter of the kinetics of the ripe pattern of parent protein and balance incorporating parametric and described parent protein.Can identify the variant amino acid of given position and for example high-affinity and slow K _OffThe special combination parameter correlation.Can be with described information and structure modeling combination (for example, the structure determination of using homology modelization, energy minimization or being undertaken) by crystallography or NMR.Therefore, the physical interaction between protein and its target being understood can formulism and be used to instruct other method of design.

Protein array.Can be fixed on the solid phase support thing presenting the polypeptide that the storehouse identifies certainly, for example, on bead or array.With regard to protein array, in the polypeptide each all is fixed on the unique address on the upholder.Usually, described address is a two-dimensional address.Protein array is described below (referring to, diagnostics for example).

Bioassay

Can use the in vitro proteoclastic inhibition of biological chemistry calibrating assessment hK1.Referring to, people such as Bourgeois for example, (1997) J.Biol.Chemistry 272:29590-29595.Can be used for monitoring the proteoclastic exemplary peptide substrates of hK1 comprises: Abz-FRSARQ-EDDnp, Abz-TFRSARQ-EDDnp, Abz-FTFRSARQ-EDDnp, Abz-AIKFFSAQ-EDDnp, Abz-VIAGRSLNPNQ-EDDnp, Abz-AMSRMSLSSFSVNRQ-EDDnp, Abz-AIKFFSRQ-EDDnp and Abz-TFFSARQ-EDDnp; wherein Abz represents that o-amino benzoyl acyl group and EDDnp represent N-(2, the 4-dinitrophenyl) quadrol.The fluorescence peptide of intramolecularly cancellation can be synthetic by classical solution; Glutamine can be located as the C-terminal residue.Referring to, people such as Hirata for example, Lett.Pept.Sci.1,299-308.Synthetic available Fmoc method uses multiple automated peptide synthesizer (PSSM-8, Shimadzu Co.) to carry out.Intramolecularly cancellation fluorogenic substrate can be prepared as in N the 2mM stock solution in the dinethylformamide and with activation damping fluid dilution.Substrate purity can (TofSpec-E Micromass) and by the 10min linear gradient of the 0-60% acetonitrile that is used for 0.075%TFA checks with the C18 tubing string reverse-phase chromatography of 2.0mL/min elution by the MALDI-TOF mass spectroscopy.

The exemplary condition of assessment enzyme kinetics is at the 50mM Tris-HCl damping fluid (pH8.3) that contains 1mM EDTA and 0.02%IGEPAL (before being Nonidet P-40) under 37 ℃.

Animal model

US5,911,988 describe the exemplary mouse model that is used for asthma.Described mouse model can be used for assessment and also discerns the protein-bonded ability of the proteic hK1 of muroid animal KLK1.Mouse also can be through transforming to express hK1.Mouse is stood long-term multiple anaphylactogen and sucks and can be used for assessing the long-time effect of anaphylactic disease in the lung and be used for describing over-reactive cell, mechanism, molecule and the amboceptor that relates to of inducing of tracheae of human lung.

Can use female BALB/c mouse (6-8 week) and it is maintained under the known condition that is used to study.

Reagent: available from Pierce Chem.Co. (Rockford, 111.) Crystalline OVA, from Sigma Chem.Co. (St.Louis, Mo.) potassium aluminium sulfate (alum), from Baxter, Healthcare Corporation (Deerfield, 111.) pyrogen-free distilled water, from lymphomed (Deerfield, 111.) 0.9% sodium-chlor (physiological saline) and from Cyclodextrin Technologies Development, Inc. (Gainesville, TRAPPSOL Fla.) ^TMHPB-L100 (moisture hydroxypropyl beta cyclodextrin; The 45 weight/volume % aqueous solution).

The malicious scheme of anaphylactogen immunity/attack.Mouse is accepted OVA and alum (the J.Exp Med.1996 of 0.2ml (100 μ g); Peritoneal injection 184:1483-1494).Can use different schemes.For example, in not accepting respectively on the same day with 100 μ gOVA noses in 0.05ml physiological saline (i.n.) administration and with 50 μ gOVA intranasal administrations in 0.05ml physiological saline before, available intraperitoneal give his life (ketamine) of the gram of 0.2ml in physiological saline (0.44mg/ml)/xylazine (6.3mg/ml) makes mouse anesthesia.HK1 is conjugated protein for assessment, (for example) before intranasal administration is attacked poison, can with protein with conjugated protein throwings of the hK1 of proof load with, and compare with the corresponding control mice of accepting not comprise the protein-bonded contrast dispensing of hK1.

Histology.Obtain tracheae and left lung (right lung is used for bronchoalveolar lavage (" BAL ")) and at room temperature it is fixed 6 to about 15 hours in 10% neutral formalin solution.After being embedded in the paraffin, tissue is cut into section and available coloured differently or the immune labeled further processing of 5 μ m.Discombe has a liking for Yihong blood cell dyeing can be used from the counting cells number with methylene blue counterstain one.Per unit tracheae area (2,200 μ m ²) Yihong blood cell number of having a liking for can be by morphometry (J.Pathol.1992; 166:395-404; Am Rev RespirDis.1993; 147:448-456) measure.Fibrosis can be differentiated with the Masson trichrome staining.Tracheal mucus can be differentiated by following staining: methylene blue, phenodin and eosin, mucoitin carminum (mucicarmine), Ai Er Xinlan (alcian blue) and Ai Er Xinlan/Guo Diansuan-Xi Fushi (periodic acid-Schiff) (PAS) react (Troyer, H., " Carbohydrates " in Principles andTechniques of Histochemistry, Little, Brown and Company, Boston, Mass., 1980:89-121; Sheehan, D.C waits the people, " Carbohydrates " in Theory and Practice of Histotechnology, BattlePress, Columbus, Ohio, 1980:159-179).Mucoitin can be dyeed with the mucoitin carmine solution; Use the tropeolin G counterstain.Acid mucoitin and sulfuric acid mucosubstance can be dyeed with Ai Er Xinlan (pH2.5); Also can use nuclear fast red counterstain.Neutral and acidic mucosubstance is differentiated by Ai Er Xinlan (pH2.5) and PAS reaction.The degree of mucus plugging tracheae (diameter is 0.5-0.8mm) also can be evaluated by morphometry.Will because of the obstruction percentage of mucous tracheal diameter by 0 to 4+ sxemiquantitative scale classification.Histology and morphometric analysis can be carried out by the individuality to conceptual design the unknown.

Pulmonary function test (pft).The 28th day, throwing and physiological saline or OVA were 24 hours afterwards in the last nose, can in vivo measure for example as discussed previously (10,1958 by the plethysmography method through the lung mechanism of intravenously input methacholine in mouse; 192:364-368; J.Appl.Physiol.1988; 64:2318-2323; J.Exp.Med.1996; 184:1483-1494) or use its modification.When pulmonary function test (pft) is finished, can be by cardiac puncture to the blood drawing of each mouse and use lung tissue to be used for further analysis with tracheae.

Bronchoalveolar lavage.After main flow segmental bronchus place is with left lung knotting, right lung can be used 0.4ml physiological saline lavation 3 times.Will from bronchoalveolar lavage (BAL) the liquid cell of the 0.05ml aliquots containig of compiling sample use the hemocytometer counting and under 4 ℃ with remaining liq under 200g centrifugal 10 minutes.Supernatant liquor is stored under-70 ℃ until carrying out the analysis of class arachic acid.With after the cell lump resuspending is in the physiological saline that contains 10% bovine serum albumin (" BSA "), can on glass slide, make the BAL cell smear.For having a liking for Yihong blood cell dyeing, with dry slide glass Discombe diluted liquid (the 0.05% eosin aqueous solution in distilled water and 5% acetone (volume/volume); J.Exp.Med.1970; 131:1271-1287) dyeing is 5-8 minute, with water rinse 0.5 minute and with 0.07% methylene blue counterstain 2 minutes.

The calibrating of tracheal mucus glycoprotein.WillMGP in the BAL liquid is by slit ink dot method and PAS staining (Anal.Biochem.1989; 182:160-164; Am.J.Respir.Cell Mol.Biol.1995; 12:296-306) calibrating.Be placed in Minifold II72-hole slit ink dot equipment (Schleicher﹠amp; Schuell) before in distilled water and subsequently in physiological saline with nitrocellulose membrane (0.2-.mu.m aperture; Schleicher﹠amp; Schuell, Keene, N.H.) wetting.Aliquots containig (0.05-0.751) (Am.J.Respir.Cell Mol.Biol.1991 with the stock solution (2.mu.m/ml) of BAL liquid sample (0.05ml) and human breathing mucoitin glycoprotein; 5:71-79) pass through water aspiration vacuum point sample on the Nitrocellulose film, and MGP is manifested by the PAS reaction.Can carry out the reflection density assay method to quantize PAS dyeing.Can the reactive bulk strength of the PAS of BAL sample be quantized by making comparisons with the typical curve of human breathing mucoitin.

Immunocytochemistry.Can use monoclonal antibody CD11c (DAB method) and Mac1 (Beringer Mannheim, ABC method with Hitomouse test kit (Zymed)) differentiates in vasculature, tracheae and the fibrosis zone/on every side inflammatory cellular type, for example dendritic cell, scavenger cell and lymphocyte.

US 6,462, and 020 description is used for arthritic exemplary mouse model, induces II Collagen Type VI sacroiliitis mouse model.It is conjugated protein to inducing the effect of the arthritic tissue of II Collagen Type VI, radiography and clinical manifestation that described mouse model can be used for assessing hK1.

Autoimmune disorders causes the morbid state and the functional impairment of waiting a moment property significantly.The sacroiliitis of various ways is represented autoimmune disorders family.In clinical field, rheumatoid arthritis (RA) is the serious joint abnormal diseases of most common form.RA is a PD.

The histopathology that is present in the sacroiliitis pathology of the histopathology of the sacroiliitis pathology among the muroid animal CIA and the RA in the human patients has great similarity.Muroid animal CIA is the applicable models of the potential therapeutic treatment of research RA.

Material and method.Mouse: DBA/1 (2) male mice (Jackson Laboratories, Bar Harbor, Me. or the B﹠amp of heavy 25gms; K Universal, Kent Wash.) is used for described research.The described strain of mouse is easy to infect CIA by injection allos II Collagen Type VI.Bovine collagen (BC), complete Freund's adjuvant (Complete Freund ' s Adjuvant) (CFA) and incomplete Freund's adjuvant (Incomplete Freund ' s Adjuvant) (ICFA) can be available from Sigma Chemical.The antigen that will be used for immunity is allocated with the 0.1M acetic acid treatment and with CFA or ICFA.

Arthritic inducing.Immunization protocol: during studying, give the II Collagen Type VI of injected in mice 100 μ g in CFA with predetermined space.

Check the development of mouse arthritis with predetermined space.The swelling and the erythema of at least one toe joint on the forward foot in a step and/or the rear foot when arthritic supposition evidence comprises twice day-night observation.

Arthritic affirmation diagnosis.The histological examination in joint: the toe joint of the animal that will put to death in appropriate intervals removes, fixes, decalcification, be embedded in paraffin, section and dyeing to observe general cell and constitutional features and to detect the cartilage matrix of the pannus in each joint suitably the time.Cellularity degree and inflammation area by the using-system photomicrography digitizing and use aforesaid standard area and the some counting technology quantitative.

The radiography assessment of carrying out toe joint is to detect the sickness rate that changes with joint after the immunity of II Collagen Type VI.Transform the mammography imaging system and be used for described research.The average area of the soft tissue in joint (pannus) is by the assay determination of computer numeral radiography, measures the variable density of adjacent sclerous tissues simultaneously by the internal standard substance comparison that is comprised with each radiography.For baseline control, can use other mouse and comparison density and area data through the identical time as sclerous tissues's variable density and pannus area.Use the significance of the difference of the density of antithesis T test evaluation control mice and experiment mice and area at each time point.

The sacroiliitis assessment.Observe the outbreak of joint of animal inflammation every day.Arthritis index draws by 0 to 4 scale classification by seriousness that each claw is involved.Based on the degree of periarticular erythema and oedema and joint deformity and score.Also quantize the swelling of hind foot by the thickness of measuring ankle from internal malleolus to external malleolus with the constant-tension capplipers gauge.

People such as Tuohy (J.Immunol. (1988) 141:1126-1130), people such as Sobel (J.Immunol. (1984) 132:2393-2401) and Traugott (Cell Immunol. (1989) 119:114-129) describe experimental autoimmune encephalitis (EAE) mouse model that is used for multiple sclerosis.It is conjugated protein that described mouse model can be used for assessing the hK1 of the symptom of regulating multiple sclerosis.

Medical composition

Feature of the present invention also is a kind of composition, for example pharmaceutically acceptable composition, and its hK1 that for example comprises (for example) hK1 binding antibody as described herein is conjugated protein.As used herein, " medical composition " contained and is used for diagnosing the compound (for example, tagged compound) of (for example, in vivo imaging) purposes and is used for the treatment of or the compound of preventive use.

As used herein, " pharmaceutically acceptable supporting agent " comprise any and all solvents, dispersion medium, dressing, antibiotic and anti-mycotic agent, etc. open and absorption delay agent and physiology on compatible analogue.In one embodiment, described supporting agent is not a water.Preferably, described supporting agent is applicable to intravenously, intramuscular, subcutaneous, non-through intestines, vertebra or epidermis dispensing (for example by injection or transfusion).According to dosing way, active compound can be coated in the material and can make the effect of the natural condition of described compound inactivation with other to protect described compound to avoid acid.

" pharmaceutically acceptable salt " refer to keep parent compound the biological activity of wanting and do not give the salt (referring to, Berge for example, S.M. waits the people, (1977) J.Pharm.Sci.66:1-19) of any improper toxicological effect.The example of described salt comprises acid salt and base addition salt.Acid salt comprises derived from the acid salt of the nontoxic mineral acid of all example hydrochloric acids, nitric acid, phosphoric acid, sulfuric acid, Hydrogen bromide, hydroiodic acid HI, phosphorous acid etc. with derived from the acid salt of the non-toxic organic acid of alkanoic acid, hydroxy alkanoic acid, aromatic acid, aliphatic sulfonic acid and the aromatic sulfonic acid etc. that replace such as mono carboxylic acid of aliphatic series and aliphatic dicarboxylic acid, through phenyl.Base addition salt comprises derived from the base addition salt such as the alkaline-earth metal of sodium, potassium, magnesium, calcium etc., with derived from such as N, the non-toxic organic amine of N '-dibenzyl-ethylenediamin, N-methylglucosamine, chloroprocaine (chloroprocaine), choline, diethanolamine, quadrol, PROCAINE HCL, PHARMA GRADE (procaine) etc. base addition salt

Composition of the present invention can take various forms.Described form is including (for example) liquid, semisolid and solid dosage form, such as liquor (for example, but injectable solution and infusion solution), dispersion liquid or suspension, lozenge, pill, pulvis, liposome and suppository.Preferred form is decided according to institute's desire dispensing pattern and treatment application.But typical preferred composition is injectable and infusion solution form, is used for throwing composition with human composition with antibody such as being similar to.The preference pattern of dispensing is non-through intestines dispensings (for example, intravenously, subcutaneous, intraperitoneal, intramuscular).In a preferred embodiment, compound through intravenous fluids or injection throw with.In another preferred embodiment, compound through intramuscular or subcutaneous injection throw with.

As used herein phrase " non-through intestines throw with " means the dispensing pattern of removing through intestines or topical administration, usually by injection, and including (but not limited to) in intravenously, intramuscular, intra-arterial, the sheath, in the capsule, in the eye socket, intracardiac, intradermal, intraperitoneal, under tracheae, subcutaneous, epidermis, under the intraarticular, capsule, under the arachnoid membrane, in the vertebra, epidural and breastbone inner injection and transfusion.

Medical composition be necessary for aseptic usually and make and condition of storage under stablize.Medical composition is management and the industry standard to guarantee that it meets dispensing after tested also.For example, according to USP 24/NF 19 methods, level of endotoxin in the preparation can use the molten born of the same parents' quality testing of king crab amoebocyte fixed (Limulus amebocyte lysate assay) (for example, using the test kit from Bio Whittaker lot#7L3790, susceptibility 0.125 EU/mL) test.The sterile state of medical composition can use thioglycolate medium to measure according to USP 24/NF 19 methods.For example, described preparation is used to inoculate described thioglycolate medium and it was cultivated 14 days or longer down at 35 ℃.The periodic test substratum is to detect microbial growth.

Composition can be allocated as solution, microemulsion, dispersion liquid, liposome or other is suitable for the ordered structure of high drug level.Sterile injectable solution can be by incorporating the active compound of aequum in the appropriate solvent into above cited a kind of composition or its combination on demand, and then filtration sterilization prepares.Usually, dispersion liquid prepares by active compound being incorporated into contain in alkaline dispersion medium and required other aseptic supporting agent from the composition in the above cited composition.Under the situation of the sterile powder that is used to prepare sterile injectable solution, the preferred preparation method is vacuum-drying and lyophilize, its powder that produces activeconstituents add any other desired from it before through the composition of the solution of sterile filtration.The adequate liquidity of solution can (for example) be kept such as the dressing of Yelkin TTS by using, under the situation of dispersion liquid, by keeping required granularity and passing through to use tensio-active agent.The prolongation of Injectable composition absorbs and can comprise the medicament that for example delay of Monostearate and gelatin absorbs in the composition and reach by making.

The exemplary methods that allotment is used for the antibody sent through suction is described below.

In one embodiment, the hK1 binding antibody through allotment with through subcutaneous delivery, for example, subcutaneous injection.Can use subcutaneous delivery to treat any illness as herein described.In one embodiment, antibody is provided as the stable freeze-dried protein composite that comprises the water-soluble protective material sugar of sucrose or trehalose (for example, such as).But freeze-drying composite rehydration with produce stable have than the protein concn in the pre-freeze-drying composite significantly high (for example, high about 2-4 doubly, preferably high 3-10 doubly and more preferably high 3-6 times) the rehydration composite of protein concn.Especially, may be when the protein concn in the pre-freeze-drying composite for 5mg/mL or when lower, the protein concn in the rehydration composite can be 50mg/mL or higher.Therefore, composite can comprise at least about the hK1 of 50mg/mL conjugated protein.Described increased protein concentration in the rehydration composite is applicable to through subcutaneous administration.

Described rehydration composite can be etc. to be opened.It can comprise sanitas (such as the injection bacteriostatic water, BWFI).The rehydration composite can be used as the multi-usage composite.Described composite be applicable to (for example) needs of patients frequently through subcutaneous throwing and protein to treat the situation of chronic medical science symptom.Water-soluble protective material and proteinic ratio can (for example) be about 100-1500 mole trehalose or sucrose: 1 mole of antibody.Damping fluid can be used for stablizing the pH value of pre-freeze-dried preparation, for example with the pH value stabilization in the 6-8 scope.For example, damping fluid can comprise one or more for example amino acid of Histidine, or other cushions or have the medicament of the pKa in the described scope in the 6-8 scope.Composite can further comprise tensio-active agent (for example, polysorbate).

Compound as herein described can by the known several different methods in affiliated field throw with.For multiple application, the approach/pattern of dispensing is intravenous injection or transfusion.For example, with regard to treatment is used, compound can by intravenous fluids with throw less than the speed of 30mg/min, 20mg/min, 10mg/min, 5mg/min or 1mg/min with to reach about 1 to 100mg/m ²Or 7 to 25mg/m ²Dosage.The approach and/or the pattern of dispensing should change according to desired result.In certain embodiments, active compound can be used the protection compound in order to avoid the supporting agent of snap-out release prepares, such as the controlled release composite that comprises implant and micro encapsulation seal delivery system.Can use biodegradable, biocompatible polymeric, such as ethene-vinyl acetate copolymer, poly-acid anhydrides, polyglycolic acid, collagen, poe and poly(lactic acid).The method of the described composite of multiple preparation has been patented or has been generally known.Referring to, Sustained and Controlled ReleaseDrug Delivery Systems for example, J.R.Robinson compiles, Marcel Dekker, Inc., New York, 1978.Medicine is allocated as many confessed technology, and is further described in following each document: Gennaro (volume), Remington:TheScience and Practice of Pharmacy, the 20th edition, Lippincott, Williams﹠amp; Wilkins (2000) (ISBN:0683306472); People such as Ansel, Pharmaceutical Dosage Forms and Drug Delivery Systems, the 7th edition, Lippincott Williams﹠amp; Wilkins Publishers (1999) (ISBN:0683305727) and Kibbe (volume), Handbook of Pharmaceutical Excipients American Pharmaceutical Association, the 3rd edition, (2000) (ISBN:091733096X).

In certain embodiments, composition can (for example) with inert diluent maybe can absorb edible supporting agent oral administration with.In compound (in case of necessity and other composition) duricrust or the soft shell gelatin capsules of also can packing into, be pressed into lozenge or directly incorporate in person under inspection's the diet.With regard to per os therapeutic dispensing, compound can merge with vehicle and use with following form: can absorb lozenge, buccal tablet, suck ingot, capsule, elixir, suspension, syrup, wafer etc.Unless be to throw and compound by the pattern beyond the intestines dispensing, compound may be coated with the material that prevents its inactivation or with compound and the material that prevents its inactivation common throw with.

Medical composition can with the known medical apparatus in affiliated field throw with.For example, in a preferred embodiment, medical composition of the present invention can with the needleless hypodermic injection unit throw with, described device is such as being exposed in United States Patent (USP) the 5th, 399, No. 163, the 5th, 383, No. 851, the 5th, 312, No. 335, the 5th, 064, No. 413, the 4th, 941, No. 880, the 4th, 790, No. 824 or the 4th, 596, the device in No. 556.The well-known example of implant of the present invention and assembly that is applicable to comprises: United States Patent (USP) the 4th, 487, and No. 603, its exposure is used for distributing with controllable rate the implantable microinfusion pump of medicine; United States Patent (USP) the 4th, 486, No. 194, its exposure is used for by the therapeutic system of skin throwing with medicine; United States Patent (USP) the 4th, 447, No. 233, it discloses the medical transfusion pump with accurate infusion rate delivering drugs; United States Patent (USP) the 4th, 447, No. 224, its exposure is used for the implantable fluid delivery apparatus of variable flow rate that continuous medicine is sent; United States Patent (USP) the 4th, 439, No. 196, its exposure has the osmotic drug delivery system of multi-cavity chamber compartment; With United States Patent (USP) the 4th, 475, No. 196, it discloses the osmotic drug delivery system.Certainly, also known multiple other described implant, delivery system and assembly.

In certain embodiments, compound can in vivo suitably distribute guaranteeing through allotment.For example, blood-brain barrier (BBB) is repelled multiple high-hydrophilic compound.By BBB (if wanting), then it can be allocated (for example) in liposome for guaranteeing treatment compound of the present invention.With regard to the method for making liposome, referring to, for example United States Patent (USP) 4,522, and 811,5,374,548 and 5,399,331.Therefore liposome can comprise that one or more selectivity are transported to the part in specific cells or the organ, strengthens targeted delivery of drugs (referring to, V.V.Ranade (1989) J.Clin.Pharmacol.29:685 for example).

Test kit is also in category of the present invention, and it comprises composition as herein described (for example, contain the protein-bonded compound of hK1 and for example treat, prevent or diagnose the composition of the working instructions of use).In one embodiment, test kit comprises (a) compound, for example comprises described compound compositions and (b) information material optionally.That described information material can be is descriptive, directiveness, sell usefulness or other is with method as herein described and/or be used for the relevant material of use of the compound of method as herein described.For example, information material is described the method for the compound of throwing or treatment or prevention hK1 associated conditions active with reduction hK1 with compound.

In one embodiment, information material can comprise with for example, and the suitable method (for example with dosage as herein described, formulation or dispensing pattern) of suitable dose, formulation or dispensing pattern is thrown the specification sheets with compound.In another embodiment, information material can comprise the specification sheets of differentiating suitable person under inspection, and described suitable person under inspection is for example human, for example has or is in the hK1 associated conditions or with the mankind of the risk of the excessive hK1 activity illness that is feature.Information material can comprise following information: about the molecular weight of the manufacturing of compound, compound, concentration, expiration date, batch or manufacturing site location information etc.The information material of test kit is not limited by its form.Under multiple situation, the information material of for example specification sheets is provided as the printed matter of printed text for example, graphic and/or photo, for example label or printing sheets.Yet information material also can provide by other form, such as braille, computer-readable material, videograph or audio recording.In another embodiment, the information material of test kit is contact details or contact information, for example actual address, e-mail address, hyperlink, network address or telephone number, wherein the user of test kit can obtain in a large number the information about compound and/or its purposes in method as herein described.Certainly, the form that information material also can any format combination provides.

Except that compound, the composition of test kit can comprise other composition, such as solvent or damping fluid, stablizer or sanitas, and/or is used for the treatment of second medicament of symptom as herein described or illness.Perhaps, other composition be can comprise in the test kit, but composition or the container different are arranged in described compound.In described embodiment, test kit can comprise the specification sheets that compound is mixed with other composition or compound is used together with other composition.

Compound can provide in any form, for example liquid, drying or lyophilized form.Preferably, described compound is pure substantially and/or aseptic.When providing compound with liquor, described liquor is preferably the aqueous solution, and wherein aseptic aqueous solution is preferred.When providing compound with dried forms, rehydration is normally by adding suitable solvent.For example sterilized water or damping fluid equal solvent optionally can be provided in test kit.

Test kit can comprise one or more containers that is used to contain compound compositions.In certain embodiments, test kit contains container, separation scraper or the compartment that independently is used for composition and information material.For example, composition can be contained in bottle, bottle or the syringe, and information material can be contained in plastic jacket or the packing.In other embodiments, the independent component of test kit is contained in the single undivided container.For example, composition comprises in bottle, bottle or the syringe of the information material that is stained with label form thereon.In certain embodiments, individual containers that test kit comprises a plurality of (for example, a bag), it respectively contains the unit dosage (for example, formulation as herein described) of one or more compounds.For example, test kit comprises a plurality of syringes, ampoule, foil packages or blister pack, and it respectively contains the single unitary dose of compound.That the container of test kit can be is airtight, (for example, can not see through the variation of moisture content or steam) of waterproof and/or lighttight.

One wherein compound contain among the embodiment of hK1 binding antibody, the specification sheets inclusion compound that is used for diagnostic use is (for example, for example the sample from the patient's who suffers from tuberculosis or other illness as herein described biopsy samples or cell in) or in vivo detect the purposes of hK1 in vitro.In another embodiment, the specification sheets that is used for the treatment of application comprises (for example suffers from tuberculosis or for example asthma as herein described, allergic asthma and non-allergic asthma), chronic obstructive pulmonary disease (COPD), multiple sclerosis, psoriasis, rheumatoid arthritis, osteopathy sacroiliitis, osteoarthritis, rhinitis, sinusitis, inflammatory bowel (such as Crohn disease and ulcerative colitis), immune-mediated diabetes, acute pancreatitis, interstitial cystitis or superfluous natural disposition illness (for example, transitivity illness or tumor-blood-vessel growth) wait the dosage and/or the dispensing pattern of being advised among the patient of other illness.Test kit can further contain at least a other reagent of taking the circumstances into consideration to allocate in one or more independent pharmaceutical preparations, such as, the diagnostic reagent for example as described herein or the diagnostic reagent of therapeutical agent or therapeutical agent and/or one or more other treat the medicament of described illness.

Treatment

Can be with the conjugated protein throwing of hK1 and for example human person under inspection's person under inspection with treatment, prevention and/or diagnosis various disease conditions, such as the hK1 associated conditions and the disease that is feature with improper or unusual hK1 activity.Exemplary illness (for example comprises asthma, allergic asthma and non-allergic asthma), chronic obstructive pulmonary disease (COPD), multiple sclerosis, psoriasis, rheumatoid arthritis, osteoarthritis, rhinitis, sinusitis, inflammatory bowel, immune-mediated diabetes, acute pancreatitis, interstitial cystitis or other hK1 associated conditions such as Crohn disease and ulcerative colitis, or superfluous natural disposition illness (for example, transitivity carcinoma of the pancreas, tumor-blood-vessel growth)." hK1 associated conditions " be wherein hK1 to the illness of small part mediation so that reduce hK1 protein level or active at least one aspect that can improve illness, for example, at least one symptom of illness.

Except that throwing, also compound can be thrown with to cell, tissue or the organ of culture (for example in vitro or exsomatize) with to the person under inspection.

As used herein, term " treatment " is defined as for the purpose of curing, cure, alleviate, alleviate, change, remedy, improve, improve or influence the inducement of the symptom of illness, illness or illness, to for example patient's person under inspection separately or with one or more other medicament combined administration or throwing and compound, or to from for example patient's person under inspection's chorista or for example the cell of cell strain is used or throwing and described medicament, described person under inspection has illness (illness for example as described herein), the symptom of illness or the inducement of illness.

As used herein, the protein-bonded amount of effective sanatory hK1 or " treatment significant quantity " refer to after person under inspection's throwing and single dose or multiple doses, is the amount of compounds effective in treatment among the person under inspection, for example at least one symptom of person under inspection's illness is cured, alleviates, alleviates or is improved to exceed the degree of expecting under the situation that does not have described treatment.For example, described illness can be tuberculosis, tuberculosis for example as herein described.

" local significant quantity " refers in detect regulating tissue target protein (for example, the amount (for example, concentration) of active aspect compounds effective hK1) of (for example being exposed in the zone of lung of hK1).The evidence of regulating can increase including (for example) the amount of substrate, and for example the proteolysis of substrate reduces.

As used herein, effectively the protein-bonded amount of hK1 or proteinic " the prevention significant quantity " of prevention illness refer to after person under inspection's throwing and single dose or multiple doses, are the protein-bonded amount of effective hK1 preventing or postponing aspect the generation of the outbreak of the illness of hK1 associated conditions for example or recurrence.

(for example) term of the quantitative difference between two states of expression " is induced ", " inhibition ", " enhancing ", " raising ", " increase ", " reduction " etc. refer to two differences between the state, significant difference (for example, P＜0.05,0.02 or 0.005) on the statistics for example.

Dosage regimen will be reacted (for example, therapeutic response) through regulating to provide best.For example, can throw and the single bolus, can throw in time and the dosage that separates for several times, or can dosage be reduced in proportion or increase as the emergency state indication of treatment situation.For ease of the homogeneity of dispensing and dosage, non-especially favourable through the intestines composition with the dosage unit form allotment.As used herein dosage unit form refers to be fit to be used for single dose the person under inspection's of institute's desire treatment the discrete unit of physics; Constituent parts contains the active compound that will treat the predetermined amount of effect as calculated with generation that is associated with required medical supporting agent.

In the treatment of compound as herein described or prevention go up effectively that the exemplary non-limiting scope of amount is 0.1-20mg/kg, more preferably be 1-10mg/kg.Compound can by intravenous fluids with throw less than the speed of 20mg/min, 10mg/min, 5mg/min or 1mg/min with to reach about 1 to 50mg/m ²Or about 5 to 20mg/m ²Dosage.Type and the seriousness that should note the symptom that dose value can be alleviated with the institute desire change.Further should be understood that for any special person under inspection, the specific administration scheme should according to individual need and throw with or the people's of the dispensing of supervision group compound professional judgement adjust in time, and dosage range as herein described only is exemplary.

Medical composition can comprise the compound as herein described of " treatment significant quantity " or " prevention significant quantity ", for example comprises the compound of the polypeptide of combination and inhibition hK1.

The treatment significant quantity of composition can be according to causing that such as morbid state, individual age, sex and weight and compound the factor of the ability that will react changes in individuality.The treatment significant quantity also is that wherein treatment is gone up useful effect better than any toxicity of composition or the amount of adverse effect.

As used herein, term " person under inspection " is intended to comprise the mankind and non-human animal.Term of the present invention " non-human animal " comprises all vertebratess, for example nonmammalian (such as chicken, Amphibians, Reptilia) and such as the Mammalss of non-human primate, sheep, dog, cow, pig etc.

Present method can be used in the culture cell of (for example in vitro or exsomatize).Described method can be used as in vivo the part of (for example, treatment or prevention) scheme the cell that is present among the person under inspection is carried out.With regard to embodiment in vivo, contact procedure is to implement and be included in effective permission protein bound to throw with hK1 conjugated protein to the person under inspection under the condition of hK1 in the person under inspection.

Throwing is described in " medical composition " with the method for compound.The suitable dose of employed compound should be decided according to age of person under inspection and weight and employed certain drug.Compound can be used as competitive medicament to suppress, to reduce the improper interaction between (for example) natural substrate and the hK1.

HK1 is conjugated protein also to can be used for sending multiple useful load, for example, comprises molecule, bioprotein and its mixture of compound, plant, fungi or bacterial origin of medicine, the emitted radiation of medicine.HK1 is conjugated protein to make the person under inspection's that useful load target hK1 is positioned at zone.

Enzyme activity toxin and its fragment for example are diphtheria toxin A fragment, the non-binding active fragments of diphtheria toxin, exotoxin A (from Pseudomonas aeruginosa (Pseudomonas aeruginosd)), ricin A chain (ricin A chain), abrin A chain (abrin A chain), the plain A chain (modeccin A chain) of enlightening not, α-sacrin, some tung oil tree albumen (Aleurites fordii protein), some carnation albumen (Dianthin protein), dyers' grapes albumen (Phytolaccaamericana protein) (PAP, PAPII and PAP-S), momordica charantia inhibitor (Morodica charantia inhibitor), Cortex jatrophae toxin (curcin), crotin (crotin), Saponaria officinalis inhibitor (Saponaria officinalis inhibitor), happy peaceful (gelonin) is situated between, NSC-69529 (mitogillin), restrictocin (restrictocin), phenomycin (phenomycin) and enomycin (enomycin).The program description of the peptide activity of preparation immunotoxin is in WO 84/03508 and WO85/03508.The example of the cytotoxin part that can engage with antibody comprises Zorubicin (adriamycin), Chlorambucil (chlorambucil), daunomycin (daunomycin), methylamine petrin (methotrexate), neocarzinostatin (neocarzinostatin) and platinum.

Under the situation of polypeptide toxin, can use recombinant nucleic acid technical construction coding to be the nucleic acid of the conjugated protein and cytotoxin (or its polypeptide fraction) of the hK1 that translates the syzygy form.Subsequently, make recombinant nucleic acid in (for example) cell, express and coded fusion polypeptide is separated.Fused protein can associate with the part physics that (for example) increases the molecular weight of compound for the stable in vivo transformation period, and is connected in part (for example, polymkeric substance) subsequently.

The previous program that protein is engaged with cytotoxic agent of having described.About Chlorambucil is engaged with protein, referring to, Flechner (1973) European Journal of Cancer for example, 9:741-745; People such as Ghose, (1972) British Medical Journal, 3:495-499 and Szekerke wait the people, (1972) Neoplasma, 19:211-215.About daunomycin and Zorubicin are engaged with protein, referring to, Hurwitz for example, people such as E., (1975) Cancer Research, people such as 35:1175-1181 and Arnon, (1982) Cancer Surveys, 1:429-449.About preparation protein-ricin joiner, referring to, U.S.4 for example, 414,148 and Osawa, T. waits (1982) Cancer Surveys, 1:373-388 and reference of wherein being quoted of people.The coupling program also is described among the EP 226 419.

In certain embodiments, the hK1 binding antibody need not to engage and can use with another part, for example, and as full length antibody.

Chronic obstructive pulmonary disease (COPD)Conjugated protein at least one symptom that also can be used for improving chronic obstructive pulmonary disease (COPD) of hK1.Pulmonary emphysema and chronic bronchitis are the part of chronic obstructive pulmonary disease (COPD).It is serious pulmonary disorder and is progressive, betides among the gerontal patient usually.COPD causes the overplumping of the structure that is called alveolar or air bag in the lung.Thereby bubble wall rupture causes the pulmonary respiration ability drop.The patient who suffers from described disease may at first experience and be short of breath and cough.The clinical index of assessment COPD is destructive index, and it measures alveolar membranes damage and emophysematous measuring, and has advised the susceptibility index as LD, and its close reflection function is unusual, especially the loss of elastic recoil.Referring to, Am Rev Respir Dis 1991 Jul for example; 144 (1): 156-9.Described compound can be used for making patient's destructive index to reduce on (for example) statistics amount significantly, for example at least 10%, 20%, 30% or 40%, or reduce at least to 50%, 40%, 30% or 20% of the normal value of the individuality of corresponding age and gender matched.

Composition can be through allotment with other pattern that is used to suck or lung is sent.Therefore, compound as herein described can be thrown with to lung tissue through sucking.Unless otherwise instructed, otherwise as used herein term " lung tissue " refer to any tissue of respiratory tract and comprise the upper respiratory tract and lower respiratory tract.

Therefore, in treatment (for example, reduce, improve) or prevent in the method for one or more symptoms that are associated with the breathing illness, can throw with hK1 conjugated protein, described breathing illness for example: asthma (for example, allergic asthma and non-allergic asthma); Chronic obstructive pulmonary disease (COPD); Relate to the tracheae inflammation, have a liking for that Yihong blood cell increases, the symptom of fibrosis and generation excessive mucus, for example cystic fibrosis and pulmonary fibrosis.

AsthmaThe symptom of asthma including (for example) stridulate, be short of breath, bronchoconstriction, tracheae overreaction, vital capacity reduction, fibrosis, tracheae inflammation and produce mucus.HK1 is conjugated protein to be can be used for improving or at least one symptom of prevention of asthma, for example in the above-mentioned symptom.HK1 is conjugated protein also can combine with another medicament that is used for the treatment of asthma throw with, described another medicament for example, steroid, leukotrienes regulator, long-acting beta-agonist, oral steroid and theophylline are received (nedocromil), sucked to anti-IgE antibodies, Sodium Cromoglicate and nedocromil.

Rheumatoid arthritis (RA).The internal layer inflammation that is characterized as joint and/or other internal of described illness.It typically is chronicly, break out but can comprise.Exemplary symptom comprises the lump (rheumatoid nodules) at (especially the zone of withstanding pressure (for example, the back side of elbow)) under joint inflammation, swelling, dyskinesia, pain, poor appetite, fever, energy loss, anaemia, the skin.HK1 is conjugated protein to be can be used for improvement or prevents at least one symptoms of rheumatoid arthritis.

HK1 is conjugated protein can combine with another medicament that is used for the treatment of rheumatoid arthritis such as NSAID and Aspirin (aspirin), pain killer and reflunomide throw with to help to reduce arthralgia, stiff and swelling.Exemplary medicament comprises disease modification antirheumatic (DMARD), such as methylamine petrin, leflunomide (leflunornide), Beracilline (D-Penicillamine), sulfasalazine (sulfasalazine), golden therapeutical agent (gold therapy), Minocycline HCl (minocycline), azathioprine (azathioprine), Plaquenil (hydroxychloroquine) (with other antimalarial drug), ciclosporin (cyclosporine), Prosorba therapy and biological agent.

