CN101300273B

CN101300273B - TRAIL recipient 2 Polypeptides and antibodies

Info

Publication number: CN101300273B
Application number: CN200680040505.4A
Authority: CN
Inventors: B·格利尼亚克; 杨晓东; S·黄-马登; I·福尔茨; 冯晓; A·费奇; S·富斯特; R·R·凯琴
Original assignee: Amgen Inc
Current assignee: Amgen Inc
Priority date: 2005-08-31
Filing date: 2006-08-28
Publication date: 2013-05-22
Anticipated expiration: 2026-08-28
Also published as: ZA200802575B; UA97096C2; CN101300273A

Abstract

Polypeptides are provided. Antibodies or antigen binding domains are provided which bind such polypeptides. Also provided are methods of obtaining an antibody that binds tumor necrosis factor (TNF)-related apoptosis-inducing ligand (''TRAIL'') Receptor-2 (TR-2) comprising administering at least one of such polypeptides to an animal and obtaining an antibody that binds TR-2 from the animal. Antibodies reactive with TR-2 are provided. Also provided are cells producing antibodies reactive with TR-2, pharmaceutical compositions comprising antibodies reactive with TR-2, methods using antibodies reactive with TR-2, and kits comprising antibodies reactive with TR-2. Also provided are methods of decreasing or preventing binding of an antibody to TR-2 by administering such a polypeptide.

Description

TRAIL receptor 2 polypeptides and antibody

The application requires the U.S. Provisional Application submitted on August 31st, 2005 number 60/713,433, and the right of priority of the U.S. Provisional Application of submitting on August 31st, 2005 number 60/713,478.U.S. Provisional Application number 60/713,433 and 60/713,478 is incorporated herein by reference for any purpose integral body.

Invention field

Polypeptide is provided.Antibody or antigen binding domains in conjunction with this type of polypeptide are provided.The method in conjunction with the antibody of apoptosis induction ligand related (" the TRAIL ") acceptor 2 of tumour necrosis factor (TNF) (TR-2) that obtains also is provided, described method comprises to animal uses at least one this type of polypeptide, and obtains the antibody in conjunction with TR-2 from animal.The antibody reacted with TR-2 is provided.Also provide the cell of producing the antibody reacted with TR-2, the pharmaceutical composition that comprises the antibody reacted with TR-2, the method for the antibody that use is reacted with TR-2, and the test kit that comprises the antibody reacted with TR-2.The method that reduces or stop antibody to be combined with TR-2 by using this type of polypeptide also is provided.

Background of invention

Interaction between TR-2 and ligand/TRAIL thereof in the inducing of apoptosis, work (referring to, for example, the people such as Almasan, Cytokine & Growth Factor Reviews 14:337-348 (2003)).TRAIL is also referred to as Apo2L/TRAIL; 4 members (TRAIL acceptor (" TR ") 1-4) with the TNF receptor superfamily; and to same several parts of relevant, solubility, opsteoprotegerin (doubting as osteoprotegerin osteoprotegerin) (" OGP ") acceptor interaction.TRAIL and TR-1 or the TR-2 combination on cell surface has triggered the apoptosis of that cell.TRAIL and TR-1 or TR-2 initial in conjunction with after, intracellular protein is recruited to death domain in the cell of acceptor, forms Signaling complex.Caspase is recruited to mixture in some cell, their autoactivations therein and activate successively other caspase and intracellular apoptosis cascade.TR-3 and TR-4 and OPG lack the cell intracellular domain of being responsible for the apoptotic signal transmission.Therefore, the combination of TRAIL and TR-3, TR-4 or OPG does not trigger apoptosis.TR-3 and TR-4 also are called as " trick " acceptor, and their overexpression has shown that Cell protection is not subject to the apoptosis induction via TRAIL.TR-2 is at various cells, comprises liver, brain, mammary gland, kidney, colon, lung, spleen, thymus gland, peripheral blood lymphocyte, prostate gland, testis, ovary, uterus and along GI various tissues.(referring to, for example, the people such as Walczak, EMBO is (1997) J.16:5386-5397; The people such as Spierings, J.Histochem.Cytochem.52:821-831 (2004)).Although TRAIL and TRAIL acceptor are wide expression, they are most active in the apoptosis induction of transformant.(referring to, for example, the people such as Daigle, Swiss Med.Wkly.131:231-237 (2001)).

Summary of the invention

In certain embodiments, provide the isolated polypeptide that comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a:

Wherein CDR1a comprises aminoacid sequence a b c d e f g h i j k l, and wherein amino acid a is glycine, and amino acid b is selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l;

Wherein CDR2a comprises aminoacid sequence m n o p q r s t u v w x y z a ' b ' c ', and wherein amino acid m is selected from tryptophane, tyrosine, Histidine, α-amino-isovaleric acid, L-glutamic acid or Serine; Amino acid n is selected from methionine(Met) or Isoleucine; Amino acid o is selected from l-asparagine, tyrosine, Serine, tryptophane or Histidine; Amino acid p is selected from proline(Pro), tyrosine, Serine, arginine, Histidine or l-asparagine; Amino acid q is selected from l-asparagine, Serine or aspartic acid; Amino acid r is selected from Serine or glycine; Amino acid s is selected from aspartic acid, Serine, Threonine or arginine; Amino acid t is selected from l-asparagine, Threonine, L-Ala, Isoleucine or tyrosine; Amino acid u is selected from Threonine, tyrosine, leucine, Methionin, l-asparagine or Isoleucine; Amino acid v is selected from glycine, tyrosine, aspartic acid or halfcystine; Amino acid w is selected from tyrosine or l-asparagine; Amino acid x is selected from L-Ala or proline(Pro); Amino acid y is selected from glutamine, Serine or aspartic acid; Amino acid z is selected from Methionin, leucine or Serine; Amino acid a ' is selected from phenylalanine, Methionin or α-amino-isovaleric acid; Amino acid b ' is selected from glutamine, Serine or Methionin; Be glycine or do not exist with amino acid c '; Wherein CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '; With

The described polypeptide of wherein being combined with light chain of antibody is in conjunction with TRAIL acceptor 2 (TR-2).

In certain embodiments, provide and comprised the isolated polypeptide that is selected from following at least one complementary determining region (CDR):

The amino acid 26-35 of SEQ ID NO:2;

The amino acid 50-66 of SEQ ID NO:2;

The amino acid 99-110 of SEQ ID NO:2;

The amino acid 26-37 of SEQ ID NO:4;

The amino acid 52-67 of SEQ ID NO:4;

The amino acid/11 00-109 of SEQ ID NO:4;

The amino acid 26-37 of SEQ ID NO:6;

The amino acid 52-67 of SEQ ID NO:6;

The amino acid/11 00-109 of SEQ ID NO:6;

The amino acid 26-37 of SEQ ID NO:8;

The amino acid 52-67 of SEQ ID NO:8;

The amino acid/11 00-109 of SEQ ID NO:8;

The amino acid 26-35 of SEQ ID NO:10;

The amino acid 50-66 of SEQ ID NO:10;

The amino acid 99-110 of SEQ ID NO:10;

The amino acid 26-35 of SEQ ID NO:12;

The amino acid 50-66 of SEQ ID NO:12;

The amino acid 99-111 of SEQ ID NO:12;

The amino acid 26-35 of SEQ ID NO:14;

The amino acid 50-65 of SEQ ID NO:14;

The amino acid 98-111 of SEQ ID NO:14;

The amino acid 26-37 of SEQ ID NO:16;

The amino acid 52-67 of SEQ ID NO:16;

The amino acid/11 00-109 of SEQ ID NO:16;

The amino acid 26-35 of SEQ ID NO:18;

The amino acid 50-66 of SEQ ID NO:18;

The amino acid 99-105 of SEQ ID NO:18;

The amino acid 26-35 of SEQ ID NO:20;

The amino acid 50-66 of SEQ ID NO:20;

The amino acid 99-118 of SEQ ID NO:20;

The amino acid 26-35 of SEQ ID NO:22;

The amino acid 50-66 of SEQ ID NO:22;

The amino acid 99-118 of SEQ ID NO:22;

The amino acid 26-35 of SEQ ID NO:24;

The amino acid 50-65 of SEQ ID NO:24;

The amino acid 98-108 of SEQ ID NO:24;

The amino acid 26-35 of SEQ ID NO:26;

The amino acid 50-66 of SEQ ID NO:26;

The amino acid 99-110 of SEQ ID NO:26;

The amino acid 26-35 of SEQ ID NO:28;

The amino acid 50-66 of SEQ ID NO:28;

The amino acid 99-117 of SEQ ID NO:28;

The amino acid 26-37 of SEQ ID NO:30;

The amino acid 52-67 of SEQ ID NO:30;

The amino acid/11 00-111 of SEQ ID NO:30;

The amino acid 26-37 of SEQ ID NO:32;

The amino acid 52-67 of SEQ ID NO:32;

The amino acid/11 00-111 of SEQ ID NO:32;

The amino acid 26-37 of SEQ ID NO:34;

The amino acid 52-67 of SEQ ID NO:34; With

The amino acid/11 00-111 of SEQ ID NO:34;

The described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.

In certain embodiments, provide the isolated polypeptide that comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b:

Wherein CDR1b comprises aminoacid sequence a1 b1 c1 d1 e1 f1 g1 h1 i1 j1 k1 l1 m1n1 o1 p1 q1, and wherein amino acid a1 is selected from arginine or Methionin; Amino acid b1 is selected from Threonine, L-Ala or Serine; Amino acid c1 is Serine; Amino acid d1 is glutamine; Amino acid e1 is selected from Serine or glycine; Amino acid f1 is selected from Isoleucine, leucine or α-amino-isovaleric acid; Amino acid g1 is selected from Serine, leucine or arginine; Amino acid h1 is selected from Threonine, Serine, Isoleucine, l-asparagine, arginine, Histidine or tyrosine; Amino acid i1 is selected from tyrosine, arginine, tryptophane, aspartic acid or Serine; J1 is selected from leucine, Isoleucine, l-asparagine, tyrosine or Serine; Amino acid k1 is selected from l-asparagine, glycine, α-amino-isovaleric acid, L-Ala or leucine; Amino acid l1 is selected from tyrosine, L-Ala or l-asparagine or does not exist; Amino acid m1 is selected from l-asparagine or Methionin or does not exist; Amino acid n1 is selected from tyrosine, l-asparagine or Isoleucine or does not exist; Amino acid o1 is selected from leucine or tyrosine or does not exist; Amino acid p1 is selected from aspartic acid or leucine or does not exist; Be selected from α-amino-isovaleric acid, L-Ala or Threonine or do not exist with amino acid q1;