Multiple sclerosis (MS).Multiple sclerosis is a central nervous system disease, it is characterized by the loss of inflammation and myelin.The patient who suffers from MS or possible clinically MS can be by confirming as MS diagnoses the standard of the diagnosis of the defined clear and definite clinically MS of symposial to differentiate people such as (, Ann.Neurol.13:227,1983) Poser.In brief, the individuality of suffering from clear and definite clinically MS has had clinical indication or a kind of clinical indication of pathology and the subclinical sign of another independent pathology of two kinds of outbreaks and two kinds of pathologies.Clear and definite MS also can be by the oligoclonal band diagnosis of IgG in the sign of two kinds of outbreaks and the celiolymph or by the combined diagnosis of the oligoclonal band of IgG in the clinical indication of outbreak, two kinds of pathologies and the celiolymph.

Can throw with the hK1 of for example hK1 binding antibody conjugated protein to the patient who suffers from MS or possible clinically MS to improve at least one symptom of MS.Conjugated protein also can the combination with another medicament of hK1 provides, described another medicament is the protein of EFN β-1 α and IFN β-1b, simulation myelin basic protein (for example, for example the synthetic protein of glatiramer acetate (glatiramer acetate) COPAXONE ), reflunomide or mitoxantrone (mitoxantrone) for example.

Effective treatment of multiple sclerosis can be checked by several different modes.Each that satisfies following standard shows that treatment effectively.Use three main standard: EDSS (the functional impairment situation scale of expansion), worsen performance or MRI (nuclear magnetic resonance).EDSS is will be owing to the clinical lesion fractionated method (Kurtzke, Neurology33:1444,1983) of MS.Type and 8 function systems of seriousness assessment with regard to nerve injury.In brief, before treatment, with regard to the lesion assessment patient in the following system: cone, cerebellum, brain stem, sense organ, intestines and bladder, vision, brain and other.Carry out follow-up investigations at the interval of determining.Scale range is that 0 (normally) is to 10 (death owing to MS).Under situation of the present invention, the decline of a full level is defined as effective treatment (Kurtzke, Ann.Neurol.36:573-79,1994).Worsen and be defined as the performance of the new symptom that is attributable to MS and follow suitably new dysautonomia (above-mentioned IFNB MS symposial).In addition, deterioration must continue 24 hours and stablize before this or improved at least 30 days at least.In brief, by the clinicist patient is carried out the standard neurologic examination.According to the variation in the neural equal (NeurologicalRating Scale) (people such as Sipe, Neurology 34:1368,1984), worsen and be slight, moderate or severe.Measure and worsen speed year and do not have the ratio that worsens the patient.If speed or do not have the ratio that worsens the patient having significant difference on the statistics between treatment group and the placebo with regard in described the measuring any one thinks that then therapy is effective.In addition, also can measure time and deterioration time length and the seriousness that worsens for the first time.As therapy, in this regard, measuring of effectiveness is significant difference on the statistics of comparing in the treatment group time of worsening for the first time or time length and seriousness with control group.MRI can be used for using gd-dtpa-Enhanced Imaging to measure active pathology people Ann.Neurol.36:14 such as (, 1994) McDonald or uses T ₂-weighting technique is measured the position and the degree of pathology.

The exemplary symptom relevant with multiple sclerosis comprises: optic neuritis, diplopia, ocular ataxy, the vision dysmetria, internuclear ophthalmoplegia, motion and sound visual illusion, import the pupil defective into, paresis, monoparesis, paraparesis, the half body paresis, / 4th paresis (quadraparesis), paralysis, posterior paralysis, hemiplegia, tetraplegia, / 4th paralysis (quadraplegia), spasticity, dysarthria, amyotrophy, spasm, twitch, a low disease, clonic spasm, myoclonus, muniple myoclonia, the not peaceful syndromes of leg, drop foot, abnormal reflection, paresthesia, numb, neurodynia, nervosa and neurogenic pain, l ' hermitte ' s, the proprioceptive function obstacle, trigeminal neuralgia, ataxia, the purpose vibration, dysmetria, vestibular ataxia, dizzy, ataxia in a minute, myodystonia, dysdiadochokinesia, frequent micturition, the cystospasm state, atonic bladder, detrusor-sphincter muscle dyssynergia, erective dysfunction, ahedonia, frigidity, constipation, just anxious, scoracratia, depressed, cognition dysfunction, dull-witted, the mood swing, emotional lability, euphoria, the bipolarity syndromes, anxiety, aphasia, speech disorder, tired, Wu Tuofu symptom (uhthoff ' s symptom), GERD and somnopathy.

OsteoarthritisOsteoarthritis is a degenerative arthropathy.It is characterized in that the cartilage destruction in the joint, therefore make bone rub each other, cause pain and lost motion.Conjugated protein at least one symptom that can be used for improving or preventing osteoarthritis of hK1.HK1 is conjugated protein can combine with another medicament that is used for the treatment of osteoarthritis such as reflunomide or NSAID throw with.

That be associated with vasculogenesis and superfluous natural disposition illness

In one embodiment, hK1 conjugated protein throw with to subject to regulate vasculogenesis or other process that is associated with tumorigenesis and tumor growth.HK1 is conjugated protein, and the hK1 that especially suppresses the hK1 enzymic activity is conjugated protein, can be used for stoping tumor-blood-vessel growth and/or cancer cells to invade and shift.HK1 inhibitor antibody is inhibitor effectively and optionally, and therefore for being used for the desirable medicament of oncotherapy.

For example, hK1 is conjugated protein (for example to can be used for reducing vasculogenesis, uncontrolled vasculogenesis or improper vasculogenesis)-such as with vascular malformation and cardiovascular disorder (for example, atherosclerosis, restenosis and arteriovenous malformotion) vasculogenesis that is associated, the chronic inflammation disease (for example, diabetes, inflammatory bowel, psoriasis and rheumatoid arthritis), unusual wound repair (for example, viewed unusual wound repair behind the excimer laser ocular surgical), circulatory disorders (for example, Raynaud's phenomenon (Raynaud ' s phenomenon)), acra sclerosis syndromes (crest syndrome) (for example, calcinosis, esophagectasis and dyomotiloty), the dermatology illness (for example, portwine stain, arterial ulcer, systemic vasculitis and scleroderma) or vision illness (for example, the blind disease that causes because of neovascular diseases, neovascular glaucoma, the cornea neovascularization, trachoma, diabetic retinopathy and myopia are degenerated).Referring to, for example Carmeliet and Jain, 2000, Nature, 407:249-257.

Particularly, the vasculogenesis that is associated of the conjugated protein tumorigenesis that can be used for reducing with for example cancer and tumor growth (for example, the growth of optimum, pernicious or metastatic tumo(u)r) of HK1.

The example of carninomatosis disease is including (but not limited to) noumenal tumour, soft tissue neoplasm and transitivity pathology.Example comprises sarcoma, gland cancer and the cancer knurl of various tracts, such as (for example influencing lung, mammary gland, lymph, stomach and intestine, colon) and urogenital tract (for example, kidney, uropoiesis epithelial cell), sarcoma, gland cancer and the cancer knurl of pharynx, prostate gland, ovary, with the gland cancer that comprises malignant tumour, described malignant tumour such as most of colorectal carcinomas, the rectum cancer, renal cell carcinoma, liver cancer, nonsmall-cell lung cancer, pharynx cancer, carcinoma of small intestine, esophagus cancer and other cancer.

Medicable exemplary noumenal tumour comprises: fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovioma, mesothelioma, Ewing's sarcoma (Ewing ' s tumor), leiomyosarcoma, rhabdosarcoma, colorectal carcinoma, carcinoma of the pancreas, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, rodent cancer, gland cancer, syringocarcinoma, sebaceous carcinoma, papillary carcinoma, papillary carcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, liver cancer, cholangiocarcinoma, syncytioma malignum, spermocytoma, embryonal carcinoma, Wilms' tumor (Wilms ' tumor), the neck cancer, tumor of testis, lung cancer, small cell lung cancer, nonsmall-cell lung cancer, bladder cancer, epithelial cancer, neurospongioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic tumor, oligodendroglioma, meningioma, melanoma, neuroblastoma and retinoblastoma.

Hk1 is conjugated protein also to can be used for treating cancer, and for example the malignant tumour of epithelium or endocrine tissue comprises respiratory system carcinoma, gastrointestinal system carcinoma, urogenital system cancer, carcinoma of testis, breast cancer, prostate cancer tumor, endocrine system cancer and melanoma.Exemplary cancer comprises gland cancer, the cancer of the tissue of uterine cervix, lung, prostate gland, mammary gland, head and neck, colon and ovary.

Hk1 is conjugated protein also to can be used for treating for example sarcoma of the malignant tumour of mesenchymal cell derivation.

Described method also can be used for suppressing the hyperplasia/neoplastic cell of hematopoietic origin (for example, derived from bone marrow system, lymphatic system or red blood corpuscle system) or the hyperplasia of its precursor cell.For example, described method can be used for treating various marrow illnesss, comprise acute preceding myeloid leukemia (APML), acute myelogenous leukemia (AML) and chronic lymphocytic leukemia (CML) and (be summarized in Vaickus, L., 1991, among the Crit.Rev.in Oncol./Hemotol.11:267-297).Can by the lymph malignant tumour of present method treatment including (but not limited to) comprise B-be ALL and T-be ALL acute lymphoblastic leukemia (ALL), lymphocytic leukemia (CLL), PL (PLL), hairy cell leukemia (HLL) and WaldenstromShi macroglobulinemia (Waldenstrom ' s macroglobulinemia) (WM).Other form of malignant lymphoma is including (but not limited to) non-Hodgkin lymphomas (non-Hodgkin ' s lymphoma) and its variant, tip t cell lymphoma, adult T cell leukemia/lymphoma (ATL), cutaneous T cell lymphoma (CTCL), large granular lymphocyte leukemia (LGF) and lymphogranulomatosis (Hodgkin ' s disease).

HK1 conjugated protein can with another medicament combination that is used for the treatment of superfluous natural disposition and/or transitivity illness throw with.The example of described other medicament comprises:

(i) other anti-angiogenic agent (for example, the inhibitor of linomide (linomide), integrin alpha v beta 3 function, angiostatin, razoxane (razoxane));

(ii) cytostatic agent, such as antiestrogen (for example, tamoxifen (tamoxifen), toremifene (toremifene), Lei Luoxifen (raloxifene), droloxifene (droloxifene), draw the former times sweet smell (iodoxifene) of trembling), gestogen (for example, megestrol (megestrol acetate)), aromatase inhibitor (Anastrozole (anastrozole) for example, letrozole (Letrozole), boron azine (borazole), Exemestane (exemestane)), hormone antagonist, antiprogestin, androgen antagonist (for example, flutamide (flutamide), Nilutamide (nilutamide), must catarrh amine (bicalutamide), acetate Sai Pulong (cyproterone acetate)), LHRH agonist and antagonist are (for example, acetate goserelin (gosereline acetate), leuproside (leuprolide)), the inhibitor of testis sterone 5 α-dihydro reductase enzyme (for example, finasteride (finasteride)), farnesyl tranfering enzyme (farnesyltransferase) inhibitor, anti-intrusion agent (for example, the inhibitor of inhibitors of metalloproteinase (as Marimastat (marimastat)) and upar function) and the inhibitor of somatomedin function, (described somatomedin is including (for example) EGF, FGF, Thr6 PDGF BB and pHGF, described inhibitor comprises growth factor antibodies, such as the growth factor receptor antibody of AVASTIN  (shellfish is cut down strain monoclonal antibody (bevacizumab)) and ERBITUX  (Cetuximab (cetuximab)), tyrosine kinase inhibitor and serine/threonine kinase inhibitor); With

(iii) be used for anti-hyperplasia/antitumor drug and its combination of medical science oncology, such as metabolic antagonist antifolate, fluorine pyrimidine, purine and neplanocin, the cytosine arabinoside (cytosinearabinoside) of methylamine petrin (for example, as) as 5 FU 5 fluorouracil; Embed antitumor antibiotics (for example, as Dx (doxorubicin) anthracycline (anthracycline), daunomycin (daunomycin), epirubicin (epirubicin) and jaundice element (idarubicin), Mitomycin-C, actinomycin (dactinomycin), Plicamycin (mithramycin)); Platinum derivatives (for example, cis-platinum (cisplatin), carboplatin (carboplatin)); Alkylating agent (for example, mustargen (nitrogen mustard), melphalan (melphalan), Chlorambucil (chlorambucil), busulfan (busulphan), endoxan, ifosfamide nitrosourea (ifosfamide nitrosoureas), thiophene are for sending (thiotepa)); Antimitotic agent (for example, as the vincaleucoblastine (vinca alkaloids) of vincristine(VCR) (vincristine) with as the taxol of TAXOL  Paclitaxel (paclitaxel), TAXOTERE  Docetaxel (docetaxel)) and such as the new microtubule agent of esperamicin (epothilone) analogue, discodermolide analogue and eleutherobin analogue); Topoisomerase enzyme inhibitor epipodophyllotoxin (epipodophyllotoxins), amsacrine (amsacrine), the Hycamtin (topotecan) of Etoposide (etoposide) and teniposide (teniposide) (for example, as); Cell cycle inhibitor (for example, yellow pyridoxol (flavopyridol)); Biological response modifier and such as the proteasome inhibitor of VELCADE  (Velcade (bortezomib)).

The exemplary conditioning agent of VEGF class somatomedin

Various medicaments can be used for regulating the activity of VEGF class somatomedin, for example regulates VEGF activity (for example, by regulating the activity of protein self, its acceptor or other signal component).Described medicament can be thrown and go to live in the household of one's in-laws on getting married with adjusting vasculogenesis and treatment natural disposition and/or transitivity illness with the conjugated protein combination of hK1.

Medicament can be micromolecular inhibitor (for example, do not comprise the compound of peptide bond and/or less than the compound of 700 Dalton molecular weights).Other medicament comprises the compound with VEGF, VEGF-C or its receptors bind, for example micromolecular compound or protein.Again, other medicament is regulated VEGF, VEGF-C or its receptor expression or activity.Exemplary medicament comprises following each thing;

Shellfish is cut down strain monoclonal antibody (AVASTIN ).The approved shellfish is cut down the strain monoclonal antibody and is used for the treatment of some cancer.US 2004-0133357 describes other antibody that can suppress VEGF.

US 2003-0120038 describes can have the active soluble VEGF-receptor fragment of inhibition.2004-0054143 describes has the little peptide fragment that suppresses active VEGF.

The active micromolecular inhibitor of known lot of V EGF.2004-0147449 describes the VEGF-R inhibitor, and it comprises a compounds, for example (4-chloro-phenyl-) [4-(4-pyridylmethyl)-dai piperazine-1-yl] succsinic acid hydrogen ammonium.

SU5416 (plug China fir (semaxanib)) is the micromolecular inhibitor of vascular endothelial growth factor (VEGF) acceptor-2 and KIT receptor tyrosine kinase.Referring to, people such as Heymach for example, Clin Cancer Res.2004 Sep1; 10 (17): 5732-40.SU11248 is another exemplary inhibitor.Referring to, Mendel (2003) Clin CancerRes.9 (1): 327-37 for example.

4-[4-(1-amino-1-methylethyl) phenyl]-2-[4-(2-morpholine-4-base-ethyl) phenyl amido] pyrimidine-5-nitrile (JNJ-17029259) but and the selectivity nanomole concentration inhibitor of other Tyrosylprotein kinase that relates in the vascular endothelial growth factor receptor-2 (VEGF-R2) that utilizes for per os of the related compound in the 5-cyanopyrimidine structured sort and the vasculogenesis such as platelet derived growth factor receptor, fibroblast growth factor receptor, VEGF-R1 and VEGF-R3.The JNJ-17029259 of nanomole concentration level can suppress the new vein in hyperplasia/migration, the vascularization that occurs and the chorioallantoic membrane calibrating and the growth of artery in the rat aorta ring model of vasculogenesis.Referring to, people such as Emanuel for example, (2004) Mol Pharmacol.2004 September; 66 (3): 635-47.

Another micromolecular inhibitor is ZD6474, though it suppresses the VEGF-R2 tyrosine kinase activity, other somatomedin is had other retarding effect.Referring to, people such as Sandstrom for example, British Journal of Cancer advanceonline publication, on August 10th, 2004; Doi:10.1038/sj.bjc.6602108 Www.bjcancer.com

PTK787/ZK 222584 is active another micromolecular inhibitor of vegf receptor tyrosine kinase.(referring to, people such as Thomas for example, (2003) Semin Oncol.2003 June; 30 (3 Suppl 6): 32-8).People such as Mohammadi are at (1998) EMBO Journal the 17th volume, and the 5896-5904 page or leaf is described pyrido [2,3-d] miazines molecule in 1998, and with its called after PD 173074, its selectivity suppresses the tyrosine kinase activity of FGF and vegf receptor.Can effectively stop vasculogenesis and not have tangible toxicity with PD 173074 to the mouse throwing.

US 2002-0165174 describes the oligonucleotide that suppresses VEGF.Can use antisense oligonucleotide to reduce VEGF and VEGF-C level.Described oligonucleotide can reduce growth of cancer cells, for example the mesothelium tumor cell growth.The antibody of also having observed vegf receptor (VEGFR-2) and VEGF-C (VEGFR-3) slows down growth of cancer cells.US2004-0138163 describes the siRNA that can suppress VEGF.Inhibitor nucleic acids, siRNA and ribozyme such as antisense oligonucleotide can be used for regulating VEGF, VEGF-C or its receptor expression.Referring to, Int J Cancer on May 1st, 2003 for example; 104 (5): 603-10.

Other method comprises the cell that kills or suppress to produce VEGF.Cell to VEGF expression has highly toxic diphtheria toxin-VEGF fusion rotein (DT-VEGF) inhibition mesothelioma hyperplasia.Referring to, 15 days September calendar year 2001 of Blood for example; 98 (6): 1904-13.

The inhibitor of VEGF quasi-molecule also is summarized in people such as (for example) Verheul, (2003) Drugs Today (Bare) .2003; Among the 39 supplementary issue C:81-93.

Suck

The composition that comprises the hK1 binding antibody can be through allotment with other pattern that is used to suck or lung is sent.Therefore, compound as herein described can be thrown with to lung tissue through sucking.Unless otherwise instructed, otherwise as used herein term " lung tissue " refer to any tissue of respiratory tract and comprise the upper respiratory tract and lower respiratory tract.The hK1 binding antibody can with one or more existing modality combinations that is used for the treatment of pulmonary disorder throw with.

In one example, compound is through allocating to be used for atomizer.In one embodiment, compound can lyophilized form store (for example, at room temperature) and before sucking rehydration in solution.

Also compound might be allocated so that suck with the medical devices of for example sucker.Referring to, U.S.6 for example, 102,035 (powder inhalers) and 6,012,454 (Diskuses).Described sucker can comprise another compartment that is in the independent compartment that is used for active compound under the pH value that is fit to store and is used for neutralization buffer and the mechanism that is used for before atomizing described compound and neutralization buffer being made up immediately.In one embodiment, sucker is a metered dose inhaler.

Be used for medicine local delivery to 3 ubiquitous systems of lung's air flue are comprised Diskus (DPI), metered dose inhaler (MDI) and atomizer.MDI, the popular approach of inhalation dosing can be used for sending the medicine that is solubilisate form or dispersion.Usually, MDI comprises freonll-11 (Freon) or other relative high atmospheric pressure propelling agent, and it forces atomization medicine to enter respiratory tract after described device starting.Be different from MDI, the DPI medicine that patient's air-breathing power will be dry powder form that places one's entire reliance upon usually is introduced in the lung.Atomizer forms the medicinal aerosol that institute's desire sucks by energy being given liquor.Also used the fluorochemicals medium to probe into the direct pulmonary delivery of medicine during liquid ventilation or the lung lavage.Described method and other method all can be used for sending the hK1 binding antibody.In one embodiment, the polymer associate of the polymkeric substance of its transformation period increase is stablized or made to the hK1 binding antibody with for example making described compound.

For example, for by inhalation dosing, the hK1 binding antibody is sent with aerosol spray form self-pressurization container or the divider or the atomizer that contain suitable propelling agent.Described compound can be drying particulate or liquid form.Comprise the particle of described compound can (for example) by spraying drying, by with the aqueous solution of the dry hK1 binding antibody of charged neutralizing agent and subsequently from dry powder produce particle or by drying in organic modification agent the aqueous solution and produce particle from dry powder subsequently and prepare.

Described compound can the aerosol spray manifestation be sent by means of suitable propelling agent (for example, Refrigerant 12, trichlorofluoromethane, dichloro tetrafluoro ethane, carbonic acid gas or other suitable gas) self-pressurization packing or atomizer easily.Under the situation of pressurised aerosol, dose unit can be determined by the valve that the amount of sending metering is provided.If particle is the allotment particle, then is used for the capsule of sucker or insufflator and cartridge case and can contains hK1 binding antibody and powder mixture through allotment such as the suitable powder matrix of lactose or starch.Except that the compound of allotment or not allotment, can mix with the hK1 binding antibody to promote to allocate or do not allocate sending and disperseing of compound such as other material of 100%DPPC or other tensio-active agent.The method for preparing drying particulate is described among the open case WO 02/32406 of (for example) PCT.

The hK1 binding antibody can be sent as dry aerosol particle aerosol to be used for (for example) through allotment, thus when throwing and the time, but its rapid absorption and can produce part or whole body therapeutic result fast.But can adjust dispensing in 2 minutes, 5 minutes, 1 hour or 3 hours of dispensing, to provide detection of radioactive.In certain embodiments, peak activity even can reach faster, for example half an hour or even 10 minutes in.Can be with the hK1 binding antibody of reaching growth transformation period more (for example through allotment, by with polymer associate such as PEG) can be used as the replacement scheme of other dispensing patterns, (for example) so that described compound enter pulmonary circulation and are distributed in other organ or the particular target organ.

In one embodiment, the hK1 binding antibody with a certain amount of send so that the polypeptide quality at least 5% be delivered to lower respiratory tract or lung depths.The lung depths has extremely abundant capillary bed.The respiratory membrane that capillary cavity and alveolar air chamber are separated is (≤6 μ m) and have perviousness as thin as a wafer.In addition, the liquid level that is lining in the alveolar surface is rich in Curosurf.In other was implemented, the composition of at least 2%, 3%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% or 80% hK1 binding antibody was delivered to lower respiratory tract or lung depths.Be delivered to effective absorption and high bioavailability that any one or both in the described tissue produce described compound.In one embodiment, compound is to use (for example) sucker or atomizer to provide with dosing.For example, compound is sent at least about 0.02mg, 0.1mg, 0.5mg, 1mg, 1.5mg, 2mg, 5mg, 10mg, 20mg, 40mg or 50mg or more dosage unit form with every dose (puff).

The bioavailability percentage can be calculated as follows: bioavailability percentage=(AUC _Noninvasive/ AUC _{Intravenously or subcutaneous}) * (dosage _{Intravenously or subcutaneous}/ dosage _Noninvasive) * 100.

Though delivery enhancer and nonessential can be used delivery enhancer such as tensio-active agent further to strengthen lung and send.As used herein " tensio-active agent " refers to have hydrophilic and compound lipophilic portion, and it is by promoting drug absorption with two immiscible interfacial interactions between mutually.For a variety of reasons, for example reduce particle coacervation, reduce macrophage phagocytic effect etc., tensio-active agent is applicable to drying particulate.When with the Curosurf coupling, because will be very beneficial for the diffusion of compound, so can reach more effective absorption of described compound such as the tensio-active agent of DPPC.Tensio-active agent in affiliated field be know and its include, but is not limited to phosphoglyceride, for example, phosphatidylcholine, L-α-phosphatidylcholine palmityl (DPPC) and diphosphatidylglycerol (DPPG); Hexadecanol; Lipid acid; Polyoxyethylene glycol (PEG); Polyoxyethylene-9-; Auryl ether; Palmitinic acid; Oleic acid; Sorbitan trioleate (Span85); The N-cholylglycine ester; The surfactivity element; Poloxamer (poloxomer); Span; Sorbitan trioleate; Tyloxypal and phosphatide.

In one embodiment, the tensio-active agent that is used to comprise the described composition of enriched material and diluted composition provided herein is high HLB (hydrophilic-lipophilic balance value) tensio-active agent.Described high HLB tensio-active agent has the HLB greater than about 10 usually.HLB is the polar arbitrary scale of tensio-active agent or surfactant mixture.In one embodiment, composition provided herein contains the high HLB tensio-active agent of 3 weight % to about 85 weight % of having an appointment.In one embodiment, high HLB tensio-active agent is the ethoxylated derivative of vitamin-E, such as cetomacrogol 1000 VitaminE Succinat (TPGS).TPGS has the HLB between about 15 and 19.Referring to, US 2004-0023935 for example.

Stabilization and confining force

In one embodiment, hK1 binding antibody and the part physics that makes its stability in the circulation of for example blood, serum, lymph, bronchoalveolar lavage fluid or other tissue and/or confining force improve (for example) at least 1.5 times, 2 times, 5 times, 10 times or 50 times are associated.

For example, the hK1 binding antibody can with for example polymer associate of the antigenic polymkeric substance of tool (such as polyoxyalkylene or polyoxyethylene) not substantially.Suitable polymers should change with weight substantially.But the usage quantity molecular-weight average is at about 200 polymkeric substance to about 35,000 (or about 1,000 to about 15,000 and 2,000 to about 12,500) scope.

For example, the water-soluble polymers of hK1 binding antibody with for example hydrophilic polyethylene polymkeric substance (for example polyvinyl alcohol and polyvinylpyrrolidone) can be engaged.The non-limiting tabulation of described polymkeric substance comprises the polyoxyalkylene homopolymer such as polyoxyethylene glycol (PEG) or polypropylene glycol, polyoxyethylene polyvalent alcohol, and its multipolymer and its segmented copolymer are as long as keep the water-soluble of segmented copolymer.Other suitable polymers comprises the segmented copolymer such as the polyoxyalkylene of polyoxyethylene, polyoxypropylene and polyoxyethylene and polyoxypropylene (Pluronics); Polymethacrylate; Carbomer (carbomer); Comprise sugar monomer (D-seminose, D-semi-lactosi and L-semi-lactosi, Fucose, fructose, the D-wood sugar, L-arabinose, the D-glucuronic acid, sialic acid, the D-galacturonic acid, D-mannuronic acid (for example polymannuronic acid or alginic acid), the D-glycosamine, D-galactosamine, D-glucose and neuraminic acid) branch or branch polysaccharide not, it comprises homopolysaccharide and mixed polysaccharide, such as lactose, amylopectin, starch, hydroxyethylamyle, amylose starch, the sulfuric acid dextran, dextran, dextrin, the glycogen or the polysaccharide subunit of hyaluronic acidic mucopolysaccharide for example; The polymkeric substance of sugar alcohol is such as poly-Sorbitol Powder and poly-N.F,USP MANNITOL; Heparin or heparitin.

Other compound also can be connected in same polymkeric substance, for example cytotoxin, mark or another target agent, for example another hK1 binding antibody or irrelevant part.For example the list of the end capped polyoxyethylene glycol of mono methoxy (mPEG) activates alkoxy end-capped polyoxyalkylene (PAO), C _1-4Alkyl-blocked polymkeric substance and dual-active polyoxyethylene (di-alcohols) can be used for crosslinked.Referring to, U.S.5 for example, 951,974.

In one embodiment, though polymkeric substance needn't be preferably water miscible crosslinked for water miscible before part.Usually, after crosslinked, product is water miscible, for example shows at least about 0.01mg/ml and more preferably at least about 0.1mg/ml and even water-soluble at least about 1mg/ml more preferably.In addition, polymkeric substance should not have hyperimmunization originality in the joiner form, if joiner be intended to by intravenous fluids, aerosol thinner or injection throw with, then it should not have and the incompatible viscosity of described approach yet.

In one embodiment, polymkeric substance only contains single reactive group that has.Its help avoids ligand molecular to be cross-linked to each other.Yet, optimizing reaction conditions with reduce crosslinked between the ligand molecular or by gel-filtration or ion exchange chromatography purification reaction product to reclaim substantially the homogeneous derivative also in the category of this paper.In other was implemented, for making a plurality of parts be connected in the purpose of polymer backbone, polymkeric substance contained two or more reactive groups.Can use gel-filtration or ion exchange chromatography to reclaim and be the derivative of wanting of homogeneous phase form substantially again.

The molecular weight of polymkeric substance can be in the scope of about 500,000 D at the most, and is preferably at least about 20,000 D or at least about 30,000 D or at least about 40 000D.Selected molecular weight can according to the character (for example, structure (such as linearity or branch)) of effective size, polymkeric substance of the joiner desiring to reach and derivatize degree and decide.

Can use covalent linkage to make the hK1 binding antibody be connected in polymkeric substance, for example, crosslinked N-terminal in part is amino and be present in ε amino and other amino, imino-, carboxyl, sulfhedryl, hydroxyl or other hydrophilic radical on the lysine residue of part.Polymkeric substance can need not to use multifunctional (difunctionality usually) linking agent with hK1 binding antibody direct covalent bonds knot.Combine by the known chemical realization based on following each group with amino covalence: (the PEG alkoxide adds the diethyl ethanoyl of bromoacetaldehyde for cyanuryl chloride, carbonyl dimidazoles, aldehyde reaction group; PEG adds phenoxide, activation succinimide ester, the activation dithiocarbonates PEG, 2,4 that DMSO and diacetyl oxide or PEG muriate add the 4-hydroxy benzaldehyde, 5-trichlorophenyl chloro-formic ester or P-chloroformate nitrophenyl ester activated PEG).Carboxyl can derive the coupling of PEG-amine by using the carbonization imide.Sulfhedryl can be by coupling in the PEG that replaces through the maleimide base (for example, alkoxyl group-PEG amine adds 4-(N-maleimide ylmethyl) hexanaphthene-1-carboxylic acid thiosuccimide ester) WO97/10847 or available from Shearwater Polymers, Inc., Huntsville, the PEG-maleimide of Ala.) derive.Perhaps, free amino group on the part (for example, ε amino on the lysine residue) available 2-imido grpup-mercaptan alkane (Traut reagent) mercaptanization, and coupling subsequently is in the PEG derivative that contains maleimide, for example, as people such as Pedley, Br.J.Cancer is described in the 70:1126-1130 (1994).

Can utilize the functionalized PEG polymkeric substance that can be connected in the hK1 binding antibody, for example from Shearwater Polymers, Inc. (Huntsville, Ala.).Described commercially available PEG derivative is including (for example) amino-PEG, the PEG amino acid ester, the PEG-hydrazides, PEG-mercaptan, the PEG-succinate, carboxymethylation PEG, the PEG-propionic acid, PEG amino acid, PEG succsinic acid succinimide ester, PEG propionic acid succinimide ester, the succinimide ester of carboxymethylation PEG, the carbonic acid succinimide ester of PEG, the succinimide ester of amino acid PEG, PEG-oxygen base carbonylic imidazole, PEG-carbonic acid nitro phenyl ester, PEG trifluoro esilate, the PEG-glycidyl ether, PEG-aldehyde, the PEG vinyl sulfone(Remzaol, the PEG-maleimide, adjacent pyridyl-the disulphide of PEG-, assorted functionalized PEG, the PEG ethenyl derivatives, PEG silane and PEG echothiophate.The reaction conditions of the described PEG derivative of coupling can according to the hK1 binding antibody, the PEGization degree of wanting and employed PEG derivative change.Some factors that relate to the selection of PEG derivative comprise: the tie point of (such as, Methionin or halfcystine R group), the stability to hydrolysis of described derivative and reactivity, stability, toxicity and the antigenicity of binding, the suitability of analysis etc.The specific working instructions of any special derivative can obtain from manufacturers.

The joiner of hK1 binding antibody and polymkeric substance can (for example) by gel-filtration or for example the ion exchange chromatography of HPLC separate with unreacted initial substance.In the same manner with the xenobiotic of joiner purifying each other.The resolution of different substances (for example, containing one or two PEG residue) also may be owing to the difference of the ionic nature of unreacted amino acid.Referring to, for example WO 96/34015.

Example

Below differentiate that for being used to hK1 is conjugated protein, especially the hK1 inhibitor exemplary strategy.

1)KLK1 cDNA

KLK1 cDNA is subcloned in the expression vector of expressing in mammalian cell, intestinal bacteria and Maas moral pichia spp (P.pastoris).Confirm the material of institute's subclone by dna sequence analysis.

2) protein expression in Maas moral pichia spp

Transform and select the clone to express hK1 with above-mentioned subclone cDNA Maas moral pichia spp, measure evaluation by gel electrophoresis of protein and enzymic activity.Measure the optimization expression condition and with big Maas moral pichia spp volume of culture (2L) induced protein.

3) protein purification

According to people such as Chan, (1998) Protein Expr Purif, 12 (3): 361-70 page or leaf purifying hK1.

4) phage presents selection and screening

Present the wedding agent that reorganization hK1 is selected in the storehouse from the Fab phage.

Use two kinds of strategies to facilitate the selection (figure) of the antibody that is incorporated into the hK1 enzyme active sites.The avtive spot wedding agent be used for hK1 bonded enzyme substrates competition and can suppress the hK1 proteolytic activity.Two kinds of phages present the purposes that the hK1 of bead is fixed in the selection strategy utilization.By mixing in the cumulative volume that 5 μ g is had phosphate buffered saline (PBS) and 0.1% polysorbas20 (Tween20), 500 μ L (PBST) through biotin labeled hK1 and 5mg through the paramagnetic bead (Dynal) that streptavidin coats and under 4 ℃, described mixture being cultivated the bead for preparing overnight.Subsequently, described mixture with PBST washing 5 times, is added 1 μ M free biotin, wash 5 times again with PBST subsequently, and blocked 1 hour with 2% skim-milk in PBST at last.Described fixed routine makes catches and produces the fixedly hK1 with enzymic activity fully through biotin labeled hK1.

In first selection strategy, with phage (titre=1 * 10 ¹³Pfu) cultivated 5 minutes with the fixedly hK1 bead of 0.25mL in PBST and cumulative volume is 1mL.Using magnetic device to extract in conjunction with the phage of fixing hK1 and with non-specific phage is washing on the dish device through 12 cycles of washing elutions (selection 1) automatically with PBST.Subsequently, by cultivating 25 μ M Trypsin inhibitor,Trasylols (the avtive spot inhibitor of hK1) 2 hours, with phage from extracting in the bead elution to obtain phage from the avtive spot elution of hK1.To respectively be adjusted to 1 * 10 according to the standard program amplification and with it by the phage of Trypsin inhibitor,Trasylol elution and the phage that still is retained on the bead ¹¹The titre of pfu and the next round that is used separately as described strategy subsequently selects and last takes turns the input phage (II wheel) of selection.

In the II wheel, described two phage-infests (are also referred to as two " groups ", use the Trypsin inhibitor,Trasylol elution for one, another after the Trypsin inhibitor,Trasylol elution, still keep in conjunction with) be added into 0.05mL respectively fixedly in the hK1 bead, produce the cumulative volume of 1mL and before carrying out 12 cycles of washing, allow cultivation 5 minutes with PBST.Make subsequently two " groups " elution from II wheel through benefiting from 25 μ M Trypsin inhibitor,Trasylols last 2 hours with produce altogether 4 be used for by ELISA screen with hK1 bonded phage-infest (being also referred to as " pond "), wherein pond 1 is to derive from the Trypsin inhibitor,Trasylol elution of I wheel and the Trypsin inhibitor,Trasylol elution of II wheel, pond 2 be derive from the Trypsin inhibitor,Trasylol elution of I wheel and II wheel still be retained in phage on the bead, pond 3 is to derive from the 1st phage that still remaines in taking turns in phage on the bead and the II wheel by the Trypsin inhibitor,Trasylol elution, and pond 4 is to derive from the 1st still to remain in the phage that still remaines in phage on the bead and the II wheel on the bead in taking turns.

Select to quote Different Strategies in 2, wherein make phage (titre=1 * 10 by phage and fixing hK1-Trypsin inhibitor,Trasylol mixture cultivation are lasted 1 hour 3 times ¹³Pfu) exhaust nonactive site hK1 wedding agent, described fixedly hK1-Trypsin inhibitor,Trasylol mixture is fixedly to prepare in the hK1 bead and make cumulative volume reach 1mL with PBST by 25 μ M Trypsin inhibitor,Trasylols being added into 0.25mL.Subsequently, will exhaust phage is added into 0.25mL and reaches fixedly that 1mL lasts 5 minutes and carry out 12 cycles of washing with PBST in the hK1 bead.Subsequently, the phage amplification with still remaining on the bead is adjusted to 1 * 10 ¹¹The titre of pfu and as the 2nd input thing of taking turns.The 2nd take turns with the 3rd take turns take turns as the 1st as described in execution, wherein utilizing 0.25mL fixedly before the selection of hK1 bead, each wheel all have utilize hK1-Trypsin inhibitor,Trasylol mixture initially exhaust step.Described selection strategy produces a phage-infest (pond 5), and it is screened with the wedding agent as hK1 by ELISA.

By the ability in conjunction with hK1 of solubility Fab ELISA calibrating from the phage in above-mentioned 5 ponds.Before carrying out described ELISA, must modify phasmid DNA and need not gene III fusion to allow Fab to express.Described dna modification is finished as follows: at first each the phasmid DNA in described 5 ponds is prepared in the e. coli tg1 cell, uses the MluI restriction enzyme to carry out dual DNA digestion subsequently and discharge the segmental DNA of encoding gene III root.After the gel-purified, carrier being connected again (the pMID21 carrier of no gene III root) and electroporation enters in the e. coli tg1 cell.Transformant is coated with shop and choosing colony and makes its about 3 hours of growth among the 100 μ l2xYT that contain 100 μ g/mL Ampicillins (ampicillin) and 0.1% glucose in 96 orifice plates under 30 ℃.Under 30 ℃,, last 16 hours adding 1mM iprotiazem base-beta galactose glycosides (IPTG) back expression Fab.Subsequently, with plate centrifugal 10min and 50 μ L supernatant liquors are used for ELISA under 600g.

Under 4 ℃, the hole execution solubility Fab ELISA that is coated with shop 96 orifice plates by the 2 μ g/mL streptavidins that at first are used for 0.1M sodium bicarbonate buffer liquid (pH9.6) spends the night.Then with the PBST washing and with the blocking-up of 1% bovine serum albumin, interpolation 10ng is through biotin labeled hK1 and at room temperature cultivated 1 hour.Subsequently, by removing with 5 hK1 that will dissociate of PBST washing.To be added among the hK1 that is fixed on the plate from the supernatant liquor (50 μ l) that above-mentioned Fab expresses and at room temperature cultivate 1 hour.Subsequently, unconjugated Fab is removed for 7 times by washing with PBST, and the anti-Fab antibody that 100 μ L horseradish peroxidases are engaged is added in the described hole.With free secondary antibody by remove and add for 7 times with PBST washing 100 μ l colorimetric peroxidase substrates (3,3 ', 5,5 '-tetramethyl benzidine/hydrogen peroxide) to allow increasing detection in conjunction with Fab by the absorption at 630nm place.Use described ELISA,, analyze 4800 Fab altogether with regard to 5 960 expressed Fab of each analysis that select in the pond.

5) dna sequence analysis

The positive solvable Fab of 1152 ELISA is carried out dna sequence analysis, and wherein 335 Fab have unique heavy chain.