Wherein CDR2b comprises aminoacid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from L-Ala, aspartic acid, leucine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid s1 is selected from Threonine, α-amino-isovaleric acid, glycine or L-Ala; Amino acid t1 is Serine; Amino acid u1 is selected from Serine, l-asparagine or Threonine; Amino acid v1 is selected from leucine, phenylalanine or arginine; Amino acid w1 is selected from glutamine, L-Ala or L-glutamic acid; Be selected from Serine, arginine or Threonine with amino acid x1;

Wherein CDR3b comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; With

The described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

The amino acid 24-34 of SEQ ID NO:36;

The amino acid 50-56 of SEQ ID NO:36;

The amino acid 89-97 of SEQ ID NO:36;

The amino acid 24-34 of SEQ ID NO:38;

The amino acid 50-56 of SEQ ID NO:38;

The amino acid 89-97 of SEQ ID NO:38;

The amino acid 24-34 of SEQ ID NO:40;

The amino acid 50-56 of SEQ ID NO:40;

The amino acid 89-97 of SEQ ID NO:40;

The amino acid 24-34 of SEQ ID NO:42;

The amino acid 50-56 of SEQ ID NO:42;

The amino acid 89-97 of SEQ ID NO:42;

The amino acid 24-34 of SEQ ID NO:44;

The amino acid 50-56 of SEQ ID NO:44;

The amino acid 89-97 of SEQ ID NO:44;

The amino acid 24-34 of SEQ ID NO:46;

The amino acid 50-56 of SEQ ID NO:46;

The amino acid 89-97 of SEQ ID NO:46;

The amino acid 24-40 of SEQ ID NO:48;

The amino acid 56-62 of SEQ ID NO:48;

The amino acid 95-103 of SEQ ID NO:48;

The amino acid 24-39 of SEQ ID NO:50;

The amino acid 55-61 of SEQ ID NO:50;

The amino acid 94-102 of SEQ ID NO:50;

The amino acid 24-40 of SEQ ID NO:52;

The amino acid 56-62 of SEQ ID NO:52;

The amino acid 95-103 of SEQ ID NO:52;

The amino acid 24-34 of SEQ ID NO:54;

The amino acid 50-56 of SEQ ID NO:54;

The amino acid 89-97 of SEQ ID NO:54;

The amino acid 24-34 of SEQ ID NO:56,

The amino acid 50-56 of SEQ ID NO:56;

The amino acid 89-97 of SEQ ID NO:56;

The amino acid 24-40 of SEQ ID NO:58;

The amino acid 56-62 of SEQ ID NO:58;

The amino acid 95-103 of SEQ ID NO:58;

The amino acid 24-34 of SEQ ID NO:60;

The amino acid 50-56 of SEQ ID NO:60;

The amino acid 89-97 of SEQ ID NO:60;

The amino acid 24-34 of SEQ ID NO:62;

The amino acid 50-56 of SEQ ID NO:62;

The amino acid 89-97 of SEQ ID NO:62;

The amino acid 24-35 of SEQ ID NO:64;

The amino acid 51-57 of SEQ ID NO:64;

The amino acid 90-88 of SEQ ID NO:64;

The amino acid 24-34 of SEQ ID NO:66;

The amino acid 50-57 of SEQ ID NO:66;

The amino acid 89-97 of SEQ ID NO:66;

The amino acid 24-34 of SEQ ID NO:68;

The amino acid 50-56 of SEQ ID NO:68; With

The amino acid 89-97 of SEQ ID NO:68;

In certain embodiments, provide the separation polynucleotide of the sequence that comprises coded polypeptide, described polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a:

Wherein CDR1a comprises aminoacid sequence a b c d e f g h i j k l, and wherein amino acid a is that glycine, amino acid b are selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l;

Wherein CDR2a comprises aminoacid sequence m n o p q r s t u v w x y z a ' b ' c ', and wherein amino acid m is selected from tryptophane, tyrosine, Histidine, α-amino-isovaleric acid, L-glutamic acid or Serine; Amino acid n is selected from methionine(Met) or Isoleucine; Amino acid o is selected from l-asparagine, tyrosine, Serine, tryptophane or Histidine; Amino acid p is selected from proline(Pro), tyrosine, Serine, arginine, Histidine or l-asparagine; Amino acid q is selected from l-asparagine, Serine or aspartic acid; Amino acid r is selected from Serine or glycine; Amino acid s is selected from aspartic acid, Serine, Threonine or arginine; Amino acid t is selected from l-asparagine, Threonine, L-Ala, Isoleucine or tyrosine; Amino acid u is selected from Threonine, tyrosine, leucine, Methionin, l-asparagine or Isoleucine; Amino acid v is selected from glycine, tyrosine, aspartic acid or halfcystine; Amino acid w is selected from tyrosine or l-asparagine; Amino acid x is selected from L-Ala or proline(Pro); Amino acid y is selected from glutamine, Serine or aspartic acid; Amino acid z is selected from Methionin, leucine or Serine; Amino acid a ' is selected from phenylalanine, Methionin or α-amino-isovaleric acid; Amino acid b ' is selected from glutamine, Serine or Methionin; Be glycine or do not exist with amino acid c ';

Wherein CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '; With

In certain embodiments, provide the separation polynucleotide of the sequence that comprises coded polypeptide, described polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b:

Wherein CDR2b comprises aminoacid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from L-Ala, aspartic acid, leucine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid s1 is selected from Threonine, α-amino-isovaleric acid, glycine or L-Ala; Amino acid t1 is Serine; Amino acid u1 is selected from Serine, l-asparagine or Threonine; Amino acid v1 is selected from leucine, phenylalanine or arginine; Amino acid w1 is selected from glutamine, L-Ala or L-glutamic acid; Be selected from Serine, arginine or Threonine with amino acid x1; Wherein CDR3 comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; With

In certain embodiments, provide and comprised separating of variable region and constant region of anti-TR-2 antibody, wherein said antibody comprises:

(i) the first polypeptide that comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a,

Wherein CDR1a comprises aminoacid sequence a b c d e f g h i j k l, and wherein amino acid a is glycine; Amino acid b is selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l;

Described the first polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2; With

(ii) the second polypeptide that comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b

Wherein CDR2b comprises aminoacid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from L-Ala, aspartic acid, leucine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid s 1 is selected from Threonine, α-amino-isovaleric acid, glycine or L-Ala; Amino acid t1 is Serine; Amino acid u1 is selected from Serine, l-asparagine or Threonine; Amino acid v1 is selected from leucine, phenylalanine or arginine; Amino acid w1 is selected from glutamine, L-Ala or L-glutamic acid; Be selected from Serine, arginine or Threonine with amino acid x1;

Wherein CDR3b comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; With

Described the second polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:2 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:36; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQID NO:4 and the second polypeptide that comprises the CDRs as shown in SEQ IDNO:38; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:6 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:40; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:8 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:42; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:10 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:44; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:12 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:46; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:14 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:48; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:16 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:50; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:18 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:52; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:20 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:54; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:22 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:56; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:24 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:58; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:26 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:60; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:28 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:62; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:30 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:64; The first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:32 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:66; Or the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ IDNO:34 and the second polypeptide that comprises the CDRs as shown in SEQ IDNO:68.

In certain embodiments, provide cell, it comprises:

(a) the first polynucleotide of the sequence that comprises coding the first polypeptide, described the first polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a, wherein CDR1a comprises aminoacid sequence a b c d e f g h i j k l, and wherein amino acid a is that glycine, amino acid b are selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l;

Wherein CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '; Described the first polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2; With

(b) the second polynucleotide of the sequence that comprises coding the second polypeptide, described the second polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b,

Wherein CDR3b comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; Described the second polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

In certain embodiments, provide and be combined separation antibody with epitope specificity, described epi-position is by being selected from least one following antibodies specific combination: Ab A, Ab B, Ab C, Ab D, Ab E, Ab F, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, Ab P and Ab Q.

In certain embodiments, provide the polypeptide that comprises at least one aminoacid sequence that is selected from SEQ ID NO:94, SEQ ID NO:95 and SEQ ID NO:96.

In certain embodiments, provide the polypeptide basically formed by least one aminoacid sequence that is selected from SEQ ID NO:94, SEQ IDNO:95 and SEQ ID NO:96.

In certain embodiments, provide in conjunction with the antibody or the antigen binding domains that are selected from least one aminoacid sequence of SEQ ID NO:94, SEQ ID NO:95 and SEQ ID NO:96.

In certain embodiments, the method in conjunction with the antibody of TR-2 that obtains is provided, described method comprises to animal uses at least one polypeptide that is selected from SEQ ID NO:94, SEQ ID NO:95 and SEQ ID NO:96, and obtains the antibody in conjunction with TR-2 from animal.

In certain embodiments, provide the method that reduces or stop antibody to be combined with TR-2, described method is by using the polypeptide that comprises at least one aminoacid sequence that is selected from SEQ ID NO:94, SEQ ID NO:95 and SEQ IDNO:96.

In certain embodiments, provide the method that reduces or stop antibody to be combined with TR-2, described method is by using the polypeptide be comprised of at least one aminoacid sequence that is selected from SEQ ID NO:94, SEQ ID NO:95 and SEQ IDNO:96.

The accompanying drawing summary

Fig. 1 has shown the immunization timetable used for the TR-2-His construct in the transgenic mice of expressing the human immunoglobulin gene in embodiment 1, via footpad inoculation (organizing 1,2 and 3) or via peritoneal injection (organizing 4 and 5).

Fig. 2 shown according to the work described in embodiment 1, measures the reactive ELISA assay method result for antigen TR-2 from some blood sample of the selected mouse of describing in Fig. 1.

Fig. 3 has shown the heavy chain (SEQ ID NO:1) of the anti-TR-2 antibody A of encoding and the nucleotide sequence of light chain (SEQ ID NO:35) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:2) and light chain (SEQ ID NO:36) variable region.

Fig. 4 has shown the heavy chain (SEQ ID NO:3) of the anti-TR-2 antibody B that encodes and the nucleotide sequence of light chain (SEQ ID NO:37) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:4) and light chain (SEQ ID NO:38) variable region.

Fig. 5 has shown the heavy chain (SEQ ID NO:5) of the anti-TR-2 antibody C that encodes and the nucleotide sequence of light chain (SEQ ID NO:39) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:6) and light chain (SEQ ID NO:40) variable region.

Fig. 6 has shown the heavy chain (SEQ ID NO:7) of the anti-TR-2 antibody D that encodes and the nucleotide sequence of light chain (SEQ ID NO:41) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:8) and light chain (SEQ ID NO:42) variable region.

Fig. 7 has shown the heavy chain (SEQ ID NO:9) of the anti-TR-2 antibody E that encodes and the nucleotide sequence of light chain (SEQ ID NO:43) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:10) and light chain (SEQ ID NO:44) variable region.

Fig. 8 has shown the heavy chain (SEQ ID NO:11) of the anti-TR-2 antibody F that encodes and the nucleotide sequence of light chain (SEQ ID NO:45) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:12) and light chain (SEQ ID NO:46) variable region.

Fig. 9 has shown the heavy chain (SEQ ID NO:13) of the anti-TR-2 antibody G that encodes and the nucleotide sequence of light chain (SEQ ID NO:47) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:14) and light chain (SEQ ID NO:48) variable region.

Figure 10 has shown the heavy chain (SEQ ID NO:15) of the anti-TR-2 antibody H that encodes and the nucleotide sequence of light chain (SEQ ID NO:49) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:16) and light chain (SEQ ID NO:50) variable region.

Figure 11 has shown the heavy chain (SEQ ID NO:17) of the anti-TR-2 antibody I of encoding and the nucleotide sequence of light chain (SEQ ID NO:51) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:18) and light chain (SEQ ID NO:52) variable region.

Figure 12 has shown the heavy chain (SEQ ID NO:19) of the anti-TR-2 antibody J that encodes and the nucleotide sequence of light chain (SEQ ID NO:53) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:20) and light chain (SEQ ID NO:54) variable region.

Figure 13 has shown the heavy chain (SEQ ID NO:21) of the anti-TR-2 antibody K that encodes and the nucleotide sequence of light chain (SEQ ID NO:55) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:22) and light chain (SEQ ID NO:56) variable region.

Figure 14 has shown the heavy chain (SEQ ID NO:23) of the anti-TR-2 antibody L that encodes and the nucleotide sequence of light chain (SEQ ID NO:57) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:24) and light chain (SEQ ID NO:58) variable region.

Figure 15 has shown the heavy chain (SEQ ID NO:25) of the anti-TR-2 antibody M that encodes and the nucleotide sequence of light chain (SEQ ID NO:59) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:26) and light chain (SEQ ID NO:60) variable region.

Figure 16 has shown the heavy chain (SEQ ID NO:27) of the anti-TR-2 antibody N that encodes and the nucleotide sequence of light chain (SEQ ID NO:61) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:28) and light chain (SEQ ID NO:62) variable region.

Figure 17 has shown the heavy chain (SEQ ID NO:29) of the anti-TR-2 antibody O that encodes and the nucleotide sequence of light chain (SEQ ID NO:63) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:30) and light chain (SEQ ID NO:64) variable region.

Figure 18 has shown the heavy chain (SEQ ID NO:31) of the anti-TR-2 antibody P that encodes and the nucleotide sequence of light chain (SEQ ID NO:65) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:32) and light chain (SEQ ID NO:66) variable region.

Figure 19 has shown the heavy chain (SEQ ID NO:33) of the anti-TR-2 antibody Q that encodes and the nucleotide sequence of light chain (SEQ ID NO:67) variable region, and the aminoacid sequence of the heavy chain of that antibody (SEQID NO:34) and light chain (SEQ ID NO:68) variable region.

Figure 20 is the aminoacid sequence comparison to the variable region of heavy chain of Q (SEQ ID NOs:2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32 and 34) about anti-TR-2 antibody A.Framework region 1 to 3 (FR1, FR2 and FR3) and complementary determining region 1 to 3 (CDR1, CDR2 and CDR3) about each sequence have been shown.

Figure 21 is the aminoacid sequence comparison to the variable region of light chain of Q (SEQ ID NOs:36,38,40,42,44,46,48,50,52,54,56,58,60,62,64,66 and 68) about anti-TR-2 antibody A.Framework region 1 to 3 (FR1, FR2 and FR3) and complementary determining region 1 to 3 (CDR1, CDR2 and CDR3) about each sequence have been shown.

The table of Figure 22 shown according to the work described in embodiment 5, according to separately with one of the anti-TR-2 of some people of ability antibody to 4 reactive group of brachymemma and chimeric N-avidin TR-2 protein bound in classification.

Figure 23 has shown according to the work described in embodiment 6,13 kinds of truncate schematic diagram of the people N-avidin-TR-2 used in epitope mapping.

The bar graph of Figure 24 has shown according to the work described in embodiment 6, the combination that the anti-TR-2 antibody of some people and N-avidin-TR-2 are truncate.

Figure 25 shown according to the work described in embodiment 6, the truncate and N-avidin-chimeric schematic diagram of cyno/ people TR-2 of the N-used in epitope mapping avidin-cyno TR-2.

Figure 26 is according to the work described in embodiment 6, the comparison of people TR-2, cyno TR-2 (short-form) and mouse TR-2 sequence.

The bar graph of Figure 27 shown according to the work described in embodiment 6, the anti-TR-2 antibody of some people and the combination that N-avidin-TR-2 is truncate, mosaic and structural domain are replaced.

The detailed description of some embodiment

Division header used herein is only for organizational goal and should not be interpreted as the theme that restriction is described.The All Files of quoting in the application or the part of file, include but not limited to, patent, patent application, article, book and paper, for any purpose integral body especially is incorporated herein by reference.

Definition

For recombinant DNA, oligonucleotide, synthetic and tissue culture and conversion (for example, electroporation, lipofection) can the Application standard technology.Enzymatic reaction and purification technique can according to the specification sheets of manufacturers or as this area, usually complete or carry out as described herein.Aforementioned techniques and operation generally can be according to ordinary methods well-known in the art with as various and the carrying out described in more concrete reference, and described reference is cited from start to finish and discusses at this specification sheets.Referring to for example, the people such as Sambrook, Molecular Cloning:A LaboratoryManual (the 2nd edition, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y. (1989)).Unless specific definition is provided, the term be combined with analytical chemistry described herein, synthetic organic chemistry and medical science and pharmaceutical chemistry, and analytical chemistry described herein, synthetic organic chemistry and medical science and pharmaceutical chemical laboratory operation and technology be well-known in the art and normally used those.For chemosynthesis, chemical analysis, medicine preparation, preparation, sending can the Application standard technology with patient treatment.

In this application, unless separately illustrated, the use of odd number comprises plural number.In this application, except as otherwise noted, the use of "or" mean " and/or ".In addition, the use that term " comprises " is not restrictive.Equally, unless separately illustrated, term for example " element " or " component " comprises element and the component that contains a unit and contains element and the component that surpasses a subunit.

As used according to present disclosure, except as otherwise noted, following term should be understood to have following implication:

As used herein, term " separation polynucleotide " should mean the polynucleotide of genome, cDNA or synthetic source or its some combination, because be not combined with all or part polynucleotide that wherein " separation polynucleotide " are found at occurring in nature its source " separation polynucleotide " (1), (2) polynucleotide that are not attached thereto at occurring in nature with it are connected, or (3) do not occur as the part of larger sequence at occurring in nature.

Term " polynucleotide " and " oligonucleotide " are used interchangeably, and mean as what this paper mentioned the Nucleotide that length is the polymerized form of at least 10 bases.In certain embodiments, base can comprise the Nucleotide of arbitrary type of at least one ribonucleotide, deoxyribonucleotide and modified forms.This term comprises the DNA of single and double chain form.Term " polynucleotide " also comprises such sequence, and it comprises one or more in SEQ ID NOs:1,3,5,7,9,11,13,15,17,19,21,23,25,27,29,31,33,35,37,39,41,43,45,47,49,51,53,55,57,59,61,63,65 and 67.In certain embodiments, polynucleotide have and about 90% or approximately 95% or approximately 96% or approximately 97% or approximately 98% or approximately 99% nucleotide sequence be equal to of the nucleotide sequence shown in Fig. 3-19.In certain embodiments, the polynucleotide of specificity polynucleotide complementation with coding some polypeptide as herein described are provided.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a, wherein CDR1a comprises aminoacid sequence a b c d e f g h i j k l, and wherein amino acid a is that glycine, amino acid b are selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l; Wherein CDR2a comprises aminoacid sequence m n o p q rs t u v w x y z a ' b ' c ', and wherein amino acid m is selected from tryptophane, tyrosine, Histidine, α-amino-isovaleric acid, L-glutamic acid or Serine; Amino acid n is selected from methionine(Met) or Isoleucine; Amino acid o is selected from l-asparagine, tyrosine, Serine, tryptophane or Histidine; Amino acid p is selected from proline(Pro), tyrosine, Serine, arginine, Histidine or l-asparagine; Amino acid q is selected from l-asparagine, Serine or aspartic acid; Amino acid r is selected from Serine or glycine; Amino acid s is selected from aspartic acid, Serine, Threonine or arginine; Amino acid t is selected from l-asparagine, Threonine, L-Ala, Isoleucine or tyrosine; Amino acid u is selected from Threonine, tyrosine, leucine, Methionin, l-asparagine or Isoleucine; Amino acid v is selected from glycine, tyrosine, aspartic acid or halfcystine; Amino acid w is selected from tyrosine or l-asparagine; Amino acid x is selected from L-Ala or proline(Pro); Amino acid y is selected from glutamine, Serine or aspartic acid; Amino acid z is selected from Methionin, leucine or Serine; Amino acid a ' is selected from phenylalanine, Methionin or α-amino-isovaleric acid; Amino acid b ' is selected from glutamine, Serine or Methionin; Be glycine or do not exist with amino acid c '; Wherein CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '; The described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.

In certain embodiments, the sequence that polynucleotide comprise the CDR2a that encodes, wherein CDR2a comprises aminoacid sequence m n o p q r s t u v w x y z a ' b ' c ', and wherein amino acid m is selected from tryptophane, tyrosine, Histidine, α-amino-isovaleric acid, L-glutamic acid or Serine; Amino acid n is selected from methionine(Met) or Isoleucine; Amino acid o is selected from l-asparagine, tyrosine, Serine, tryptophane or Histidine; Amino acid p is selected from proline(Pro), tyrosine, Serine, arginine, Histidine or l-asparagine; Amino acid q is selected from l-asparagine, Serine or aspartic acid; Amino acid r is selected from Serine or glycine; Amino acid s is selected from aspartic acid, Serine, Threonine or arginine; Amino acid t is selected from l-asparagine, Threonine, L-Ala, Isoleucine or tyrosine; Amino acid u is selected from Threonine, tyrosine, leucine, Methionin, l-asparagine or Isoleucine; Amino acid v is selected from glycine, tyrosine, aspartic acid or halfcystine; Amino acid w is selected from tyrosine or l-asparagine; Amino acid x is selected from L-Ala or proline(Pro); Amino acid y is selected from glutamine, Serine or aspartic acid; Amino acid z is selected from Methionin, leucine or Serine; Amino acid a ' is selected from phenylalanine, Methionin or α-amino-isovaleric acid; Amino acid b ' is selected from glutamine, Serine or Methionin; Be glycine or do not exist with amino acid c '.