6) Fab binding affinity and enzyme suppress the mensuration of usefulness

Measure the binding affinity of 335 Fab by surface plasma resonance (SPR).Binding constant (table 1) in 0.4 to the 100nM above scope of acquisition.Be under the 120nM all 335 Fab screening enzymic activitys to be suppressed at first, then measure the IC of the Fab of selected number in single Fab concentration ₅₀Value (table 1).The IC of Fab no matter ₅₀Whether be measured to, Fab all may be or may not be enzyme inhibitors.Inhibitor can be present in the active ability of bronchial tissue's kallidinogenase in bronchoalveolar lavage (BAL) liquid to measure its inhibition through further assessment.

In addition, check to find to suppress among the high-affinity Fab of hK1 24 in further detail.In the table 1, use asterisk to indicate described Fab.Initial high-throughput SPR of use and enzyme suppress being used in combination of screening and utilize a small amount of Fab of novel high throughput method purifying to differentiate described Fab.A large amount of described 24 Fab of use standard antibody method of purification purifying allow more accurate and at length check its biochemical property.It is execution in the reaction buffer (20mM Tris pH7.5,150mM NaCl, 1mM EDTA, 0.1%PEG, 0.1%TritonX-100) of the Fab that contains 5nM hK1, varied concentration of 100uL and 100 μ M Pro-Phe-Arg-AMC substrates at cumulative volume in black, 96 holes, the little quantitative plate of round bottom that the stable state enzyme suppresses calibrating.Before the interpolation substrate is with initial action, under 30 ℃ hK1 and Fab were being cultivated 1 hour in the hole of assaying table.Subsequently, adding substrate and use fluorescent plate reader (Gemini XS, Molecular Devices) uses the excitation wavelength of 360nm and the emission wavelength of 460nm to monitor described reaction.The standard Hyperbolic Equation that the initial rate of substrate hydrolysis is suppressed to the inhibitor concentration mapping and by the nonlinear regression and fitting enzyme is to provide IC ₅₀Estimated value.

Particular exemplary antibody is appointed as the M0103-A01 form.All with " M0 " beginning, therefore leading by removing sometimes " M0 " simplifies all titles.Antibody title " 103-A01 " is meant same antibody with " M0103-A01 ".The HC called after HC-103-A01 of M0103-A01 and LC called after " LC-103-A01 ".

7) mouse model of tracheae inflammation research

Can just regulate the ability of tracheae inflammation and assess Fab in the mouse model of asthma, for example as people such as Kips, (2003) " Murine models of asthma " Eur.Respir.J.22 is described in the 374-382.But Fab also reformatting is the IgG molecule and uses one or more calibrating assessments as herein described.

Table 1: binding data

Title	IC50(nM)	Kon(M-1s-1)	Koff(s-1)	Kd(M)
					*098-E09 *131-F07 *139-F04 *139-E12 *097-F03 *102-G12 *102-E09 *106-G12 *102-H11 *138-C09 *098-G05 *133-E02 *112-D03 *098-F02 136-E10 136-A07 *097-G01 138-G11	2.5±3.3 0.35+0.58 20.7±8.7 2.9±0.49 1.1+0.54 3.3±0.34 0.89±0.25 5.1±0.4 4.5±1.7 10.9±3.9 4.0±2.2 2.4±1.1 6.0±0.3 6.9±1.6 22 25 7.2±3.0 28	8.36E+05 2.46E+05 1.84E+05 1.25E+05 8.37E+04 2.29E+06 1.75E+05 2.27E+05 1.04E+05 5.36E+05 1.10E+05 3.23E+05 1.73E+05 2.60E+05	1.59E-03 2.09E-04 1.34E-03 2.33E-04 4.42E-04 7.41E-03 1.53E-03 1.46E-03 9.88E-04 6.22E-04 1.06E-03 2.58E-03 8.32E-04 1.61E-03	1.91E-09 8.47E-10 7.27E-09 1.87E-09 5.28E-09 3.24E-09 8.78E-09 6.42E-09 9.51E-09 1.16E-09 9.66E-09 7.98E-09 4.80E-09 6.19E-09

139-A09 *114-G06 *139-C02 *093-F09 110-F08 *117-F04 111-D11 112-D07 131-B05 131-D01

34 1.2±0.53 1.0±0.30 2.5±2.13 44 5.2±1.5 160 ＞120

1.94E+05 1.67E+05 6.00E+05 2.98E+05 1.34E+05 2.55E+05 3.11E+05

1.06E-03 1.66E-04 2.79E-03 1.97E-03 7.81E-04 1.17E-04 3.25E-04

5.48E-09 9.99E-10 4.66E-09 6.61E-09 5.81E-09 4.60E-10 1.04E-09

Title	IC50(nM)	Kon(M-1s-1)	Koff(s-1)	Kd(M)
					*137-E01 132-C08 114-G09 138-A03 135-H01 108-D11 136-E12 136-D07 138-E02 131-F09 134-B03 104-H12 135-F03 *134-D07 135-C12 097-F04 111-C10 094-E08 138-B10 139-F09 104-A12 132-D02 138-C04 132-H11 132-D08 121-A07 126-F11 098-H04 093-C02 115-G12 135-B05 136-F08 110-H11 110-D11 114-H07 098-E01 135-C11 095-A11 092-F08	0.4±0.25 10.1+5.1	1.23E+05 3.42E+05 4.24E+05 1.90E+05 1.95E+05 3.30E+05 4.81E+05 1.76E+05 6.14E+05 3.11E+05 3.99E+05 5.67E+05 2.35E+05 5.13E+04 1.21E+05 1.06E+05 1.04E+06 1.59E+05 2.65E+05 3.01E+05 6.73E+05 7.17E+05 1.71E+05 4.40E+05 1.95E+05 9.22E+05 2.85E+05 4.61E+05 1.80E+05 1.51E+05 1.11E+05 4.76E+05 9.75E+05 2.10E+05 1.10E+06 1.48E+05 3.37E+05 5.25E+05 2.18E+05	1.58E-04 4.44E-04 6.20E-04 3.25E-04 3.96E-04 6.90E-04 1.07E-03 3.97E-04 1.38E-03 7.34E-04 9.60E-04 1.38E-03 6.25E-04 1.38E-04 3.76E-04 3.46E-04 3.41E-03 5.30E-04 8.90E-04 1.08E-03 2.46E-03 3.03E-03 7.28E-04 1.98E-03 9.23E-04 4.38E-03 1.44E-03 2.55E-03 1.03E-03 8.65E-04 6.55E-04 2.84E-03 6.01E-03 1.34E-03 7.03E-03 1.01E-03 2.31E-03 3.79E-03 1.58E-03	1.29E-09 1.30E-09 1.46E-09 1.71E-09 2.03E-09 2.09E-09 2.23E-09 2.26E-09 2.26E-09 2.36E-09 2.41E-09 2.43E-09 2.66E-09 2.69E-09 3.10E-09 3.26E-09 3.29E-09 3.34E-09 3.36E-09 3.60E-09 3.66E-09 4.22E-09 4.26E-09 4.50E-09 4.73E-09 4.75E-09 5.05E-09 5.53E-09 5.72E-09 5.73E-09 5.90E-09 5.97E-09 6.17E-09 6.38E-09 6.39E-09 6.80E-09 6.85E-09 7.22E-09 7.27E-09

104-B12 109-A11 131-G03 135-E10

2.30E+05 4.70E+05 3.41E+05 2.72E+05

1.71E-03 3.50E-03 2.56E-03 2.06E-03

7.41E-09 7.45E-09 7.49E-09 7.57E-09

Title	IC50(nM)	Kon(M-1s-1)	Koff(s-1)	Kd(M)
					130-B02 097-D04 093-G06 108-E11 127-C06 105-B06 107-D01 107-F11 101-E01 138-E12 109-E08 104-B03 096-E11 103-A03 099-A09 105-F02 112-C02 125-C04 122-H04 092-B12 139-A04 108-C10 138-B06 137-B01 111-B02 129-B09 099-C12 099-E11 103-F07 102-A04 119-B09 097-E07 117-A12 126-C10 127-D05 126-B12 122-G06 107-D05 127-H05 112-C12 098-C10 095-H10 117-C04 094-F03		3.94E+05 2.03E+05 3.72E+05 4.18E+05 6.62E+05 4.09E+05 8.96E+05 6.80E+05 9.66E+04 1.48E+05 8.80E+05 7.07E+05 8.27E+05 9.31E+05 2.67E+05 9.47E+05 4.81E+05 4.78E+05 6.82E+04 1.41E+05 4.11E+05 3.53E+05 6.64E+04 1.92E+05 9.29E+10 1.73E+06 3.54E+05 6.05E+05 5.64E+05 3.27E+05 3.52E+05 5.85E+05 2.43E+05 4.51E+05 6.39E+04 3.85E+05 7.08E+06 1.95E+05 2.87E+05 2.90E+05 1.20E+05 5.56E+05 2.01E+05	3.07E-03 1.68E-03 3.13E-03 3.55E-03 5.75E-03 3.60E-03 8.12E-03 6.60E-03 9.41E-04 1.49E-03 8.89E-03 7.11E-03 8.36E-03 9.88E-03 2.85E-03 1.03E-02 5.23E-03 5.36E-03 8.02E-04 1.71E-03 5.03E-03 4.31E-03 8.29E-04 2.40E-03 1.17E+03 2.20E-02 4.61E-03 7.96E-03 7.43E-03 4.35E-03 4.83E-03 8.15E-03 3.45E-03 6.48E-03 9.21E-04 5.79E-03 1.07E-01 3.00E-03 4.39E-03 4.65E-03 1.93E-03 9.18E-03 3.34E-03 6.44E-03	7.80E-09 8.28E-09 8.40E-09 8.50E-09 8.69E-09 8.81E-09 9.06E-09 9.71E-09 9.74E-09 1.00E-08 1.01E-08 1.01E-08 1.01E-08 1.06E-08 1.07E-08 1.09E-08 1.09E-08 1.12E-08 1.18E-08 1.21E-08 1.22E-08 1.22E-08 1.25E-08 1.25E-08 1.27E-08 1.27E-08 1.30E-08 1.32E-08 1.32E-08 1.33E-08 1.37E-08 1.39E-08 1.42E-08 1.44E-08 1.44E-08 1.50E-08 1.52E-08 1.53E-08 1.53E-08 1.60E-08 1.61E-08 1.65E-08 1.66E-08 1.67E-08

Title	IC50(nM)	Kon(M-1s-1)	Koff(s-1)	Kd(M)
					124-G12 131-A03 099-D05 131-C08 116-C09 128-F11 131-D12 128-A06 095-G09 137-G10 106-E12 124-H10 122-D01 093-C10 106-C06 117-B07 126-H09 093-G09 114-E02 102-G11 132-C01 123-E02 096-D03 131-G06 094-D08 128-E07 114-F04 122-A05 102-H02 092-G06 113-E03 118-E07 093-E11 124-C12 115-F08 121-H07 136-B06 097-F08 107-B05 124-A01 102-C12 103-E09		3.72E+05 2.65E+06 3.31E+05 2.64E+05 5.20E+05 7.80E+05 1.05E+05 3.32E+05 5.01E+06 2.72E+05 1.23E+08 5.91E+04 2.05E+05 3.33E+05 6.28E+05 3.62E+05 1.80E+07 4.58E+05 4.03E+05 4.67E+05 1.37E+05 2.29E+05 3.37E+05 7.03E+04 2.37E+05 4.83E+05 3.48E+05 1.47E+05 3.40E+05 2.15E+05 1.59E+10 2.74E+06 1.34E+06 3.89E+06 4.84E+05 1.68E+05 2.00E+05 3.53E+05 6.00E+05 3.36E+05 7.39E+07 2.43E+05	6.40E-03 4.60E-02 5.78E-03 4.66E-03 9.47E-03 1.43E-02 1.93E-03 6.12E-03 9.34E-02 5.13E-03 2.44E+00 1.20E-03 4.27E-03 7.52E-03 1.42E-02 8.26E-03 4.15E-01 1.05E-02 9.36E-03 1.16E-02 3.50E-03 6.13E-03 9.45E-03 2.09E-03 7.07E-03 1.47E-02 1.07E-02 4.53E-03 1.05E-02 6.63E-03 5.00E+02 8.71E-02 4.36E-02 1.26E-01 1.58E-02 5.50E-03 6.61E-03 1.17E-02 2.03E-02 1.14E-02 2.62E+00 8.84E-03	1.72E-08 1.74E-08 1.75E-08 1.77E-08 1.82E-08 1.83E-08 1.84E-08 1.84E-08 1.86E-08 1.88E-08 1.99E-08 2.03E-08 2.08E-08 2.26E-08 2.26E-08 2.28E-08 2.30E-08 2.30E-08 2.32E-08 2.48E-08 2.56E-08 2.68E-08 2.81E-08 2.97E-08 2.98E-08 3.04E-08 3.07E-08 3.08E-08 3.08E-08 3.09E-08 3.15E-08 3.18E-08 3.24E-08 3.25E-08 3.26E-08 3.28E-08 3.30E-08 3.31E-08 3.38E-08 3.39E-08 3.54E-08 3.64E-08

Title	IC50(nM)	Kon(M-1s-1)	Koff(s-1)	Kd(M)
					131-F12 093-C08 115-F04 113-G11 102-C02 112-D04 108-E10 104-D12 116-E08 102-D07 103-H07 116-A08 106-D06 113-D05 110-D06 124-C04 *111-C12 *107-D12 *109-F02	2.9±1.1 1.7±3.2 2.8+0.2	1.28E+05 1.42E+05 3.14E+05 2.86E+05 1.44E+05 1.76E+05 2.30E+05 3.14E+05 2.25E+05 1.55E+05 1.00E+05 1.13E+05 1.54E+05 1.41E+04 7.56E+04 7.68E+05	4.72E-03 5.78E-03 1.31E-02 1.21E-02 6.28E-03 9.56E-03 1.34E-02 1.92E-02 1.64E-02 1.22E-02 8.60E-03 9.94E-03 1.90E-02 2.14E-03 1.75E-02 2.86E-01	3.70E-08 4.07E-08 4.16E-08 4.25E-08 4.35E-08 5.42E-08 5.84E-08 6.13E-08 7.27E-08 7.86E-08 8.57E-08 8.83E-08 1.23E-07 1.52E-07 2.32E-07 3.72E-07

Exemplary antibody comprises following sequence: the aminoacid sequence of classifying exemplary variable region of light chain down as:

＞LC-092-B12 (SEQ ID NO：1022)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISNYLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPYTF

GQGTKLEVRR TVAAP

＞LC-092-F08 (SEQ ID NO：1023)

AQDIQMTQSP ATLSLSPGER ATLSCGASQS VSSSYLAWYQ QKPGLAPRLL

IYDASSRATG IPDRFSGSGS GTDFTLTISS LQPEDFATYY CLHDYNFPFT

FGPGTKVDIK RTVAAP

＞LC-092-G06 (SEQID NO：1024)

AQSELTQPRS VSGSPGQSVT ISCTGTSGEV ANYNYVSWYH QDPGLVPKLK

IYDVSRRPSG VPDRFSGAKS GDTASLTISG LQAEDEGDYY CASYVGNDKL

VFGGGTKLTV LGQPKAAP

＞LC-093-C02 (SEQ ID NO：1025)

AQDIQMTQSP GTLSLSPGDR ATLSCRASQS VGSDYLAWYQ QKPGQAPRLL

IFAASTRATG IPDRFSGSGS ATDFTLTISS LEPEDFAVYF CQQYASPPRT

FGQGTKVEIK RTVAAP

＞LC-093-C08 (SEQ ID NO：1026)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IFAASTRATG IPDRFSGSGS ATDFTLTISS LEPEDFAVYY CQQRSNWPPE

LTFSGGTKVE IKRTVAAP

＞LC-093-C10 (SEQ ID NO：1027)

AQDIQITQSP SSLSASVGDR VTITCRASQG ISNYLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC EQLNSFPHAF

GQGTKVEIKR TVAAP

＞LC-093-E11(SEQ ID NO：1028)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSSYLAWYQQ KPGQAPRLLI

YDASNRATGI PARFSGSGSG TDFTLTISSL EPEDFAVYYC QQRSNWLTFG

GGTKVEIKRT VAAP

＞LC-093-F09 (SEQ ID NO：1029)

AQSALTQPAS VSGSPGQSIT ISCTGTSTDD VGGYNYVSWY QQHPGKAPKL

MIYDVINRPS GVSNRFSGSK SGNTASLTIS GLQAEDEADY YCSSYTSRGT

RVFGTGTKVT VLGQPKANP

＞LC-093-G06 (SEQ ID NO：1030)

AQSALTQPRS VSGSLGQSVT ISCTGSTSDV GGYTYVSWYQ QEPGKAPKLM

IHDVSKRPSG VPDRFSGSKS GNTASLIISG LQAEDEADYY CCSYAGSYSY

VFGTGTKVTV LGQPKANP

＞LC-093-G09 (SEQ ID NO：1031)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSFLNWYQQ KPGKAPKLLL

MYAASSLQSG VPSRFSGSGS GTDFTLTISS LQPEDFATYY CQQSYSTPYT

FGQGTKLEIK RTVAAP

＞LC-094-D08 (SEQ ID NO：1032)

AQSALTQPPS ASGTPGQRVT ISCSGSSSNI GSNNVNWYQQ LPGTAPKLLI

YSNDQRPSGV PDRFSGSKSA TSASLAISGL QSEDEADYHC AAWDDSLNGP

VFGGGTKLTV LGQPKAAP

＞LC-094-E08 (SEQ ID NO：1033)

AQSVLTQPAS VSGSPGQSIT ISCTGTSSDV GAYNYVSWYQ QHPGKVPELM

IYDVSNRPSG VSHRFSGSKS GNTASLTISG LQAEDDADYY CSSFTSRKTW

VFGTGTKVTV LGQPKANP

＞LC-094-F03 (SEQ ID NO：1034)

AQDIQMTQSP GTLSLSLGET ATLSCRASQT VGGSYLAWYR QKPGQAPSLL

IYAASNRAPG IPDRFSGSGS GTDFTLTISS LQSEDFAVYY CQQYGSSMYT

FGQGTILEIK RTVAAP

＞LC-095-A11 (SEQ ID NO：1035)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSSYLAWYQQ KPGQAPRLLI

YDASNRATGI PARFSGSGSG TDFTLTISSL EPEDFAVYYC QQRSNWPPYT

FGQGTKLEIK RTVAAP

＞LC-095-G09 (SEQ ID NO：1036)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISNWLAWYQQ KPGKAPKLLI

YKASTLQTGV PSRFSGSGSG TEFSLTISSL QPDDFATYYC QQTYSAPFNF

GPGTKVDIKR TVAAP

＞LC-095-H10 (SEQ ID NO：1037)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISSWLAWYQQ KPGKAPKLLI

YTASSLESGV PSRFSAGGSG TEFTLTISSL QPDDFGTYYC QQYNSYSLTF

GGGTKVEIKR TVAAP

＞LC-096-D03 (SEQ ID NO：1038)

AQSELTQPRS VSGSPGQSVT ISCTGTTRDV GAYKYVSWYQ QYPGKAPKLM

LSDVSKRPSG VPDRFSGSKS GNTASLTISG LQSEDEADYY CCSFAGSYTW

IFGGGTKVTV LGQPKAAP

＞LC-096-D09 (SEQ ID NO：1039)

AQSALTQPPS ASGTPGQRVT ISCSGSSSNI GTNRVNWYQQ IPGTAPKLLI

YSNNQRPSGV PDRFSDSKSG TSASLAISGL qSEDEADYYC AAWDDSLNGV

VFGGGTKLTV LGQPKAAP

＞LC-096-E11 (SEQ ID NO：1040)

AQDIQMTQSP LSLSASVGDR VTMTCRASQT ISSYLNWYQQ KPGKAPKLLI

YTTSSLQSGV PSRFSGSGSG TEFTLTISSL QPEDFATYYC QQSHSSPTFG

GGTKVEIKRT VAAP

＞LC-097-D04(SEQ ID NO：1041)

AQSELTQPPS ASGTPGQRVT ISCSGSSSNI GSNYVYWYQQ LPGMAPKLLI

YRNNQRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSMSGV

VFGGGTKLTV LSQPKAAP

＞LC-097-E07 (SEQ ID NO：1042)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLSWFQQ RPGKAPKLLI

YSASNLQSGV PLRFSGSGSG TDFTLTISSL QPEDFATYYC QQIYRTPLTF

GGGTKVEIKR TVAAP

＞LC-097-F03 (SEQ ID NO：1043)

AQDIQMTQSP ATLSVSPGGR ATLSCRASQS VRKNVAWYQQ KPGQPPRLLI

YGASTRATGV PARFSGSGSG TEFTLTISRM QPEDFVVYHC QQYSSWPAFG

QGTMVEINRT VAAP

＞LC-097-F04 (SEQ ID NO：1044)

AQYELTQPPS ASGTPGQRVT ISCSGSSSNI GSNYVHWYQQ LPGTAPKLLI

YRNNRRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSLSGL

VVFGGGTKLT VLGQPKAAP

＞LC-097-F08 (SEQ ID NO：1045)

AQDIQMTQSP STLSASVGDR VTISCRASQG IGTHLNWYQQ KLGNVPKLLI

YAASGLQSGV PPRFSGSGSG TDFTLTISSL HPEDSATYFC QQSYSVPRTF

GQGTKVEIKR TVAAP

＞LC-097-G01(SEQ ID NO：1046)

AQDIQMTQSP SSLSASVGDR VTITCRASQG IGNYLVWYQH KPGNVPRVLI

YAASTLQSGV PSRFRGSGSG TDFTLTISGL QPEDVATYYC QKYDGAPFTF

GPGTKVDLKR TVAAP

＞LC-097-G07 (SEQ ID NO：1047)

AQDIQMTQSP FLPVCILqET ESPSLAGQVR PLAGILNWYQ QKPGTAPKLL

IYAASSLQSG VPSRFSGDGS GTDFTLTISS LQPEDFGIYF CQQSYSAPRT

FGQGTKVEIK RTVAAP

＞LC-098-C10 (SEQ ID NO：1048)

AQDIQMTQSP SSLSASVGDR VTITCRASHN IYTSLNWLQQ KPGKAPKLLI

YGASTLENGV PSRFSGSASG TDFTLTISSL QPEDSATYYC QQSYTSVTFG

QGTKLEIRRT VAAP

＞LC-098-E01 (SEQ ID NO：1049)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSGFLAWYQQ KPGQAPRLLI

YDASNRATGI PARFSGSGSG TDFTLTITRL EPEDFAVYYC QQYGDSSPIT

FGPGTRLEIK RTVAAP

＞LC-098-E09 (SEQ ID NO：1050)

AQDIQMTQSP SSVSASVGDS VTISCWTIYD ISSWLAWYQQ RPGQAPKLLI

YAASRLATGV PSRFRGSGSG TDFTLTITNL QPEDVATYYC QQTKDFPLTF

GGGTRVDLKR TVAAP

＞LC-098-G05 (SEQ ID NO：1051)

AQDIQMTQSP SSLSASVGDR VTITCRASQT ISRYLNWYQQ KPGTAPKLLI

YAASSLQSGV PSRFSGDGSG TDFTLTISSL QPEDFGIYFC QQSYSAPRTF

GQGTKVEIKR TVAAP

＞LC-098-H04 (SEQ ID NO：1052)

AQSELTQPPS ASGSPGQSLT ISCTGGRRDI GNYNYVSWYQ QHPGKAPRLI

IYDVRKRPSG VPDRFSGSKS GNVAFLTVSG LQTDDEADYY CGSYTGTSNV

FGSGTTVTVL GQPKANP

＞LC-131-G03 (SEQ ID NO:1052) LC-098-H04 is identical with LC-131-G03

AQSELTQPPS ASGSPGQSLT ISCTGGRRDI GNYNYVSWYQ QHPGKAPRLI

IYDVRKRPSG VPDRFSGSKS GNVAFLTVSG LQTDDEADYY CGSYTGTSNV

FGSGTTVTVL GQPKANP

＞LC-099-A09 (SEQ ID NO：1053)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VYSSYLAWYQ QKPGQAPRLL

IYAASNRAIG IPDRFSGSGS GTDFTLTISS MQPEDFATYY CQQSYSTPRF

GQGTKLEIKR TVAAP

＞LC-099-C12(SEQ ID NO：1054)

AQSVLTQPAS VSGSPGQSIT ISCTGTTSDV GGYKYVSWYQ HHPGKVPKLI

IYEVNHRPSG VSHRFSGSKS GNTASLIISA LQAEDEADYY CYSYTSDSTP

YVFGTGTKVT VLGQPKANP

＞LC-099-D05(SEQ ID NO：1055)

AQDIQMTQSP GTLSLSPGER ATLSCRASQT VSSNYLAWYQ QKPGLAPRLL

IYGVSNRAAG IPDRFSGRGS GTDFTLIINR LEPEDFAVYY CQHYGSSAFT

FGRGTKLEIE RTVAAP

＞LC-099-E11 (SEQ ID NO：1056)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISNWLAWYQQ KPGRAPKLLI

YKASTLESGV PSRFSGSGSG TEFTLTISSL QPDDFATYYC QHYNSYSLFG

QGTKLEIKRT VAAP

＞LC-101-E01 (SEQ ID NO：1057)

AQDIQMTQSP SSVSASVGDR VTITCRAGQN IYYWLAWYQQ KPGKAPQLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQAKSFPVTF

GGGTKVEIKR TVAAP

＞LC-102-A04(SEQ ID NO：1058)

AQSELTQPPS ASGTPGQRVT ISCSGSSSNI GSNYVYWYQQ YPGTVPKLLI

HSNNQRPSGV PDRFSGPKSG TSASLAISGL RSEDEADYYC ATWDNSLSAW

VFGGGTKLTV LRQPKAAP

＞LC-102-C02 (SEQ ID NO：1059)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISSSLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPWTF

GQGTKVEIKR TVAAP

＞LC-102-C12 (SEQ ID NO：1060)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISNYLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDVATYYC QKYNSAPWTF

GQGTKVEIKR TVAAP

＞LC-102-D07 (SEQ ID NO：1061)

AQDIQMTQSP GTLSLSPGER ATLSCGASQS VSTTYIAWYQ HKPGQPPRLL

IYGASNRATG IPDRFRGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSPYT

FGQGTKLEIK RTVAAP

＞LC-102-E09 (SEQ ID NO：1062)

AQDIQMTQSP DSLSLSPGER ATLSCRASQS ISSSYLAWYQ QTPGQAPRLL

IYHASSRATG VPARFSGSGS GTDFTLTISS LEPEDFAVYY CQQRNNWPPS

FTFGGGTKVE TKRTVAP

＞LC-102-G11 (SEQ ID NO：1063)

AQDIQMTQSP DTLSLSPGER ATLSCRASES VSSSYFAWYQ QKRGQAPRLL

IYGASRRVTG IPDRFSGSGS GTDFTLTITR LEPEDFAVYY CQQYGGSPRS

FGQGTKVEIK RTVAAP

＞LC-102-G12(SEQ ID NO：1064)

AQSELTQPPS ASGTPGQRVT ISCSGTLSNI GTNIVSWFQQ LPGTAPKLLI

YNDHRRPSGV PDRFSGSKSA TSASLAISGL QSEDEADYYC AAWDDSLNGV

VFGGGTKLTV LSQPKAAP

＞LC-133-E08 (SEQ ID NO:1064) LC-102-G12 is consistent with LC-133-E08

AQSELTQPPS ASGTPGQRVT ISCSGTLSNI GTNIVSWFQQ LPGTAPKLLI

YNDHRRPSGV PDRFSGSKSA TSASLAISGL QSEDEADYYC AAWDDSLNGV

VFGGGTKLTV LSQPKAAP

＞LC-102-H02(SEQ ID NO：1065)

AQDIQMTQSP ATLSLSPGER AALSCRASQS VSNFLAWYQQ KPGQAPRLLI

YGASNRATDI PARFSGSGSG TDFTLTISSL EPEDFATYYC QQANSFPLTF

GGGTKVEIKR TVAAP

＞LC-102-H11(SEQ ID NO：1066)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSSYLAWYQQ KPGQAPRLLI

YGASSRATGI PDRFSGSGSG TDFTLTISSL EPEDFAVYYC QQRGGWPLTF

GGGTKVEIRR TVAAP

＞LC-103-A01 (SEQ ID NO：1067)

AQSALTQPPS ASGTPGQTVT ISCSGSSSNI GTNFVYWYQQ LPGTAPKLLI

YRSIKRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSLSGV

VFGGGTKLTV LGQPKAAP

＞LC-103-A03 (SEQ ID NO：1068)

AQDIQMTQSP SSVSASVGDR VTITCRASQD VRRFLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPITF

GQGTRLEIKR TVAAP

＞LC-103-E09 (SEQ ID NO：1069)

AQYELTQPAS VSGSPGQSIT ISCTGTNTDV GGYNFVSWYQ QYPGKAPKLI

IFDVTNRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CCSYTNTNTL

VFAGGTKVTV LGQPKANP

＞LC-103-F07 (SEQ ID NO：1070)

AQSALTQPAS VSGSPGQSIT ISCTGTSNDI GRTNYVSWYR QDPGRAPKLI

LFEVSNRPSG ISNRFSASKS GSTASLTISG LQADDESDYY CSSCTTAPVC

VFGNGTRVTV LGQPKANP

＞LC-103-H07(SEQ ID NO：1071)

AQDIQMTQSP SSLSASVGDR VAITCRASQS IDTYLNWYQQ KPGKAPKLLI

YAASKLEDGV PSRFSGSGTG TDFTLTIRSL QPEDFATYYC QPYNTYPITF

GQGTRLEIKR TVAAP

＞LC-104-A12 (SEQ ID NO：1072)

AQDIQMTQSP SSLSASVGDR VTITCRASQS IGASLNWYQQ KPQKAPKLLI

YTASNLQSGV PSRFSGSGSG TDFTLTISSL LPEDFATYYC QQNYRGRTFG

QGTKLEIKRT VAAP

＞LC-104-B03 (SEQ ID NO：1073)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSRYT

FGQGTKLEIK RTVAAP

＞LC-104-B12(SEQ ID NO：1074)

AQDIQMTQSP SSLSASVGDR VTITCRASQA ISNYLAWYQQ QPGKVPKLLI

SAASTLQSGV PSRFSGSGSG TDFTLSISSL QPEDVATYYC QTYYSVPFTF

GPGTKVDFKR TVAAP

＞LC-104-D12(SEQ ID NO：1075)

AQDIQMTQSP SSLSASVGDR VTITCRASLS VSGYLNWYQH KPGRAPNLLI

YATSSLQSGV PSRFSGSGSG TDFTLTVSSL QPEDLATYYC QQSYSSPYTF

GQGTKVEIKG TVAAP

＞LC-104-H12(SEQ ID NO：1076)

AQSELTQPPS ASGSPGQSVT ISCTGTSSDV GAYNYVSWYQ QHPGKAPKLI

IYEVNKRPSG VPDRFSASKS GNTASLTVSG LQAEDEADYY CNSYAGSNSL

IFGGGTKLTV LGQPKAAP

＞LC-105-B06(SEQ ID NO：1077)

AQDIQMTQSP SSLSASVGDR VTLTCRASQS IANYLNWYQQ KPGKAPKLLV

YAASRLHSGV PSRFSGRGSG TDFTLTITSL EPDDLATYYC QQSHASPLTF

GGGTKVEIKR TVAAP

＞LC-105-F02 (SEQ ID NO：1078)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISSWLAWYQQ KPGEAPKLLI

YAASSLRNGV PSRFIGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPPTF

GGGTKVEIKR TVAAP

＞LC-106-C06 (SEQ ID NO：1079)

AQDIQMTQSP ATLSASVGDR VTITCRASQS VSRWLAWYQQ KPGKAPKLLI

YKASTLESGV PSRFSGSGSG TEFTLTISSL QPDDFATYYC QQYGAFGQGT

KVEIRRTVAA P

＞LC-106-D06 (SEQ ID NO：1080)

AQDIQMTQSP SSVSASVGDR VTITCRASQG ISSWLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPYTF

GQGTKLEIKR TVAP

＞LC-106-E12 (SEQ ID NO：1081)

AQDIQMTQSP GTLSLSPGER ATLSCRPSQS VYSNYLAWYQ QKPGQAPRLL

IYGASTRATG IPDRFSGSAS GTDFTLTISR LEPEDFAVYY CQQYGNSYTF

GPGTKVDIKR TVAAP

＞LC-106-G12 (SEQ ID NO：1082)

AQDIQMTQSP SFVSASIGDR VTITCRASQS IGTLLNWYQH KPGTVPSLLI

YGASNLRGGV PARFSGSGSG TDFTLTISSL QPEDFATYYC QHDTFGGGTK

VDIKRTVAAP

＞LC-107-B05 (SEQ ID NO：1083)

AQSELTQPAS VSGSPGQSIT ISCTGTSSDV GAYNYVSWYQ QHPGKVPKLM

IYEVSNRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CNSYTTSATL

VFGGGTKLTV LSQPKAAP

＞LC-107-D01 (SEQ ID NO：1084)

AQSELTQPAS VSGPPGQSIT ISCTGTSSDV GGYNYVSWYQ QHPGKAPKLI

IYEVSNRPSG VSYRFSASKS DNTASLTISG VQAEDEADYY CSSYKRGGTY

VFGTGTTVIV LGQPKANP

＞LC-107-D05 (SEQ ID NO：1085)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISSWLAWYQQ KPGKVPKLLI

HAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDVATYYC QKYNSAPLTF

GGGTKVEIKR TVAAP

＞LC-107-D12 (SEQ ID NO：1086)

AQYELTQPPS ASGTPGQRVT ISCSGSSSDI GSNTVNWYQQ LPGTAPKLLI

YSNNQRPSGV PDRFSGSKSG TSASLAISGL QSEDEADYYC AAWDDSLNGY

VFGTGTKVTV LGQPKANP

＞LC-107-F11(SEQ ID NO：1087)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSGSYLAWYQ QKPGQAPSLL

IYGAYSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQHYGSSPRT

FGGGTKVEIK RTVAAP

＞LC-108-C10 (SEQ ID NO：1088)

AQSVLTQPAS VSGSPGQSIT ISCTGTSSDV GGYNYVSWYQ QHPGQPPKLI

IYEVSNRASG VSNRFSGSKF GNTASLTISG LQSEDEADYH CSSYTSSTTL

LFGGGTRLTV LGQPKAAP

＞LC-108-D11 (SEQ ID NO：1089)

AQSVLTQPAS VSGSPGQSIT ISCTGTNTDV GGYNLVSWYQ QHPGKAPKLI

IYEVSNRPSG VSNRFSGSKS GNTASLTISG LQAEDEVDYY CGSYTSSSTH

VFGSGTKVTV LGQPKANP

＞LC-108-E10 (SEQ ID NO：1090)

AQDIQMTQSP GTLSLSPGER ATLSCRASQT VSSTYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGGGS GTDFTLTISR LEPEDFAVYY CQQYGSSPPR

YTFGQGTKLE IKRTVAAP

＞LC-108-E11 (SEQ ID NO：1091)

AQYELTQPAS VSGSPGQSIT ISCTATSSDL GSYNFVSWYQ QHPDKAPKLM

IFEVSRRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CCSYAGSNTY

VFGTGTKVTV LGQPKANP

＞LC-109-A11 (SEQ ID NO：1092)

AQSELTQPAS VSGSPGQSIT ISCTGTSSDV GGYNYLSWYQ QHPGKAPKLM

IYGVSNRPSG VSTRFSGSKS GNTASLTISG LQAEDEADYY CSSYTSTGTR

VFGGGTRLTV LGQPKAAP

＞LC-109-E08 (SEQ ID NO：1093)

AQDIQMTQSP ATLSVSPGER ATLSCRASQS VSSNLAWYQQ KPGQAPRLLI

YGASTRATGI PARFSGSGSG TEFTLTISSL QSEDFAVYYC QQNNNWPPSF

TFGPGTKVDI KRTVAAP

＞LC-110-D06(SEQ ID NO：1094)

AQDIQMTQSP STLSASLGDR VIITCRASQG IRSWLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPWTF

GQGTKVEIKR TVAAP

＞LC-110-D11(SEQID NO：1095)

AQSELTQPAS VSGSPGQSIT ISCTGTSTDV GGYNYVSWYQ KHPGKAPKLM

IYDVSNRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CSSYTNTITV

VFGGGTKLTV LGQPKAAP

＞LC-110-F08(SEQ ID NO：1096)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VVSNFLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFILTISR LEPEDFAVYY CQQYGSSPYS

FGQGTKLEIK RTVAAP

＞LC-110-G01 (SEQ ID NO：1097)

AQYELTQPPS ASGTPGQRVT ISCSGSSSNI GSNTVNWYQQ LPGTAPKLLI

YSNNQRPSGV PDRFSGSKSG TSASLAISGL QSEDEADYYC AAWDDSLNGY

VFGTGTKVTV LGQPKANP

＞LC-110-H11(SEQ ID NO：1098)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL HPEDFATYYC QQTYSTLFTF

GPGTKVHIKR TVAAP

＞LC-111-B02(SEQ ID NO：1099)

AQYELTQDPA VSVALGQTVR ITCQGDSLRS YYASWYQQKP GQAPVLVIYS

KSNRPSGIPD RFSGSSSGST ASLTITGAQA EDEADYYCNS RDSSGNHLVF

GGGTKLTVLG QPKAAP

＞LC-111-C10 (SEQ ID NO：1100)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPEKAPQLLI

FAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYNAPWTF

GQGTKVEIRR TVAAP

＞LC-111-D11 (SEQID NO：1101)

AQDIQMTQSP ATLSLSPGDR ATLSCRASQR VSSTFLAWYQ QRPGQAPRLV

IFGTSSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CHQYGSSPRT

FGQGTKVEIK RTVAAP

＞LC-112-C02 (SEQ ID NO：1102)

AQDIQMTQSP GTLSLSPGDR ATLSCRASQS VSSNYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFGVYY CQQFGSSLFT

FGPGTKVNIK RTVAAP

＞LC-112-C12 (SEQID NO：1103)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTEFTLTISS LQSEDSAVYY CQQYNNWPPL

TFGGGTKVEI KRTVAAP

＞LC-112-D04 (SEQID NO：1104)

AQSALTQPLS VSVALGQTAR ITCGGNNIGS KNVHWYQQKP GQAPVLVIYR

DSNRPSGIPE RFSGSNSGNT ATLTISGAQA GDEADYYCQV WDSSTVFGGG

TKLTVLGQPK AAP

＞LC-112-D07(SEQ ID NO：1105)

AQDIQMTQSP SSLSASVGDR VTITCRASQT IRTYLNWYQQ KPGIAPKFLI

YDASNLQTGV PSRFSGSGSG THFTLTISSL QPEDFGTYYC QQSYGGPPTF

GRGTKIEIKR TVAAP

＞LC-113-D05(SEQ ID NO：1106)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSSFT

FGPGTKVDIK RTVAAP

＞LC-113-E03 (SEQ ID NO：1107)

AQDIQMTQSP SSLSASVGDR VTITCRASQS INSYLMWYQQ KPGKAPNLLI

YAASSLQNGV PSRFSGSGSG TDFTLTISSL QPEDFAAYYC QQSYSTPLTF

GGGTKVEIKR TVAAP

＞LC-113-G11 (SEQ ID NO：1108)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTLVTF

GQGTKLEIKR TVAAP

＞LC-114-D02 (SEQ ID NO：1109)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISNYLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDVATYYC QKYNSAPWTF

GQGTKVEIKR TVAAP

＞LC-114-E02 (SEQ ID NO：1110)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VGGYLAWYQQ KPGQAPRLLI

YDASKRATGI PARFSGSGSG TDFTLTISSL EPEDFAVYYC QQRSKWPPYT

FGQGTKLEIK RTVAAP

＞LC-114-F04 (SEQ ID NO：1111)

AQSELTQPAS VSGSPGQSIA ISCTGTSSDI GAYPFVSWYQ QHPGKAPKLL

IYGVTTRPFG VSDRFSGSKS GSTASLTISG LQAEDEADYY CSSYAGGRNL

PYVFGTGTTV TVLGQPKANP

＞LC-114-G06 (SEQ ID NO：1112)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISSWLAWYQQ RPERAPKSLI

YAASSLERGV PSRFRGSGSG TDFTLTISSL QPEDFGTYYC QQYHNFPLTF

GGGTRVEINR TVAAP

＞LC-114-G09 (SEQ ID NO：1113)

AQSALTQPAS VSGSPGQSIT ISCTGTSSDV GSYKLVSWYQ QHPGKAPKLM

IYEGSKRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CCSYAGSSTW

VFGGGTKLTV LGQPKAAP

＞LC-114-H07(SEQ ID NO：1114)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSPFT

FGPGTKVDIK RTVAAP

＞LC-115-F04 (SEQ ID NO：1115)

AQSALTQPPS VSVSPGQTVS ITCSGDDLGG RHVSWFQQLS GQSPVLVIYQ

DDKRPSGIPE RFSGSNSGNT ATLTISGTQS VDEGDYYCLA WHNYKYVFGS

GTTVTVLRQP KANP

＞LC-115-F08(SEQ ID NO：1116)

AQSELTQPAS VSGSPGQSIT ISCTGTSSDV GGYNYVSWYQ QHPGKAPKLM

IYEVSNRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CSSYTSSSTW

VFGGGTKLTV LGQPKAAP

＞LC-115-G12 (SEQ ID NO：1117)

AQDIQMTQSP VTLSLSPGDR ATLSCRASQS VSFNLAWYQH KPGQAPRLLM

FDASNRATGI PDRFSGSGSG TDFTLTIKRL EPEDFAVYYC QQYGTSPFTF

GPGTNVDVKR TVAAP

＞LC-116-A08 (SEQ ID NO：1118)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSNSYLAWYQ QRPGQAPRLL

IYGASSRATG IPDRFSGSGS GTEFTLTISR LEPEDFAVYY CQQFGSSTGY

TFGQGTKLEL KRTVAAP

＞LC-116-C09(SEQ ID NO：1119)

AQSALTQPPS VSVSPGQTAS ITCFGDKLGD KYGSWYQQKP GQSPVLVLYQ

YWKPPSGIPE RFSGSNSGNT ATLTINVTQT MDEADYYCQA WDSNTVVFGG

GTKLTVLGQP KAAP

＞LC-116-E08 (SEQ ID NO：1120)

AQDIQMTQSP SSLSASVGDR VTITCLASQS ISSYLNWYQQ KPGKAPKLLI

YDASSLESGV PSRFSGSGSG TDFTLTISSL QPEDFAIYYC QQFNGYPPIT

FGQGTRLEIK RTVAAP

＞LC-117-A12 (SEQ ID NO：1121)

AQDIQMTQSP GTLSLSPGDR ATLSCRASQS VGSDYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSLYT

FGQGTKLEIK RTVAAP

＞LC-117-B07 (SEQ ID NO：1122)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSSYLAWYQQ KPGQAPRLLI

YDASNRATGI PARFSGSGSG TDFTLTISSL EPEDFAVYYC QQRSNFGGGT

KVEIKRTVAA P

＞LC-117-C04 (SEQ ID NO：1123)

AQDIQMTQSP LSLSASVGDR VTITCRASQT FNNYLNWYQQ KPGKAPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPFTF

GPGTKVDIKR TVAAP

＞LC-117-F04(SEQ ID NO：1124)

AQDIQMTQSP SSLSASVGDR VTITCRASQG IGNYLAWYQQ KPGKAPNLLI

YKTSNLQSGV PSRFRGSGSG TEFTLTITSV QPDDFATYFC QRYDSYSQYI

FGQGTKLETK RTVAAP

＞LC-117-F08(SEQ ID NO：1125)

AQYELTQPPS VSVSPGQTAT ITCSGDKLEE KYVCWYQQKP GQSPAVVIYQ

DTKRPSGVPD RFSGSKSGTS ASLAISGLRS EDEADYYCAT WDDSLSGPVF

GGGTKLTVLG QPKAAP

＞LC-118-E07 (SEQ ID NO：1126)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISNYLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDVATYYC QKYNSAPLTF

GGGTKVEIKR TVAAP

＞LC-119-B09(SEQ ID NO：1127)

AQDIQMTQSP ATLSVSPGEG ATLSCRASQS VGNSLAWYQQ KPGQAPRVLV

YGASTRASGI PARFSGSGSV TEFTLTISSL QSEDFAVYYC QEYNKWPITF

GQGTRLERKR TVAAP

＞LC-121-A07 (SEQ ID NO：1128)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSNYLAWYQ QKPGQAPRLL

MYGASYRATG IPDRFSGSGS GTDFTLTISS LQPEDFATYY CQQSYSTPWT

FGQGTKVEIK RTVAAP

＞LC-121-H07(SEQ ID NO：1129)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISRYLNWYQQ KPGKAPKLLI

YAASNLQSGV PSRFSGSGSG RDYTLTISSL QPEDFVTYYC QQSYSTPWTF

GQGTKVEIKR TVAAP

＞LC-122-A05 (SEQ ID NO：1130)

AQDIQMTQSP SSFSASTGDR VTITCRASQS ISRWLAWYQQ KPGKAPKLLI

YEASTLESGV PSTFSGSGSG TEFTLTISSL QPDDFATYYC QQYNSYPLTF

GGGTKVEIKR TVAAP

＞LC-122-D01 (SEQ ID NO：1131)

AQDIQMTQSP SSVSASVGDR VTITCRASQD IRTWLAWYQQ KSGKAPKLLI

YSSSSLQSGI SSRFSGSGSG TDFTLTISNL QPEDSAIYYC QQATTFPWTF

GQGTKVEIKR TVAAP

＞LC-122-G06 (SEQ ID NO：1132)

AQSALTQPAS VSGSPGQSIT ISCTGTSSDV GGYNYVSWYQ QHPGKAPKLM

IYDVSKRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CSSYTSSITL

VVFGGGTKLT VLSQPKAAP

＞LC-122-H04 (SEQ ID NO：1133)

AQYELTQPRS VSGSPGQSVT ISCTGTSSDV GGFNYVSWYQ QHPGKAPKLM

IYDVSKRPSG VPDRFSGSKS GTTASLTISG LQADDEADYY CCSYTGNYTY

VFGTGTKVTV LGQPKANP

＞LC-123-E02(SEQ ID NO：1134)

AQSALTQPAS VSGSPGQSIT ISCSGTSSDV GAYYHVSWYQ QHPGKAPKLM

IYDVSNRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CSLYIGTSTP

WVFGGGTKLT VLGQPKAAP

＞LC-124-A01 (SEQ ID NO：1135)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ITGYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLSISSL QPEDFATYYC QQSYSTPYTF

GQGTKLEIKR TVAAP

＞LC-124-C04 (SEQ ID NO：1136)

AQDIQMTQSP SSLSASVGDR VTITCRASQT ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGGGSG TDFTLTISSL QPEDFATYYC QQSYSTPMYT

FGQGTKLYIK RTVAAP

＞LC-124-C12 (SEQ ID NO：1137)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSNYLAWYQ QKPGKAPKLL

IYAASSLQSG VPSRFSGSGS GTDFTLTISS LQPEDFATYY CQQRWTFGQG

TKVEIKRTVA AP

＞LC-124-G12 (SEQ ID NO：1138)

AQDIQMTQSP SSVSASVGDR VTITCRASQG ISSWLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPLTF

GGGTKVEIKR TVAAP

＞LC-124-H10 (SEQ ID NO：1139)

AQDIQMTQSP ATLSLSAGER ATLSCRASQS VNIDVGWYQQ KPGQAPRLLI

YDASKRATGI PTRFSGSGSG TDFTLTIANL EPEDFAVYYC QQRARWLTFG

GGTRLEIKRT VAAP

＞LC-125-C04 (SEQ ID NO：1140)

AQSELTQPAS VSGSPGQSIT ISCAGTSSDL GGYDYVSWYQ QYPGKAPKLI

IYQVGRRPSG VSNRFSGSKS GNTASLTISG LQTEDEADYY CSSYTSSRTR

VFGGGTRVTV LGQPKAAP

＞LC-126-B12 (SEQ ID NO：1141)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSSYT

FGQGTKLEIK RTVAAP

＞LC-126-C10 (SEQ ID NO：1142)

AQDIQMTQSP SSLSASVGDR VTITCRAGQT INNYLNWYQQ KPGKAPNLLI

YAASNLQSGV PSRFSGSKSG TDFTLTISSL QPEDFATYHC QQTYRTPFTF

GGGTRVEIKG TVAAP

＞LC-126-F11 (SEQ ID NO：1143)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISSWLAWYQQ KPGKAPKLLI

YKASSLESGV PSRFSGSGSG TEFTLTISSL QPDDFATYYC QQYNSYRYTF

GQGTKLEIKR TVAAP

＞LC-126-H09 (SEQ ID NO：1144)

AQSVLTQDPA VSVALGQTVR FTCQGDSLRN YHPSWYQQKP RQAPVLVMFG

RNNRPSGIPD RFSGSTSGGT ASLTITATQA DDEADYFCSS RDGSGNFLFG

GGTKLTVLGQ PKAAP

＞LC-127-C06 (SEQ ID NO：1145)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISSWLAWYQQ KPGKAPKLLI

YKASSLGSGV PSRFSGSGSG TEFTLTISSL QPDDFATYYC QQYYSYSQTF

GQGTKVEIKR TVAAP

＞LC-127-D05 (SEQ ID NO：1146)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPRTF

GPGTKVDIKR TVAAP

＞LC-127-H05 (SEQ ID NO：1147)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKVPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPDDFATYYC QQTFSTWTFG

QGTKVEIRRT VAAP

＞LC-128-A06 (SEQ ID NO：1148)

AQSALTQPPS LSVSAGQTAT ITCSPSGGGL RNKYVSWYQQ RPGQSPFLVI

YKDAERPSGI PERFSGSNFG NTATLTIGGT QAMDEADFYC LAWDSTTRVF

GGGTKLTVLS QPKAAP

＞LC-128-E07 (SEQ ID NO：1149)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISSWLAWYQQ KPGKAPKLLI

YKASSLESGV PSRFSGSGSG TEFTLTISSL QPDDFATYYC QQYNSYPVTF

GQGTKLEIKR TVAAP

＞LC-128-F11 (SEQ ID NO：1150)

AQDIQMTQSP SSLSASVGDR VTITCRASQY ISTYLAWYQQ KPGKAPKLLI

YKASDLESGV PSRFSGSGSG TEFTLTINSL QPDDFATYYC QQYNTYWTFG

HGTKVEIKRT VAAP

＞LC-129-B09 (SEQ ID NO：1151)

AQDIQMTQSP SFLSASVGDR VTITCRASQG ISSYLAWYQQ KPGKAPKLLI

YAASTLQSGV PSRFSGSGSG TEFTLTISSL QPEDFATYYC QQLNSYPRTF

GQGTKVEIKR TVAAP

＞LC-130-B02 (SEQ ID NO：1152)

AQSALTQPAS VSGSPGQSIT ISCTDTSGNV GSYNLVSWYQ QHPDKAPKLM

IFEVSRRPSG VSDRFSGSKS GNTASLTISG LQAEDEADYY CCSYAGSNTY

VFGTGTKVTV LGQPKANP

＞LC-131-A03 (SEQ ID NO：1153)

AQYELTQPAS VSGSPGQSIT ISCTGTNTDV GGYNYVSWYQ QYPGKAPKLM

IYEVNHRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CSSYTYRNTY

VFGTGTKVTV LGQPKANP

＞LC-131-B05 (SEQ ID NO：1154)

AQYELTQPAS VSGSPGQSIT ISCTGTSSDI GAYNYVSWYQ QHPGKAPKLM

IYDVSNRPSG VSNRFSGSKS GNTASLTISG LQAEDEADYY CSSYRSSSLM

FGGGTKLTVL GQPKAAP

＞LC-131-C08 (SEQ ID NO：1155)

AQDIQMTQSP SSVSASVGDR VTITCRASQD VLVSFAWYQQ RPGTAPKLLI

YAASHLHPGV PSRFSASGSG TDFTLTINGL QPEDFATYYC QQARSFPHTF

GQGTRLEKKR TVAAP

＞LC-131-C09 (SEQ ID NO：1156)

AQYELTQPPS ASGTPGQRVT ISCSGSSSNI GTNRVNWYQQ IPGTAPKLLI

YSNNQRPSGV PDRFSDSKSG TSASLAISGL QSEDEADYYC AAWDDSLTGP

VFGGGTKVTV LRQPKAAP

＞LC-131-D01 (SEQ ID NO：1157)

AQDIQMTQSP GTLSLSPGER ASLSCRASQS VSSNYLSWYQ QKPGQAPRLL

IYGTSNRASG IPVRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGGAPLF

IFGPGTRVDI KRTVAAP

＞LC-131-D12 (SEQ ID NO：1158)

AQSALTQPPS VSVAPGQTAS ISCSGNILDN SYASWFQQKP GQSPVMVIHR

DNKRPSGIPE RFSGSTSGNT ATLTISGTQA VDEADYYCQA WDRTTGVFGT

GTRLTVLRQP KAAP

＞LC-131-F07 (SEQ ID NO：1159)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPLTF

GGGTKVEIKR TVAAP

＞LC-131-F09 (SEQ ID NO：1160)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSSYLAWYQQ KPGQAPRLLI

YDASNRATGI PARFSGSGSG TDFTLTISSL EPEDFAVYYC QQRSNWLWTF

GQGTKVEIKR TVAAP

＞LC-131-F12 (SEQ ID NO：1161)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPRTF

GQGTKVEIKR TVAAP

＞LC-131-G06 (SEQ ID NO：1162)

AQYELTQAPS ASGTPGQRVT ISCSGSRSNI GTNPVNWYQH VPGTAPKLLI

LVNNQRPSGV PDRFSGSKSG ASASLAISGL QSEDEAEYYC ATWDGSLNGP

VFGGGTKLTV LRQPKAAP

＞LC-132-C01 (SEQ ID NO：1163)

AQSALTQPRS VSGSPGQSVT ISCTGTGSNI DGYNYVSWYQ QYPGNAPKLI

IYDVGKRPSG VPNRFSGSKS GNTASLTISG LQAEDEADYY CCSYAGSYSY

VFGVGTKVTV LGQPKANP

＞LC-132-C08 (SEQ ID NO：1164)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPRTF

GQGTKLEIKR TVAAP

＞LC-132-D02 (SEQ ID NO：1165)

AQSALTQPRS VSGSPGQSVT ISCTGTSSDV NYVSWYQQHP GKAPKLMIYA

VTKRPSGVPD RFSGSKSGNT ASLTVSGLQA EDEADYYCSS YAGSNNLVFG

GGTKLTVLGQ PKAAP

＞LC-132-D08 (SEQ ID NO：1166)

AQDIQMTQSP STLSASVGDR VTITCRASQS ISNWLAWYQQ KPGKAPKLLI

YMASTLESGV PSRFSGSGSG TEFTLTISSL QPADFATYYC QQYNSYSVTF

GGGTKVEIKR TVAAP

＞LC-132-H11 (SEQ ID NO：1167)

AQYELTQPPS ASGTPGQRVT ISCSGSSSNI GSNYVYWYQQ LPGTAPKLLI

YRNNQRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSLSGH

AVFGGGTQLT VLGQPKAAP

＞LC-133-E02 (SEQ ID NO：1168)

AQSVLTQPPS ASGTPGQRVT VSCSGSSSNI GSNIVSWYRQ LPGTAPKLLI

YSNNRRPSGV PDRFSGSKSG TSASLAISGL QSEDEADYYC AAWDDSLNGH

VFGGGTKLTV LRQPKAAP LC-133-E02 is identical with LC-138-G11

＞LC-138-G11(SEQ ID NO：1168)

AQSVLTQPPS ASGTPGQRVT VSCSGSSSNI GSNIVSWYRQ LPGTAPKLLI

YSNNRRPSGV PDRFSGSKSG TSASLAISGL QSEDEADYYC AAWDDSLNGH

VFGGGTKLTV LRQPKAAP

＞LC-134-B03 (SEQ ID NO：1169)

AQSALTQPPS ASGSPGQSVT ISCTGTSSDV GAYNYVSWYQ QHPGKAPKLI

IYEVNKRPSG VPDRFSASKS GNTASLTVSG LQAEDEADYY CNSYAGSNSL

IFGGGTKLTV LGQPKAAP

＞LC-134-D07 (SEQ ID NO：1170)

AQDIQMTQSP SSLSASVGDR VTITCRASQG IRNDLGWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC LQDYNYPRTF

GQGTKVEIKR TVAAP

＞LC-135-B05 (SEQ ID NO：1171)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISTYLNWYQQ KPGKAPNLLI

YAASSLQSGV PSRFSGSGSG TDFTLTVSSL QPEDFGTYYC QQYNSFPFSF

GQGTRLEINR TVAAP

＞LC-135-C11 (SEQ ID NO：1172)

AQDIQMTQSP SSVSASVGDR VTITCRASQG ISSWLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPSLT

FGGGTKVEIK RTVAAP

＞LC-135-C12 (SEQ ID NO：1173)

AQSELTQPPS MSGTPGQRVI ISCSGSNSNI GNNFVYWYQQ VAGSAPKLLI

FRNNQRPSGV PDRFTVSKSG ASASLAIGGL RSEDEADYYC AAWDDSLSGV

LFGGGTKVTV LGQPKAAP

＞LC-135-E10 (SEQ ID NO：1174)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTLYTF

GQGTKLEIKR TVAAP

＞LC-135-F03 (SEQ ID NO：1175)

AQDIQMTQSP ATLSLSPGER ATLSCRASQS VSSYLAWYQQ KPGQAPRLLI

YDASNRATGI PARFSGSGSG TDFTLTISSL EPEDFAVYYC QQRSNWPSPI

AFGQGTRLEI KRTVAAP

＞LC-136-A07 (SEQ ID NO：1176)

AQSVLTQPPS ASGTPGQRVT ISCSGGSSNI GSNRVNWYQQ VPGTAPKLLI

DSNNQRPSGV PDRFSGSKSG TSASLAISGL QSEDEAGYYC AAWDDNLIGP

VFGGGTKLTV LGQPKAAP

＞LC-136-B06 (SEQ ID NO：1177)

AQDIQMTQSP SSLSASVGDR VTITCRASQS ISSYLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPWTF

GQGTKVEIKR TVAAP

＞LC-136-D07 (SEQ ID NO：1178)

AQDIQMTQSP SSVSASVGDR VTITCRASQD IASWLAWYQQ KPEKVPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTINSL QPEDFATYYC QQANSFPFTF

GPGTKVDFKR TVAAP

＞LC-136-E10 (SEQ ID NO：1179)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VTTFLAWYQR KPGQAPRLLI

YGASSRAADI PDRFSGSGSG TDFTLTISRL EPEDLAVYYC QQYRFSPPTF

GPGTTVDIRR TVAAP

＞LC-136-E12 (SEQ ID NO：1180)

AQDIQMTQSP SSLSASVGDR VTITCRASHV INIDLGWYQQ KPGKAPKLLI

YGASHLQRGV PSRFSGSGSG TVFTLTISGL QPEDFATYYC LQDSFYPRTF

GQGTRLEIKR TVAAP

＞LC-136-F08 (SEQ ID NO：1181)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGqAPRLL

IYDASNRATG IPARFSGSGS GTDFTLTISS LEPEDFAVYY CQQWDTFGQG

TKLEIKRTVA AP

＞LC-137-B01 (SEQ ID NO：1182)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSGWT

FGQGTKVEIK RTVAAP

＞LC-137-E01 (SEQ ID NO：1183)

AQSELTQPPS ASGTPGQRVT ISCSGTNSNI GSNSVFWYQQ LSGTAAPKVL

ILRNSQRPSG VSDRFSGSKS GTSASLAISG LRSEDEADYY CATWDDSLRS

PVFGGGTKLT VLGQPKAAP

＞LC-137-G10 (SEQID NO：1184)

AQDIQMTQSP SSLSASVGDR VTITCRTSQT VSTFLNWYQQ KPGTAPKLLI

YAASRLQSGV PSRFSGSGSE TDFTLTISRL QPEDFATYYC QQSFTSPRTF

GLGTKVEIKR TVAAP

＞LC-138-A03 (SEQID NO：1185)

AQDIQMTQSP SSVSASVGDR VTITCRASQG ISSWLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPHTF

GQGTKLEIKR TVAAP

＞LC-138-B04 (SEQID NO：1186)

AQDIQMTQSP SSLSASVGDR VAITCRASQS IDTYLNWYQQ KPGKAPKLLI

YAASKLEDGV PSRFSGSGTG TDFTLTIRSL QPEDFASYFC QQSYSSPGIT

FGPGTKVEIK RTVAAP

＞LC-138-B06 (SEQID NO：1187)

AQDIQMTQSP SSVSASVGDR VTITCRASQG ISSWLAWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQANSFPLTF

GGGTKVEIKR TVAAP

＞LC-138-B10 (SEQ ID NO：1188)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISR LEPEDFAVYY CQQYGSSLTF

GQGTRLEIKR TVAAP

＞LC-138-C04 (SEQ ID NO：1189)

AQYELTQPAS VSGSPGQSIT ISCTGTSSDV GAYNYVSWYQ HHPGKAPKLL

IYDVSNRPSG ISSRFSGSKS GNTASLTISG LQAEDEADYY CSSYTSSYTW

VFGGGTKLTV LSQPKAAP

＞LC-138-C09 (SEQ ID NO：1190)

AQSELTQPAS VAGSPGQSIT ISCTGTSSDV GLYNFVSWYQ QHPGKAPKLM

IYDVSRRPSG VSNRFSASKS GTRASLTVSG LQAEDEADYY CSSYAGSNNY

VFGTGTKVTV LGQPKANP

＞LC-138-E02 (SEQ ID NO：1191)

AQDIQMTQSP SSVSASVGDR VTITCRASQG ISSWLVWYQQ KPGKAPKLLI

YAASSLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQAKTFPLTF

GGGTKVEIKR TVAAP

＞LC-138-E12 (SEQ ID NO：1192)

AQSALTQPAS VSGSPGQSIT ISCSGTSSDV GAYYHVSWYQ QHPGKAPKLM

IYEVTNRPSG ISNRFSGSKS GNTASLTISG LQADDEADYF CSSYTTSNTL

VFGGGTKVTV LGQPKAAP

＞LC-139-A04 (SEQ ID NO：1193)

AQDIQMTQSP GTLSLSPGDR ATLSCRASQS VSSSYLAWYQ QKAGQAPRLL

IYGAASRATG IPDRFSGSGS GTDFTLTISR LEREDFAVYY CQQYGSSPLI

TFGQGTRLEI KRTVAAP

＞LC-139-A09 (SEQ ID NO：1194)

AQDIVMTQTP PSLPVNPGEP ASISCRSSQS LVYSDGNTYL NWFQQRPGQS

PRRLIYKVSN RDSGVPDRFS GSGSGTDFTL KISRVEAEDV GVYYCMQGTH

WQRLTFGGGT KVEIKRTVAA P

＞LC-139-C02 (SEQ ID NO：1195)

AQDIQMTQSP SSLSASVGDR VTITCRASQG IRRALAWYQQ KPGKPPKLLI

NDASSLESGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPPWT

FGQGTKVEIK RTVAAP

＞LC-139-E12 (SEQ ID NO：1196)

AQDIQMTQSP SSVSASVGDR VTMTCRASRG ISNWLAWYQQ KPGKAPELLI

HSASTLHAGV PSRFSGSGSG TDFTLTISSL QPGDFATYYC QEGNSFPYTF

GQGTNLQIKR TVAAP

＞LC-139-F04 (SEQ ID NO：1197)

AQDIQMTQSP SSLSASVGDR VTITCRASQA ISTNLNWYQQ KPGKAPKLLI

YAASSLQSGV PSRFIGSGSG TDFTLTISSL HPEDFATYHC QQTFSPPHTF

GQGTKVEIQR TVAAP

＞LC-139-F09 (SEQ ID NO：1198)

AQDIQMTQSP GTLSLSPGER ATLSCRASQS VSSSYLAWYQ QKPGQAPRLL

IYGASSRATG IPDRFSGSGS GTDFTLTISS LQLEDFGTYF CQQSYRSPYT

FGQGTKVDIK RTVAAP

＞LC-135-H01 (SEQ ID NO：1376)

AQSVLTQPPS ASGTPGQRVT ISCSGSSSNI GSNYVYWYQQ LPGTAPKLLI

YRNNQRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSLSGV

VFGGGTKLTV LGQPKAAP

Above-mentioned light chain variable territory is from following light chain variable type:

LC-092-B12κA20

LC-092-F08κA11

LC-092-G06λ2e

LC-093-C02κA27

LC-093-C08κA27

LC-093-C10κA20

LC-093-E11κL6

LC-093-F09λ2a2

LC-093-G06λ2e

LC-093-G09κ012

LC-094-D08λ1c

LC-094-E08λ2a2

LC-094-F03κA27

LC-095-A11κL6

LC-095-G09κL12

LC-095-H10κL12

LC-096-D03λ2e

LC-096-D09λ1c

LC-096-E11κ012

LC-097-D04λ1g

LC-097-E07κ012

LC-097-F03κL2

LC-097-F04λ1g

LC-097-F08κ012

LC-097-G01κA20

LC-097-G07κ012

LC-098-C10κ012

LC-098-E01κA27

LC-098-E09κL19

LC-098-G05κ012

LC-098-H04λ2c

LC-099-A09κA27

LC-099-C12λ2a2

LC-099-D05κA27

LC-099-E11κL12

LC-101-E01κL5

LC-102-A04λ1g

LC-102-C02κA20

LC-102-C12κA20

LC-102-D07κA27

LC-102-E09κL6

LC-102-G11κA27

LC-102-G12λ1c

LC-102-H02κL6

LC-102-H11κL6

LC-103-A01λ1g

LC-103-A03κL5

LC-103-E09λ2a2

LC-103-F07λ2a2

LC-103-H07κ012

LC-104-A12κ012

LC-104-B03κA27

LC-104-B12κA20

LC-104-D12κ012

LC-104-H12λ2c

LC-105-B06κ012

LC-105-F02κL11

LC-106-C06κL12

LC-106-D06κL5

LC-106-E12κA27

LC-106-G12κ012

LC-107-B05λ2a2

LC-107-D01λ2a2

LC-107-D05κA20

LC-107-D12λ1c

LC-107-F11κA27

LC-108-C10λ2a2

LC-108-D11λ2a2

LC-108-E10κA27

LC-108-E11λ2b2

LC-109-A11λ2a2

LC-109-E08κL2

LC-110-D06κ012

LC-110-D11λ2a2

LC-110-F08κA27

LC-110-G01λ1c

LC-110-H11κ012

LC-111-B02λ31

LC-111-C10κ012

LC-111-D11κA27

LC-112-C02κA27

LC-1 12-C12κL16

LC-112-D04λ3j

LC-112-D07κ012

LC-113-D05κA27

LC-113-E03κ012

LC-113-G11κ012

LC-114-D02κA20

LC-114-E02κL6

LC-114-F04λ2a2

LC-114-G06κL15

LC-114-G09λ2b2

LC-114-H07κA27

LC-115-F04λ3r

LC-115-F08λ2a2

LC-115-G12κA27

LC-116-A08κA27

LC-116-C09λ3r

LC-116-E08κL5

LC-117-A12κA27

LC-117-B07κL6

LC-117-C04κ012

LC-117-F04κL12

LC-117-F08λ1g

LC-118-E07κA20

LC-119-B09κL2

LC-121-A07κL16

LC-121-H07κ012

LC-122-A05κL12

LC-122-D01κL19

LC-122-G06λ2a2

LC-122-H04λ2e

LC-123-E02λ2a2

LC-124-A01κ012

LC-124-C04κ012

LC-124-C12κ012

LC-124-G12κL5

LC-124-H10κL6

LC-125-C04λ2a2

LC-126-B12κA27

LC-126-C10κ012

LC-126-F11κL12

LC-126-H09λ31

LC-127-C06κL12

LC-127-D05κ012

LC-127-H05κ012

LC-128-A06λ3r

LC-128-E07κL12

LC-128-F11κL12

LC-129-B09κL8

LC-130-B02λ2b2

LC-131-A03λ2a2

LC-131-B05λ2a2

LC-131-C08κL5

LC-131-C09λ1c

LC-131-D01κA27

LC-131-D12λ3r

LC-131-F07κ012

LC-131-F09κL6

LC-131-F12κ012

LC-131-G03 λ 2c is identical with LC-098-H04

LC-131-G06λ1c

LC-132-C01λ2e

LC-132-C08κ012

LC-132-D02λ2c

LC-132-D08κL12

LC-132-H11λ1g

LC-133-E02λ1c

LC-133-E08 λ 1c is identical with LC-102-G12

LC-134-B03λ2c

LC-134-D07κL11

LC-135-B05κ012

LC-135-C11κL5

LC-135-C12λ1g

LC-135-E10κ012

LC-135-F03κL6

LC-136-A07λ1c

LC-136-B06κ012

LC-136-D07κL5

LC-136-E10κA27

LC-136-E12κL11

LC-136-F08κL6

LC-137-B01κA27

LC-137-E01λ1g

LC-137-G10κ012

LC-138-A03κL5

LC-138-B04κ012

LC-138-B06κL5

LC-138-B10κA27

LC-138-C04λ2a2

LC-138-C09λ2a2

LC-138-E02κL5

LC-138-E12λ2a2

LC-138-G11 λ 1c is identical with LC-133-E02

LC-139-A04κA27

LC-139-A09κA17

LC-139-C02κL4

LC-139-E12κL5

LC-139-F04κ012

LC-139-F09κA27

Classify the aminoacid sequence of exemplary variable region of heavy chain down as.

＞HC-092-B12(SEQ ID NO：1199)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS QYLMAWVRQA PGKGLEWVSS

IYPSGGNTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR

SIVPYSSSWY MPRDYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-092-F08 (SEQID NO：1200)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYPMIWVRQA PGKGLEWVSG

IYSSGGTTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-092-G06 (SEQID NO：1201)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYGMNWVRQA PGKGLEWVSS

IWSSGGYTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

IWYSMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-093-C02 (SEQ ID NO：1202)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYAMHWVRQA PGKGLEWVSW

ISPSGGQTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAK

VLRYFDWLLG GDAFDIWGQG TMVTVSSAST KGPSVFPLAP

＞HC-093-C08 (SEQID NO：1203)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYPMGWVRQA PGKGLEWVSS

IYPSGGSTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-093-C10 (SEQID NO：1204)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYTMEWVRQA PGKGLEWVSV

IRPSGGTTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED MAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-093-E11 (SEQID NO：1205)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYEMAWVRQA HGKGLEWVSV

IYPSGGATRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVA

QYYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-093-F09 (SEQ ID NO：1206)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS DYRMWWVRQA PGKGLEWVSY

IVPSGGQTSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGS

AYQRRTYSSG WYSASGRRGT AEYFQHWGQG TLVTVSSAST KGPSVFPLAP

＞HC-093-G06(SEQ ID NO：1207)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYHMDWVRQA PGKGLEWVSV

IGPSGGFTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAL

GGSLDYWGQG TLVTVSSAST KGPSVFPLAP

＞HC-093-G09 (SEQ ID NO：1208)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYSMTWVRQA PGKGLEWVSS

ISPSGGATKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

AASLPFDYWG QGTLVTVSSA STKGPSVFPL AP

＞HC-094-D08 (SEQ ID NO：1209)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMVWVRQA PGKGLEWVSG

IVPSGGYTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTRAS

YSSFGLDYWG QGTLVTVSSA STKGPSVFPL AP

＞HC-094-E08 (SEQ ID NO：1210)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS FYAMVWVRQA PGKGLEWVSY

ISPSGGATWY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKPS

RPRYYYDSSG YYSSAFDIWG QGTMVTVSSA STKGPSVFPL AP

＞HC-094-F03 (SEQ ID NO：1211)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYQMSWVRQA PGKGLEWVSS

IYPSGGATKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARMG

LHHSFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-095-A11 (SEQ ID NO：1212)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYMMGWNRQA PGKGLEWVSS

IRSSGGATAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-095-G09 (SEQ ID NO：1213)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYNMHWVRQA PGKGLEWVSV

IYPSGGYTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

NWGGIDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-095-H10 (SEQ ID NO：1214)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYKMQWVRQA PGKGLEWVSS

IYSSGGKTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARTP

GYNYFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-096-D03 (SEQ ID NO：1215)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYWMVWVRQA PGKGLEWVSW

IGPSGGGTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGN

GGFDSWGRGT LVTVSSASTK GPSVFPLAP

＞HC-096-D09 (SEQ ID NO：1216)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYHMYWVRQA PGKGLEWVSY

IRPSGGNTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARDR

RGYSSSKGYY YYGMDVWGQG TTVTVSSAST KGPSVFPLAP

＞HC-096-E11 (SEQ ID NO：1217)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMYWVRQA PGKGLEWVSS

IYPSGGFTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

VYGTFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-097-D04 (SEQ ID NO：1218)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYFMHWVRQA PGKGLEWVSY

ISSSGGLTGY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

PVYYYDSSGS HSAFDIWGQG TMVTVSKRLH QGPIGLPAST LLQEH

＞HC-097-E07 (SEQ ID NO：1219)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYQMMWVRKA PGKGLEWVSY

IRSSGGKTDY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-097-F03 (SEQ ID NO：1220)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYLMYWVRQA PGKGLEWVSS

IRPSGGNTLY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGR

GILTGYYWGY YYYMDVWGKG TTVTVSSAST KGPSVFPLAP

＞HC-097-F04 (SEQ ID NO：1221)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYQMWWVRQA PGKGLEWVSS

ISSSGGLTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED MAVYYCARVK

LNYYGSGSYS LDAFDIWGQG TNGHRLKRLH QGPIGLPAST LLQEH

＞HC-097-F08 (SEQ ID NO：1222)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYPMVWVRQA PGKGLEWVSV

IGPSGGKTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-097-G01 (SEQ ID NO：1223)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYYMQWVRQA PGKGLEWVSG

ISSSGGSTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARSQ

RRTYYPNFGD AFDIWGQGTM VTVSSASTKG PSVFPLAP

＞HC-097-G07 (SEQ ID NO：1224)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS KYKMGWVRQA PGKGLEWVSY

IRPSGGMTFY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTREH

YQASYSSSAW FRMVPAGAFD IWGQGTMVTV SSASTKGPSV FPLAP

＞HC-098-C10 (SEQ ID NO：1225)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS FYSMMWVRQA PGKGLEWVSG

ISSSGGTTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

TNYDYVWGSY RSHYYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-098-E01 (SEQ ID NO：1226)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS MYGMTWVRQA PGKGLEWVSG

ISPSGGRTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKHR

RDYVWWTYGM DVWGQGTTVT VSSASTKGPS VFPLAP

＞HC-098-E09 (SEQ ID NO：1227)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYGMQWVRQA PGKGLEWVSY

IYPSGGGTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRV

AVAGSWYYYY YGMDVWGQGT TVTVSSASTK GPSVFPLAP

＞HC-098-H04 (SEQ ID NO：1228)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMSWVRQA PGKGLEWVSR

IYPSGGQTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGA

GWFDPWGQGT LVTVSSASTK GPSVFPLAP

＞HC-099-A09 (SEQ ID NO：1229)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYAMFWVRQA PGKGLEWVSS

ISPSGGKTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

GTALDYWGQG TLVTVSSAST KGPSVFPLAP

＞HC-099-C12 (SEQ ID NO：1230)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYAMHWVRQA PGKGLEWVSS

IWPSGGATFY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATSP

TLNAFHIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-099-D05 (SEQ ID NO：1231)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYIMSWVRQA PGKGLEWVSS

IWPSGGHTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAG

SYYAGDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-099-E11 (SEQ ID NO：1232)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYTMVWVRQA PGKGLEWVSS

IYSSGGRTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARIV

VVPSAQFYFY YGMDVWGQGT TVTVSSASTK GPSVFPLAP

＞HC-101-E01 (SEQ ID NO：1233)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYIMSWVRQA PGKGLEWVSS

IVSSGGVTLY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTKNI

NLRYDILTGY FDIWGQGTTV TVSSASTKGP SVFPLAP

＞HC-102-A04 (SEQ ID NO：1234)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMNWVRQA PGKGLEWVSR

IYPSGGVTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGY

GMDVWGQGTM VTVSSASTKG PSVFPLAP

＞HC-102-C02 (SEQ ID NO：1235)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMHWVRQA PGKGLEWVSS

IYPSGGKTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVF

GYYDFWSGYP GAFDYWGQGT LVTVSSASTK GPSVFPLAP

＞HC-102-C12 (SEQ ID NO：1236)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMGWVRQA PGKGLEWVSV

IWPSGGITKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGN

GGFDSWGQGT LVTVSSASTK GPSVFPLAP

＞HC-102-D07 (SEQ ID NO：1237)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYWMWWVRQA PGKGLEWVSS

IGPSGGATRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTTGS

AGNWGQGTLV TVSSASTKGP SVFPLAP

＞HC-102-E09 (SEQ ID NO：1238)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS LYLMEWVRQA PGKGLEWVSS

IGSSGGATWY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCATDT

SRVAGTRLRN YYYYYGMDVW GQGTTVTVSS ASTKGPSVFP LAP

＞HC-102-G11 (SEQ ID NO：1239)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYMMEWVRQA PGKGLEWVSG

ISPSGGTTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTRGG

YNNYYYALDV WGQGTTVTVS SASTKGPSVF PLAP

＞HC-102-G12 (SEQ ID NO：1240)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS KYKMVWVRQA PGKGLEWVSS

IYPSGGITAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCAKDI

TPGGGSGFRL PKNYYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-102-H02 (SEQ ID NO：1241)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS MYRMFWVRQA PGKGLEWVSV

IGPSGGQTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-102-H11 (SEQ ID NO：1242)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS KYPMTWVRQA PGKGLEWVSS

ISSSGGKTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTRGG

SSTLYFMDVW GQGTTVTVSS ASTKGPSVFP LAP

＞HC-103-A03 (SEQ ID NO：1243)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYPMSWVRQA PGKGLEWVSY

IRSGGYTTYA DSVKGRFTIS RDNSKNTLYL QMNSLRAEDT AVYYCARERV

FCSGGRCGSY FDYWGQGTLV TVSSASTKGP SVFPLAP

＞HC-103-E09 (SEQ ID NO：1244)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYSMLWVRQA PGKGLEWVSS

IRPSGGFTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

VWDAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-103-F07(SEQ ID NO：1245)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS LYAMHWVRQA PGKGLEWVSS

IRPSGGLTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-103-H07 (SEQ ID NO：1246)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYGMYWVRQA PGKGLEWVSS

IYPSGGWTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

SWGAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-104-A12 (SEQ ID NO：1247)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYNMGWVRQA PGKGLEWVSR

IGSSGGKTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKTN

YDFWSGYLPN PNPYPLDYWG QGTLVTVSSA STKGPSVFPL AP

＞HC-104-B03 (SEQ ID NO：1248)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYTMFWVRQA PGKGLEWVSS

IYPSGGITRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVS

FWNAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-104-B12 (SEQ ID NO：1249)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS IYAMIWVRQA PGKGLEWVSS

IYSSGGPTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGR

RTYYYDSSGY YKYAAFDIWG QGTMVTVSSA STKGPSVFPL AP

＞HC-104-D12 (SEQ ID NO：1250)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYAMFWVRQA PGKGLEWVSS

IWPSGGKTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARGG

SYLAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-104-H12 (SEQ ID NO：1251)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS YYQMSWVRQA PGKGLEWVSR

IGPSGGLTSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-105-B06 (SEQ ID NO：1252)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYAMGWVRQA PGKGLEWVSR

IWPSGGHTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCATGY

SSTWGAFDIW GQGTMVTVSS ASTKGPSVFP LAP

＞HC-105-F02 (SEQ ID NO：1253)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYWMAWVRQA PGKGLEWVSS

IGPSGGSTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARTA

RWLSFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-106-C06 (SEQ ID NO：1254)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYTMQWVRQA PGKGLEWVSS

IYPSGGATKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARSG

YYYGLDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-106-D06 (SEQ ID NO：1255)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS YYLMGWVRQA PGKGLEWVSS

IGPSGGTTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

LGAAFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-106-E12 (SEQ ID NO：1256)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYNMTWVRQA PGKGLEWVSS

IWPSGGATRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARTS

RFYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-106-G12 (SEQ ID NO：1257)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYRMHWVRQA PGKGLEWVSS

IWPSGGKTHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAL

QYGSGSYFYA PKSYYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-107-B05 (SEQ ID NO：1258)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMMWVRQA PGKGLEWVSS

IRSSGGITRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

YYYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-107-D01 (SEQ ID NO：1259)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYGMGWVRQA PGKGLEWVSV

IRPSGGTTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

RYYSLDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-107-D05 (SEQ ID NO：1260)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYHMQWVRQA PGKGLEWVSS

IYPSGGFTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDG

GQGGWGQGTL VTVSSASTKG PSVFPLAP

＞HC-107-D12 (SEQ ID NO：1261)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS MYKMGWVRQA PGKGLEWVSS

IVPSGGKTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKDI

TPGGGFGVPL A.KLLLL

＞HC-107-F11 (SEQ ID NO：1262)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYQMHWVRQA PGKGLEWVSS

IRPSGGRTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-108-C10 (SEQ ID NO：1263)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYVMAWVRQA PGKGLEWVSV

IRPSGGKTLY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVA

GRVGVPATKK NWFDPWGQGT LVTVSSASTK GPSVFPLAP

＞HC-108-D11 (SEQ ID NO：1264)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYVMTWVRQA PGKGLEWVSV

IYPSGGATKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

AGGMDVWGQG TTVTVSSAST KGPSVFPLAP

＞HC-108-E10 (SEQ ID NO：1265)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYRMVWVRQA PGKGLEWVSS

IYPSGGYTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVI

TYNNFDSWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-108-E11 (SEQ ID NO：1266)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYSMMWVRQA PGKGLEWVSS

IYPSGGFTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

FYGGFVWGQG TTVTVSSAST KGPSVFPLAP

＞HC-109-A11 (SEQ ID NO：1267)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYTMTWVRQA PGKGLEWVSV

IYPSGGHTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARGG

RWFSLDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-109-E08 (SEQ ID NO：1268)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYTMLWVRQA PGKGLEWVSS

IYSSGGSTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-110-D06 (SEQ ID NO：1269)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYRMAWVRQA PGKGLEWVSY

IGPSGGSTSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARVG

TFYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-110-D11 (SEQ ID NO：1270)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMYWVRQA PGKGLEWVSR

IGPSGGWTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-110-F08 (SEQ ID NO：1271)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYVMYWVRQA PGKGLEWVSG

IGPSGGFTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTRDR

YFGSGYYMAA YYYYGMDVWG QGTTVTVSSA STKGPSVFPLAP

＞HC-110-G01 (SEQ ID NO：1272)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS MYKMGWVRQA PGKGLEWVSS

IVPSGGKTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKDI

TPGGGSGFRL PKNYYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-110-H11 (SEQ ID NO：1273)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYKMEWVRQA PGKGLEWVSR

IRPSGGVTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTRGG

LWDAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-111-B02 (SEQ ID NO：1274)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS KYWMAWVRQA PGKGLEWVSV

IYPSGGQTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTLMR

YGDRFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-111-C10 (SEQ ID NO：1275)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYVMSWVRQA PGKGLEWVSS

IRSSGGRTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-111-D11 (SEQID NO：1276)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYAMMWVRQA PGKGLEWVSS

IWPSGGHTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATDQ

GYFWGQGTLV TVSSASTKGP SVFPLAP

＞HC-112-C02 (SEQ ID NO：1277)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYYMLWVRQA PGKGLEWVSY

IRSSGGSTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKPT

VDGAFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-112-C12 (SEQID NO：1278)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYMMYWVRQA PGKGLEWVSS

IYPSGGKTLY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-112-D04 (SEQID NO：1279)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMVWVRQA PGKGLEWVSG

IRSSGGVTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-112-D07 (SEQID NO：1280)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS FYSMSWVRQA PGKGLEWVSR

IRPSGGRTDY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR

LYYYGSGSYY YGAFDIWGQG TMVTVSSAST KGPSVFPLAP

＞HC-113-D05 (SEQID NO：1281)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYIMMWVRQA PGKGLEWVSS

IYPSGGSTIY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCASAT

EGYYYGMDVW GQGTTVTVSS ASTKGPSVFP LAP

＞HC-113-E03 (SEQID NO：1282)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYTMFWVRQA PGKGLEWVSG

IYPSGGKTIY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATGG

VFGVVDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-113-G11 (SEQID NO：1283)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYMMQWVRQA PGKGLEWVSG

IGPSGGWTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TALYYCARLN

YGDYVWGQGT LVTVSSASTK GPSVFPLAP

＞HC-114-E02 (SEQID NO：1284)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYKMMWVRQA PGKGLEWVSY

IYPSGGWTGY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

PWDAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-114-F04 (SEQID NO：1285)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYYMAWVRQA PGKGLEWVSY

ISPSGGSTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARGG

GTGTFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-114-G06 (SEQ ID NO：1286)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS YYEMEWVRQA PGKGLEWVSS

IRPSGGRTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKGR

SNYYGSGSYS PKYFQHWGQG TLVTVSSAST KGPSVFPLAP

＞HC-114-G09 (SEQ ID NO：1287)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYAMDWVRQA PGKGLEWVSV

IYSSGGKTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-114-H07 (SEQ ID NO：1288)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMHWVRQA PGKGLEWVSV

IYPSGGTTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARGN

SGSHDYWGQG TLVTVSSAST KGPSVFPLAP

＞HC-115-F04(SEQ ID NO：1289)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYLMGWVRQA PGKGLEWVSS

IGPSGGYTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGT

GTTFFWGQGT LVTVSSASTK GPSVFPLAP

＞HC-115-F08 (SEQ ID NO：1290)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMVWVRQA PGKGLEWVSS

IYPSGGMTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

IANSFNIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-115-G12 (SEQ ID NO：1291)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYFMEWVRQA PGKGLEWVSG

ISSSGGNTLY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR

FGNYYGSGSK LQHDAFDIWG QGTMVTVSSA STKGPSVFPL AP

＞HC-116-A08 (SEQ ID NO：1292)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS YYMMGWVRQA PGKGLEWVSS

IYTSGGYTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAS

IYYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-116-C09 (SEQ ID NO：1293)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYKMHWVRQA PGKGLEWVSS

IYPSGGWTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATGY

SSTWGAFDIW GQGTMVTVSS ASTKGPSVFP LAP

＞HC-116-E08 (SEQ ID NO：1294)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYPMIWVRQA PGKGLEWVSV

IYPSGGMTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

SYGSLGYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-117-A12 (SEQ ID NO：1295)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYKMSWVRQA PGKGLEWVSG

IYPSGGLTQY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-117-B07 (SEQ ID NO：1296)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYTMWWVRQA PGKGLEWVSR

IWPSGGTTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

SYLAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-117-C04 (SEQ ID NO：1297)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMFWVRQA PGKGLEWVSS

IWPSGGNTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARPA

YYYAMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-117-F04 (SEQ ID NO：1298)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS MYVMNWVRQA PGKGLEWVSR

IGSSGGGTLY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVS

VYRIRNYYYY AMDVWGQGTT VTVSSASTKG PSVFPLAP

＞HC-118-E07 (SEQ ID NO：1299)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYPMDWVRQA PGKGLEWVSR

IYPSGGATKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARGG

GAFDIWGQGT MVTVSSASTK GPSVFPLAP

＞HC-119-B09 (SEQ ID NO：1300)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYWMYWVRQA PGKGLEWVSV

IGSSGGVTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARGP

YRGYFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-121-A07 (SEQ ID NO：1301)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMYWVRQA PGKGLEWVSR

IYPSGGATSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKSG

SWYSFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-121-H07 (SEQ ID NO：1302)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS KYYMVWVRQA PGKGLEWVSV

IGPSGGWTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVD

YSNYFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-122-A05 (SEQ ID NO：1303)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYPMHWVRQA PGKGLEWVSS

IWPSGGHTSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARLS

QPIWGQGTLV TVSSASTKGP SVFPLAP

＞HC-122-D01 (SEQ ID NO：1304)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYIMHWVRQA PGKGLEWVSG

IYPSGGSTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAL

VGALDYWGQG TLVTVSSAST KGPSVFPLAP

＞HC-122-G06(SEQ ID NO：1305)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYAMHWVRQA PGKGLEWVSV

ISPSGGATRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCVRAS

TVTTLFQHWG QGTLVTVSSA STKGPSVFPL AP

＞HC-122-H04 (SEQ ID NO：1306)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYAMAWVRQA PGKGLEWVSS

IGSSGGVTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAAPR

GRYYYDSSGY YYGGVFDYWG QGTLVTVSSA STKGPSVFPL AP

＞HC-123-E02 (SEQ ID NO：1307)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYRMDWVRQA PGKGLEWVSG

IYPSGGHTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR

GWYGADYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-124-A01 (SEQ ID NO：1308)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYPMYWVRQA PGKGLEWVSR

IGPSGGHTDY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAG

VWSGLDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-124-C04 (SEQ ID NO：1309)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYLMGWVRQA PGKGLEWVSV

IGPSGGLTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARFR

GYYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-124-C12 (SEQ ID NO：1310)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYTMQWVRQA PGKGLEWVSR

IYSSGGGTYY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARVG

PWGAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-124-G12 (SEQ ID NO：1311)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYWMRWVRQA PGKGLEWVSS

IGSSGGMTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAL

RWRAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-124-H10 (SEQ ID NO：1312)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYFMNWVRQA PGKGLEWVSS

IGSSGGYTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-125-C04 (SEQ ID NO：1313)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMDWVRQA PGKGLEWVSS

IYPSGGWTRY ADSVKGRFTI SRDNSRNTLY LQMNSLRAED TAVYYCARRG

QWLVLDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-126-B12 (SEQ ID NO：1314)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMGWVRQA PGKGLEWVSS

IYPSGGKTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCASPS

TGSWSAFDIW GQGTMVTVSS ASTKGPSVFP LAP

＞HC-126-C10 (SEO ID NO：1315)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYKMHWVRQA PGKGLEWVSS

IWPSGGGTFY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARTS

GGFGAFDIWG QGTMVTVSSA STKGPSVFPL AP

＞HC-126-F11 (SEQ ID NO：1316)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYWMMWVRQA PGKGLEWVSW

IGSSGGFTWY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGN

GGFDSWGQGT LVTVSSASTK GPSVFPLAP

＞HC-126-H09 (SEQ ID NO：1317)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYTMMWVRQA PGKGLEWVSG

IYPSGGQTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

SWYAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-127-C06 (SEQ ID NO：1318)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYPMHWVRQA PGKGLEWVSY

IGPSGGWTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAP

SGYWGQGTLV TVSSASTKGP SVFPLAP

＞HC-127-D05 (SEQ ID NO：1319)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYYMMWVRQA PGKGLEWVSS

IGPSGGMTDY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATGG

TASLDYWGQG TLVTVSSAST KGPSVFPLAP

＞HC-127-H05 (SEQ ID NO：1320)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMGWVRQA PGKGLEWVSS

IWPSGGHTSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARHY

GMDVWGQGTT VTVSSASTKG PSVFPLAP

＞HC-128-A06 (SEQ ID NO：1321)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYKMNWVRQA PGKGLEWVSY

IGSSGGKTGY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TALYYCARVG

VWYGMDVWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-128-E07 (SEQ ID NO：1322)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS FYPMSWVRQA PGKGLEWVSY

IWPSGGATRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGN

GGFDSWGQGT LVTVSSASTK GPSVFPLAP

＞HC-128-F11 (SEQ ID NO：1323)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS FYVMGWVRQA PGKGLEWVSW

IGPSGGRTWY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR

GWYGIDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-129-B09 (SEQ ID NO：1324)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYWMHWVRQA PGKGLEWVSS

IGPSGGVTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARHG

SWGGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-130-B02 (SEQ ID NO：1325)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYSMVWVRQA PGKGLEWVSR

IYSSGGSTHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-131-A03 (SEQ ID NO：1326)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYTMGWVRQA PGKGLEWVSV

IYPSGGMTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARTS

GGTPWGFWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-131-B05 (SEQ ID NO：1327)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMSWVRQA PGKGLEWVSY

IYPSGGKTHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-131-D01 (SEQ ID NO：1328)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYNMNWVRQA PGKGLEWVSV

IYPSGGQTHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

IWHSFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-131-D12 (SEQ ID NO：1329)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYNMYWVRQA PGKGLEWVSY

IVPSGGATHY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAL

RGYSYGPRGY YYYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-131-F09 (SEQ ID NO：1330)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMQWVRQA PGKGLEWVSG

IGPSGGSTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-131-F12 (SEQ ID NO：1331)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYAMGWVRQA PGKGLEWVSY

IGPSGGNTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDV

GGSGSYYMLS YYYYGMDVWG QGTMVTVSSA STKGPSVFPL AP

＞HC-131-G03 (SEQ ID NO：1332)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMQWVRQA PGKGLEWVSS

IYPSGGSTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCARFN

VGFDLWGRGT LVTVSSASTK GPSVFPLAP

＞HC-131-G06 (SEQ ID NO：1333)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYAMTWVRQA PGKGLEWVSS

ISSSGGDTAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATER

HYIWGSYRYS WFDPWGQGTL VTVSSASTKG PSVFPLAP

＞HC-132-C01 (SEQ ID NO：1334)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYLMWWVRQA PGKGLEWVSV

IGPSGGWTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARHP

GDYWGQGTLV TVSSASTKGP SVFPLAP

＞HC-132-C08 (SEQ ID NO：1335)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS LYFMTWVRQA PGKGLEWVSS

ISSSGGSTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

QWLAFDYWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-132-D02 (SEQ ID NO：1336)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYPMDWVRQA PGKGLEWVSY

IGPSGGSTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARHV

PPGTWGQGTL VTVSSASTKG PSVFPLAP

＞EC-132-D08 (SEQ ID NO：1337)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMYWVRQA PGKGLEWVSW

IGPSGGHTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARLQ

QGLDYWGQGT LVTVSSASTK GPSVFPLAP

＞HC-132-H11 (SEQ ID NO：1338)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYQMHWVRQA PGKGLEWVSR

IRSSGGATSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARGG

GYSYYFDYWG QGTLVTVSSA STKGPSVFPL AP

＞HC-133-E02 (SEQ ID NO：1339)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYTMTWVRQA PGKGLEWVSS

IYPSGGHTSY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKDT

RVGPWLVRAP LDYWGQGTLV TVSSASTKGP SVFPLAP

＞HC-133-E08 (SEQ ID NO：1340)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS KYKMVWVRQA PGKGLEWVSS

IYPSGGITAY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCAKDI

TPGGGSGFRL PKNYYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-134-B03 (SEQ ID NO：1341)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYSMFWVRQA PGKGLEWVSS

IYPSGGQTDY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

LLWSFDSWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-134-D07 (SEQ ID NO：1342)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYFMIWVRQA PGKGLEWVSS

IVPSGGPTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARRG

PVYYYDSSGS HSAFDIWGQG TMVTVSSAST KGPSVFPLAP

＞HC-135-B05 (SEQ ID NO：1343)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYLMDWVRQA PGKGLEWVSV

IYPSGGHTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-135-C11 (SEQ ID NO：1344)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYSMQWVRQA PGKGLEWVSG

IRPSGGSTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAVST

GGYYYGMDVW GQGTTVTVSS ASTKGPSVFP LAP

＞HC-135-C12 (SEQ ID NO：1345)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYHMSWVRQA PGKGLEWVSS

IRPSGGSTIY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAREP

RRYYYDSSGS YDAFDIWGQG TMVTVSSAST KGPSVFPLAP

＞HC-135-E10 (SEQ ID NO：1346)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYRMSWVRQA PGKGLEWVSS

ISPSGGPTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARPS

GDSSVYRPFA SWGQGTLVTV SSASTKGPSV FPLAP

＞HC-135-F03 (SEQ ID NO：1347)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMIWVRQA PGKGLEWVSW

IYPSGGNTIY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TATYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-136-B06 (SEQ ID NO：1348)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS SYIMIWVRQA PGKGLEWVSY

IRPSGGYTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVI

NAGPGRGYYW RGYSYSDAFD IWGQGTMVTV SSASTKGPSV FPLAP

＞HC-136-D07 (SEQ ID NO：1349)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYKMGWVRQA PGKGLEWVSS

IYPSGGVTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTKNI

NLRYDILTGY FDIWGQGTTV TVSSASTKGP SVFPLAP

＞HC-136-E10 (SEQ ID NO：1350)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYNMAWVRQA PGKGLEWVSS

IWPSGGRTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKGV

RSSGLLERGR VYDAFDIWGQ GTMVTVSSAS TKGPSVFPLA P

＞HC-136-E12 (SEQ ID NO：1351)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS NYSMQWVRQA PGKGLEWVSS

IGPSGGRTWY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-136-F08 (SEQ ID NO：1352)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYKMGWVRQA PGKGLEWVSS

IRPSGGATKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

IWDAFDIWGQ GAMVTVSSAS TKGPSVFPLA P

＞HC-137-B01 (SEQ ID NO：1353)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS YYRMTWVRQA PGKGLEWVSS

IYPSGGYTIY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARLG

QWLALDRWGQ GTLVTVSSAS TKGPSVFPLA P

＞HC-137-E01 (SEQ ID NO：1354)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYSMTWVRQA PGKGLEWVSS

IYPSGGKTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCASMV

APYYYDSSGY PPAEYFQHWG QGTLVTVSSA STKGPSVFPL AP

＞HC-137-G10 (SEQ ID NO：1355)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYVMHWVRQA PGKGLEWVSY

ISPSGGVTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKAL

YPGMGYYYGM DVWGQGTTVT VSSASTKGPS VFPLAP

＞HC-138-A03 (SEQ ID NO：1356)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS FYLMFWVRQA PGKGLEWVSG

IYPSGGQTVY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARAI

GAGSSFWGQG TLVTVSSAST KGPSVFPLAP

＞HC-138-B06 (SEQ ID NO：1357)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYTMSWVRQA PGKGLEWVSG

IYPSGGETWY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKNI

NLRYDILTGY FDFWGQGTMV TVSSASTKGP SVFPLAP

＞HC-138-B10 (SEQ ID NO：1358)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYSMTWVRQA PGKGLEWVSR

IYPSGGRTGY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCATDS

GGYYYGMDVW GQGTTVTVSS ASTKGPSVFP LAP

＞HC-138-C04 (SEQ ID NO：1359)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYHMQWVRQA PGKGLEWVSG

IRSSGGVTGY SDSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAREV

TMVRGVYYYY YGMDVWGQGT TVTVSSASTK GPSVFPLAP

＞HC-138-C09 (SEQ ID NO：1360)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS PYRMSWVRQA PGKGLEWVSV

ISPSGGITEY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKHE

RGSGYYVTPP AGAFDIWGQG TMVTVSSAST KGPSVFPLAP

＞HC-138-E02 (SEQ ID NO：1361)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYMMHWVRQA PGKGLEWVSG

IGPSGGKTPY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-138-E12 (SEQ ID NO：1362)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYVMRWVRQA PGKGLEWVSS

IYPSGGQTRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAMYYCATGY

SSTWGAFDIW GQGTMVTVSS ASTKGPSVFP LAP

＞HC-138-G11 (SEQ ID NO：1363)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYTMTWVRQA PGKGLEWVSS

IYPSGGHTSY ADSVKGRFTI SRDNSKNTFY LQMNSLRAED TAVYYCAKDT

RVGPWLVRAP LDYWGQGTLV TVSSASTKGP SVFPLAP

＞HC-139-A04 (SEQ ID NO：1364)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYPMTWVRQA PGKGLEWVSS

IRPSGGNTGY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARLT

GYSSGWFWAY GMDVWGQGTT VTVSSASTKG PSVFPLAP

＞HC-139-A09 (SEQ ID NO：1365)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYFMVWVRQA PGKGLEWVSG

IGPSGGLTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARSD

WLNYWGQGTL VTVSSASTKG PSVFPLAP

＞HC-139-C02 (SEQ ID NO：1366)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS YYKMHWVRQA PGKGLEWVSV

IYPSGGKTTY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTREH

YQASYSSSAW FRMVPAGAFD IWGQGTMVTV SSASTKGPSV FPLAP

＞HC-139-E12 (SEQ ID NO：1367)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS WYSMNWVRQA PGKGLEWVSS

IGPSGGGTKY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCASRG

QRKQWLPPNG AFDIWGQGTM VTVSSASTKG PSVFPLAP

＞HC-139-F04 (SEQ ID NO：1368)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYYMEWVRQA PGKGLEWVSS

ISSSGGSTEY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCAKDI

TPGGGSGFRL PKNYYYYGMD VWGQGTTVTV SSASTKGPSV FPLAP

＞HC-139-F09 (SEQ ID NO：1369)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS HYVMAWVRQA PGKGLEWVSR

IGPSGGYTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA P

＞HC-135-H01 (SEQ ID NO：1377)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS AYLMAWVRQA PGKGLEWVSS

IRPSGGETRY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCTRDQ

KGYPDSSSFR RYYYYYGMDV WGQGTTVTVS SASTKGPSVF PLAP

Antibody 098-E09 has at the CDR1 place and comprises sequence TGTSSDIGAYNYVS (SEQ ID NO:1), comprises sequence D VSNRPS (SEQ ID NO:2) at the CDR2 place, and comprises the LC variable domain of sequence SSYRSSSLM (SEQID NO:3) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMS (SEQ ID NO:4), comprises sequence YIYPSGGKTHYADSVKG (SEQ ID NO:5) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:6) at the CDR3 place.

Antibody 131-F07 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:7), comprises sequence A ASSLQS (SEQ ID NO:8) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPLT (SEQ IDNO:9) at the CDR3 place.It has and comprises sequence KYKMV (SEQ ID NO:10) at the CDR1 place, comprise sequence SIYPSGGITAYADSVKG (SEQ ID NO:11) at the CDR2 place, and comprise the HC variable domain of sequence D ITPGGGSGFRLPKNYYYYGMDV (SEQ ID NO:12) at the CDR3 place.

Antibody 139-F04 has at the CDR1 place and comprises sequence RASQGISSWLA (SEQ ID NO:13), comprises sequence A ASSLER (SEQ ID NO:14) at the CDR2 place, and comprises the LC variable domain of sequence QQYHNFPLT (SEQ IDNO:15) at the CDR3 place.It has at the CDR1 place and comprises sequence YYEME (SEQ ID NO:16), comprises sequence SIRPSGGRTVYADSVKG (SEQ ID NO:17) at the CDR2 place, and comprises the HC variable domain of sequence GRSNYYGSGSYSPKYFQH (SEQ ID NO:18) at the CDR3 place.

Antibody 139-E12 has at the CDR1 place and comprises sequence RASQSVSSNYLS (SEQ ID NO:19), comprises sequence GTSNRAS (SEQ ID NO:20) at the CDR2 place, and comprises the LC variable domain of sequence QQYGGAPLFI (SEQID NO:21) at the CDR3 place.It has at the CDR1 place and comprises sequence HYNMN (SEQ ID NO:22), comprises sequence VIYPSGGQTHYADSVKG (SEQ ID NO:23) at the CDR2 place, and comprises the HC variable domain of sequence RGIWHSFDI (SEQ ID NO:24) at the CDR3 place.

Antibody 097-F03 has at the CDR1 place and comprises sequence SGTNSNIGSNSVF (SEQ ID NO:25), comprises sequence RNSQRPS (SEQ ID NO:26) at the CDR2 place, and comprises the LC variable domain of sequence A TWDDSLRSPV (SEQ ID NO:27) at the CDR3 place.It has and comprises sequence RYSMT (SEQ ID NO:28) at the CDR1 place, comprise sequence SIYPSGGKTTYADSVKG (SEQ ID NO:29) at the CDR2 place, and comprise the HC variable domain of sequence MVAPYYYDSSGYPPAEYFQH (SEQ ID NO:30) at the CDR3 place.

Antibody 102-G12 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:3 1), comprises sequence A ASSLQS (SEQ ID NO:32) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPRT (SEQ IDNO:33) at the CDR3 place.It has at the CDR1 place and comprises sequence LYFMT (SEQ ID NO:34), comprises sequence SISSSGGSTKYADSVKG (SEQ ID NO:35) at the CDR2 place, and comprises the HC variable domain of sequence GGQWLAFDY (SEQ ID NO:36) at the CDR3 place.

Antibody 102-E09 has at the CDR1 place and comprises sequence TGTSSDVGSYKLVS (SEQ ID NO:37), comprises sequence EGSKRPS (SEQ ID NO:38) at the CDR2 place, and comprises the LC variable domain of sequence C SYAGSSTWV (SEQ ID NO:39) at the CDR3 place.It has at the CDR1 place and comprises sequence HYAMD (SEQ ID NO:40), comprises sequence VIYSSGGKTTYADSVKG (SEQ ID NO:41) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:42) at the CDR3 place.

Antibody 106-G12 has at the CDR1 place and comprises sequence RASQGISSWLA (SEQ ID NO:43), comprises sequence A ASSLQS (SEQ ID NO:44) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPHT (SEQ IDNO:45) at the CDR3 place.It has at the CDR1 place and comprises sequence FYLMF (SEQ ID NO.46), comprises sequence GIYPSGGQTVYADSVKG (SEQ ID NO:47) at the CDR2 place, and comprises the HC variable domain of sequence A IGAGSSF (SEQ ID NO:48) at the CDR3 place.

Antibody 102-H11 has at the CDR1 place and comprises sequence RASRGISNWLA (SEQ ID NO:49), comprises sequence SASTLHA (SEQ ID NO:50) at the CDR2 place, and comprises the LC variable domain of sequence QEGNSFPYT (SEQ IDNO:51) at the CDR3 place.It has at the CDR1 place and comprises sequence WYSMN (SEQ ID NO:52), comprises sequence SIGPSGGGTKYADSVKG (SEQ ID NO:53) at the CDR2 place, and comprises the HC variable domain of sequence RGQRKQWLPPNGAFDI (SEQ ID NO:54) at the CDR3 place.

Antibody 138-C09 has at the CDR1 place and comprises sequence WTIYDISSWLA (SEQ ID NO:55), comprises sequence A ASRLAT (SEQ ID NO:56) at the CDR2 place, and comprises the LC variable domain of sequence QQTKDFPLT (SEQ IDNO:57) at the CDR3 place.It has at the CDR1 place and comprises sequence RYGMQ (SEQ ID NO:58), comprises sequence YIYPSGGGTVYADSVKG (SEQ ID NO:59) at the CDR2 place, and comprises the HC variable domain of sequence RVAVAGSWYYYYYGMDV (SEQ ID NO:60) at the CDR3 place.

Antibody 098-G05 has at the CDR1 place and comprises sequence SGSSSNIGSNYVY (SEQ ID NO:61), comprises sequence RNNQRPS (SEQ ID NO:62) at the CDR2 place, and comprises the LC variable domain of sequence A AWDDSLSGVV (SEQ ID NO:63) at the CDR3 place.It has and comprises sequence A YLMA (SEQ ID NO:64) at the CDR1 place, comprise sequence SIRPSGGETRYADSVKG (SEQ ID NO:65) at the CDR2 place, and comprise the HC variable domain of sequence D QKGYPDSSSFRRYYYYYGMDV (SEQ ID NO:66) at the CDR3 place.

Antibody 136-E10 has at the CDR1 place and comprises sequence TGTNTDVGGYNLVS (SEQ ID NO:67), comprises sequence EVSNRPS (SEQ ID NO:68) at the CDR2 place, and comprises the LC variable domain of sequence GSYTSSSTHV (SEQ ID NO:69) at the CDR3 place.It has at the CDR1 place and comprises sequence WYVMT (SEQ ID NO:70), comprises sequence VIYPSGGATKYADSVKG (SEQ ID NO:71) at the CDR2 place, and comprises the HC variable domain of sequence RGAGGMDV (SEQ ID NO:72) at the CDR3 place.

Antibody 136-A07 has at the CDR1 place and comprises sequence RASHVINIDLG (SEQ ID NO:73), comprises sequence GASHLQR (SEQ ID NO:74) at the CDR2 place, and comprises the LC variable domain of sequence LQDSFYPRT (SEQ IDNO:75) at the CDR3 place.It has at the CDR1 place and comprises sequence NYSMQ (SEQ ID NO:76), comprises sequence SIGPSGGRTWYADSVKG (SEQ ID NO:77) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:78) at the CDR3 place.

Antibody 097-G01 has at the CDR1 place and comprises sequence RASQDIASWLA (SEQ ID NO:79), comprises sequence A ASSLQS (SEQ ID NO:80) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPFT (SEQ IDNO:81) at the CDR3 place.It has at the CDR1 place and comprises sequence PYKMG (SEQ ID NO:82), comprises sequence SIYPSGGVTTYADSVKG (SEQ ID NO:83) at the CDR2 place, and comprises the HC variable domain of sequence NINLRYDILTGYFDI (SEQ ID NO:84) at the CDR3 place.

Antibody 138-G11 has at the CDR1 place and comprises sequence RASQGISSWLV (SEQ ID NO:85), comprises sequence A ASSLQS (SEQ ID NO:86) at the CDR2 place, and comprises the LC variable domain of sequence QQAKTFPLT (SEQ IDNO:87) at the CDR3 place.It has at the CDR1 place and comprises sequence HYMMH (SEQ ID NO:88), comprises sequence GIGPSGGKTPYADSVKG (SEQ ID NO.89) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:90) at the CDR3 place.

Antibody 139-A09 has at the CDR1 place and comprises sequence RASQSVSSYLA (SEQ ID NO.91), comprises sequence D ASNRAT (SEQ ID NO:92) at the CDR2 place, and comprises the LC variable domain of sequence QQRSNWLWT (SEQID NO:93) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMQ (SEQ ID NO:94), comprises sequence GIGPSGGSTTYADSVKG (SEQ ID NO:95) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:96) at the CDR3 place.

Antibody 114-G06 has at the CDR1 place and comprises sequence TGTSSDVGAYNYVS (SEQ ID NO:97), comprises sequence EVNKRPS (SEQ ID NO:98) at the CDR2 place, and comprises the LC variable domain of sequence NSYAGSNSLI (SEQ ID NO:99) at the CDR3 place.It has at the CDR1 place and comprises sequence HYSMF (SEQ ID NO:100), comprises sequence SIYPSGGQTDYADSVKG (SEQ ID NO:101) at the CDR2 place, and comprises the HC variable domain of sequence RGLLWSFDS (SEQ ID NO:102) at the CDR3 place.

Antibody 139-C02 has and comprises sequence TGTSSDVGAYNYVS (SEQ ID NO:103) at the CDR1 place, comprise sequence EVNKRPS (SEQ ID NO:104) at the CDR2 place, and comprise the LC variable domain of sequence NSYAGSNSLI (SEQ ID NO:105) at the CDR3 place.It has at the CDR1 place and comprises sequence YYQMS (SEQ ID NO:106), comprises sequence RIGPSGGLTSYADSVKG (SEQ ID NO:107) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:108) at the CDR3 place.

Antibody 093-F09 has at the CDR1 place and comprises sequence RASQSVSSYLA (SEQ ID NO:109), comprises sequence D ASNRAT (SEQ ID NO:110) at the CDR2 place, and comprises the LC variable domain of sequence QQRSNWPSPIA (SEQID NO:111) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMI (SEQ ID NO:112), comprises sequence WIYPSGGNTIYADSVKG (SEQ ID NO:113) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:114) at the CDR3 place.

Antibody 110-F08 has at the CDR1 place and comprises sequence RASQGIRNDLG (SEQ ID NO:115), comprises sequence A ASSLQS (SEQ ID NO:116) at the CDR2 place, and comprises the LC variable domain of sequence LQDYNYPRT (SEQID NO:117) at the CDR3 place.It has and comprises sequence RYFMI (SEQ ID NO:118) at the CDR1 place, comprise sequence SIVPSGGPTRYADSVKG (SEQ ID NO:119) at the CDR2 place, and comprise the HC variable domain of sequence RGPVYYYDSSGSHSAFDI (SEQ ID NO:120) at the CDR3 place.

Antibody 117-F04 has and comprises sequence SGSNSNIGNNFVY (SEQ ID NO:121) at the CDR1 place, comprise sequence RNNQRPS (SEQ ID NO:122) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSLSGVL (SEQ ID NO:123) at the CDR3 place.It has and comprises sequence RYHMS (SEQ ID NO:124) at the CDR1 place, comprise sequence SIRPSGGSTIYADSVKG (SEQ ID NO:125) at the CDR2 place, and comprise the HC variable domain of sequence EPRRYYYDSSGSYDAFDI (SEQ ID NO:126) at the CDR3 place.

Antibody 111-D11 has at the CDR1 place and comprises sequence RASQSVSSYLA (SEQ ID NO:127), comprises sequence GASSRAT (SEQ ID NO:128) at the CDR2 place, and comprises the LC variable domain of sequence QQRGGWPLT (SEQID NO:129) at the CDR3 place.It has at the CDR1 place and comprises sequence KYPMT (SEQ ID NO:130), comprises sequence SISSSGGKTQYADSVKG (SEQ ID NO:131) at the CDR2 place, and comprises the HC variable domain of sequence GGSSTLYFMDV (SEQ ID NO:132) at the CDR3 place.

Antibody 112-D07 has and comprises sequence SGSSSNIGSNYVH (SEQ ID NO:133) at the CDR1 place, comprise sequence RNNRRPS (SEQ ID NO:134) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSLSGLVV (SEQ ID NO:135) at the CDR3 place.It has and comprises sequence PYQMW (SEQ ID NO:136) at the CDR1 place, comprise sequence SISSSGGLTRYADSVKG (SEQ ID NO:137) at the CDR2 place, and comprise the HC variable domain of sequence VKLNYYGSGSYSLDAFDI (SEQ ID NO:138) at the CDR3 place.

Antibody 131-B05 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:139), comprises sequence A ASSLQS (SEQ ID NO:140) at the CDR2 place, and comprises the LC variable domain of sequence QQSYNAPWT (SEQID NO:141) at the CDR3 place.It has at the CDR1 place and comprises sequence RYVMS (SEQ ID NO:142), comprises sequence SIRSSGGRTMYADSVKG (SEQ ID NO:143) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:144) at the CDR3 place.

Antibody 131-DO1 has and comprises sequence TGTSSDVGAYNYVS (SEQ ID NO.145) at the CDR1 place, comprise sequence D VSNRPS (SEQ ID NO:146) at the CDR2 place, and comprise the LC variable domain of sequence SSFTSRKTWV (SEQ ID NO:147) at the CDR3 place.It has and comprises sequence FYAMV (SEQ ID NO:148) at the CDR1 place, comprise sequence YISPSGGATWYADSVKG (SEQ ID NO:149) at the CDR2 place, and comprise the HC variable domain of sequence PSRPRYYYDSSGYYSSAFDI (SEQ ID NO:150) at the CDR3 place.

Antibody 137-E01 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:151), comprises sequence GASSRAT (SEQ ID NO:152) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSLT (SEQ ID NO:153) at the CDR3 place.It has at the CDR1 place and comprises sequence HYSMT (SEQ ID NO:154), comprises sequence RIYPSGGRTGYADSVKG (SEQ ID NO:155) at the CDR2 place, and comprises the HC variable domain of sequence D SGGYYYGMDV (SEQ ID NO:156) at the CDR3 place.

Antibody 132-C08 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:157), comprises sequence GASSRAT (SEQ ID NO:158) at the CDR2 place, and comprises the LC variable domain of sequence QQSYRSPYT (SEQ ID NO:159) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMA (SEQ ID NO:160), comprises sequence RIGPSGGYTMYADSVKG (SEQ ID NO:161) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:162) at the CDR3 place.

Antibody 114-G09 has at the CDR1 place and comprises sequence RASQSIGASLN (SEQ ID NO:163), comprises sequence TASNLQS (SEQ ID NO:164) at the CDR2 place, and comprises the LC variable domain of sequence QQNYRGRT (SEQ IDNO:165) at the CDR3 place.It has and comprises sequence HYNMG (SEQ ID NO:166) at the CDR1 place, comprise sequence RIGSSGGKTAYADSVKG (SEQ ID NO:167) at the CDR2 place, and comprise the HC variable domain of sequence TNYDFWSGYLPNPNPYPLDY (SEQ ID NO:168) at the CDR3 place.

Antibody 138-A03 has at the CDR1 place and comprises sequence TGTSSDVNYVS (SEQ ID NO:169), comprises sequence A VTKRPS (SEQ ID NO:170) at the CDR2 place, and comprises the LC variable domain of sequence SSYAGSNNLV (SEQ ID NO:171) at the CDR3 place.It has at the CDR1 place and comprises sequence HYPMD (SEQ ID NO:172), comprises sequence YIGPSGGSTRYADSVKG (SEQ ID NO:173) at the CDR2 place, and comprises the HC variable domain of sequence HVPPGT (SEQ ID NO:174) at the CDR3 place.

Antibody 135-H01 has and comprises sequence TGTSSDVGAYNYVS (SEQ ID NO:175) at the CDR1 place, comprise sequence D VSNRPS (SEQ ID NO:176) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSSYTWV (SEQ ID NO:177) at the CDR3 place.It has and comprises sequence HYHMQ (SEQ ID NO:178) at the CDR1 place, comprise sequence SIRPSGGETRYADSVKG (SEQ ID NO:179) at the CDR2 place, and comprise the HC variable domain of sequence EVTMVRGVYYYYYGMDV (SEQ ID NO:180) at the CDR3 place.

Antibody 108-D11 has and comprises sequence SGSSSNIGSNYVY (SEQ ID NO:181) at the CDR1 place, comprise sequence RNNQRPS (SEQ ID NO:182) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSLSGHAV (SEQ ID NO:183) at the CDR3 place.It has at the CDR1 place and comprises sequence HYQMH (SEQ ID NO:184), comprises sequence RIRSSGGATSYADSVKG (SEQ ID NO:185) at the CDR2 place, and comprises the HC variable domain of sequence GGGYSYYFDY (SEQ ID NO:186) at the CDR3 place.

Antibody 136-E12 has and comprises sequence TGTSTDDVGGYNYVS (SEQ ID NO:187) at the CDR1 place, comprise sequence D VINRPS (SEQ ID NO:188) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSRGTRV (SEQ ID NO:189) at the CDR3 place.It has and comprises sequence D YRMW (SEQ ID NO:190) at the CDR1 place, comprise sequence YIVPSGGQTSYADSVKG (SEQ ID NO:191) at the CDR2 place, and comprise the HC variable domain of sequence GSAYQRRTYSSGWYSASGRRGTAEYFQH (SEQIDNO:192) at the CDR3 place.

Antibody 136-D07 has at the CDR1 place and comprises sequence RASQSISNWLA (SEQ ID NO:193), comprises sequence MASTLES (SEQ ID NO:194) at the CDR2 place, and comprises the LC variable domain of sequence QQYNSYSVT (SEQID NO:195) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMY (SEQ ID NO:196), comprises sequence WIGPSGGHTMYADSVKG (SEQ ID NO:197) at the CDR2 place, and comprises the HC variable domain of sequence LQQGLDY (SEQ ID NO:198) at the CDR3 place.

Antibody 138-E02 has at the CDR1 place and comprises sequence RASQSVSSNYLA (SEQ ID NO:199), comprises sequence YGASYRAT (SEQ ID NO:200) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPWT (SEQ ID NO:201) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMY (SEQ IDNO:202), comprises sequence RIYPSGGATSYADSVKG (SEQ ID NO:203) at the CDR2 place, and comprises the HC variable domain of sequence SGSWYSFDY (SEQ ID NO:204) at the CDR3 place.

Antibody 131-F09 has at the CDR1 place and comprises sequence RASQGIGNYLV (SEQ ID NO:205), comprises sequence A ASTLQS (SEQ ID NO:206) at the CDR2 place, and comprises the LC variable domain of sequence QKYDGAPFT (SEQID NO:207) at the CDR3 place.It has at the CDR1 place and comprises sequence PYYMQ (SEQ ID NO:208), comprises sequence GISSSGGSTQYADSVKG (SEQ ID NO:209) at the CDR2 place, and comprises the HC variable domain of sequence SQRRTYYPNFGDAFDI (SEQ ID NO:210) at the CDR3 place.

Antibody 1 34-B03 has at the CDR1 place and comprises sequence RASQSISSWLA (SEQ ID NO:211), comprises sequence KASSLES (SEQ ID NO:212) at the CDR2 place, and comprises the LC variable domain of sequence QQYNSYRYT (SEQID NO:213) at the CDR3 place.It has at the CDR1 place and comprises sequence HYWMM (SEQ ID NO:214), comprises sequence WIGSSGGFTWYADSVKG (SEQ ID NO:215) at the CDR2 place, and comprises the HC variable domain of sequence GNGGFDS (SEQ ID NO.216) at the CDR3 place.

Antibody 104-H12 has and comprises sequence SGTLSNIGTNIVS (SEQ ID NO:217) at the CDR1 place, comprise sequence NDHRRPS (SEQ ID NO:218) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSLNGVV (SEQ ID NO:219) at the CDR3 place.It has and comprises sequence KYKMV (SEQ ID NO:220) at the CDR1 place, comprise sequence SIYPSGGITAYADSVKG (SEQ ID NO:221) at the CDR2 place, and comprise the HC variable domain of sequence D ITPGGGSGFRLPKNYYYYGMDV (SEQ ID NO:222) at the CDR3 place.

Antibody 135-F03 has and comprises sequence RSSQSLVYSDGNTYLN (SEQ ID NO:223) at the CDR1 place, comprise sequence KVSNRDS (SEQ ID NO:224) at the CDR2 place, and comprise the LC variable domain of sequence MQGTHWQRLT (SEQ ID NO:225) at the CDR3 place.It has at the CDR1 place and comprises sequence WYFMV (SEQ ID NO:226), comprises sequence GIGPSGGLTYADSVKG (SEQ ID NO:227) at the CDR2 place, and comprises the HC variable domain of sequence SDWLNY (SEQ ID NO:228) at the CDR3 place.

Antibody 134-D07 has and comprises sequence TGGRRDIGNYNYVS (SEQ ID NO:229) at the CDR1 place, comprise sequence D VRKRPS (SEQ ID NO:230) at the CDR2 place, and comprise the LC variable domain of sequence GSYTGTSNV (SEQ ID NO:231) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMS (SEQ ID NO:232), comprises sequence RIYPSGGQTYYADSVKG (SEQ ID NO:233) at the CDR2 place, and comprises the HC variable domain of sequence GAGWFDP (SEQ ID NO:234) at the CDR3 place.

Antibody 135-C12 has at the CDR1 place and comprises sequence RASQSVGSDYLA (SEQ ID NO:235), comprises sequence A ASTRAT (SEQ ID NO:236) at the CDR2 place, and comprises the LC variable domain of sequence QQYASPPRT (SEQ ID NO:237) at the CDR3 place.It has and comprises sequence WYAMH (SEQ ID NO:238) at the CDR1 place, comprise sequence WISPSGGQTKYADSVKG (SEQ ID NO:239) at the CDR2 place, and comprise the HC variable domain of sequence A KVLRYFDWLLGGDAFDI (SEQ ID NO:240) at the CDR3 place.

Antibody 097-F04 has at the CDR1 place and comprises sequence RASQSVSFNLA (SEQ ID NO:241), comprises sequence FDASNRAT (SEQ ID NO:242) at the CDR2 place, and comprises the LC variable domain of sequence QQYGTSPFT (SEQID NO:243) at the CDR3 place.It has and comprises sequence PYFME (SEQ ID NO:244) at the CDR1 place, comprise sequence GISSSGGNTLYADSVKG (SEQ ID NO.245) at the CDR2 place, and comprise the HC variable domain of sequence D RFGNYYGSGSKLQHDAFDI (SEQ ID NO:246) at the CDR3 place.

Antibody 111-C10 has at the CDR1 place and comprises sequence RASQTIRTYLN (SEQ ID NO:247), comprises sequence D ASNLQT (SEQ ID NO:248) at the CDR2 place, and comprises the LC variable domain of sequence QQSYGGPPT (SEQID NO:249) at the CDR3 place.It has and comprises sequence FYSMS (SEQ ID NO:250) at the CDR1 place, comprise sequence RIRPSGGRTDYADSVKG (SEQ ID NO:251) at the CDR2 place, and comprise the HC variable domain of sequence D RLYYYGSGSYYYGAFDI (SEQ ID NO:252) at the CDR3 place.

Antibody 094-E08 has at the CDR1 place and comprises sequence RASQSISTYLN (SEQ ID NO:253), comprises sequence A ASSLQS (SEQ ID NO:254) at the CDR2 place, and comprises the LC variable domain of sequence QQYNSFPFS (SEQ IDNO:255) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMD (SEQ ID NO.256), comprises sequence VIYPSGGHTNYADSVKG (SEQ ID NO:257) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:258) at the CDR3 place.

Antibody 138-B10 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:259), comprises sequence D ASNRAT (SEQ ID NO:260) at the CDR2 place, and comprises the LC variable domain of sequence QQWDT (SEQID NO:261) at the CDR3 place.It has at the CDR1 place and comprises sequence HYKMG (SEQ ID NO:262), comprises sequence SIRPSGGATKYADSVKG (SEQ ID NO:263) at the CDR2 place, and comprises the HC variable domain of sequence VGIWDAFDI (SEQ ID NO:264) at the CDR3 place.

Antibody 139-F09 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:265), comprises sequence A ASSLQS (SEQ ID NO:266) at the CDR2 place, and comprises the LC variable domain of sequence QQTYSTLFT (SEQ IDNO:267) at the CDR3 place.It has at the CDR1 place and comprises sequence HYKME (SEQ ID NO:268), comprises sequence RIRPSGGVTKYADSVKG (SEQ ID NO:269) at the CDR2 place, and comprises the HC variable domain of sequence GGLWDAFDI (SEQ ID NO:270) at the CDR3 place.

Antibody 104-A12 has and comprises sequence SGSSSNIGSNIVS (SEQ ID NO:271) at the CDR1 place, comprise sequence SNNRRPS (SEQ ID NO:272) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSLNGHV (SEQ ID NO:273) at the CDR3 place.It has at the CDR1 place and comprises sequence WYTMT (SEQ ID NO:274), comprises sequence SIYPSGGHTSYADSVKG (SEQ ID NO:275) at the CDR2 place, and comprises the HC variable domain of sequence D TRVGPWLVRAPLDY (SEQ ID NO:276) at the CDR3 place.

Antibody 132-D02 has and comprises sequence TGTSTDVGGYNYVS (SEQ ID NO:277) at the CDR1 place, comprise sequence D VSNRPS (SEQ ID NO:278) at the CDR2 place, and comprise the LC variable domain of sequence SSYTNTITW (SEQ ID NO:279) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMY (SEQ ID NO:280), comprises sequence RIGPSGGWTKYADSVKG (SEQ ID NO:281) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:282) at the CDR3 place.

Antibody 138-C04 has and comprises sequence RASQSVSSSYLA (SEQ ID NO at the CDR1 place; 283), comprise sequence GASSRAT (SEQ ID NO:284) at the CDR2 place, and comprise the LC variable domain of sequence QQYGSSPFT (SEQ ID NO:285) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMH (SEQ ID NO:286), comprises sequence VIYPSGGTTKYADSVKG (SEQ ID NO:287) at the CDR2 place, and comprises the HC variable domain of sequence GNSGSHDY (SEQ ID NO:288) at the CDR3 place.

Antibody 132-H11 has at the CDR1 place and comprises sequence RASQGIGNYLA (SEQ ID NO:289), comprises sequence KTSNLQS (SEQ ID NO:290) at the CDR2 place, and comprises the LC variable domain of sequence QRYDSYSQYI (SEQID NO:291) at the CDR3 place.It has at the CDR1 place and comprises sequence MYVMN (SEQ ID NO:292), comprises sequence RIGSSGGGTLYADSVKG (SEQ ID NO:293) at the CDR2 place, and comprises the HC variable domain of sequence VSVYRIRNYYYYAMDV (SEQ ID NO:294) at the CDR3 place.

Antibody 132-D08 has at the CDR1 place and comprises sequence RASQSVSGFLA (SEQ ID NO:295), comprises sequence D ASNRAT (SEQ ID NO:296) at the CDR2 place, and comprises the LC variable domain of sequence QQYGDSSPIT (SEQID NO:297) at the CDR3 place.It has at the CDR1 place and comprises sequence MYGMT (SEQ ID NO:298), comprises sequence GISPSGGRTYYADSVKG (SEQ ID NO:299) at the CDR2 place, and comprises the HC variable domain of sequence HRRDYVWWTYGMDV (SEQ ID NO:300) at the CDR3 place.

Antibody 121-A07 has at the CDR1 place and comprises sequence RASQGISSWLA (SEQ ID NO:301), comprises sequence A ASSLQS (SEQ ID NO:302) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPSLT (SEQID NO:303) at the CDR3 place.It has at the CDR1 place and comprises sequence NYSMQ (SEQ ID NO:304), comprises sequence GIRPSGGSTRYADSVKG (SEQ ID NO:305) at the CDR2 place, and comprises the HC variable domain of sequence STGGYYYGMDV (SEQ ID NO:306) at the CDR3 place.

Antibody 126-F11 has at the CDR1 place and comprises sequence RASQSVSSYLA (SEQ ID NO:307), comprises sequence D ASNRAT (SEQ ID NO:308) at the CDR2 place, and comprises the LC variable domain of sequence QQRSNWPPYT (SEQID NO:309) at the CDR3 place.It has at the CDR1 place and comprises sequence PYMMG (SEQ ID NO:310), comprises sequence SIRSSGGATAYADSVKG (SEQ ID NO:311) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:312) at the CDR3 place.

Antibody 098-H04 has at the CDR1 place and comprises sequence RASQAISTNLN (SEQ ID NO:313), comprises sequence A ASSLQS (SEQ ID NO:314) at the CDR2 place, and comprises the LC variable domain of sequence QQTFSPPHT (SEQ IDNO.315) at the CDR3 place.It has and comprises sequence RYYME (SEQ ID NO:316) at the CDR1 place, comprise sequence SISSSGGSTEYADSVKG (SEQ ID NO:317) at the CDR2 place, and comprise the HC variable domain of sequence D ITPGGGSGFRLPKNYYYYGMDV (SEQ ID NO:318) at the CDR3 place.

Antibody 093-C02 has at the CDR1 place and comprises sequence GASQSVSSSYLA (SEQ ID NO:319), comprises sequence D ASSRAT (SEQ ID NO:320) at the CDR2 place, and comprises the LC variable domain of sequence LHDYNFPFT (SEQ ID NO.321) at the CDR3 place.It has at the CDR1 place and comprises sequence HYPMI (SEQ ID NO:322), comprises sequence GIYSSGGTTKYADSVKG (SEQ ID NO:323) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:324) at the CDR3 place.

Antibody 115-G12 has at the CDR1 place and comprises sequence RASQAISNYLA (SEQ ID NO:325), comprises sequence A ASTLQS (SEQ ID NO:326) at the CDR2 place, and comprises the LC variable domain of sequence QTYYSVPFT (SEQID NO:327) at the CDR3 place.It has and comprises sequence IYAMI (SEQ ID NO:328) at the CDR1 place, comprise sequence SIYSSGGPTQYADSVKG (SEQ ID NO:329) at the CDR2 place, and comprise the HC variable domain of sequence GRRTYYYDSSGYYKYAAFDI (SEQ ID NO:330) at the CDR3 place.

Antibody 135-B05 has and comprises sequence TGTSSDVGGYNYLS (SEQ ID NO:331) at the CDR1 place, comprise sequence GVSNRPS (SEQ ID NO:332) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSTGTRV (SEQ ID NO:333) at the CDR3 place.It has at the CDR1 place and comprises sequence NYTMT (SEQ ID NO:334), comprises sequence VIYPSGGHTTYADSVKG (SEQ ID NO:335) at the CDR2 place, and comprises the HC variable domain of sequence GGRWFSLDY (SEQ ID NO:336) at the CDR3 place.

Antibody 136-F08 has and comprises sequence TGGRRDIGNYNYVS (SEQ ID NO:337) at the CDR1 place, comprise sequence D VRKRPS (SEQ ID NO:338) at the CDR2 place, and comprise the LC variable domain of sequence GSYTGTSNV (SEQ ID NO:339) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMQ (SEQ ID NO:340), comprises sequence SIYPSGGSTRYADSVKG (SEQ ID NO:341) at the CDR2 place, and comprises the HC variable domain of sequence FNVGFDL (SEQ ID NO:342) at the CDR3 place.

Antibody 110-H11 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:343), comprises sequence A ASSLQS (SEQ ID NO:344) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTLYT (SEQID NO:345) at the CDR3 place.It has at the CDR1 place and comprises sequence WYRMS (SEQ ID NO.346), comprises sequence SISPSGGPTKYADSVKG (SEQ ID NO:347) at the CDR2 place, and comprises the HC variable domain of sequence PSGDSSVYRPFAS (SEQ ID NO:348) at the CDR3 place.

Antibody 110-D11 has and comprises sequence TDTSGNVGSYNLVS (SEQ ID NO.349) at the CDR1 place, comprise sequence EVSRRPS (SEQ ID NO:350) at the CDR2 place, and comprise the LC variable domain of sequence C SYAGSNTYV (SEQ ID NO:351) at the CDR3 place.It has at the CDR1 place and comprises sequence NYSMV (SEQ ID NO:352), comprises sequence RIYSSGGSTHYADSVKG (SEQ ID NO:353) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:354) at the CDR3 place.

Antibody 114-H07 has and comprises sequence SGGSSNIGSNRVN (SEQ ID NO:355) at the CDR1 place, comprise sequence SNNQRPS (SEQ ID NO:356) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDNLIGPV (SEQ ID NO:357) at the CDR3 place.It has and comprises sequence WYHMY (SEQ ID NO:358) at the CDR1 place, comprise sequence YIRPSGGNTNYADSVKG (SEQ ID NO:359) at the CDR2 place, and comprise the HC variable domain of sequence D RRGYSSSKGYYYYGMDV (SEQ ID NO:360) at the CDR3 place.

Antibody 098-E01 has and comprises sequence SGSSSNIGSNYVY (SEQ ID NO:361) at the CDR1 place, comprise sequence RNNQRPS (SEQ ID NO:362) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSMSGVV (SEQ ID NO:363) at the CDR3 place.It has and comprises sequence HYFMH (SEQ ID NO:364) at the CDR1 place, comprise sequence YISSSGGLTGYADSVKG (SEQ ID NO:365) at the CDR2 place, and comprise the HC variable domain of sequence RGPVYYYDSSGSHSAFDI (SEQ ID NO:366) at the CDR3 place.

Antibody 135-C11 has and comprises sequence TGSTSDVGGYTYVS (SEQ ID NO:367) at the CDR1 place, comprise sequence D VSKRPS (SEQ ID NO:368) at the CDR2 place, and comprise the LC variable domain of sequence C SYAGSYSYV (SEQ ID NO:369) at the CDR3 place.It has at the CDR1 place and comprises sequence NYHMD (SEQ ID NO:370), comprises sequence VIGPSGGFTRYADSVKG (SEQ ID NO:371) at the CDR2 place, and comprises the HC variable domain of sequence A LGGSLDY (SEQ ID NO:372) at the CDR3 place.

Antibody 095-A11 has and comprises sequence TATSSDLGSYNFVS (SEQ ID NO.373) at the CDR1 place, comprise sequence EVSRRPS (SEQ ID NO:374) at the CDR2 place, and comprise the LC variable domain of sequence C SYAGSNTYV (SEQ ID NO:375) at the CDR3 place.It has at the CDR1 place and comprises sequence NYSMM (SEQ ID NO:376), comprises sequence SIYPSGGFTKYADSVKG (SEQ ID NO:377) at the CDR2 place, and comprises the HC variable domain of sequence VGFYGGFV (SEQ ID NO:378) at the CDR3 place.

Antibody 092-F08 has at the CDR1 place and comprises sequence RASQSISSWLA (SEQ ID NO:379), comprises sequence KASSLGS (SEQ ID NO:380) at the CDR2 place, and comprises the LC variable domain of sequence QQYYSYSQT (SEQID NO:381) at the CDR3 place.It has at the CDR1 place and comprises sequence NYPMH (SEQ ED NO:382), comprises sequence YIGPSGGWTRYADSVKG (SEQ ID NO:383) at the CDR2 place, and comprises the HC variable domain of sequence A PSGY (SEQ ID NO:384) at the CDR3 place.

Antibody 104-B12 has and comprises sequence TGTSSDVGLYNFVS (SEQ ID NO:385) at the CDR1 place, comprise sequence D VSRRPS (SEQ ID NO:386) at the CDR2 place, and comprise the LC variable domain of sequence SSYAGSNNYV (SEQ ID NO:387) at the CDR3 place.It has and comprises sequence PYRMS (SEQ ID NO:388) at the CDR1 place, comprise sequence VISPSGGITEYADSVKG (SEQ ID NO:389) at the CDR2 place, and comprise the HC variable domain of sequence HERGSGYYVTPPAGAFDI (SEQ ID NO:390) at the CDR3 place.

Antibody 109-A11 has at the CDR1 place and comprises sequence RASQSIANYLN (SEQ ID NO:391), comprises sequence A ASRLHS (SEQ ID NO:392) at the CDR2 place, and comprises the LC variable domain of sequence QQSHASPLT (SEQID NO:393) at the CDR3 place.It has at the CDR1 place and comprises sequence HYAMG (SEQ ID NO:394), comprises sequence RIWPSGGHTQYADSVKG (SEQ ID NO:395) at the CDR2 place, and comprises the HC variable domain of sequence GYSSTWGAFDI (SEQ ID NO:396) at the CDR3 place.

Antibody 131-G03 has and comprises sequence TGTSSDVGGYNYVS (SEQ ID NO:397) at the CDR1 place, comprise sequence EVSNRPS (SEQ ID NO:398) at the CDR2 place, and comprise the LC variable domain of sequence SSYKRGGTYV (SEQ ID NO:399) at the CDR3 place.It has at the CDR1 place and comprises sequence WYGMG (SEQ ID NO:400), comprises sequence VIRPSGGTTRYADSVKG (SEQ ID NO:401) at the CDR2 place, and comprises the HC variable domain of sequence GGRYYSLDY (SEQ ID NO:402) at the CDR3 place.

Antibody 135-E10 has at the CDR1 place and comprises sequence RASQSVTTFLA (SEQ ID NO:403), comprises sequence GASSRAA (SEQ ID NO:404) at the CDR2 place, and comprises the LC variable domain of sequence QQYRFSPPT (SEQID NO:405) at the CDR3 place.It has and comprises sequence PYNMA (SEQ ID NO:406) at the CDR1 place, comprise sequence SIWPSGGRTRYADSVKG (SEQ ID NO:407) at the CDR2 place, and comprise the HC variable domain of sequence GVRSSGLLERGRVYDAFDI (SEQ ID NO:408) at the CDR3 place.

Antibody 130-B02 has at the CDR1 place and comprises sequence RASQSVSGSYLA (SEQ ID NO:409), comprises sequence GAYSRAT (SEQ ID NO:410) at the CDR2 place, and comprises the LC variable domain of sequence QHYGSSPRT (SEQ ID NO:411) at the CDR3 place.It has at the CDR1 place and comprises sequence HYQMH (SEQ ID NO:412), comprises sequence SIRPSGGRTAYADSVKG (SEQ ID NO:413) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:414) at the CDR3 place.

Antibody 097-D04 has at the CDR1 place and comprises sequence RAGQMYYWLA (SEQ ID NO:415), comprises sequence A ASSLQS (SEQ ID NO:416) at the CDR2 place, and comprises the LC variable domain of sequence QQAKSFPVT (SEQ ID NO:417) at the CDR3 place.It has at the CDR1 place and comprises sequence PYBVIS (SEQ ID NO:418), comprises sequence SIVSSGGVTLYADSVKG (SEQ ID NO:419) at the CDR2 place, and comprises the HC variable domain of sequence NINLRYDILTGYFDI (SEQ ID NO:420) at the CDR3 place.

Antibody 093-G06 has and comprises sequence SGTSSDVGAYYHVS (SEQ ID NO:421) at the CDR1 place, comprise sequence EVTNRPS (SEQ ID NO:422) at the CDR2 place, and comprise the LC variable domain of sequence SSYTTSNTLV (SEQ ID NO:423) at the CDR3 place.It has at the CDR1 place and comprises sequence WYVMR (SEQ ID NO:424), comprises sequence SIYPSGGQTRYADSVKG (SEQ ID NO.425) at the CDR2 place, and comprises the HC variable domain of sequence GYSSTWGAFDI (SEQ ID NO:426) at the CDR3 place.

Antibody 108-E11 has at the CDR1 place and comprises sequence RASQSVSSNLA (SEQ ID NO.427), comprises sequence GASTRAT (SEQ ID NO:428) at the CDR2 place, and comprises the LC variable domain of sequence QQNNNWPPSFT (SEQID NO:429) at the CDR3 place.It has at the CDR1 place and comprises sequence HYTML (SEQ ID NO:430), comprises sequence SIYSSGGSTYYADSVKG (SEQ ID NO:431) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:432) at the CDR3 place.

Antibody 127-C06 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:433), comprises sequence GASSRAT (SEQ ID NO:434) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSRYT (SEQ ID NC-.435) at the CDR3 place.It has at the CDR1 place and comprises sequence NYTMF (SEQ ID NO:436), comprises sequence SIYPSGGITRYADSVKG (SEQ ID NO:437) at the CDR2 place, and comprises the HC variable domain of sequence VSFWNAFDI (SEQ ID NO:438) at the CDR3 place.

Antibody 105-B06 has at the CDR1 place and comprises sequence RASQTISSYLN (SEQ ID NO:439), comprises sequence TTSSLQS (SEQ ED NO:440) at the CDR2 place, and comprises the LC variable domain of sequence QQSHSSPT (SEQ IDNO:441) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMY (SEQ ID NO:442), comprises sequence SIYPSGGFTAYADSVKG (SEQ ID NO:443) at the CDR2 place, and comprises the HC variable domain of sequence RGVYGTFDI (SEQ ID NO:444) at the CDR3 place.

Antibody 107-D01 has at the CDR1 place and comprises sequence RASQDVRRFLA (SEQ ID NO:445), comprises sequence A ASSLQS (SEQ ID NO:446) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPIT (SEQ ID NO:447) at the CDR3 place.It has at the CDR1 place and comprises sequence HYPMS (SEQ ID NO:448), comprises sequence YIRSGGYTTYADSVKG (SEQ ID NO:449) at the CDR2 place, and comprises the HC variable domain of sequence ERVFCSGGRCGSYFDY (SEQ ID NO:450) at the CDR3 place.

Antibody 107-F11 has at the CDR1 place and comprises sequence RASQSVYSSYLA (SEQ ID NO:451), comprises sequence A ASNR (SEQ ID NO:452) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPR (SEQID NO:453) at the CDR3 place.It has at the CDR1 place and comprises sequence RYAMF (SEQ ID NO:454), comprises sequence SISPSGGKTRYADSVKG (SEQ ID NO:455) at the CDR2 place, and comprises the HC variable domain of sequence GGGTALDY (SEQ ID NO.456) at the CDR3 place.

Antibody 101-E01 has at the CDR1 place and comprises sequence RASQSISSWLA (SEQ ID NO.457), comprises sequence A ASSLRN (SEQ ID NO:458) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPPT (SEQ IDNO:459) at the CDR3 place.It has at the CDR1 place and comprises sequence RYWMA (SEQ ID NO:460), comprises sequence SIGPSGGSTKYADSVKG (SEQ ID NO:461) at the CDR2 place, and comprises the HC variable domain of sequence TARWLSFDY (SEQ ID NO:462) at the CDR3 place.

Antibody 138-E12 has at the CDR1 place and comprises sequence RASQSVSSNYLA (SEQ ID NO:463), comprises sequence GASSRAT (SEQ ID NO:464) at the CDR2 place, and comprises the LC variable domain of sequence QQFGSSLFT (SEQ ID NO:465) at the CDR3 place.It has at the CDR1 place and comprises sequence NYYML (SEQ ID NO:466), comprises sequence YIRSSGGSTRYADSVKG (SEQ ID NO:467) at the CDR2 place, and comprises the HC variable domain of sequence PTVDGAFDY (SEQ ID NO:468) at the CDR3 place.

Antibody 109-E08 has and comprises sequence A GTSSDLGGYDYVS (SEQ ID NO:469) at the CDR1 place, comprise sequence QVGRRPS (SEQ ID NO:470) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSSRTRV (SEQ ID NO:471) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMD (SEQ ID NO:472), comprises sequence SIYPSGGWTRYADSVKG (SEQ ID NO:473) at the CDR2 place, and comprises the HC variable domain of sequence RGQWLVLDY (SEQ ID NO:474) at the CDR3 place.

Antibody 104-B03 has and comprises sequence TGTSSDVGGFNYVS (SEQ ID NO:475) at the CDR1 place, comprise sequence D VSKRPS (SEQ ID NO:476) at the CDR2 place, and comprise the LC variable domain of sequence C SYTGNYTYV (SEQ ID NO:477) at the CDR3 place.It has and comprises sequence RYAMA (SEQ ID NO:478) at the CDR1 place, comprise sequence SIGSSGGVTKYADSVKG (SEQ ID NO:479) at the CDR2 place, and comprise the HC variable domain of sequence PRGRYYYDSSGYYYGGVFDY (SEQ ID NO:480) at the CDR3 place.

Antibody 096-E11 has at the CDR1 place and comprises sequence RASQGISNYLA (SEQ ID NO:481), comprises sequence A ASTLQS (SEQ ID NO:482) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPYT (SEQID NO:483) at the CDR3 place.It has and comprises sequence QYLMA (SEQ ID NO:484) at the CDR1 place, comprise sequence SIYPSGGNTNYADSVKG (SEQ ID NO:485) at the CDR2 place, and comprise the HC variable domain of sequence D RSIVPYSSSWYMPRDYYYGMDV (SEQ ID NO:486) at the CDR3 place.

Antibody 103-A03 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:487), comprises sequence GAASRAT (SEQ ID NO:488) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSPLIT (SEQ ID NO:489) at the CDR3 place.It has at the CDR1 place and comprises sequence HYPMT (SEQ ID NO:490), comprises sequence SIRPSGGNTGYADSVKG (SEQ ID NO:491) at the CDR2 place, and comprises the HC variable domain of sequence LTGYSSGWFWAYGMDV (SEQ ID NO:492) at the CDR3 place.

Antibody 099-A09 has and comprises sequence TGTSSDVGGYNYVS (SEQ ID NO:493) at the CDR1 place, comprise sequence EVSNRAS (SEQ ID NO:494) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSSTTLL (SEQ ID NO:495) at the CDR3 place.It has and comprises sequence RYVMA (SEQ ID NO:496) at the CDR1 place, comprise sequence VIRPSGGKTLYADSVKG (SEQ ID NO:497) at the CDR2 place, and comprise the HC variable domain of sequence VAGRVGVPATKKNWFDP (SEQ ID NO:498) at the CDR3 place.

Antibody 105-F02 has at the CDR1 place and comprises sequence RASQGISSWLA (SEQ ID NO:499), comprises sequence A ASSLQS (SEQ ID NO.500) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPLT (SEQ IDNO:501) at the CDR3 place.It has at the CDR1 place and comprises sequence PYTMS (SEQ ID NO:502), comprises sequence GIYPSGGETWYADSVKG (SEQ ID NO:503) at the CDR2 place, and comprises the HC variable domain of sequence NINLRYDILTGYFDF (SEQ ID NO:504) at the CDR3 place.

Antibody 112-C02 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:505), comprises sequence GASSRAT (SEQ ID NO:506) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSGWT (SEQ ID NO.507) at the CDR3 place.It has at the CDR1 place and comprises sequence YYRMT (SEQ ID NO:508), comprises sequence SIYPSGGYTIYADSVKG (SEQ ID NO.509) at the CDR2 place, and comprises the HC variable domain of sequence LGQWLALDH (SEQ ID NO:510) at the CDR3 place.

Antibody 125-C04 has at the CDR1 place and comprises sequence QGDSLRSYYAS (SEQ ID NO:511), comprises sequence SKSNRPS (SEQ ID NO-.512) at the CDR2 place, and comprises the LC variable domain of sequence NSRDSSGNHLV (SEQID NO:513) at the CDR3 place.It has at the CDR1 place and comprises sequence KYWMA (SEQ ID NO:514), comprises sequence VIYPSGGQTKYADSVKG (SEQ ID NO:515) at the CDR2 place, and comprises the HC variable domain of sequence MRYGDRFDY (SEQ ID NO:516) at the CDR3 place.

Antibody 122-H04 has at the CDR1 place and comprises sequence RASQGISSYLA (SEQ ID NO:517), comprises sequence A ASTLQS (SEQ ID NO:518) at the CDR2 place, and comprises the LC variable domain of sequence QQLNSYPRT (SEQID NO:519) at the CDR3 place.It has at the CDR1 place and comprises sequence RYWMH (SEQ ID NO:520), comprises sequence SIGPSGGVTKYADSVKG (SEQ ID NO:521) at the CDR2 place, and comprises the HC variable domain of sequence HGSWGGMDV (SEQ ID NO:522) at the CDR3 place.

Antibody 092-B12 has and comprises sequence TGTTSDVGGYKYVS (SEQ ID NO:523) at the CDR1 place, comprise sequence EVNHRPS (SEQ ID NO:524) at the CDR2 place, and comprise the LC variable domain of sequence YSYTSDSTPYV (SEQ ID NO:525) at the CDR3 place.It has at the CDR1 place and comprises sequence RYAMH (SEQ ID NO:526), comprises sequence SIWPSGGATFYADSVKG (SEQ ID NO:527) at the CDR2 place, and comprises the HC variable domain of sequence SPTLNAFHI (SEQ ID NO:528) at the CDR3 place.

Antibody 139-A04 has at the CDR1 place and comprises sequence RASQSISNWLA (SEQ ID NO:529), comprises sequence KASTLES (SEQ ID NO:530) at the CDR2 place, and comprises the LC variable domain of sequence QHYNSYSL (SEQ IDNO:531) at the CDR3 place.It has at the CDR1 place and comprises sequence HYTMV (SEQ ID NO:532), comprises sequence SIYSSGGRTNYADSVKG (SEQ ID NO:533) at the CDR2 place, and comprises the HC variable domain of sequence IVWPSAQFYFYYGMDV (SEQ ID NO:534) at the CDR3 place.

Antibody 108-C10 has and comprises sequence TGTSNDIGRTNYVS (SEQ ID NO:535) at the CDR1 place, comprise sequence EVSNRPS (SEQ ID NO:536) at the CDR2 place, and comprise the LC variable domain of sequence SSCTTAPVCV (SEQ ID NO:537) at the CDR3 place.It has at the CDR1 place and comprises sequence LYAMH (SEQ ID NO:538), comprises sequence SIRPSGGLTKYADSVKG (SEQ ID NO:539) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:540) at the CDR3 place.

Antibody 138-B06 has and comprises sequence SGSSSNIGSNYVY (SEQ ID NO:541) at the CDR1 place, comprise sequence SNNQRPS (SEQ ID NO:542) at the CDR2 place, and comprise the LC variable domain of sequence A TWDNSLSAWV (SEQ ID NO:543) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMN (SEQ ID NO:544), comprises sequence RIYPSGGVTVYADSVKG (SEQ ID NO:545) at the CDR2 place, and comprises the HC variable domain of sequence GYGMDV (SEQ ID NO:546) at the CDR3 place.

Antibody 137-B01 has at the CDR1 place and comprises sequence RASQSVGNSLA (SEQ ID NO:547), comprises sequence GASTRAS (SEQ ID NO:548) 5 and at the CDR2 place, comprises the LC variable domain of sequence QEYNKWPIT (SEQ ID NO.549) at the CDR3 place.It has at the CDR1 place and comprises sequence WYWMY (SEQ ID NO:550), comprises sequence VIGSSGGVTKYADSVKG (SEQ ID NO:551) at the CDR2 place, and comprises the HC variable domain of sequence GPYRGYFDY (SEQ ID NO:552) at the CDR3 place.

Antibody 111-B02 has at the CDR1 place and comprises sequence RASQSISSYLS (SEQ ID NO:553), comprises sequence SASNLQS (SEQ ID NO:554) at the CDR2 place, and comprises the LC variable domain of sequence QQIYRTPLT (SEQ IDNO:555) at the CDR3 place.It has at the CDR1 place and comprises sequence WYQMM (SEQ ID NO.556), comprises sequence SYIRSSGGKTDYADSVKG (SEQ ID NO:557) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:558) at the CDR3 place.

Antibody 129-B09 has at the CDR1 place and comprises sequence RASQSVGSDYLA (SEQ ID NO:559), comprises sequence GASSRAT (SEQ ID NO:560) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSLYT (SEQ ID NO:561) at the CDR3 place.It has at the CDR1 place and comprises sequence NYKMS (SEQ ID NO:562), comprises sequence GIYPSGGLTQYADSVKG (SEQ ID NO:563) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:564) at the CDR3 place.

Antibody 099-C12 has at the CDR1 place and comprises sequence RAGQTINNYLN (SEQ ID NO:565), comprises sequence A ASNLQS (SEQ ID NO:566) at the CDR2 place, and comprises the LC variable domain of sequence QQTYRTPFT (SEQ ID NO:567) at the CDR3 place.It has at the CDR1 place and comprises sequence HYKMH (SEQ ID NO:568), comprises sequence SIWPSGGGTFYADSVKG (SEQ ID NO:569) at the CDR2 place, and comprises the HC variable domain of sequence TSGGFGAFDI (SEQ ID NO:570) at the CDR3 place.

Antibody 099-E11 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:571), comprises sequence A ASSLQS (SEQ ID NO:572) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPRT (SEQ IDNO:573) at the CDR3 place.It has at the CDR1 place and comprises sequence HYYMM (SEQ ID NO.574), comprises sequence SIGPSGGMTDYADSVKG (SEQ ID NO.575) at the CDR2 place, and comprises the HC variable domain of sequence GGTASLDY (SEQ ID NO:576) at the CDR3 place.

Antibody 103-F07 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:577), comprises sequence GASSRAT (SEQ ID NO:578) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSSYT (SEQ ID NO:579) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMG (SEQ ID NO:580), comprises sequence SIYPSGGKTTYADSVKG (SEQ ID NO:58 1) at the CDR2 place, and comprises the HC variable domain of sequence PSTGSWSAFDI (SEQ ID NO:582) at the CDR3 place.

Antibody 102-A04 has and comprises sequence TGTSSDVGGYNYVS (SEQ ID NO:583) at the CDR1 place, comprise sequence D VSKRPS (SEQ ID NO:584) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSSITLVV (SEQ ID NO:585) at the CDR3 place.It has at the CDR1 place and comprises sequence WYAMH (SEQ ID NO:586), comprises sequence VISPSGGATRYADSVKG (SEQ ID NO:587) at the CDR2 place, and comprises the HC variable domain of sequence A STVTTLFQH (SEQ ID NO:588) at the CDR3 place.

Antibody 119-B09 has at the CDR1 place and comprises sequence RASQSISSWLA (SEQ ID NO:589), comprises sequence A ASTLQS (SEQ ID NO:590) at the CDR2 place, and comprises the LC variable domain of sequence QKYNSAPLT (SEQID NO:591) at the CDR3 place.It has at the CDR1 place and comprises sequence NYHMQ (SEQ ID NO:592), comprises sequence SIYPSGGFTRYADSVKG (SEQ ID NO:593) at the CDR2 place, and comprises the HC variable domain of sequence D GGQGG (SEQ ID NO:594) at the CDR3 place.

Antibody 097-E07 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:595), comprises sequence A ASSLQS (SEQ ID NO:596) at the CDR2 place, and comprises the LC variable domain of sequence QQTFSTWT (SEQ IDNO:597) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMG (SEQ ID NO:598), comprises sequence SIWPSGGHTSYADSVKG (SEQ ID NO:599) at the CDR2 place, and comprises the HC variable domain of sequence HYGMDV (SEQ ID NO:600) at the CDR3 place.