In certain embodiments, the sequence that polynucleotide comprise the CDR3a that encodes, described CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least 2 complementary determining regions (CDRs) that are selected from CDR1a, CDR2a and CDR3a, and the described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises CDR1a, CDR2a and CDR3a, and the described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises antibody heavy chain variable region.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises the human antibody heavy chain variable region.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises CH.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises people's CH.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises the IDNO:2 as SEQ; SEQ ID NO:4, SEQ ID NO:6; SEQ ID NO:8, SEQ ID NO:10; SEQ ID NO:12, SEQ ID NO:14; SEQ ID NO:16, SEQ ID NO:18; Aminoacid sequence shown in SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32 or SEQ ID NO:34.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises inhuman CH.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the CH that described polypeptide comprises the kind except the people.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises and is selected from following at least one complementary determining region (CDR): the amino acid 26-35 of SEQ ID NO:2; The amino acid 50-66 of SEQ ID NO:2; The amino acid 99-110 of SEQ ID NO:2; The amino acid 26-37 of SEQ ID NO:4; The amino acid 52-67 of SEQ ID NO:4; The amino acid/11 00-109 of SEQ ID NO:4; The amino acid 26-37 of SEQ ID NO:6; The amino acid 52-67 of SEQ ID NO:6; The amino acid/11 00-109 of SEQ ID NO:6; The amino acid 26-37 of SEQ ID NO:8; The amino acid 52-67 of SEQ ID NO:8; The amino acid/11 00-109 of SEQ ID NO:8; The amino acid 26-35 of SEQ ID NO:10; The amino acid 50-66 of SEQ ID NO:10; The amino acid 99-110 of SEQ ID NO:10; The amino acid 26-35 of SEQ ID NO:12; The amino acid 50-66 of SEQ ID NO:12; The amino acid 99-111 of SEQ ID NO:12; The amino acid 26-35 of SEQ ID NO:14; The amino acid 50-65 of SEQ ID NO:14; The amino acid 98-111 of SEQ ID NO:14; The amino acid 26-37 of SEQ ID NO:16; The amino acid 52-67 of SEQ ID NO:16; The amino acid/11 00-109 of SEQ ID NO:16; The amino acid 26-35 of SEQ ID NO:18; The amino acid 50-66 of SEQ ID NO:18; The amino acid 99-105 of SEQ ID NO:18; The amino acid 26-35 of SEQ ID NO:20; The amino acid 50-66 of SEQ ID NO:20; The amino acid 99-118 of SEQ ID NO:20; The amino acid 26-35 of SEQ ID NO:22; The amino acid 50-66 of SEQ ID NO:22; The amino acid 99-118 of SEQ ID NO:22; The amino acid 26-35 of SEQ ID NO:24; The amino acid 50-65 of SEQ ID NO:24; The amino acid 98-108 of SEQ ID NO:24; The amino acid 26-35 of SEQ ID NO:26; The amino acid 50-66 of SEQ ID NO:26; The amino acid 99-110 of SEQ ID NO:26; The amino acid 26-35 of SEQ ID NO:28; The amino acid 50-66 of SEQ ID NO:28; The amino acid 99-117 of SEQ ID NO:28; The amino acid 26-37 of SEQ ID NO:30; The amino acid 52-67 of SEQ ID NO:30; The amino acid/11 00-111 of SEQ ID NO:30; The amino acid 26-37 of SEQ ID NO:32; The amino acid 52-67 of SEQ ID NO:32; The amino acid/11 00-111 of SEQ ID NO:32; The amino acid 26-37 of SEQ ID NO:34; The amino acid 52-67 of SEQ ID NO:34; With the amino acid/11 00-111 of SEQ ID NO:34, the described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least 2 CDRs in SEQ IDNOS.2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32 or 34.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least 3 CDRs in SEQ ID NOS.2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32 or 34.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:2, SEQ ID NO:2 and the amino acid 99-110 of SEQ ID NO:2.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQ ID NO:4, SEQ ID NO:4 and the amino acid/11 00-109 of SEQ ID NO:4.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQID NO:6, SEQ ID NO:6 and the amino acid/11 00-109 of SEQ IDNO:6.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQ ID NO:8, SEQ ID NO:8 and the amino acid/11 00-109 of SEQ ID NO:8.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:10, SEQ ID NO:10 and the amino acid 99-110 of SEQ ID NO:10.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:12, SEQ ID NO:12 and the amino acid 99-111 of SEQ ID NO:12.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-65 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:14, SEQ ID NO:14 and the amino acid 98-111 of SEQ ID NO:14.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQ ID NO:16, SEQ ID NO:16 and the amino acid/11 00-109 of SEQ ID NO:16.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:18, SEQ ID NO:18 and the amino acid 99-105 of SEQ ID NO:18.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:20, SEQ ID NO:20 and the amino acid 99-118 of SEQ ID NO:20.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:22, SEQ ID NO:22 and the amino acid 99-118 of SEQ ID NO:22.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-65 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:24, SEQ ID NO:24 and the amino acid 98-108 of SEQ ID NO:24.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:26, SEQ ID NO:26 and the amino acid 99-110 of SEQ ID NO:26.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-66 of the amino acid 26-35 that described polypeptide comprises SEQ ID NO:28, SEQ ID NO:28 and the amino acid 99-117 of SEQ ID NO:28.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQ ID NO:30, SEQ ID NO:30 and the amino acid/11 00-111 of SEQ ID NO:30.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQ ID NO:32, SEQ ID NO:32 and the amino acid/11 00-111 of SEQ ID NO:32.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 52-67 of the amino acid 26-37 that described polypeptide comprises SEQ ID NO:34, SEQ ID NO:34 and the amino acid/11 00-111 of SEQ ID NO:34.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b, wherein CDR1b comprises aminoacid sequence a1 b1 c1 d1 e1 f1 g1 h1 i1 j1 k1 l1 m1 n1 o1p1 q1, and wherein amino acid a1 is selected from arginine or Methionin; Amino acid b1 is selected from Threonine, L-Ala or Serine; Amino acid c1 is Serine; Amino acid d1 is glutamine; Amino acid e 1 is selected from Serine or glycine; Amino acid f1 is selected from Isoleucine, leucine or α-amino-isovaleric acid; Amino acid g1 is selected from Serine, leucine or arginine; Amino acid h1 is selected from Threonine, Serine, Isoleucine, l-asparagine, arginine, Histidine or tyrosine; Amino acid i1 is selected from tyrosine, arginine, tryptophane, aspartic acid or Serine; J1 is selected from leucine, Isoleucine, l-asparagine, tyrosine or Serine; Amino acid k1 is selected from l-asparagine, glycine, α-amino-isovaleric acid, L-Ala or leucine; Amino acid l1 is selected from tyrosine, L-Ala or l-asparagine or does not exist; Amino acid m1 is selected from l-asparagine or Methionin or does not exist; Amino acid n1 is selected from tyrosine, l-asparagine or Isoleucine or does not exist; Amino acid o1 is selected from leucine or tyrosine or does not exist; Amino acid p1 is selected from aspartic acid or leucine or does not exist; Be selected from α-amino-isovaleric acid, L-Ala or Threonine or do not exist with amino acid q 1; Wherein CDR2b comprises aminoacid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from L-Ala, aspartic acid, leucine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid s1 is selected from Threonine, α-amino-isovaleric acid, glycine or L-Ala; Amino acid t1 is Serine; Amino acid u1 is selected from Serine, l-asparagine or Threonine; Amino acid v1 is selected from leucine, phenylalanine or arginine; Amino acid w1 is selected from glutamine, L-Ala or L-glutamic acid; Be selected from Serine, arginine or Threonine with amino acid x1; Wherein CDR3b comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; The described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least 2 complementary determining regions (CDRs) that are selected from CDR1b, CDR2b and CDR3b, and the described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises CDR1b, CDR2b and CDR3b, and the described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises antibody chain variable region.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises the human antibody light chain variable region.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises constant region of light chain.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises people's constant region of light chain.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises the aminoacid sequence as shown in SEQ IDNO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ IDNO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ IDNO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ IDNO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66 or SEQ IDNO:68.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises inhuman constant region of light chain.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the constant region of light chain that described polypeptide comprises the kind except the people.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises and is selected from following at least one complementary determining region (CDR): the amino acid 24-34 of SEQ ID NO:36; The amino acid 50-56 of SEQ ID NO:36; The amino acid 89-97 of SEQ ID NO:36; The amino acid 24-34 of SEQ ID NO:38; The amino acid 50-56 of SEQ ID NO:38; The amino acid 89-97 of SEQ ID NO:38; The amino acid 24-34 of SEQ ID NO:40; The amino acid 50-56 of SEQ ID NO:40; The amino acid 89-97 of SEQ ID NO:40; The amino acid 24-34 of SEQ ID NO:42; The amino acid 50-56 of SEQ ID NO:42; The amino acid 89-97 of SEQ ID NO:42; The amino acid 24-34 of SEQ ID NO:44; The amino acid 50-56 of SEQ ID NO:44; The amino acid 89-97 of SEQ ID NO:44; The amino acid 24-34 of SEQ ID NO:46; The amino acid 50-56 of SEQ ID NO:46; The amino acid 89-97 of SEQ ID NO:46; The amino acid 24-40 of SEQ ID NO:48; The amino acid 56-62 of SEQ ID NO:48; The amino acid 95-103 of SEQ ID NO:48; The amino acid 24-39 of SEQ ID NO:50; The amino acid 55-61 of SEQ ID NO:50; The amino acid 94-102 of SEQ ID NO:50; The amino acid 24-40 of SEQ ID NO:52; The amino acid 56-62 of SEQ ID NO:52; The amino acid 95-103 of SEQ ID NO:52; The amino acid 24-34 of SEQ ID NO:54; The amino acid 50-56 of SEQ ID NO:54; The amino acid 89-97 of SEQ ID NO:54; The amino acid 24-34 of SEQ ID NO:56; The amino acid 50-56 of SEQ ID NO:56; The amino acid 89-97 of SEQ ID NO:56; The amino acid 24-40 of SEQ ID NO:58; The amino acid 56-62 of SEQ ID NO:58; The amino acid 95-103 of SEQ ID NO:58; The amino acid 24-34 of SEQ ID NO:60; The amino acid 50-56 of SEQ ID NO:60; The amino acid 89-97 of SEQ ID NO:60; The amino acid 24-34 of SEQ ID NO:62; The amino acid 50-56 of SEQ ID NO:62; The amino acid 89-97 of SEQ ID NO:62; The amino acid 24-35 of SEQ ID NO:64; The amino acid 51-57 of SEQ ID NO:64; The amino acid 90-88 of SEQ ID NO:64; The amino acid 24-34 of SEQ ID NO:66; The amino acid 50-57 of SEQ ID NO:66; The amino acid 89-97 of SEQ ID NO:66; The amino acid 24-34 of SEQ ID NO:68; The amino acid 50-56 of SEQ ID NO:68; Amino acid 89-97 with SEQ ID NO:68; The described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least 2 CDRs in SEQ IDNOS.36,38,40,42,44,46,48,50,52,54,56,58,60,62,64 or 68.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, described polypeptide comprises at least 3 CDRs in SEQ ID NOS.36,38,40,42,44,46,48,50,52,54,56,58,60,62,64 or 68.

In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:36, SEQ ID NO:36 and the amino acid 89-97 of SEQ ID NO:36.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:38, SEQ ID NO:38 and the amino acid 89-97 of SEQ ID NO:38.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQID NO:40, SEQ ID NO:40 and the amino acid 89-97 of SEQID NO:40.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 89-97 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:42, the amino acid 50-56 of SEQ IDNO:42 and SEQ ID NO:42.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ IDNO:44, SEQ ID NO:44 and the amino acid 89-97 of SEQ IDNO:44.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:46, SEQ ID NO:46 and the amino acid 89-97 of SEQ ID NO:46.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 56-62 of the amino acid 24-40 that described polypeptide comprises SEQ ID NO:48, SEQ ID NO:48 and the amino acid 95-103 of SEQ ID NO:48.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 55-61 of the amino acid 24-39 that described polypeptide comprises SEQ ID NO:50, SEQ ID NO:50 and the amino acid 94-102 of SEQ ID NO:50.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 56-62 of the amino acid 24-40 that described polypeptide comprises SEQ ID NO:52, SEQ ID NO:52 and the amino acid 95-103 of SEQ ID NO:52.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the 24-34 that described polypeptide comprises SEQ ID NO:54, SEQ ID NO:54 and the amino acid 89-97 of SEQ ID NO:54.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:56, SEQ ID NO:56 and the amino acid 89-97 of SEQ ID NO:56.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 56-62 of the amino acid 24-40 that described polypeptide comprises SEQ ID NO:58, SEQ ID NO:58 and the amino acid 95-103 of SEQ ID NO:58.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:60, SEQ ID NO:60 and the amino acid 89-97 of SEQ ID NO:60.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:62, SEQ ID NO:62 and the amino acid 89-97 of SEQ ID NO:62.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 51-57 of the amino acid 24-35 that described polypeptide comprises SEQ ID NO:64, SEQ ID NO:64 and the amino acid 90-88 of SEQ ID NO:64.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-57 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:66, SEQ ID NO:66 and the amino acid 89-97 of SEQ ID NO:66.In certain embodiments, the sequence that polynucleotide comprise coded polypeptide, the amino acid 50-56 of the amino acid 24-34 that described polypeptide comprises SEQ ID NO:68, SEQ ID NO:68 and the amino acid 89-97 of SEQ ID NO:68.

In certain embodiments, the application has discussed encoding antibody heavily and some polynucleotide of light chain.In certain embodiments, the application has discussed some polynucleotide of encoding antibody variable region of heavy chain.In certain embodiments, the application has discussed some polynucleotide of encoding human antibody heavy chain variable region.In certain embodiments, the application has discussed some polynucleotide of encoding antibody variable region of light chain.In certain embodiments, the application has discussed some polynucleotide of encoding human antibody chain variable region.In certain embodiments, the application has discussed some polynucleotide of encoding antibody CH.In certain embodiments, the application has discussed some polynucleotide of encoding human heavy chain of antibody constant region.In certain embodiments, the application has discussed some polynucleotide of the heavy chain of antibody constant region of the kind of coding except the people.In certain embodiments, the application has discussed some polynucleotide of encoding antibody constant region of light chain.In certain embodiments, the application has discussed some polynucleotide of encoding human antibody light chain constant region.In certain embodiments, the application has discussed some polynucleotide of the antibody light chain constant region of the kind of coding except the people.In certain embodiments, the application has discussed some polynucleotide of coding single-chain antibody.

In certain embodiments, these antibody weights and light chain polynucleotide and polypeptide are that people's antibody weighs and light chain polynucleotide and polypeptide.In certain embodiments, polynucleotide comprise the nucleotide sequence as shown in SEQ IDNOS.SEQ ID NOs:1,3,5,7,9,11,13,15,17,19,21,23,25,27,29,31,33,35,37,39,41,43,45,47,49,51,53,55,57,59,61,63,65 or 67.In certain embodiments, polynucleotide comprise such nucleotide sequence, and it has one or more disappearances, interpolation and/or the displacement of one or more Nucleotide of those sequences.In certain embodiments, the nucleotide sequence that polynucleotide comprise encoding amino acid sequence, described aminoacid sequence comprises the aminoacid sequence as shown in SEQID NOS:2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32,34,36,38,40,42,44,46,48,50,52,54,56,58,60,62,64,66 or 68.In certain embodiments, provide and comprised for example variable region sequences of CDR1 to CDR3 of complementary determining region (CDRs).In certain embodiments, variable region polynucleotide and polypeptide are people variable region polynucleotide and polypeptide.

Term " naturally occurring Nucleotide " comprises deoxyribonucleotide and ribonucleotide.Deoxyribonucleotide includes but not limited to, adenosine, guanine, cytosine(Cyt) and thymidine.Ribonucleotide includes but not limited to, adenosine, cytosine(Cyt), thymidine and uridylic.Term " Nucleotide of modification " includes but not limited to, has the Nucleotide of glycosyl group modification or displacement etc.Term " polynucleotide key " includes but not limited to, polynucleotide key such as thiophosphatephosphorothioate, phosphorodithioate, seleno phosphoric acid ester, two seleno phosphoric acid ester, phosphoroanilothioate, phoshoraniladate, phosphoramidate etc.Referring to, for example, the people such as LaPlanche, Nucl.Acids Res.14:9081 (1986); The people such as Stec, J.Am.Chem.Soc.106:6077 (1984); The people such as Stein, Nucl.AcidsRes.16:3209 (1988); The people such as Zon, Anti-Cancer Drug Design 6:539 (1991); The people such as Zon, Oligonucleotides and Analogues:A Practical Approach, 87-108 page (F.Eckstein, editor, Oxford University Press, Oxford England (1991)); The people such as Stec, U.S. Patent number 5,151,510; Uhlmann and Peyman Chemical Reviews90:543 (1990).In certain embodiments, polynucleotide can comprise for detection of mark.

Term " isolated polypeptide " refers to any polypeptide, its (1) is not containing its common found at least some protein therewith, (2) be substantially free of from identical source for example from other protein of identical type, (3) by from different types of cell expressing, or (4) do not exist at occurring in nature.

Term " polypeptide ", " peptide " and " protein " are used interchangeably in this article, and refer to by the peptide bond of peptide bond or modification interconnected 2 or the more polymkeric substance of amino acids, i.e. peptide isostere.This term application is in comprising naturally occurring amino acid whose aminoacid polymers, and the aminoacid polymers of the naturally occurring amino acid of wherein one or more amino-acid residue right and wrong or naturally occurring amino acid whose chemical analog.Aminoacid polymers can comprise for example translates by one or more natural processes one or more amino-acid residues that post-treatment is modified, and/or one or more amino-acid residues of having been modified by one or more chemical modification technologies known in the art.

" fragment " of reference polypeptide refers to the amino acid adjacent segments from any part of reference polypeptide.Fragment can be any length that is less than reference polypeptide length.

" variant " of reference polypeptide refers to respect to reference polypeptide to have the polypeptide of one or more amino-acid substitutions, deletion or insertion.In certain embodiments, the variant of reference polypeptide has the posttranslational modification site (that is, glycosylation site) of change.In certain embodiments, the variant of reference polypeptide and reference polypeptide is all specific-binding agent.In certain embodiments, the variant of reference polypeptide and reference polypeptide is all antibody.

The variant of reference polypeptide includes but not limited to, glycosylation variants.Glycosylation variants comprises and reference polypeptide comparison, the wherein reformed variant of the number of glycosylation site and/or type.In certain embodiments, the glycosylation variants of reference polypeptide comprises the N linked glycosylation site than the more or less number of reference polypeptide.In certain embodiments, N linked glycosylation site is characterised in that sequence A sn-X-Ser or Asn-X-Thr, and the amino-acid residue of wherein being appointed as X can be any amino-acid residue except proline(Pro).In certain embodiments, the rearrangement that the glycosylation variants of reference polypeptide comprises N connection carbohydrate chain, wherein one or more N linked glycosylation sites (be generally naturally occurring those) is eliminated, and one or more new N connection site is produced.

The variant of reference polypeptide includes but not limited to, the halfcystine variant.In certain embodiments, the halfcystine variant comprises such variant, and wherein one or more cysteine residues of reference polypeptide are replaced by one or more non-cysteine residues; And/or the one or more non-cysteine residues of reference polypeptide is replaced by one or more cysteine residues.In certain embodiments, the halfcystine variant may be useful, when specific polypeptide must refolding becomes the biological activity conformation, for example, after insoluble occlusion body separates.In certain embodiments, the halfcystine variant of reference polypeptide has than reference polypeptide cysteine residues still less.In certain embodiments, the halfcystine variant of reference polypeptide has the halfcystine of even number, so that the interaction caused by azygous halfcystine drops to is minimum.In certain embodiments, the halfcystine variant has than the more cysteine residues of natural protein.

" derivative " of reference polypeptide refers to: polypeptide: (1) has one or more modifications of one or more amino-acid residues of reference polypeptide; And/or (2) wherein one or more peptide base keies are replaced with one or more non-peptide base keies; And/or (3) wherein N-terminal and/or C-terminal are modified.Some exemplary modification includes but not limited to, acetylize, acidylate, the ADP-ribosylation, amidation, biotinylation, the covalent attachment of riboflavin, the covalent attachment of heme moiety, the covalent attachment of Nucleotide or nucleotide derivative, the covalent attachment of lipid or lipid derivate, the covalent attachment of phosphatidylinositols, crosslinked, cyclisation, disulfide linkage forms, demethylation, the formation of covalent cross-linking, the formation of Gelucystine, the formation of pyroglutamate, formyloxy, gamma-carboxylation, glycosylation, the GPI anchor forms, hydroxylation, iodate, methylate, myristoylation, oxidation, proteolysis processing, phosphorylation, isoprenylation, racemization, selenizing (selenoylation), sulfation, the amino acid of transfer RNA (tRNA) mediation is to the interpolation of protein for example arginyl and ubiquitin.In certain embodiments, the derivative of reference polypeptide and reference polypeptide is all specific-binding agent.In certain embodiments, the derivative of reference polypeptide and reference polypeptide is all antibody.