Antibody 117-A12 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:601), comprises sequence GASSRAT (SEQ ID NO:602) at the CDR2 place, and comprises the LC variable domain of sequence QQYNNWPPLT (SEQ ID NO:603) at the CDR3 place.It has at the CDR1 place and comprises sequence RYMMY (SEQ ID NO:604), comprises sequence SIYPSGGKTLYADSVKG (SEQ ID NO:605) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:606) at the CDR3 place.

Antibody 126-C10 has at the CDR1 place and comprises sequence RASHNIYTSLN (SEQ ID NO:607), comprises sequence GASTLEN (SEQ ID NO:608) at the CDR2 place, and comprises the LC variable domain of sequence QQSYTSVT (SEQ IDNO:609) at the CDR3 place.It has and comprises sequence FYSMM (SEQ ID NO:610) at the CDR1 place, comprise sequence GISSSGGTTKYADSVKG (SEQ ID NO:611) at the CDR2 place, and comprise the HC variable domain of sequence GGTNYDYVWGSYRSHYYYYGMDV (SEQ ID NO:612) at the CDR3 place.

Antibody 127-D05 has at the CDR1 place and comprises sequence RASQSISSWLA (SEQ ID NO:613), comprises sequence TASSLES (SEQ ID NO:614) at the CDR2 place, and comprises the LC variable domain of sequence QQYNSYSLT (SEQ IDNO:615) at the CDR3 place.It has at the CDR1 place and comprises sequence NYKMQ (SEQ ID NO:616), comprises sequence SIYSSGGKTVYADSVKG (SEQ ID NO:617) at the CDR2 place, and comprises the HC variable domain of sequence TPGYNYFDY (SEQ ID NO:618) at the CDR3 place.

Antibody 126-B12 has at the CDR1 place and comprises sequence RASQTFNNYLN (SEQ ID NO:619), comprises sequence A ASTLQS (SEQ ID NO:620) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPFT (SEQ ID NO:621) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMF (SEQ ID NO:622), comprises sequence SIWPSGGNTKYADSVKG (SEQ ID NO:623) at the CDR2 place, and comprises the HC variable domain of sequence PAYYYAMDV (SEQ ID NO:624) at the CDR3 place.

Antibody 122-G06 has at the CDR1 place and comprises sequence RASQTVGGSYLA (SEQ ID NO:625), comprises sequence A ASNRAP (SEQ ID NO:626) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSMYT (SEQ ID NO:627) at the CDR3 place.It has at the CDR1 place and comprises sequence WYQMS (SEQ ID NO:628), comprises sequence SIYPSGGATKYADSVKG (SEQ ID NO:629) at the CDR2 place, and comprises the HC variable domain of sequence MGLHHSFDY (SEQ ID NO:630) at the CDR3 place.

Antibody 107-D05 has at the CDR1 place and comprises sequence RASQGISSWLA (SEQ ID NO:63 1), comprises sequence A ASSLQS (SEQ ID NO:632) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPLT (SEQ IDNO:633) at the CDR3 place.It has at the CDR1 place and comprises sequence HYWMR (SEQ ID NO:634), comprises sequence SIGSSGGMTKYADSVKG (SEQ ID NC-.635) at the CDR2 place, and comprises the HC variable domain of sequence A LRWRAFDI (SEQ ID NO:636) at the CDR3 place.

Antibody 127-H05 has and comprises sequence TGTNTDVGGYNYVS (SEQ ID NO:637) at the CDR1 place, comprise sequence EVNHRPS (SEQ ID NO:638) at the CDR2 place, and comprise the LC variable domain of sequence SSYTYRNTYV (SEQ ID NO:639) at the CDR3 place.It has at the CDR1 place and comprises sequence WYTMG (SEQ ID NO:640), comprises sequence VIYPSGGMTKYADSVKG (SEQ ID NO:641) at the CDR2 place, and comprises the HC variable domain of sequence TSGGTPWGF (SEQ ID NO:642) at the CDR3 place.

Antibody 112-C12 has at the CDR1 place and comprises sequence RASQTVSSNYLA (SEQ ID NO:643), comprises sequence GVSNRAA (SEQ ID NO:644) at the CDR2 place, and comprises the LC variable domain of sequence QHYGSSAFT (SEQ ID NO:645) at the CDR3 place.It has at the CDR1 place and comprises sequence NYMS (SEQ ID NO:646), comprises sequence SIWPSGGHTRYADSVKG (SEQ ID NO:647) at the CDR2 place, and comprises the HC variable domain of sequence A GSYYAGDY (SEQ ID NO:648) at the CDR3 place.

Antibody 098-C10 has at the CDR1 place and comprises sequence RASQDVLVSFA (SEQ ID NO:649), comprises sequence A ASHLHP (SEQ ID NO:650) at the CDR2 place, and comprises the LC variable domain of sequence QQARSFPHT (SEQID NO.651) at the CDR3 place.It has at the CDR1 place and comprises sequence WYSMT (SEQ ID NO:652), comprises sequence SISPSGGATKYADSVKG (SEQ ID NO:653) at the CDR2 place, and comprises the HC variable domain of sequence GGAASLPFDY (SEQ ID NO:654) at the CDR3 place.

Antibody 095-H10 has at the CDR1 place and comprises sequence FGDKLGDKYGS (SEQ ID NO:655), comprises sequence QYWKRPS (SEQ ID NO:656) at the CDR2 place, and comprises the LC variable domain of sequence QAWDSNTVV (SEQ ID NO:657) at the CDR3 place.It has at the the.sequence NYKMH of CDR1 place (SEQ ID NO:658), comprises sequence SIYPSGGWTVYADSVKG (SEQ ID NO:659) at the CDR2 place, and comprises the HC variable domain of sequence GYSSTWGAFDI (SEQ ID NO:660) at the CDR3 place.

Antibody 117-C04 has at the CDR1 place and comprises sequence RASQYISTYLA (SEQ ID NO:661), comprises sequence KASDLES (SEQ ID NO:662) at the CDR2 place, and comprises the LC variable domain of sequence QQYNTYWT (SEQID NO:663) at the CDR3 place.It has at the CDR1 place and comprises sequence FYVMG (SEQ ID NO:664), comprises sequence WIGPSGGRTWYADSVKG (SEQ ID NO.665) at the CDR2 place, and comprises the HC variable domain of sequence D RGWYGIDV (SEQ ID NO:666) at the CDR3 place.

Antibody 094-F03 has at the CDR1 place and comprises sequence SGNILDNSYAS (SEQ ID NO:667), comprises sequence RDNKRPS (SEQ ID NO:668) at the CDR2 place, and comprises the LC variable domain of sequence QAWDRTGV (SEQID NO:669) at the CDR3 place.It has and comprises sequence WYNMY (SEQ ID NO:670) at the CDR1 place, comprise sequence YIVPSGGATHYADSVKG (SEQ ID NO:671) at the CDR2 place, and comprise the HC variable domain of sequence A LRGYSYGPRGYYYYGMDV (SEQ ID NO.672) at the CDR3 place.

Antibody 124-G12 has at the CDR1 place and comprises sequence SPSGGGLRNKYVS (SEQ ID NO:673), comprises sequence KDAERPS (SEQ ID NO:674) at the CDR2 place, and comprises the LC variable domain of sequence LAWDSTTRV (SEQ ID NO:675) at the CDR3 place.It has at the CDR1 place and comprises sequence HYKMN (SEQ ID NO:676), comprises sequence YIGSSGGKTGYADSVKG (SEQ ID NO:677) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:678) at the CDR3 place.

Antibody 131-A03 has at the CDR1 place and comprises sequence RASQSISNWLA (SEQ ID NO.679), comprises sequence KASTLQT (SEQ ID NO:680) at the CDR2 place, and comprises the LC variable domain of sequence QQTYSAPFN (SEQID NO:681) at the CDR3 place.It has at the CDR1 place and comprises sequence NYNMH (SEQ ID NO:682), comprises sequence VIYPSGGYTVYADSVKG (SEQ ID NO:683) at the CDR2 place, and comprises the HC variable domain of sequence RGNWGGIDY (SEQ ID NO:684) at the CDR3 place.

Antibody 099-D05 has at the CDR1 place and comprises sequence RTSQTVSTFLN (SEQ ID NO:685), comprises sequence A ASRLQS (SEQ ID NO:686) at the CDR2 place, and comprises the LC variable domain of sequence QQSFTSPRT (SEQ IDNO.687) at the CDR3 place.It has at the CDR1 place and comprises sequence RYVMH (SEQ ID NO:688), comprises sequence YISPSGGVTKYADSVKG (SEQ ID NO.689) at the CDR2 place, and comprises the HC variable domain of sequence A LYPGMGYYYGMDV (SEQ ID NO:690) at the CDR3 place.

Antibody 131-C08 has at the CDR1 place and comprises sequence RPSQSVYSNYLA (SEQ ID NO:691), comprises sequence GASTRAT (SEQ ID NO:692) at the CDR2 place, and comprises the LC variable domain of sequence QQYGNSYT (SEQ ID NO:693) at the CDR3 place.It has at the CDR1 place and comprises sequence HYNMT (SEQ ID NO:694), comprises sequence SIWPSGGATRYADSVKG (SEQ ID NO:695) at the CDR2 place, and comprises the HC variable domain of sequence TSRFYGMDV (SEQ ID NO:696) at the CDR3 place.

Antibody 116-C09 has at the CDR1 place and comprises sequence RASQSVNIDVG (SEQ ID NO:697), comprises sequence D ASKRAT (SEQ ID NO:698) at the CDR2 place, and comprises the LC variable domain of sequence QQRARWLT (SEQ IDNO:699) at the CDR3 place.It has at the CDR1 place and comprises sequence NYFMN (SEQ ID NO:700), comprises sequence SIGSSGGYTRYADSVKG (SEQ ID NO:701) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:702) at the CDR3 place.

Antibody 128-F11 has at the CDR1 place and comprises sequence RASQDIRTWLA (SEQ ID NO.703), comprises sequence SSSSLQS (SEQ ID NO:704) at the CDR2 place, and comprises the LC variable domain of sequence QQATTFPWT (SEQ IDNO:705) at the CDR3 place.It has at the CDR1 place and comprises sequence HYIMH (SEQ ID NO:706), comprises sequence GIYPSGGSTKYADSVKG (SEQ ID NO:707) at the CDR2 place, and comprises the HC variable domain of sequence A LVGALDY (SEQ ID NO:708) at the CDR3 place.

Antibody 131-D12 has at the CDR1 place and comprises sequence RASQGISNYLA (SEQ ID NO:709), comprises sequence A ASTLQS (SEQ ED NO:710) at the CDR2 place, and comprises the LC variable domain of sequence EQLNSFPHA (SEQ IDNO:711) at the CDR3 place.It has at the CDR1 place and comprises sequence RYTME (SEQ ED NO:712), comprises sequence VERPSGGTTMYADSVKG (SEQ ID NO:713) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:714) at the CDR3 place.

Antibody 128-A06 has at the CDR1 place and comprises sequence RASQSVSRWLA (SEQ ID NO:715), comprises sequence KASTLES (SEQ ED NO:716) at the CDR2 place, and comprises the LC variable domain of sequence QQYGA (SEQID NO:717) at the CDR3 place.It has at the CDR1 place and comprises sequence HYTMQ (SEQ ID NO:718), comprises sequence SIYPSGGATKYADSVKG (SEQ ID NO:719) at the CDR2 place, and comprises the HC variable domain of sequence SGYYYGLDV (SEQ ED NO:720) at the CDR3 place.

Antibody 095-G09 has at the CDR1 place and comprises sequence RASQSVSSYLA (SEQ ID NO:721), comprises sequence D ASNRAT (SEQ ID NO:722) at the CDR2 place, and comprises the LC variable domain of sequence QQRSN (SEQ ID NO:723) at the CDR3 place.It has at the CDR1 place and comprises sequence HYTMW (SEQ ED NO:724), comprises sequence RTWPSGGTTRYADSVKG (SEQ ID NO:725) at the CDR2 place, and comprises the HC variable domain of sequence GGSYLAFDI (SEQ ID NO:726) at the CDR3 place.

Antibody 137-G10 has at the CDR1 place and comprises sequence QGDSLRNYHPS (SEQ ED NO:727), comprises sequence FGRNNRPS (SEQ ED NO:728) at the CDR2 place, and comprises the LC variable domain of sequence SSRDGSGNFL (SEQ ED NO:729) at the CDR3 place.It has at the CDR1 place and comprises sequence NYTMM (SEQ ID NO:730), comprises sequence GIYPSGGQTAYADSVKG (SEQ ID NO:731) at the CDR2 place, and comprises the HC variable domain of sequence GGSWYAFDI (SEQ ID NO:732) at the CDR3 place.

Antibody 106-E12 has at the CDR1 place and comprises sequence RASQSISSFLN (SEQ ID NO:733), comprises sequence YAASSLQS (SEQ ID NO:734) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPYT (SEQID NO:735) at the CDR3 place.It has at the CDR1 place and comprises sequence WYSMT (SEQ ID NO:736), comprises sequence SISPSGGATKYADSVKG (SEQ ID NO:737) at the CDR2 place, and comprises the HC variable domain of sequence GGAASLPFDY (SEQ ID NO:738) at the CDR3 place.

Antibody 124-H10 has at the CDR1 place and comprises sequence RASQSVGGYLA (SEQ ID NO:739), comprises sequence D ASKRAT (SEQ ID NO:740) at the CDR2 place, and comprises the LC variable domain of sequence QQRSKWPPYT (SEQ ID NO:741) at the CDR3 place.It has at the CDR1 place and comprises sequence HYKMM (SEQ ID NO:742), comprises sequence YIYPSGGWTGYADSVKG (SEQ ID NO:743) at the CDR2 place, and comprises the HC variable domain of sequence VGPWDAFDI (SEQ ID NO:744) at the CDR3 place.

Antibody 122-D01 has at the CDR1 place and comprises sequence RASESVSSSYFA (SEQ ID NO:745), comprises sequence GASRRVT (SEQ ID NO:746) at the CDR2 place, and comprises the LC variable domain of sequence QQYGGSPRS (SEQ ID NO:747) at the CDR3 place.It has at the CDR1 place and comprises sequence RYMME (SEQ IDNO-.748), comprises sequence GISPSGGTTKYADSVKG (SEQ ID NO:749) at the CDR2 place, and comprises the HC variable domain of sequence GGYNNYYYALDV (SEQ ID NO:750) at the CDR3 place.

Antibody 093-C10 has and comprises sequence TGTGSNIDGYNYVS (SEQ ID NO:751) at the CDR1 place, comprise sequence D VGKRPS (SEQ ID NO:752) at the CDR2 place, and comprise the LC variable domain of sequence C SYAGSYSYV (SEQ ID NO:753) at the CDR3 place.It has at the CDR1 place and comprises sequence RYLMW (SEQ ID NO:754), comprises sequence VIGPSGGWTRYADSVKG (SEQ ID NO:755) at the CDR2 place, and comprises the HC variable domain of sequence HPGDY (SEQ ID NO:756) at the CDR3 place.

Antibody 106-C06 has and comprises sequence SGTSSDVGAYYHVS (SEQ ID NO:757) at the CDR1 place, comprise sequence D VSNRPS (SEQ ID NO.758) at the CDR2 place, and comprise the LC variable domain of sequence SLYIGTSTPWV (SEQ ID NO:759) at the CDR3 place.It has at the CDR1 place and comprises sequence HYRMD (SEQ ID NO:760), comprises sequence GIYPSGGHTNYADSVKG (SEQ ID NO:761) at the CDR2 place, and comprises the HC variable domain of sequence D RGWYGADY (SEQ ID NO:762) at the CDR3 place.

Antibody 117-B07 has and comprises sequence TGTTRDVGAYKYVS (SEQ ID NO:763) at the CDR1 place, comprise sequence D VSKRPS (SEQ ID NO:764) at the CDR2 place, and comprise the LC variable domain of sequence C SFAGSYTWI (SEQ ID NO:765) at the CDR3 place.It has at the CDR1 place and comprises sequence HYWMV (SEQ ID NO:766), comprises sequence WIGPSGGGTVYADSVKG (SEQ ID NO.767) at the CDR2 place, and comprises the HC variable domain of sequence GNGGFDS (SEQ ID NO:768) at the CDR3 place.

Antibody 126-H09 has and comprises sequence SGSRSNIGTNPVN (SEQ ID NO:769) at the CDR1 place, comprise sequence VNNQRPS (SEQ ID NO:770) at the CDR2 place, and comprise the LC variable domain of sequence A TWDGSLNGPV (SEQ ID NO:771) at the CDR3 place.It has at the CDR1 place and comprises sequence SYAMT (SEQ ID NO:772), comprises sequence SISSSGGDTAYADSVKG (SEQ ID NO:773) at the CDR2 place, and comprises the HC variable domain of sequence ERHYIWGSYRYSWFDP (SEQ ID NO:774) at the CDR3 place.

Antibody 093-G09 has and comprises sequence SGSSSNIGSNNVN (SEQ ID NO:775) at the CDR1 place, comprise sequence SNDQRPS (SEQ ID NO:776) at the CDR2 place, and comprise the LC variable domain of sequence A AWDDSLNGPV (SEQ ID NO.777) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMV (SEQ ID NO:778), comprises sequence GIVPSGGYTMYADSVKG (SEQ ID NO:779) at the CDR2 place, and comprises the HC variable domain of sequence A SYSSFGLDY (SEQ ID NO:780) at the CDR3 place.

Antibody 114-E02 has at the CDR1 place and comprises sequence RASQSISSWLA (SEQ ID NO:781), comprises sequence KASSLES (SEQ ID NO:782) at the CDR2 place, and comprises the LC variable domain of sequence QQYNSYPVT (SEQID NO:783) at the CDR3 place.It has at the CDR1 place and comprises sequence FYPMS (SEQ ID NO.784), comprises sequence YIWPSGGATRYADSVKG (SEQ ID NO:785) at the CDR2 place, and comprises the HC variable domain of sequence GNGGFDS (SEQ ID NO:786) at the CDR3 place.

Antibody 102-G11 has and comprises sequence TGTSSDIGAYPFVS (SEQ ID NO:787) at the CDR1 place, comprise sequence GVTTRPF (SEQ ID NO:788) at the CDR2 place, and comprise the LC variable domain of sequence SSYAGGRNLPYV (SEQ ID NO:789) at the CDR3 place.It has at the CDR1 place and comprises sequence HYYMA (SEQ ID NO:790), comprises sequence YISPSGGSTKYADSVKG (SEQ ID NO:791) at the CDR2 place, and comprises the HC variable domain of sequence GGGTGTFDI (SEQ ID NO:792) at the CDR3 place.

Antibody 132-C01 has at the CDR1 place and comprises sequence RASQSISRWLA (SEQ ID NO:793), comprises sequence EASTLES (SEQ ID NO:794) at the CDR2 place, and comprises the LC variable domain of sequence QQYNSYPLT (SEQ IDNO:795) at the CDR3 place.It has at the CDR1 place and comprises sequence HYPMH (SEQ ID NO:796), comprises sequence SIWPSGGHTSYADSVKG (SEQ ID NO:797) at the CDR2 place, and comprises the HC variable domain of sequence LSQPI (SEQ ID NO:798) at the CDR3 place.

Antibody 123-E02 has at the CDR1 place and comprises sequence RASQSVSNFLA (SEQ ID NO:799), comprises sequence GASNRAT (SEQ ID NO:800) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPLT (SEQID NO:801) at the CDR3 place.It has at the CDR1 place and comprises sequence MYRMF (SEQ ID NO:802), comprises sequence VIGPSGGQTAYADSVKG (SEQ ID NO:803) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:804) at the CDR3 place.

Antibody 096-D03 has and comprises sequence TGTSGEVANYNYVS (SEQ ID NO:805) at the CDR1 place, comprise sequence D VSRRPS (SEQ ID NO:806) at the CDR2 place, and comprise the LC variable domain of sequence A SYVGNDKLV (SEQ ID NO-.807) at the CDR3 place.It has at the CDR1 place and comprises sequence HYGMN (SEQ ID NO:808), comprises sequence SIWSSGGYTTYADSVKG (SEQ ID NO:809) at the CDR2 place, and comprises the HC variable domain of sequence VGIWYSMDV (SEQ ID NO:810) at the CDR3 place.

Antibody 131-G06 has at the CDR1 place and comprises sequence RASQSINSYLN (SEQ ID NO:811), comprises sequence A ASSLQN (SEQ ID NO:812) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPLT (SEQID NO:813) at the CDR3 place.It has at the CDR1 place and comprises sequence RYTMF (SEQ ID NO:814), comprises sequence GIYPSGGKTIYADSVKG (SEQ ID NO:815) at the CDR2 place, and comprises the HC variable domain of sequence GGVFGVVDY (SEQ ID NO:816) at the CDR3 place.

Antibody 094-D08 has at the CDR1 place and comprises sequence RASQGISNYLA (SEQ ID NO:817), comprises sequence A ASTLQS (SEQ ID NO:818) at the CDR2 place, and comprises the LC variable domain of sequence QKYNSAPLT (SEQID NO:819) at the CDR3 place.It has at the CDR1 place and comprises sequence HYPMD (SEQ ID NO:820), comprises sequence RIYPSGGATKYADSVKG (SEQ ID NO.821) at the CDR2 place, and comprises the HC variable domain of sequence GGGAFDI (SEQ ID NO:822) at the CDR3 place.

Antibody 128-E07 has at the CDR1 place and comprises sequence RASQSVSSYLA (SEQ ID NO:823), comprises sequence D ASNRAT (SEQ ID NO:824) at the CDR2 place, and comprises the LC variable domain of sequence QQRSNWLT (SEQ IDNO:825) at the CDR3 place.It has at the CDR1 place and comprises sequence HYEMA (SEQ ID NO:826), comprises sequence SVIYPSGGATRYADSVKG (SEQ ID NO:827) at the CDR2 place, and comprises the HC variable domain of sequence VAQYYGMDV (SEQ ID NO:828) at the CDR3 place.

Antibody 114-F04 has at the CDR1 place and comprises sequence RASQSVSSNYLA (SEQ ID NO:829), comprises sequence A ASSLQS (SEQ ID NO:830) at the CDR2 place, and comprises the LC variable domain of sequence QQRWT (SEQID NO:831) at the CDR3 place.It has at the CDR1 place and comprises sequence WYTMQ (SEQ ID NO:832), comprises sequence RIYSSGGGTYYADSVKG (SEQ ID NO.833) at the CDR2 place, and comprises the HC variable domain of sequence VGPWGAFDI (SEQ ID NO:834) at the CDR3 place.

Antibody 122-A05 has and comprises sequence TGTSSDVGGYNYVS (SEQ ID NO:835) at the CDR1 place, comprise sequence EVSNRPS (SEQ ID NO:836) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSSSTWV (SEQ ID NO:837) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMV (SEQ ID NO:838), comprises sequence SIYPSGGMTVYADSVKG (SEQ ID NO:839) at the CDR2 place, and comprises the HC variable domain of sequence RGIANSFNI (SEQ ID NO:840) at the CDR3 place.

Antibody 102-H02 has at the CDR1 place and comprises sequence RASQSISRYLN (SEQ ID NO:841), comprises sequence A ASNLQS (SEQ ID NO:842) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPWT (SEQID NO:843) at the CDR3 place.It has at the CDR1 place and comprises sequence KYYMV (SEQ ID NO:844), comprises sequence VIGPSGGWTTYADSVKG (SEQ ID NO:845) at the CDR2 place, and comprises the HC variable domain of sequence VDYSNYFDY (SEQ ID NO:846) at the CDR3 place.

Antibody 092-G06 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:847), comprises sequence A ASSLQS (SEQ ID NO:848) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPWT (SEQID NO:849) at the CDR3 place.It has and comprises sequence SYMI (SEQ ID NO:850) at the CDR1 place, comprise sequence YIRPSGGYTKYADSVKG (SEQ ID NO:851) at the CDR2 place, and comprise the HC variable domain of sequence VINAGPGRGYYWRGYSYSDAFDI (SEQ ID NO:852) at the CDR3 place.

Antibody 113-E03 has at the CDR1 place and comprises sequence RASQGIGTHLN (SEQ ID NO:853), comprises sequence A ASGLQS (SEQ ID NO:854) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSVPRT (SEQID NO:855) at the CDR3 place.It has at the CDR1 place and comprises sequence NYPMV (SEQ ID NO:856), comprises sequence VIGPSGGKTKYADSVKG (SEQ ID NO:857) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:858) at the CDR3 place.

Antibody 118-E07 has and comprises sequence TGTSSDVGAYNYVS (SEQ ID NO:859) at the CDR1 place, comprise sequence EVSNRPS (SEQ ID NO:860) at the CDR2 place, and comprise the LC variable domain of sequence NSYTTSATLV (SEQ ID NO:861) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMM (SEQ ID NO:862), comprises sequence SIRSSGGITRYADSVKG (SEQ ID NO.863) at the CDR2 place, and comprises the HC variable domain of sequence VGYYYGMDV (SEQ ID NO.864) at the CDR3 place.

Antibody 093-E11 has at the CDR1 place and comprises sequence RASQSITGYLN (SEQ ID NO:865), comprises sequence A ASSLQS (SEQ ID NO:866) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPYT (SEQID NO:867) at the CDR3 place.It has at the CDR1 place and comprises sequence NYPMY (SEQ ID NO:868), comprises sequence RIGPSGGHTDYADSVKG (SEQ ID NO:869) at the CDR2 place, and comprises the HC variable domain of sequence A GVWSGLDY (SEQ ID NO.870) at the CDR3 place.

Antibody 124-C12 has at the CDR1 place and comprises sequence RASQGISNYLA (SEQ ID NO:871), comprises sequence A ASTLQS (SEQ ID NO:872) at the CDR2 place, and comprises the LC variable domain of sequence QKYNSAPWT (SEQID NO:873) at the CDR3 place.It has at the CDR1 place and comprises sequence HYLMG (SEQ ID NO:874), comprises sequence VIWPSGGITKYADSVKG (SEQ ID NO:875) at the CDR2 place, and comprises the HC variable domain of sequence GNGGFDS (SEQ ID NO.876) at the CDR3 place.

Antibody 115-F08 has and comprises sequence TGTNTDVGGYNFVS (SEQ ID NO:877) at the CDR1 place, comprise sequence D VTNRPS (SEQ ID NO:878) at the CDR2 place, and comprise the LC variable domain of sequence SSYTSSSTWV (SEQ ID NO:879) at the CDR3 place.It has at the CDR1 place and comprises sequence HYSML (SEQ ID NO:880), comprises sequence SIRPSGGFTKYADSVKG (SEQ ID NO:881) at the CDR2 place, and comprises the HC variable domain of sequence RGVWDAFDI (SEQ ID NO:882) at the CDR3 place.

Antibody 121-H07 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:883), comprises sequence A ASSLQS (SEQ ID NO:884) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPRT (SEQ IDNO:885) at the CDR3 place.It has and comprises sequence RYAMG (SEQ ID NO:886) at the CDR1 place, comprise sequence YIGPSGGNTTYADSVKG (SEQ ID NO.887) at the CDR2 place, and comprise the HC variable domain of sequence D VGGSGSYYMLSYYYYGMDV (SEQ ID NO:888) at the CDR3 place.

Antibody 136-B06 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:889), comprises sequence A ASTRAT (SEQ ID NO:890) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPWT (SEQ ID NO:891) at the CDR3 place.It has at the CDR1 place and comprises sequence RYPMG (SEQ ID NO:892), comprises sequence SIYPSGGSTYYADSVKG (SEQ ID NO:893) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:894) at the CDR3 place.

Antibody 097-F08 has at the CDR1 place and comprises sequence SGDDLGGRHVS (SEQ ID NO:895), comprises sequence QDDKRPS (SEQ ID NO:896) at the CDR2 place, and comprises the LC variable domain of sequence LAWHNYKYV (SEQ ID NO:897) at the CDR3 place.It has at the CDR1 place and comprises sequence NYLMG (SEQ ID NO:898), comprises sequence SIGPSGGYTKYADSVKG (SEQ ID NO:899) at the CDR2 place, and comprises the HC variable domain of sequence GTGTTFF (SEQ ID NO:900) at the CDR3 place.

Antibody 107-B05 has at the CDR1 place and comprises sequence RASQSISSYLN (SEQ ID NO:901), comprises sequence A ASSLQS (SEQ ID NO:902) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTLVT (SEQID NO:903) at the CDR3 place.It has at the CDR1 place and comprises sequence HYMMQ (SEQ ID NO:904), comprises sequence GIGPSGGWTKYADSVKG (SEQ ID NO:905) at the CDR2 place, and comprises the HC variable domain of sequence LNYGDYV (SEQ ID NO:906) at the CDR3 place.

Antibody 124-A01 has at the CDR1 place and comprises sequence RASQGISSSLA (SEQ ID NO:907), comprises sequence A ASTLQS (SEQ ID NO:908) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPWT (SEQID NO:909) at the CDR3 place.It has and comprises sequence HYLMH (SEQ ID NO.910) at the CDR1 place, comprise sequence SIYPSGGKTQYADSVKG (SEQ ID NO:911) at the CDR2 place, and comprise the HC variable domain of sequence VFGYYDFWSGYPGAFDY (SEQ ID NO:912) at the CDR3 place.

Antibody 102-C12 has at the CDR1 place and comprises sequence GGNNIGSKNVH (SEQ ID NO:913), comprises sequence RDSNRPS (SEQ ID NO:914) at the CDR2 place, and comprises the LC variable domain of sequence QVWDSSTV (SEQ ID NO:915) at the CDR3 place.It has at the CDR1 place and comprises sequence HYVMV (SEQ ID NO:916), comprises sequence GIRSSGGVTAYADSVKG (SEQ ID NO:917) at the CDR2 place, and comprises the HC variable domain of sequence VGVWYGMDV (SEQ ID NO:918) at the CDR3 place.

Antibody 103-E09 has at the CDR1 place and comprises sequence RASQTVSSTYLA (SEQ ID NO.919), comprises sequence GASSRAT (SEQ ID NO:920) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSPPRYT (SEQ ID NO:921) at the CDR3 place.It has at the CDR1 place and comprises sequence RYRMV (SEQ ID NO:922), comprises sequence SIYPSGGYTKYADSVKG (SEQ ID NO:923) at the CDR2 place, and comprises the HC variable domain of sequence VITYNNFDS (SEQ ID NO:924) at the CDR3 place.

Antibody 13 1-F12 have at the CDR1 place and comprise sequence RASLSVSGYLN (SEQ ID NO:925), comprise sequence A TSSLQS (SEQ ID NO:926) at the CDR2 place, and comprise the LC variable domain of sequence QQSYSSPYT (SEQ IDNO:927) at the CDR3 place.It has at the CDR1 place and comprises sequence HYAMF (SEQ ID NO:928), comprises sequence SIWPSGGKTMYADSVKG (SEQ ID NO.929) at the CDR2 place, and comprises the HC variable domain of sequence GGSYLAFDI (SEQ ID NO:930) at the CDR3 place.

Antibody 093-C08 has at the CDR1 place and comprises sequence LASQSISSYLN (SEQ ID NO:931), comprises sequence D ASSLES (SEQ ID NO:932) at the CDR2 place, and comprises the LC variable domain of sequence QQFNGYPPIT (SEQID NO:933) at the CDR3 place.It has at the CDR1 place and comprises sequence RYPMI (SEQ ID NO:934), comprises sequence VIYPSGGMTKYADSVKG (SEQ ID NO:935) at the CDR2 place, and comprises the HC variable domain of sequence VGSYGSLGY (SEQ ID NO:936) at the CDR3 place.

Antibody 115-F04 has at the CDR1 place and comprises sequence GASQSVSTTYIA (SEQ ID NO:937), comprises sequence GASNRAT (SEQ ID NO:938) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSPYT (SEQ ID NO:939) at the CDR3 place.It has at the CDR1 place and comprises sequence RYWMW (SEQ ID NO:940), comprises sequence SIGPSGGATRYADSVKG (SEQ ID NO:941) at the CDR2 place, and comprises the HC variable domain of sequence GSAGN (SEQ ID NO:942) at the CDR3 place.

Antibody 113-G11 has at the CDR1 place and comprises sequence RASQSIDTYLN (SEQ ID NO:943), comprises sequence A ASKLED (SEQ ID NO:944) at the CDR2 place, and comprises the LC variable domain of sequence QPYNTYPIT (SEQID NO:945) at the CDR3 place.It has at the CDR1 place and comprises sequence HYGMY (SEQ ID NO:946), comprises sequence SIYPSGGWTNYADSVKG (SEQ ID NO:947) at the CDR2 place, and comprises the HC variable domain of sequence RGSWGAFDI (SEQ ID NO:948) at the CDR3 place.

Antibody 102-C02 has at the CDR1 place and comprises sequence RASQSVSNSYLA (SEQ ID NO:949), comprises sequence GASSRAT (SEQ ID NO:950) at the CDR2 place, and comprises the LC variable domain of sequence QQFGSSTGYT (SEQ ID NO:951) at the CDR3 place.It has at the CDR1 place and comprises sequence YYMMG (SEQ ID NO:952), comprises sequence SIYTSGGYTKYADSVKG (SEQ ID NO:953) at the CDR2 place, and comprises the HC variable domain of sequence A SIYYGMDV (SEQ ID NO.954) at the CDR3 place.

Antibody 112-D04 has at the CDR1 place and comprises sequence RASQGISSWLA (SEQ ID NO:955), comprises sequence A ASSLQS (SEQ ID NO:956) at the CDR2 place, and comprises the LC variable domain of sequence QQANSFPYT (SEQID NC-.957) at the CDR3 place.It has at the CDR1 place and comprises sequence YYLMG (SEQ ID NO:958), comprises sequence SIGPSGGTTKYADSVKG (SEQ ID NO:959) at the CDR2 place, and comprises the HC variable domain of sequence RGLGAAFDY (SEQ ID NO:960) at the CDR3 place.

Antibody 108-E10 has at the CDR1 place and comprises sequence RASQSVSSSYLA (SEQ ID NO:961), comprises sequence GASSRAT (SEQ ID NO:962) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSSFT (SEQ ID NO:963) at the CDR3 place.It has at the CDR1 place and comprises sequence HYMM (SEQ ID NO:964), comprises sequence SIYPSGGSTIYADSVKG (SEQ ID NO.965) at the CDR2 place, and comprises the HC variable domain of sequence A TEGYYYGMDV (SEQ ID NO:966) at the CDR3 place.

Antibody 104-D12 has at the CDR1 place and comprises sequence RASQGIRSWLA (SEQ ID NO:967), comprises sequence A ASSLQS (SEQ ID NO:968) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPWT (SEQ ID NO:969) at the CDR3 place.It has at the CDR1 place and comprises sequence WYRMA (SEQ ID NO:970), comprises sequence YIGPSGGSTSYADSVKG (SEQ ID NO.971) at the CDR2 place, and comprises the HC variable domain of sequence VGTFYGMDV (SEQ ID NO:972) at the CDR3 place.

Antibody 116-E08 has at the CDR1 place and comprises sequence RASQTISSYLN (SEQ ID NO:973), comprises sequence A ASSLQS (SEQ ID NO:974) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSTPMYT (SEQID NO:975) at the CDR3 place.It has at the CDR1 place and comprises sequence WYLMG (SEQ ID NO:976), comprises sequence VIGPSGGLTKYADSVKG (SEQ ID NO.977) at the CDR2 place, and comprises the HC variable domain of sequence FRGYYGMDV (SEQ ID NO:978) at the CDR3 place.

Antibody 102-D07 has at the CDR1 place and comprises sequence RASQSVRKNVA (SEQ ID NO:979), comprises sequence GASTRAT (SEQ ID NO:980) at the CDR2 place, and comprises the LC variable domain of sequence QQYSSWPA (SEQ ID NO.981) at the CDR3 place.It has and comprises sequence NYLMY (SEQ ID NO:982) at the CDR1 place, comprise sequence SIRPSGGNTLYADSVKG (SEQ ID NO:983) at the CDR2 place, and comprise the HC variable domain of sequence GRGILTGYYWGYYYYMDV (SEQ ID NO:984) at the CDR3 place.

Antibody 103-H07 has and comprises sequence RASQSISSSYLA (SEQ ID NO:985) at the CDR1 place, comprise sequence HASSRAT (SEQ ID NO:986) at the CDR2 place, and comprise the LC variable domain of sequence QQRNNWPPSFT (SEQ ID NO:987) at the CDR3 place.It has and comprises sequence LYLME (SEQ ID NO:988) at the CDR1 place, comprise sequence SIGSSGGATWYADSVKG (SEQ ID NO:989) at the CDR2 place, and comprise the HC variable domain of sequence D TSRVAGTRLRNYYYYYGMDV (SEQ ID NO:990) at the CDR3 place.

Antibody 116-A08 has at the CDR1 place and comprises sequence RASQSIGTLLN (SEQ ID NO:991), comprises sequence GASNLRG (SEQ ID NO:992) at the CDR2 place, and comprises the LC variable domain of sequence QHDT (SEQ ID NO:993) at the CDR3 place.It has and comprises sequence WYRMH (SEQ ID NO:994) at the CDR1 place, comprise sequence SIWPSGGKTHYADSVKG (SEQ ID NO:995) at the CDR2 place, and comprise the HC variable domain of sequence A LQYGSGSYFYAPKSYYYYGMDV (SEQ ID NO:996) at the CDR3 place.

Antibody 106-D06 has at the CDR1 place and comprises sequence RASQTISRYLN (SEQ ID NO:997), comprises sequence A ASSLQS (SEQ ID NO:998) at the CDR2 place, and comprises the LC variable domain of sequence QQSYSAPRT (SEQID NO:999) at the CDR3 place.It has and comprises sequence KYKMG (SEQ ID NO.1000) at the CDR1 place, comprise sequence YIRPSGGMTFYADSVKG (SEQ ID NO:1001) at the CDR2 place, and comprise the HC variable domain of sequence EHYQASYSSSAWFRMVPAGAFDI (SEQ ID NO:1002) at the CDR3 place.

Antibody 113-D05 has and comprises sequence RASQGIRRALA (SEQ ID NO:1003) at the CDR1 place, comprise sequence D ASSLES (SEQ ID NO.1004) at the CDR2 place, and comprise the LC variable domain of sequence QQSYSTPPWT (SEQ ID NO:1005) at the CDR3 place.It has and comprises sequence YYKMH (SEQ ID NO:1006) at the CDR1 place, comprise sequence VIYPSGGKTTYADSVKG (SEQ ID NO:1007) at the CDR2 place, and comprise the HC variable domain of sequence EHYQASYSSSAWFRMVPAGAFDI (SEQ ID NO:1008) at the CDR3 place.

Antibody 110-D06 has at the CDR1 place and comprises sequence RASQSWSNFLA (SEQ ID NO.1009), comprises sequence GASSRAT (SEQ ID NO:1010) at the CDR2 place, and comprises the LC variable domain of sequence QQYGSSPYS (SEQ ID NO:1011) at the CDR3 place.It has and comprises sequence NYVMY (SEQ ID NO:1012) at the CDR1 place, comprise sequence GIGPSGGFTTYADSVKG (SEQ ID NO:1013) at the CDR2 place, and comprise the HC variable domain of sequence D RYFGSGYYMAAYYYYGMDV (SEQ ID NO:1014) at the CDR3 place.