Polypeptide includes but not limited to, the aminoacid sequence of for example translating post-treatment or modifying by chemical modification technology well-known in the art by natural process.In certain embodiments, the generation Anywhere that modification can be in polypeptide, comprise peptide main chain, amino acid side chain and amino or C-terminal.In some this type of embodiment, modification can be present in identical or different degree several site of given polypeptide.In certain embodiments, given polypeptide comprises is permitted eurypalynous modification, for example one or more amino acid whose deletion, interpolation and/or the displacement of native sequences.In certain embodiments, polypeptide can be branch and/or ring-type.The ring type polypeptide of ring-type, branch and branch can be produced by the natural process (including but not limited to ubiquitin) after translation, maybe can be prepared by synthetic method.As described below, term " polypeptide " also comprises such sequence, the heavy chain that it comprises antibody and/or the aminoacid sequence of light chain, described antibody is selected from Ab A, Ab B, Ab C, Ab D, Ab E, Ab F, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, Ab P and Ab Q (referring to SEQ ID NOS:2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32,34,36,38,40,42,44,46,48,50,52,54,56,58,60,62,64,66 and 68).Term " polypeptide " also comprises the one or more amino acid whose sequence that one or more are deleted, add and/or replace with those sequences.In certain embodiments, some peptide sequence comprises at least one complementary determining region (CDR).

In certain embodiments, polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a, wherein CDR1a comprises aminoacid sequence a b c d e f gh i j k l, wherein amino acid a is glycine, and amino acid b is selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l; Wherein CDR2a comprises aminoacid sequence m n o p q r s t u v w x y z a ' b ' c ', and wherein amino acid m is selected from tryptophane, tyrosine, Histidine, α-amino-isovaleric acid, L-glutamic acid or Serine; Amino acid n is selected from methionine(Met) or Isoleucine; Amino acid o is selected from l-asparagine, tyrosine, Serine, tryptophane or Histidine; Amino acid p is selected from proline(Pro), tyrosine, Serine, arginine, Histidine or l-asparagine; Amino acid q is selected from l-asparagine, Serine or aspartic acid; Amino acid r is selected from Serine or glycine; Amino acid s is selected from aspartic acid, Serine, Threonine or arginine; Amino acid t is selected from l-asparagine, Threonine, L-Ala, Isoleucine or tyrosine; Amino acid u is selected from Threonine, tyrosine, leucine, Methionin, l-asparagine or Isoleucine; Amino acid v is selected from glycine, tyrosine, aspartic acid or halfcystine; Amino acid w is selected from tyrosine or l-asparagine; Amino acid x is selected from L-Ala or proline(Pro); Amino acid y is selected from glutamine, Serine or aspartic acid; Amino acid z is selected from Methionin, leucine or Serine; Amino acid a ' is selected from phenylalanine, Methionin or α-amino-isovaleric acid; Amino acid b ' is selected from glutamine, Serine or Methionin; Be glycine or do not exist with amino acid c '; Wherein CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '; The described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.

In certain embodiments, polypeptide comprises at least 2 complementary determining regions (CDR) that are selected from CDR1a, CDR2a and CDR3a, and the described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.In certain embodiments, polypeptide comprises CDR1a, CDR2a and CDR3a, and the described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.

In certain embodiments, polypeptide comprises antibody heavy chain variable region.In certain embodiments, polypeptide comprises the human antibody heavy chain variable region.In certain embodiments, polypeptide comprises CH.In certain embodiments, polypeptide comprises people's CH.In certain embodiments, polypeptide comprises the aminoacid sequence as shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32 or SEQ ID NO:34.In certain embodiments, polypeptide comprises inhuman CH.In certain embodiments, the CH that polypeptide comprises the kind except the people.

In certain embodiments, polypeptide comprises and is selected from following at least one complementary determining region (CDR): the amino acid 26-35 of SEQ ID NO:2; The amino acid 50-66 of SEQ ID NO:2; The amino acid 99-110 of SEQ ID NO:2; The amino acid 26-37 of SEQ ID NO:4; The amino acid 52-67 of SEQ ID NO:4; The amino acid/11 00-109 of SEQ ID NO:4; The amino acid 26-37 of SEQ ID NO:6; The amino acid 52-67 of SEQ ID NO:6; The amino acid/11 00-109 of SEQ ID NO:6; The amino acid 26-37 of SEQ ID NO:8; The amino acid 52-67 of SEQ ID NO:8; The amino acid/11 00-109 of SEQ ID NO:8; The amino acid 26-35 of SEQ ID NO:10; The amino acid 50-66 of SEQ ID NO:10; The amino acid 99-110 of SEQ ID NO:10; The amino acid 26-35 of SEQ ID NO:12; The amino acid 50-66 of SEQ ID NO:12; The amino acid 99-111 of SEQ ID NO:12; The amino acid 26-35 of SEQ ID NO:14; The amino acid 50-65 of SEQ ID NO:14; The amino acid 98-111 of SEQ ID NO:14; The amino acid 26-37 of SEQ ID NO:16; The amino acid 52-67 of SEQ ID NO:16; The amino acid/11 00-109 of SEQ ID NO:16; The amino acid 26-35 of SEQ ID NO:18; The amino acid 50-66 of SEQ ID NO:18; The amino acid 99-105 of SEQ ID NO:18; The amino acid 26-35 of SEQ ID NO:20; The amino acid 50-66 of SEQ ID NO:20; The amino acid 99-118 of SEQ ID NO:20; The amino acid 26-35 of SEQ ID NO:22; The amino acid 50-66 of SEQ ID NO:22; The amino acid 99-118 of SEQ ID NO:22; The amino acid 26-35 of SEQ ID NO:24; The amino acid 50-65 of SEQ ID NO:24; The amino acid 98-108 of SEQ ID NO:24; The amino acid 26-35 of SEQ ID NO:26; The amino acid 50-66 of SEQ ID NO:26; The amino acid 99-110 of SEQ ID NO:26; The amino acid 26-35 of SEQ ID NO:28; The amino acid 50-66 of SEQ ID NO:28; The amino acid 99-117 of SEQ ID NO:28; The amino acid 26-37 of SEQ ID NO:30; The amino acid 52-67 of SEQ ID NO:30; The amino acid/11 00-111 of SEQ ID NO:30; The amino acid 26-37 of SEQ ID NO:32; The amino acid 52-67 of SEQ ID NO:32; The amino acid/11 00-111 of SEQ ID NO:32; The amino acid 26-37 of SEQ ID NO:34; The amino acid 52-67 of SEQ ID NO:34; Amino acid/11 00-111 with SEQ ID NO:34; The described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.In certain embodiments, polypeptide comprises at least 2 CDRs in SEQID NOS.2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32 or 34.In certain embodiments, polypeptide comprises at least 3 CDRs in SEQ ID NOS.2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32 or 34.

In certain embodiments, the amino acid 99-110 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:2, the amino acid 50-66 of SEQ ID NO:2 and SEQ ID NO:2.In certain embodiments, the amino acid 52-67 of polypeptide comprises SEQ ID NO:4 amino acid 26-37, SEQ IDNO:4 and the amino acid/11 00-109 of SEQ ID NO:4.In certain embodiments, the amino acid/11 00-109 of the amino acid 26-37 that polypeptide comprises SEQ ID NO:6, the amino acid 52-67 of SEQ ID NO:6 and SEQ ID NO:6.In certain embodiments, the amino acid/11 00-109 of the amino acid 26-37 that polypeptide comprises SEQ ID NO:8, the amino acid 52-67 of SEQ ID NO:8 and SEQ ID NO:8.In certain embodiments, the amino acid 99-110 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:10, the amino acid 50-66 of SEQ ID NO:10 and SEQ ID NO:10.In certain embodiments, the amino acid 99-111 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:12, the amino acid 50-66 of SEQ ID NO:12 and SEQ ID NO:12.In certain embodiments, the amino acid 98-111 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:14, the amino acid 50-65 of SEQ ID NO:14 and SEQ ID NO:14.In certain embodiments, the amino acid/11 00-109 of the amino acid 26-37 that polypeptide comprises SEQ ID NO:16, the amino acid 52-67 of SEQ ID NO:16 and SEQ ID NO:16.In certain embodiments, the amino acid 99-105 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:18, the amino acid 50-66 of SEQ ID NO:18 and SEQ ID NO:18.In certain embodiments, the amino acid 99-118 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:20, the amino acid 50-66 of SEQ ID NO:20 and SEQ ID NO:20.In certain embodiments, the amino acid 99-118 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:22, the amino acid 50-66 of SEQ ID NO:22 and SEQ ID NO:22.In certain embodiments, the amino acid 98-108 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:24, the amino acid 50-65 of SEQ ID NO:24 and SEQ ID NO:24.In certain embodiments, the amino acid 99-110 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:26, the amino acid 50-66 of SEQ ID NO:26 and SEQ ID NO:26.In certain embodiments, the amino acid 99-117 of the amino acid 26-35 that polypeptide comprises SEQ ID NO:28, the amino acid 50-66 of SEQ ID NO:28 and SEQ ID NO:28.In certain embodiments, the amino acid/11 00-111 of the amino acid 26-37 that polypeptide comprises SEQ ID NO:30, the amino acid 52-67 of SEQ ID NO:30 and SEQ ID NO:30.In certain embodiments, the amino acid/11 00-111 of the amino acid 26-37 that polypeptide comprises SEQ ID NO:32, the amino acid 52-67 of SEQ ID NO:32 and SEQ ID NO:32.In certain embodiments, the amino acid/11 00-111 of the amino acid 26-37 that polypeptide comprises SEQ ID NO:34, the amino acid 52-67 of SEQ ID NO:34 and SEQ ID NO:34.

In certain embodiments, polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b, wherein CDR1b comprises aminoacid sequence a1 b1 c1 d1e1 f1 g1 h1 i1 j1 k1 l1 m1 n1 o1 p1 q1, and wherein amino acid a1 is selected from arginine or Methionin; Amino acid b1 is selected from Threonine, L-Ala or Serine; Amino acid c1 is Serine; Amino acid d1 is glutamine; Amino acid e1 is selected from Serine or glycine; Amino acid f1 is selected from Isoleucine, leucine or α-amino-isovaleric acid; Amino acid g1 is selected from Serine, leucine or arginine; Amino acid h1 is selected from Threonine, Serine, Isoleucine, l-asparagine, arginine, Histidine or tyrosine; Amino acid i1 is selected from tyrosine, arginine, tryptophane, aspartic acid or Serine; J1 is selected from leucine, Isoleucine, l-asparagine, tyrosine or Serine; Amino acid k1 is selected from l-asparagine, glycine, α-amino-isovaleric acid, L-Ala or leucine; Amino acid l1 is selected from tyrosine, L-Ala or l-asparagine or does not exist; Amino acid m1 is selected from l-asparagine or Methionin or does not exist; Amino acid n1 is selected from tyrosine, l-asparagine or Isoleucine or does not exist; Amino acid o1 is selected from leucine or tyrosine or does not exist; Amino acid p1 is selected from aspartic acid or leucine or does not exist; Be selected from α-amino-isovaleric acid, L-Ala or Threonine or do not exist with amino acid q1; Wherein CDR2b comprises aminoacid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from L-Ala, aspartic acid, leucine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid s1 is selected from Threonine, α-amino-isovaleric acid, glycine or L-Ala; Amino acid t1 is Serine; Amino acid u1 is selected from Serine, l-asparagine or Threonine; Amino acid v1 is selected from leucine, phenylalanine or arginine; Amino acid w1 is selected from glutamine, L-Ala or L-glutamic acid; Be selected from Serine, arginine or Threonine with amino acid x1; Wherein CDR3b comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; The described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

In certain embodiments, polypeptide comprises at least 2 complementary determining regions (CDR) that are selected from CDR1b, CDR2b and CDR3b, and the described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.In certain embodiments, polypeptide comprises CDR1b, CDR2b and CDR3b, and the described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.

In certain embodiments, polypeptide comprises antibody chain variable region.In certain embodiments, polypeptide comprises the human antibody light chain variable region.In certain embodiments, polypeptide comprises constant region of light chain.In certain embodiments, antibody comprises people's constant region of light chain.In certain embodiments, polypeptide comprises the aminoacid sequence as shown in SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66 or SEQ ID NO:68.In certain embodiments, polypeptide comprises inhuman constant region of light chain.In certain embodiments, the constant region of light chain that polypeptide comprises the kind except the people.

In certain embodiments, polypeptide comprises and is selected from following at least one complementary determining region (CDR): the amino acid 24-34 of SEQ ID NO:36; The amino acid 50-56 of SEQ ID NO:36; The amino acid 89-97 of SEQ ID NO:36; The amino acid 24-34 of SEQ ID NO:38; The amino acid 50-56 of SEQ ID NO:38; The amino acid 89-97 of SEQ ID NO:38; The amino acid 24-34 of SEQ ID NO:40; The amino acid 50-56 of SEQ ID NO:40; The amino acid 89-97 of SEQ ID NO:40; The amino acid 24-34 of SEQ ID NO:42; The amino acid 50-56 of SEQ ID NO:42; The amino acid 89-97 of SEQ ID NO:42; The amino acid 24-34 of SEQ ID NO:44; The amino acid 50-56 of SEQ ID NO:44; The amino acid 89-97 of SEQ ID NO:44; The amino acid 24-34 of SEQ ID NO:46; The amino acid 50-56 of SEQ ID NO:46; The amino acid 89-97 of SEQ ID NO:46; The amino acid 24-40 of SEQ ID NO:48; The amino acid 56-62 of SEQ ID NO:48; The amino acid 95-103 of SEQ ID NO:48; The amino acid 24-39 of SEQ ID NO:50; The amino acid 55-61 of SEQ ID NO:50; The amino acid 94-102 of SEQ ID NO:50; The amino acid 24-40 of SEQ ID NO:52; The amino acid 56-62 of SEQ ID NO:52; The amino acid 95-103 of SEQ ID NO:52; The amino acid 24-34 of SEQ ID NO:54; The amino acid 50-56 of SEQ ID NO:54; The amino acid 89-97 of SEQ ID NO:54; The amino acid 24-34 of SEQ ID NO:56, the amino acid 50-56 of SEQ ID NO:56; The amino acid 89-97 of SEQ ID NO:56; The amino acid 24-40 of SEQ ID NO:58; The amino acid 56-62 of SEQ ID NO:58; The amino acid 95-103 of SEQ ID NO:58; The amino acid 24-34 of SEQ ID NO:60; The amino acid 50-56 of SEQ ID NO:60; The amino acid 89-97 of SEQ ID NO:60; The amino acid 24-34 of SEQ ID NO:62; The amino acid 50-56 of SEQ ID NO:62; The amino acid 89-97 of SEQ ID NO:62; The amino acid 24-35 of SEQ ID NO:64; The amino acid 51-57 of SEQ ID NO:64; The amino acid 90-88 of SEQ ID NO:64; The amino acid 24-34 of SEQ ID NO:66; The amino acid 50-57 of SEQ ID NO:66; The amino acid 89-97 of SEQ ID NO:66; The amino acid 24-34 of SEQ ID NO:68; The amino acid 50-56 of SEQ ID NO:68; Amino acid 89-97 with SEQ ID NO:68; The described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.In certain embodiments, polypeptide comprises at least 2 CDRs in SEQID NOS.36,38,40,42,44,46,48,50,52,54,56,58,60,62,64,66 or 68.In certain embodiments, polypeptide comprises at least 3 CDRs in SEQID NOS.36,38,40,42,44,46,48,50,52,54,56,58,60,62,64,66 or 68.

In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:36, the amino acid 50-56 of SEQ ID NO:36 and SEQ ID NO:36.In certain embodiments, the amino acid 50-56 of polypeptide comprises SEQ ID NO:38 amino acid 24-34, SEQID NO:38 and the amino acid 89-97 of SEQ ID NO:38.In certain embodiments, the amino acid 50-56 of polypeptide comprises SEQ ID NO:40 amino acid 24-34, SEQ IDNO:40 and the amino acid 89-97 of SEQ ID NO:40.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:42, the amino acid 50-56 of SEQ ID NO:42 and SEQ ID NO:42.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:44, the amino acid 50-56 of SEQ ID NO:44 and SEQ ID NO:44.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:46, the amino acid 50-56 of SEQ ID NO:46 and SEQ ID NO:46.In certain embodiments, the amino acid 95-103 of the amino acid 24-40 that polypeptide comprises SEQ ID NO:48, the amino acid 56-62 of SEQ ID NO:48 and SEQ ID NO:48.In certain embodiments, the amino acid 94-102 of the amino acid 24-39 that polypeptide comprises SEQ ID NO:50, the amino acid 55-61 of SEQ ID NO:50 and SEQ ID NO:50.In certain embodiments, the amino acid 95-103 of the amino acid 24-40 that polypeptide comprises SEQ ID NO:52, the amino acid 56-62 of SEQ ID NO:52 and SEQ ID NO:52.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:54, the amino acid 50-56 of SEQ ID NO:54 and SEQ ID NO:54.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:56, the amino acid 50-56 of SEQ ID NO:56 and SEQ ID NO:56.In certain embodiments, the amino acid 95-103 of the amino acid 24-40 that polypeptide comprises SEQ ID NO:58, the amino acid 56-62 of SEQ ID NO:58 and SEQ ID NO:58.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:60, the amino acid 50-56 of SEQ ID NO:60 and SEQ ID NO:60.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:62, the amino acid 50-56 of SEQ ID NO:62 and SEQ ID NO:62.In certain embodiments, the amino acid 90-88 of the amino acid 24-35 that polypeptide comprises SEQ ID NO:64, the amino acid 51-57 of SEQ ID NO:64 and SEQ ID NO:64.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:66, the amino acid 50-57 of SEQ ID NO:66 and SEQ ID NO:66.In certain embodiments, the amino acid 89-97 of the amino acid 24-34 that polypeptide comprises SEQ ID NO:68, the amino acid 50-56 of SEQ ID NO:68 and SEQ ID NO:68.

Term " naturally occurring " means the object that can find at occurring in nature when being applied to object.For example, that can from the source of occurring in nature, separate is present in biology (comprising virus), and not in laboratory or otherwise by manually polypeptide or the polynucleotide of modification are naturally occurring intentionally.

As used herein, term " is operably connected " and refers in allowing the component of the relation that they work in its expection mode.For example, in the background of polynucleotide sequence, when control sequence and encoding sequence interosculate by this way, when making the expression of encoding sequence compatible condition being issued in the function with encoding sequence, control sequence can " be operably connected " with encoding sequence.

Term " control sequence " refers to affect their expression of the encoding sequence of combination and polynucleotide sequences of processing with it.The character of this type of control sequence can be different according to host living beings.Include but not limited to promotor, ribosome bind site and transcription termination sequence about procaryotic some exemplary control sequence.Include but not limited to promotor, enhanser and transcription termination sequence about Eukaryotic some exemplary control sequence.In certain embodiments, " control sequence " can comprise homing sequence and/or fusion partner sequence.

In certain embodiments, when the first and the second polynucleotide encoding sequence are transcribed into single during in abutting connection with mRNA, describedly singlely in abutting connection with mRNA, can be translated into singlely in abutting connection with polypeptide, the first polynucleotide encoding sequence is operably connected with the second polynucleotide encoding sequence.

In the background of polypeptide, if the polypeptide of every kind of connection can work in its expection mode, two or more polypeptide " are operably connected " so.The polypeptide that can work in its expection mode when being operably connected with another kind of polypeptide may be able to or can not work in its expection mode when not being operably connected with another kind of polypeptide.For example, in certain embodiments, when connecting, the first polypeptide may not work in its expection mode, but, by with the second polypeptide, being connected and may being stablized, thereby make it become, can work in its expection mode.Alternately, in certain embodiments, when connecting, the first polypeptide may be able to not work in its expection mode, and may retain that activity when being operably connected with the second polypeptide.

As used herein, single in abutting connection with peptide sequence or single while being connected in abutting connection with peptide sequence by they are synthesized by they are translated into when two or more polypeptide, two or more polypeptide are " fusions ".In certain embodiments, two or more fusion polypeptide can be translated by two or more polynucleotide encoding sequences that are operably connected in vivo.In certain embodiments, two or more fusion polypeptide can be translated by two or more polynucleotide encoding sequences that are operably connected in vitro.

As used herein, if the polypeptide of every kind of connection can work in its expection mode, two or more polypeptide " operationally merge " so.

In certain embodiments, the first polypeptide that comprises 2 or more different polypeptide units is regarded as being connected with the second polypeptide, as long as at least one different polypeptide units of the first polypeptide are connected with the second polypeptide.As non-limitative example, in certain embodiments, in all following situations, antibody is regarded as being connected with the second polypeptide: (a) one of heavy chain polypeptide of the second polypeptide and antibody is connected; (b) one of light chain polypeptide of the second polypeptide and antibody is connected; (c) one of heavy chain polypeptide of first molecule of the second polypeptide and antibody is connected, and with one of light chain polypeptide of second molecule of the second and antibody, is connected; (d) first of first and second molecule and antibody of the second polypeptide is connected with second heavy chain polypeptide, and the 3rd of the second polypeptide is connected with second light chain polypeptide with first of the 4th molecule and antibody.

In certain embodiments, word " the first polypeptide is connected with the second polypeptide " comprises following situation: (a) unique one minute sub-connection of unique molecule of the first polypeptide and the second polypeptide; (b) unique molecule of the first polypeptide and the second polypeptide surpasses a minute sub-connection; (c) unique one minute sub-connection that surpasses a molecule and the second polypeptide of the first polypeptide; (d) minute sub-connection that surpasses that surpasses a molecule and the second polypeptide of the first polypeptide.In certain embodiments, when unique one minute period of the day from 11 p.m. to 1 a.m that surpasses a molecule and the second polypeptide that connects molecule and comprise the first polypeptide, the first polypeptide all or be less than all molecules can with the second polypeptid covalence or non-covalent the connection.In certain embodiments, when minute period of the day from 11 p.m. to 1 a.m that surpasses that the connection molecule comprises the first polypeptide, one or more molecules of the first polypeptide can covalently or non-covalently be connected with other molecules of the first polypeptide.