Antibody 124-C04 has and comprises sequence RASQRVSSTFLA (SEQ ID NO:1015) at the CDR1 place, comprise sequence GTSSRAT (SEQ ID NO:1016) at the CDR2 place, and comprise the LC variable domain of sequence HQYGSSPRT (SEQ ID NO.1017) at the CDR3 place.It has at the CDR1 place and comprises sequence WYAMM (SEQ ID NO:1018), comprises sequence SIWPSGGHTKYADSVKG (SEQ ID NO.1019) at the CDR2 place, and comprises the HC variable domain of sequence D QGYF (SEQ ID NO:1020) at the CDR3 place.

Antibody 097-B12 has at the CDR1 place and comprises sequence RASQPISTSLA (SEQ ID NO:1380), comprises sequence KASILET (SEQ ID NO:1381) at the CDR2 place, and comprises the LC variable domain of sequence QQYASPSYT (SEQID NO:1382) at the CDR3 place.It has and comprises sequence KYVMW (SEQ ID NO:1383) at the CDR1 place, comprise sequence SIVSSGGRTSYADSVKG (SEQ ID NO:1384) at the CDR2 place, and comprise the HC variable domain of sequence A YDYVWGSYRYKGRPVGAFDI (SEQ ID NO:1385) at the CDR3 place.As seen described antibody suppresses mK1 and hK1.

Some exemplary antibody described herein have the sequence internal relation.Two antibody of shared LC, HC CDR3 or HC CDR1 and HC CDR2 may be gone up roughly the same site with hK1 and combine.Total HC CDR3 especially indicates shared binding site.Therefore, the inhibition activity of two antibody of shared sequence designated area may be similar.Particularly, if the inhibitor that antibody is hK1, then any antibody of sharing the sequence of described first antibody might also be inhibitor.For example:

M0098-G05 has consistent HC variable domain with M0097-G07, but its LC variable domain difference.

M0103-A01 has consistent HC variable domain with M0096-D09, but its LC variable domain difference.

M0114-D02 has consistent HC variable domain with M0097-G01, but its LC variable domain difference.

M0117-F08 has consistent HC variable domain with M0102-G12, but its LC variable domain difference.Known M0102-G12 suppresses hK1.

M0131-C08 has consistent HC variable domain with M0093-G09, but its LC variable domain difference.

M0131-C09 has consistent HC variable domain with M0096-D09, but its LC variable domain difference.

M0131-F07 has consistent HC variable domain with M0102-G12, but its LC variable domain difference.Known M0131-F07 and M0102-G12 suppress hK1.

M0131-G03 has consistent LC variable domain with M0098-H04.

M0133-E08 has consistent LC variable domain with M0102-G12.Known M0102-G12 suppresses hK1.

M0136-A07 has consistent HC variable domain with M0096-D09, but its LC variable domain difference.

M0138-B04 has consistent HC variable domain with M0102-G12, but its LC variable domain difference.Known M0102-G12 suppresses hK1.

M0138-G11 has consistent LC variable domain with M0133-E02.

Following HC variable domain has identical CDR3 sequence (VGVWYGMDV): M0092-F08, M0093-C08, M0093-C10,14 (M0095-A11, M0097-E07, M0097-F08, M0102-H02, M0103-F07, M0104-H12, M0107-F11, M0109-E08, M0110-D11, M0111-C10, M0112-C12, M0112-D04, M0114-G09, M0117-A12, M0124-H10, M0128-A06, M0130-B02, M0131-B05, M0131-F09, M0135-B05, M0135-F03, M0136-E12, M0138-E02, M0139-F09).

The HC variable domain of M0096-D03, M0102-C12, M0126-F11 and M0128-E07 is shared cdr3=GNGGFDS.The HC variable domain of M0097-D04 and M0134-D07 is shared cdr3=RGPVYYYDSSGSHSAFDI.

The HC variable domain of M0097-G07 and M0139-C02 is shared cdr3=EHYQASYSSSAWFRMVPAGAFDI.The HC variable domain of M0101-E01 and 1M0136-D07 is shared cdr3=NINLRYDILLTTLTGYFDI.The HC variable domain of M0102-G12 and M0110-G01 is shared cdr3=DITPGGGSGFRLPKNYYYYGMDV.M0102-G12 suppresses hK1.The HC variable domain of M0102-G12 and M0133-E08 is shared cdr3=DITPGGGSGFRLPKNYYYYGMDV.M0102-G12 suppresses hK1.Its LC variable domain is identical.The HC variable domain of M0102-G12 and M0139-F04 is shared cdr3=DITPGGGSGFRLPKNYYYYGMDV; M0139-F04 and M0102-G12 suppress hK1.Its LC variable domain difference.The HC variable domain of M0104-D12 and M0117-B07 is shared cdr3=GGSYLAFDI, LC difference.M0105-B06 and M0116-C09 share cdr3=GYSSTWGAFDI.M0105-B06 and M0138-E12 share cdr3=GYSSTWGAFDI.M0133-E02 and M0138-G11 share cdr3=DTRVGPWLVRAPLDY.

M0102-G12 and M0133-E08 share HC CDR1=KyKmV (SEQ ID NO:10) and HC CDR2=SiYPsggItAyadsvkg (SEQ ID NO:11).M0102-G12 suppresses hK1.M0107-D12 and M0110-G01 share HC CDR1=MyKmG and HC CDR2=SiVPsggKtQyadsvkg.M0133-E02 and M0138-G11 share HC CDR1=WyTmT and HC CDR2=SiYPsggHtSyadsvkg.

Sequence with exemplary antibody of at least one CDR the same with discriminating hK1 inhibitor might also be inhibitor.

M0103-A03 does not have C at the 96th place.

M0107-D12 contains terminator.

M0107-D12 does not have WG main structure (motif) at the section start of FR4.

The following is the sequence (for example, extending to the end of CH1) of the C-terminal of the sequence of exemplary HC variable domain and described variable domain.

＞HC-092-B12-CH1(SEQ ID NO：1370)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS QYLMAWVRQA PGKGLEWVSS

IYPSGGNTNY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED TAVYYCARDR

SIVPYSSSWY MPRDYYYGMD VWGQGTTVTV SSASTKGPSV FPLAPSSKST

SGGTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV

VTVPSSSLGT QTYICNVNHK PSNTKVDKKV EPKSC

In eukaryotic cell, yeast cell or bacterial cell, express and have suitably folding VL-light chain constant domain and the combination results Fab of VH-CH1.Fab is monomeric and has the molecular mass of about 50 kDa usually.

Following sequence illustration extends to the HC variable domain of CH1, hinge area, CH2 and the CH3 of IgG 1:

＞HC-093-C10-CH1-H-CH2-CH3 (SEQID NO：1371)

EVQLLESGGG LVQPGGSLRL SCAASGFTFS RYTMEWVRQA PGKGLEWVSV

IRPSGGTTMY ADSVKGRFTI SRDNSKNTLY LQMNSLRAED MAVYYCARVG

VWYGMDVWGQ GTTVTVSSAS TKGPSVFPLA PSSKSTSGGT AALGCLVKDY

FPEPVTVSWN SGALTSGVHT FPAVLQSSGL YSLSSVVTVP SSSLGTQTYI

CNVNHKPSNT KVDKKVEPKS CDKTHTCPPC PAPELLGGPS VFLFPPKPKD

TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT KPREEQYNST

YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY

TLPPSRDELT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD

SDGSFFLYSK LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGK

Comprise VH::CH1:: hinge area:: the HC of CH2::CH3 is together with comprising the light chain variable territory:: the expression of the LC of light chain constant domain produces IgG.In giving example, should obtain IgG1.Other IgG type can be replaced coding CH1: by the DNA with coding (for example) IgG4: hinge area:: the DNA of CH2::CH3 or use modified Fc sequence to obtain.

The following is the sequence (for example, extending to the terminal of C κ) of the C-terminal of the sequence of exemplary LC variable domain and described variable domain:

＞LC-092-B12-Cκ(SEQID NO：1372)

AQDIQMTQSP SSLSASVGDR VTITCRASQG ISNYLAWYQQ KPGKVPKLLI

YAASTLQSGV PSRFSGSGSG TDFTLTISSL QPEDFATYYC QQSYSTPYTF

GQGTKLEVRR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN FYPREAKVQW

KVDNALQSGN SQESVTEQDS KDSTYSLSST LTLSKADYEK HKVYACEVTH

QGLSSPVTKS FNRGEC

Following LC sequence has λ 1 constant region:

＞LC-103-F07-Cλ(SEQID NO：1373)

AQSALTQPAS VSGSPGQSIT ISCTGTSNDI GRTNYVSWYR QDPGRAPKLI

LFEVSNRPSG ISNRFSASKS GSTASLTISG LQADDESDYY CSSCTTAPVC

VFGNGTRVTV L GQPKANPTV TLFPPSSEEL QANKATLVCL ISDFYPGAVT

VAWKADGSPV KAGVETTKPS KQSNNKYAAS SYLSLTPEQW KSHRSYSCQV

THEGSTVEKT VAPTECS

Following LC sequence has λ 2 constant regions:

＞LC-131-D12- Cλ2 (SEQ ID NO：1374)

AQSALTQPPS VSVAPGQTAS ISCSGNILDN SYASWFQQKP GQSPVMVIHR

DNKRPSGIPE RFSGSTSGNT ATLTISGTQA VDEADYYCQA WDRTTGVFGT

GTRLTVLRQP KAAPSVTLFP PSSEELQANK ATLVCLISDF YPGAVTVAWK

ADSSPVKAGV ETTTPSKQSN NKYAASSYLS LTPEQWKSHR SYSCQVMHEG

STVEKTVAPA ECS

Following LC sequence has λ 7 constant regions:

＞LC-132-H11- Cλ7 (SEQ ID NO：1375)

AQYELTQPPS ASGTPGQRVT ISCSGSSSNI GSNYVYWYQQ LPGTAPKLLI

YRNNQRPSGV PDRFSGSKSG TSASLAISGL RSEDEADYYC AAWDDSLSGH

AVFGGGTQLT VLGQPKAAPS VTLFPPSSEE LQANKATLVC LVSDFYPGAV

TVAWKADGSP VKVGVETTKP SKQSNNKYAA SSYLSLTPEQ WKSHRSYSCR

VTHEGSTVEK TVAPTECS

Lambda light chain presents a few types: for example λ 1, λ 2 and λ 7, decide according to the sequence of C λ.Antibody with λ LC can be classified as follows.

λ7

LC-132-H11

λ2

LC-131-D12

LC-131-G06

LC-132-D02

LC-134-B03

LC-136-A07

LC-104-H12

LC-109-A11

LC-122-G06

LC-125-C04

LC-102-G12

LC-092-G06

LC-110-D11

LC-108-C10

LC-128-A06

LC-135-H01

LC-123-E02

λ1

LC-116-C09

LC-108-E11

LC-099-C12

LC-131-A03

LC-108-D11

LC-114-F04

LC-132-C01

LC-122-H04

LC-094-E08

LC-093-F09

LC-093-G06

LC-103-E09

LC-103-F07

LC-098-H04

LC-131-G03

LC-097-F04

LC-135-C12

LC-131-B05

LC-112-D04

LC-096-D03

LC-126-H09

LC-114-G09

LC-094-D08

LC-102-A04

In no case wish C λ by (for example) C λ 1 be changed to C λ 2 change expressed antibody in conjunction with character.Therefore, we change the λ constant region and do not change to change expression or pharmacokinetics in conjunction with character (such as suppressing hK1).

The proteolytic enzyme specificity of selected inhibitor

The selected antibody of test suppresses the ability of other proteolytic enzyme.Be showed in the table 2 is the results of 3 antibody of test at 13 serine proteases except that hK1.Obviously, under the full test concentration (1 μ M) of antibody, we can not observe the inhibition to any other proteolytic enzyme.

Program.Calibrating is to carry out in little quantitative plate at the bottom of 96 hole circles of black with the cumulative volume of 100 μ L.Enzyme, substrate, damping fluid and experiment condition are listed in table 3 and 4.In the dark at room temperature, enzyme and Fab inhibitor were cultivated 1 hour, added the substrate of indicatrix thereafter.Subsequently, use Spectramax Gemini XS fluorescent plate reader to use the speed of wavelength monitoring enzymatic substrate hydrolysis listed in the table 3.

The specificity of table 2.hK1 antibody inhibition

Proteolytic enzyme	DX-2300	Ki(nM) M0093-F09	M0137-E0
				The human chymotrypsin human prothrombin of the human plasmin human urokinase of the hK1 hK2 hK3 hK4 hK5 hK8 human plasma callicrein human insulin human PKC of human elastin enzyme	0.039-0.06 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000	1.9 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000	0.9 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000 ＞1000

Table 3. is used for the general introduction of the test condition of proteolytic enzyme specificity measurement

Enzyme	Substrate	[S] (μM)	[E] (nM)	Buffering λ Ex λ Em liquid
					The human neutrophil elastoser of the human pancreas trypsase of restructuring (Pichia pastoris) callicrein human urine callicrein (hK1) human tissue callicrein 2 (hK2) plasma kallikrein human tissue callicrein 3 (hK3, PSA) human tissue callicrein 4 (hK4) human tissue callicrein 5 (hK5) human tissue callicrein 8 (hK8) plasmin chymotrypsin protein kinase C urokinase fibrin ferment	Pro-Phe-Arg-AMC Pro-Phe-Arg-AMC Pro-Phe-Arg-AMC Pro-Phe-Arg-AMC Mu-His-Ser-Ser-Lys-Leu-Gln-AFC Boc-Val-Pro-Arg-AMC Boc-Val-Pro-Arg-AMC Boc-Val-Pro-Arg-AMC N-Suc-Ala-Phe-Lys-AMC Pro-Phe-Arg-AMC N-MS-Ala-Ala-Pro-Val-AMC Trp-AMC Boc-Leu-Ser-Thr-Arg-AMC Z-Gly-Gly-Arg-AMC benzoyl-Phe-Va1-Arg-AMC	100 100 100 100 100 100 100 100 100 100 100 100 100 100 500	5 5 5 5 20 10 20 40 5 5 5 5 5 5 5	KAL 360 460 KAL 360 460 KAL 360 460 KAL 360 460 PSA 400 505 KAL 380 460 KAL 380 460 KAL 380 460 PLA 360 460 KAL 360 460 KAL 360 460 PSA 360 460 PKC 360 460 PSA 360 460 KAL 350 470

The damping fluid component of table 4. table 3

KAL	PSA	PLA	PKC
				20mM Tris pH7.5 150mM NaCl 1mM EDTA 0.1％PEG 0.1％Triton X-100	50mM HEPES pH7.5 100mM NaCl， 0.2％BSA	50mM HEPES pH7.5 150mM NaCl 2mM CaCl 2 0.1％Triton X-100	20mM Tris-Cl pH8.0 100mM NaCl 1mM CaCl 2 0.1％Triton X-100

Selected antibody inhibition is to the active effect of hK1 kininogenaseCheck that selected antibody inhibition is to measure it to the active effect of hK1 kininogenase.SDS-PAGE shows that all 9 antibody of being checked suppress the kininogenase activity of hK1 to high molecular weight kininogen (HMWK).Complete HMWK is the migration of 100kDa protein, and it produces the fragment (swimming lane 1 and 2) of moving with about 60kDa through hK1 digestion.9 hK1 inhibitor of all tests (swimming lane 4 to 12) stop HMWK digestion.By under 37 ℃, 10nM hK1 being cultivated 3 hours with 1 μ M Ab, add 0.1mg/mL HMWK thereafter and carry out experiment.Under 37 ℃, make HMWK and hK1 reaction 3 hours.To irreducibility SDS-PAGE gel (10% acrylamide) loading 1 μ g HMWK and according to the standard program electrophoresis.Swimming lane 1 contains indigested HMWK, swimming lane 2 contains the HMWK of digestion, swimming lane 3 contains the molecular weight marker from Invitrogen, swimming lane 4 contains HMWK+hK1+DX-2300, swimming lane 5 contains HMWK+hK1+M0106-G12, swimming lane 6 contains HMWK+hK1+M0111-C12, swimming lane 7 contains HMWK+hK1+M0098-G05, swimming lane 8 contains HMWK+hK1+M0137-E01, swimming lane 9 contains HMWK+hK1+M0112-D03, swimming lane 10 contains HMWK+hK1+M0102-H11, and swimming lane 11 contains HMWK+hK1+M0114-G06, and swimming lane 12 contains HMWK+hK1+M0098-E09.

Obviously all 9 Fab suppress the degree of digestion LMWK.Yet as if only DX-2300 (swimming lane 4) and M0137-E01 (swimming lane 8) suppress LMWK digestion fully.With regard to HMWK, reaction is carried out as mentioned above and except that replace HMWK with LMWK, swimming lane contains identical above-mentioned inclusion.

The epitope grouping of selected hK1 binding antibody

Use surface plasma resonance (SPR) method (Fig. 1, table 5) according to whether the epitope upward identical with DX-2300 identification hK1 will be selected the antibody grouping.This type of information can make discovery be used for the pairing antibody of the hK1 of quantitative measurment biological sample.Described information is to also having value with antibody with the reserve inhibitor of main inhibitor (DX-2300) comparison and selection and DX-2300 shared same antigen decision base or different epitopes.Fig. 1 shows and to share the Fab (M0112-D07) (figure A) of same antigen decision base with DX-2300 and in conjunction with two representative influence charts of the Fab (scheming B) of different epitopes.Provide what person Fab and DX-2300 to share the basic indication of same antigen decision divided by the resulting ratio of described signal (R2) of Fab only the spr signal (R1) of the Fab of injection after the DX-2300.Because the R2/R1 ratio that the second time of DX-2300 injection produces is 0.1, so have apparent the Fab indication greater than 0.1 R2/R1 ratio in conjunction with the epitope different with DX-2300.Among 12 Fab that checked, only M0111-C12, M0137-E01 do not share identical epitope (table 5) with DX-2300 with M0139-A09.The epitope of some Fab (M0109-F02 and M0098-E09) can be overlapped with the epitope of DX-2300, because observe M signal response (table 5).

Table 5. carries out the epitope grouping by surface plasma resonance

Fab	Signal (R1) according to DX-2300	Fab is the signal of Fab (R2) only	Ratio R 1/R2	Whether share the DX-2300 epitope
					M0093-F09 M0111-C12 M0137-E01 M0109-F02 M0114-G06 M0098-E09 M0139-A09 M0134-D07 M0117-F04 M0098-E09 M0136-A07 M0112-D07 M0131-F07 (DX-2300)	5.9 38.8 20.8 9.5 4.8 8.5 25.7 2.3 6.7 8.5 5.0 3.0 3.8	32.7 65.1 49.1 30.2 27.5 34.8 40.0 14.7 29.7 29.9 27.6 16.1 27.6	0.18 0.60 0.42 0.31 0.17 0.24 0.64 0.16 0.23 0.28 0.18 0.19 0.1	Whether whether no part is that part is

The mechanism that hK1 suppresses and the K of selected inhibitor _iMeasure

Because the optimum antibody of being found is the inhibitor of combining closely of hK1, do not measure enzyme inhibition mechanism so do not use classical Michaelis-Menten method.The combine closely method of inhibition mechanism of inhibitor of mensuration is described (R.A.Copeland, " Enzymes ", the 2nd edition, Wiley, Toronto, 2000).The inhibitor of combining closely refers to have the inhibition constant (K near can be used for measuring active minimum enzyme concn _iValue) inhibitor, with regard to hK1, suppressing constant is about 1nM.The inhibition method of combining closely need be measured the IC of the inhibitor in the different concentration of substrate scopes ₅₀Under the described conditions, being that the inhibitor of emulative mode desmoenzyme should be showed the IC that responds concentration of substrate with respect to substrate ₅₀Linearity increase.On the contrary, the IC that does not change with concentration of substrate ₅₀Value indication inhibitor is being noncompetitive mode desmoenzyme with respect to substrate.Noncompetitive inhibitor is not blocked the ability of substrate desmoenzyme with combining of enzyme, but may be by making amino acid distortion prevention substrate hydrolysis step subsequently important in the catalysis.

DX-2300 shows IC ₅₀Value is to the linear dependence (Fig. 2) of concentration of substrate.Therefore, DX-2300 is for suppressing the inhibitor of combining closely of hK1 in competitive mode, and it indicates it to stop the substrate combination with combining of hK1.The understanding that suppresses mechanism is also allowed to measure inhibition constant (K _i).DX-2300 has the K of 60pM as Fab _iValue and have the K of about 39pM as IgG _iValue.

Also measure the inhibition mechanism and the K of selected reserve antibody (M0093-F09 and M0137-E01) _iValue.As shown in Figure 3A, M0093-F09 suppresses hK1 in the noncompetitive mode, wherein K _iBe 1.9nM.Fig. 3 B proof reserve antibody inhibition M0137-E01 suppresses hK1 in competitive mode, wherein K _iBe 0.9nM.

DX-2300 carries out the curve inhibition analysis

Used march line method (R.A.Copeland, " Enzymes ", the 2nd edition, Wiley, Toronto, 2000) research DX-2300 to suppress the time-dependent manner of hK1.Described method is used to characterize the interactional step between kinetics step or description inhibitor and the enzyme.Be showed among Fig. 4 A carry out that curve proof DX-2300 suppresses the time-dependent manner of hK1 and will be for the data fitting that inhibitor obtained of each concentration to provide a description the interactional dynamic (dynamical) index rate constant between enzyme and the inhibitor.When inhibitor concentration is mapped, described rate constant (k _ObsValue) show hyperbolic line dependency (Fig. 4 B), the initial recombination thing experience isomerization between its indication DX-2300 and the hK1 has with formation even the new mixture of high-affinity more.

The active DX-2300 of class kallidinogenase suppresses in the animal urineBefore showed tissue kallikrein (K1) secrete in urine in.Kizuki, people such as K., Adv Exp Med Biol, 1986.198 Pt A-.329-337; Takaoka, people such as M., Biochem Biophys Res Commun, 1984.122 (3): 1282-1288; Murthy, people such as K.K., Arch BiochemBiophys, 1986.244 (2): 563-571; Stella, people such as R.C., Adv Exp Med Biol, 1989.247B:195-200.Use synthetic peptide substrates, can measure the activity of the K1 in urine.Because the major part of secreted K1 is nonactive alternative form usually, so essential with another protease activated described enzyme such as trypsinase or thermolysin.Before the activity of measuring the K1 in urine, use specific inhibitor (trypsinase is phosphoramidon as Trypsin inhibitor SBTI or for thermolysin) to suppress activating enzymes.

By Fig. 5 obviously as can be known DX-2300 suppress to be present in class kallidinogenase activity in the urine of the mankind, sheep, dog, monkey, rabbit and cow.Yet it does not suppress viewed or from the class kallidinogenase activity of pig urine in the rodent (mouse, rat, guinea pig or hamster).The measured active DX-2300 IC of class kallidinogenase during sheep urine is urinated with the people ₅₀Value quite is respectively 2.9 ± 1.6nM and 3.0 ± 0.6nM (Fig. 6).

Program.At first animal urine is cushioned by the 1M HEPES (pH7.7) that adds 1/10 volume, and centrifugal to remove solid.Subsequently, under 31 ℃, supernatant liquor (1mL) is activated 30 minutes by adding 50 μ L trypsinase agarose resins.Subsequently, deactivate with 10 μ M Trypsin inhibitor SBTIs with any remaining trypsinase that removes the trypsinase agarose and may be leach sample is centrifugal from resin.Exist or do not exist under the situation of 0.5 μ M DX-2300, in the Kal of 96 orifice plates damping fluid (table 4) hole (100 μ L cumulative volume), measuring activation urine (9 μ L) medium vessels and releive plain synthesizing the activity of substrate Pro-Phe-Arg-AMC (100 μ M[are final]).At first, under 37 ℃, will activate urine and cultivate 30 minutes, add substrate thereafter with DX-2300.

The active inhibition of class kallidinogenase in the human bronchoalveolar lavage fluid

As measured in bronchoalveolar lavage fluid (BAL), proved with healthy individual and compared that tissue kallikrein 1 (hK1) increases in the tracheae of asthmatic patient.Proud, people such as D., Am JRespir CellMolBiol, 1993.8 (1): 16-19; Christiansen, people such as S.C., Am Rev Respir Dis, 1992.145 (4Pt 1): 900-905; Christiansen, people such as S.C., JClin Invest, 1987.79 (1): 188-197.As using synthetic substrate Pro-Phe-Arg-AMC measured, visible DX-2300 suppresses from the class kallidinogenase activity (referring to Fig. 7) among 4 different mild asthma patients' the BAL.Described synthetic substrate can by other protease hydrolysis among the BAL and therefore described substrate be not the active selective substrate of kininogenase.Because having showed DX-2300 is the high degree of specificity inhibitor of K1, it can not suppress described other proteolytic enzyme among the BAL.Therefore, do not wish that DX-2300 suppresses the protease activity among the BAL fully.In BAL, produce observations that the proteolytic enzyme of real mass suppresses and confirm that K1 is active in the asthmatic patient and raise having the active described specific inhibitor of K1.

The effect of DX-2300 in the sheep model of asthma

The sheep model of asthma before had been used to confirm potential asthmatic medicament target and test medicament.Forteza, people such as R., AmJRespir Crit Care Med, 1996.154 (1): 36-42; Rosen, people such as S.D., Am JPathol, 2005.166 (3): 935-944; Scuri, people such as M., JAppl Physiol, 2002.93 (6): 1900-1906.In described model, the measurement that the supersensitivity sheep is attacked poison and lung resistance with ascaris suum antigen provides the indication of following bronchoconstriction.Shown in Fig. 8 A, DX-2300 reduces early stage bronchoconstriction slightly and suppresses the late period bronchoconstriction and reaches more than 80%.

The effect in human patients is also indicated in the measurement of the tracheae overreaction (AHR) in the sheep model of asthma.Shown in Fig. 9 B, after anaphylactogen was attacked poison, DX-2300 blocked AHR fully.According to make bronchoconstriction increase baseline more than 400% (PC400) need how many carbachols (a kind of cholinergic agonist) to measure AHR.Anaphylactogen is attacked after the poison, and tracheae is to described agonist overreaction, so induce the required amount of the bronchoconstriction of same degree to reduce.Fig. 9 B proof DX-2300 blocking-up AHR development, because attack after the poison, the bronchoconstriction that reaches the PC400 degree needs the carbachol of same amount.

Measured as increasing by lung resistance, high molecular weight kininogen (HMWK) (K1 substrate) is thrown and caused the moderate bronchoconstriction to sheep by sucking.Described increase is the generation owing to the kassinin kinin that is caused by the tracheae kininogenase.DX-2300 suppresses the observations (Figure 10) of HMWK inductive bronchoconstriction and supports that K1 is the observations of main kininogenase in the tracheae.Christiansen, people such as S.C., Am Rev Respir Dis, 1992.145 (4Pt1): 900-905; Lauredo, people such as IT., Am JPhysiol Lung Cell Mol Physiol, 2004.286 (4): L734-40.

The heavy chain of encoding D X-2300 and the DNA's of light chain is codon optimized

By GENEART, expert in the field uses proprietary algorithm that both of the DNA of the heavy chain of encoding D X-2300 and light chain are all optimized to express in Chinese hamster ovary (CHO) cell.During codon optimized, adjust use according to the codon bias of Cricetus gene.Avoid high (＞80%) or low (＜30%) GC content district as far as possible.In addition, with regard to heavy chain, remove 4 protokaryons and suppress the consistent hidden donor splicing site of main structure with 7.With regard to light chain, remove single intron.

DX-2300 light chain (SBQ ID NO:1378) through optimizing

AAGCTTATGGGCTGGTCCTGTATCATCCTGTTTCTGGTGGCCACCGCCACCTCTGGCGTG

AACTCCTCCAGACTGGAGGTGTGGACCTACATCTGGGTGACCATGACCTCCACCCTGCCT

TTCTCTCCAGGCGTGCACTCCGACATCCAGATGACCCAGTCCCCCTCTTCTCTGTCTGCC

TCTGTGGGCGACAGAGTGACCATCACCTGTCGGGCCTCCCAGTCCATCTCCTCCTACCTG

AACTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGCTGCTGATCTACGCCGCTTCCTCT

CTGCAGTCTGGCGTGCCTTCCAGATTCTCCGGCTCTGGCTCTGGCACCGATTTCACCCTG

ACCATCTCCAGCCTGCAGCCTGAGGATTTCGCCACCTACTACTGCCAGCAGTCCTACTCT

ACCCCTCTGACCTTTGGCGGCGGAACAAAGGTGGAGATCAAGAGGACCGTGGCCGCTCCT

TCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCTGGCACCGCCTCTGTGGTG

TGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCC

CTGCAGTCCGGCAATTCCCAGGAGTCTGTGACCGAGCAGGACTCCAAGGACAGCACCTAC

TCCCTGTCCTCTACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCC

TGTGAGGTGACCCACCAGGGCCTGTCTAGCCCTGTGACCAAGTCCTTCAACCGGGGCGAG

TGCTGATGAGAATTC

DX-2300 heavy chain (SEQ ID NO:1379) through optimizing

AAGCTTATGGGCTGGTCCTGTATCATCCTGTTTCTGGTGGCCACCGCCACCGGCGCTCAC

TCTGAGGTGCAGCTGCTGGAGTCTGGCGGCGGACTGGTGCAGCCTGGCGGCTCTCTGAGA

CTGTCTTGTGCCGCCTCCGGCTTCACCTTCTCCAAGTACAAGATGGTGTGGGTGAGGCAG

GCCCCTGGCAAGGGCCTGGAGTGGGTGTCCTCCATCTACCCATCTGGCGGCATCACCGCC

TACGCCGATTCTGTGAAGGGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTG

TACCTGCAGATGAACTCCCTGAGAGCCGAGGATACCGCCATGTACTACTGTGCCAAGGAC

ATCACCCCTGGCGGAGGATCTGGCTTCCGGCTGCCCAAGAATTACTACTACTACGGCATG

GATGTGTGGGGCCAGGGCACCACCGTGACCGTGTCCTCTGCTTCTACCAAGGGCCCTTCC

GTGTTTCCTCTGGCCCCTTCCTCCAAGTCTACCTCCGGCGGCACCGCCGCTCTGGGCTGC

CTGGTGAAGGACTACTTCCCTGAGCCCGTGACAGTGTCTTGGAACTCTGGCGCTCTGACC

TCTGGCGTGCACACCTTCCCTGCTGTGCTGCAGTCCTCCGGCCTGTACTCTCTGTCCTCC

GTGGTGACCGTGCCTTCTTCTTCTCTGGGCACCCAGACCTACATCTGTAACGTGAACCAC

AAGCCCTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGTGACAAGACCCAC

ACCTGCCCTCCCTGTCCTGCCCCTGAGCTGCTGGGCGGACCTTCTGTGTTCCTGTTCCCC

CCCAAGCCTAAGGACACCCTGATGATCTCCAGGACCCCTGAGGTGACCTGTGTGGTGGTG

GACGTGTCTCACGAGGATCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTG

CACAACGCCAAGACCAAGCCTAGGGAGGAGCAGTACAACTCCACCTACCGGGTGGTGTCT

GTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAATACAAGTGTAAGGTGTCC

AACAAGGCCCTGCCTGCCCCCATCGAGAAAACCATCTCCAAGGCCAAGGGCCAGCCTCGG

GAGCCTCAGGTGTACACCCTGCCTCCTAGCAGGGAGGAGATGACCAAGAACCAGGTGTCC

CTGACCTGTCTGGTGAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAAT

GGCCAGCCCGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCTGACGGCTCCTTC

TTCCTGTACTCCAAGCTGACCGTGGACAAGTCCAGATGGCAGCAGGGCAACGTGTTCTCC

TGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCC

CCCGGCAAGTGATGAGAATTC

The those skilled in the art should only use normal experiment can recognize a plurality of equivalents that maybe can confirm specific embodiment of the present invention as herein described.Described equivalent is intended to be covered by in the following claims.

Claims (33)

1. protein, it comprises heavy chain immunoglobulin (HC) variable domain sequence and light chain immunoglobulin (LC) variable domain sequence, and wherein said HC variable domain sequence and described LC variable domain sequence form the antigen binding site that combines and suppress the enzymic activity of hK1 with hK1.

2. protein according to claim 1, wherein said protein has K less than 10nM to hK1 _i

3. protein according to claim 1, wherein said protein has K less than 1nM to hK1 _i

4. protein according to claim 1, wherein said protein has K less than 0.1nM to hK1 _i

5. protein according to claim 1, wherein said protein have in one or more following character:

(i) described heavy chain variable domain sequence comprises:

(a) comprise aminoacid sequence: the CDR1 of Xaa1-Tyr-Xaa2-Met-Xaa3, wherein Xaa1 is selected from Lys and His; Xaa2 is selected from Lys, Va1 and Ser; Xaa3 is selected from Va1, Ile and Thr;

(b) comprise aminoacid sequence: the CDR2 of Xaa1-Ile-Tyr-Pro-Ser-Gly-Gly-Xaa2-Thr-Xaa3-Tyr-Ala-Asp-S er-Val-Lys-Gly, wherein Xaa1 is selected from Ser, Trp and Arg; Xaa2 is selected from Ile, Asn and Arg; Xaa3 is selected from Ala, Ile and Gly; And/or

(c) comprise at least 8 amino acid whose from

The CDR3 of DITPGGGSGFRLPKNYYYYGMDV (SEQ ID NO:12), VGVWYGMDV (SEQ IDNO:114) or DSGGYYYGMDV (SEQ ID NO:156);

(ii) described light chain variable territory sequence comprises: (a) comprise aminoacid sequence:

The CDR1 of Arg-Ala-Ser-Gln-Ser-Xaa1-Ser-Ser-Xaa2-Xaa3-Xaa4-Xaa5 (SEQ ID NO:1380), wherein Xaa1 is selected from Ile and Val; Xaa2 is selected from Tyr and Ser; Xaa3 is selected from Leu and Tyr; Xaa4 is that to be selected from Asn, Ala and Leu and Xaa5 be Ala or does not exist;

(b) comprise aminoacid sequence: the CDR2 of Xaa1-Ala-Ser-Xaa2-Xaa3-Xaa4, wherein Xaa1 is selected from Ala, Asp and Gly; Xaa2 is selected from Ser and Asn; Xaa3 is selected from Leu and Arg; Xaa4 is selected from Gln and Ala; And Xaa4 is selected from Ser and Thr; And/or

(c) comprise at least 5 amino acid whose CDR3 from QQSYSTPLT (SEQ ID NO:9), QQRSNWPSPIA (SEQ ID NO:111) and QQYGSSLT (SEQ ID NO:153);

(iii) described heavy chain variable domain sequence comprises with SEQ ID NO:1206,1245 or 1354 heavy chain variable domain sequence having at least 85% conforming sequence;

(iv) described light chain variable territory sequence comprises with SEQ ID NO:1029,1070 or 1183 light chain variable territory sequence having at least 85% conforming sequence;

(v) described heavy chain variable domain sequence comprise by under stringent condition with the sequence of the nucleic acid encoding of the sequence hybridization of coding SEQ ID NO:1206,1245 or 1354 heavy chain variable domain sequence;

(vi) described light chain variable territory sequence comprise by under stringent condition with the sequence of the nucleic acid encoding of the sequence hybridization of coding SEQ ID NO:1029,1070 or 1183 light chain variable territory sequence; And/or

(vii) described protein combines with hK1 with antibody DX-2300, M093-F09 or M137-E01 competition.

6. protein according to claim 1, it comprises the CDR district of described DX-2300 antibody.

7. protein according to claim 1, it comprises the CDR district of described M093-F09 antibody.

8. protein according to claim 1, it comprises the CDR district of described M137-E01 antibody.

9. protein according to claim 1, the corresponding variable domain sequence of wherein said heavy chain and light chain variable territory sequence and SEQ ID NO:1245 and 1070 has at least 90% consistence.

10. protein according to claim 1, the corresponding variable domain sequence of wherein said heavy chain and light chain variable territory sequence and SEQ ID NO:1206 and 1029 has at least 90% consistence.

11. protein according to claim 1, the corresponding variable domain sequence of wherein said heavy chain and light chain variable territory sequence and SEQ ID NO:1183 and 1354 has at least 90% consistence.

12. protein according to claim 1, wherein at least 80% FR district is consistent with the FR sequence that from the human reproduction is the FR sequence of sequence or DX-2300, M093-F09 or M137-E01.

13. protein according to claim 1, it does not have immunogenicity in human body.

14. protein according to claim 1, it is a total length IgG antibody.

15. protein according to claim 1, it is for antigen-binding fragments of antibodies and do not comprise the Fc territory.

16. a medical composition, it comprises protein according to claim 1 and pharmaceutically acceptable supporting agent.

17. a treatment or the method for preventing the hK1 associated conditions, described method comprises:

With protein according to claim 1 with the amount of effective treatment or prevention hK1 associated conditions throw with to the person under inspection.

18. method according to claim 17, wherein said illness are to be selected from by following each sick group that forms: asthma, chronic obstructive pulmonary disease (COPD), multiple sclerosis, psoriasis, rheumatoid arthritis, osteoarthritis, rhinitis, sinusitis, inflammatory bowel, immune-mediated diabetes, acute pancreatitis, interstitial cystitis or superfluous natural disposition illness.

19. method according to claim 18, wherein said illness are asthma and described asthma is allergic asthma or non-allergic asthma.

20. method according to claim 18, wherein said illness is transitivity carcinoma of the pancreas or tumor-blood-vessel growth for go to live in the household of one's in-laws on getting married natural disposition illness and described superfluous natural disposition illness.

21. method according to claim 19, wherein said medical composition be by suck to throw with.

22. method according to claim 21, wherein said medical composition be to use sucker throw with.

23. method according to claim 20, it further comprises throws and second medicament of regulating vasculogenesis.

24. method according to claim 23, wherein said second medicament are VEGF antibody or its Fab.

25. regulate the active method of hK1 for one kind, described method comprises:

Provide hK1 according to claim 1 conjugated protein; With described protein is contacted with hK1 to be enough to the regulating active amount of hK1.

26. method according to claim 25, wherein said contact is in vitro.

27. method according to claim 25, wherein said contact is in vivo.

28. the method that in vitro hK1 albumen exists in the test sample, described method comprises:

(i) allowing the conjugated protein and described hK1 albumen of hK1 according to claim 1 to take place under the interactional condition, described sample is contacted with described hK1 is conjugated protein; With

(ii) detect the interaction between the conjugated protein and sample of described hK1.

29. method according to claim 28 is wherein fixed at least one among the conjugated protein or described hK1 of described hK1.

30. one kind is detected the method that hK1 exists in vivo, described method comprises:

(i) allowing the conjugated protein and described hK1 albumen of hK1 according to claim 1 to take place under the interactional condition, with the conjugated protein throwing of described hK1 and to the person under inspection; With

(ii) detect the conjugated protein formation of the mixture between the conjugated protein and hK1 of described hK1 in the intravital position of described person under inspection or described person under inspection's body of described hK1.

31. method according to claim 30, wherein said person under inspection is human person under inspection.

32. method according to claim 31, wherein said detection comprises subject imaging.

33. method according to claim 32 is wherein given described hK1 conjugated protein in addition mark with the MRI detectable label.

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