As used herein, " nonrigid connector " refers to according to its chemical structure, and prediction is not fixed on any linker in three-dimensional space.Those skilled in the art can predict whether specific linker is elastic in its expection background.In certain embodiments, comprise 3 or more the peptide linker of amino acids be nonrigid connector.

As used herein, conventional usage is followed in 20 kinds of conventional amino acid and abbreviation thereof.Referring to Immunology--A Synthesis (the 2nd edition, E.S.Golub and D.R.Gren, editor, Sinauer Associates, Sunderland, Mass. (1991)).In certain embodiments, one or more unconventional amino acid can be introduced in polypeptide.Term " unconventional amino acid " refers to not be any amino acid of one of 20 kinds of conventional amino acid.Term " amino acid that non-natural exists " refers to the amino acid can not find at occurring in nature.The amino acid that non-natural exists is unconventional amino acid whose subgroup.Unconventional amino acid includes but not limited to, 20 kinds of amino acid whose steric isomers of routine (for example, D-amino acid), alpha-non-natural amino acid for example α-, α-bis-substituted amino acids, the N-alkyl amino acid, lactic acid, homoserine, same halfcystine, 4-Hydroxyproline, Gla, ε-N, N, the N-trimethyl lysine, ε-N-acetyllysine, the O-phosphoserine, the N-acetylserine, N-formylmethionine, 3-Methyl histidine, the 5-hydroxylysine, σ-N-methylarginine, and other similar amino acid known in the art and imino-acid are (for example, 4-Hydroxyproline).In polypeptide symbol used herein, secundum legem usage and convention, left direction is that N-terminal direction and right direction are the C-terminal directions.

In certain embodiments, conservative amino acid replacement comprises the displacement with one or more unconventional amino-acid residues.In certain embodiments, synthetic by chemical peptide rather than by the unconventional amino-acid residue of synthetic introducing in biosystem.

Term " acidic residues " refers to when introducing in polypeptide between 2 other identical or different amino-acid residues, the amino-acid residue of the D-that comprises at least one acidic-group or L-form.In certain embodiments, acidic residues comprises the side chain that contains at least one acidic-group.Exemplary acid residue includes but not limited to, aspartic acid (D) and L-glutamic acid (E).In certain embodiments, acidic residues can be unconventional amino acid.

The D-that term " aromatic residue " refers to comprise at least one aromatic group or the amino-acid residue of L-form.In certain embodiments, aromatic residue comprises the side chain that contains at least one aromatic group.Exemplary aromatic residue includes but not limited to, phenylalanine (F), tyrosine (Y) and tryptophane (W).In certain embodiments, aromatic residue can be unconventional amino acid.

Term " alkaline residue " refers to can comprise the F-of at least one basic group or the amino-acid residue of L-form when the identical or different one or more amino-acid residues of next-door neighbour are introduced in polypeptide.In certain embodiments, alkaline residue comprises the side chain that contains at least one basic group.Exemplary alkaline residue includes but not limited to, Histidine (H), Methionin (K) and arginine (R).In certain embodiments, alkaline residue can be unconventional amino acid.

Term " neutral hydrophilic residue " refers to comprise at least one wetting ability and/or polar group but do not comprise the D-of acidity or basic group or the amino-acid residue of L-form when the identical or different one or more amino-acid residues of next-door neighbour are introduced in polypeptide.In certain embodiments, the neutral hydrophilic residue comprises the side chain that contains at least one neutral hydrophilic group.Exemplary neutral hydrophilic residue includes but not limited to, L-Ala (A), halfcystine (C), Serine (S), Threonine (T), l-asparagine (N) and glutamine (Q).In certain embodiments, the neutral hydrophilic residue can be unconventional amino acid.

Term " lipotropy residue " and " Laa " refer to have that at least one is uncharged, aliphatics and/or the D-of aromatic group or the amino-acid residue of L-form.In certain embodiments, the lipotropy residue comprise and contain that at least one is uncharged, the side chain of aliphatics and/or aromatic group.Exemplary lipotropy residue includes but not limited to, L-Ala (A), phenylalanine (F), Isoleucine (I), leucine (L), nor-leucine (Nile), methionine(Met) (M), α-amino-isovaleric acid (V), tryptophane (W) and tyrosine (Y).In certain embodiments, the lipotropy residue can be unconventional amino acid.

Term " amphipathic residue " refers to become the D-of wetting ability or lipotropy residue or the amino-acid residue of L-form.Exemplary amphipathic residue includes but not limited to, L-Ala (A).In certain embodiments, amphipathic residue can be unconventional amino acid.

Term " NOT-function residue " refers to lack acidity, alkalescence and the D-of aromatic group or the amino-acid residue of L-form when the identical or different one or more amino-acid residues of next-door neighbour are introduced in polypeptide.In certain embodiments, the NOT-function residue comprises the side chain that contains at least one NOT-function group.Exemplary NOT-function residue includes but not limited to, methionine(Met) (M), glycine (G), L-Ala (A), α-amino-isovaleric acid (V), Isoleucine (I), leucine (L) and nor-leucine (Nle).In certain embodiments, the NOT-function residue can be unconventional amino acid.

In certain embodiments, glycine (G) and proline(Pro) (P) are the considered amino-acid residues that can affect the polypeptide chain direction.

In certain embodiments, conservative substitution can relate to the member who replaces a kind of residue type with the member of identical residue type.As non-limitative example, in certain embodiments, conservative substitution can relate to different acidic residues for example E replace for example D of acidic residues.In certain embodiments, non-conservative displacement can relate to the member who replaces a kind of residue type with the member of different residue types.As non-limitative example, in certain embodiments, non-conservative displacement can relate to alkaline residue for example K replace for example D of acidic residues.In certain embodiments, the another kind of radical amino acid replacement of cysteine residues, with the disulfide linkage formation of that position in prevention and polypeptide.

In the preparation of conservative or non-conservative displacement, according to some embodiment, hydrophilic index that can considered amino acid.Can specify the hydrophilic index of every seed amino acid based on its hydrophobicity and charge characteristic.20 kinds of naturally occurring amino acid whose hydrophilic indexes are: Isoleucine (+4.5); α-amino-isovaleric acid (+4.2); Leucine (+3.8); Phenylalanine (+2.8); Halfcystine/Gelucystine (+2.5); Methionine(Met) (+1.9); L-Ala (+1.8); Glycine (0.4); Threonine (0.7); Serine (0.8); Tryptophane (0.9); Tyrosine (1.3); Proline(Pro) (1.6); Histidine (3.2); L-glutamic acid (3.5); Glutamine (3.5); Aspartic acid (3.5); L-asparagine (3.5); Methionin (3.9); And arginine (4.5).

The importance of hydrophile amino acid number in to protein, giving mutual biological function is known in the art.The people such as Kyte, J.Mol.Biol., 157:105-131 (1982).Known can have with some amino acid substitution other amino acid of similar hydrophilic index or score in some cases, and still retain similar biological activity.In the change of carrying out based on hydrophilic index, in certain embodiments, comprised the amino-acid substitution of its hydrophilic index in ± 2.In certain embodiments, be included in ± those in 1, and in certain embodiments, be included in ± those in 0.5.

This area it should also be understood that similar amino acid whose displacement can carry out effectively based on wetting ability, particularly when consequent biological function protein or peptide expection for when the immunology embodiment is used, as in the situation that this paper.In certain embodiments, as determined by its contiguous amino acid whose wetting ability, the maximum local average wetting ability of protein is relevant to its immunogenicity and antigenicity, relevant to the biological property of polypeptide.

Specified following hydrophilicity value to these amino-acid residues: arginine (+3.0); Methionin (+3.0); Aspartic acid (+3.0 ± 1); L-glutamic acid (+3.0 ± 1); Serine (+0.3); L-asparagine (+0.2); Glutamine (+0.2); Glycine (0); Threonine (0.4); Proline(Pro) (0.5 ± 1); L-Ala (0.5); Histidine (0.5); Halfcystine (1.0); Methionine(Met) (1.3); α-amino-isovaleric acid (1.5); Leucine (1.8); Isoleucine (1.8); Tyrosine (2.3); Phenylalanine (2.5) and tryptophane (3.4).In the change of carrying out at the hydrophilicity value based on similar, in certain embodiments, comprised the amino-acid substitution of its hydrophilicity value in ± 2.In certain embodiments, be included in ± those in 1, and in certain embodiments, be included in ± those in 0.5.In some cases, can also identify the epi-position from primary amino acid sequence based on wetting ability.These zones are also referred to as " epi-position core area ".

Exemplary amino acid displacement is listed in table 1.

Table 1: amino-acid substitution

Original residue	Exemplary displacement	Exemplary displacement more specifically
			Ala	Val，Leu，Ile	Val
Arg	Lys，Gln，Asn	Lys
			Asn	Gln	Gln
Asp	Glu	Glu
			Cys	Ser，Ala	Ser
Gln	Asn	Asn
			Glu	Asp	Asp
Gly	Pro，Ala	Ala
			His	Asn，Gln，Lys，Arg	Arg
Ile	Leu, Val, Met, Ala, Phe, nor-leucine	Leu
			Leu	Nor-leucine, Ile, Val, Met, Ala, Phe	Ile

Original residue	Exemplary displacement	Exemplary displacement more specifically
			Lys	Arg, Isosorbide-5-Nitrae diamino-butyric acid, Gln, Asn	Arg
Met	Leu，Phe，Ile	Leu
			Phe	Leu，Val，Ile，Ala， Tyr	Leu
Pro	Ala	Gly
			Ser	Thr，Ala，Cys	Thr
Thr	Ser	Ser
			Trp	Tyr，Phe	Tyr
Tyr	Trp，Phe，Thr，Ser	Phe
			Val	Ile, Met, Leu, Phe, Ala, nor-leucine	Leu

Similarly, as used herein, except as otherwise noted, the left distal end of strand polynucleotide sequence is 5 ' end; The left direction of double-stranded polynucleotide sequence is called as 5 ' direction.5 ' to 3 ' of nascent RNA transcript adds direction and is called as in this article transcriptional orientation; There is the sequence identical with RNA and be that the sequence area of 5 ' to 5 ' end of rna transcription thing is called as " upstream sequence " in this article on the DNA chain; There is the sequence identical with RNA and be that the sequence area of 3 ' to 3 ' end of rna transcription thing is called as " downstream sequence " in this article on the DNA chain.

In certain embodiments, conservative amino acid replacement comprises the amino-acid residue that non-natural exists, and it is generally synthetic by chemical peptide rather than pass through the synthetic introducing in biosystem.The amino-acid residue that those non-naturals exist includes but not limited to, intends the amino acid moiety of peptide and other reverses or reverse form.

The technician can use well-known technology to determine the suitable displacement variant of reference polypeptide as described herein.In certain embodiments, by target, be considered to the unessential target region of activity, those skilled in the art can identify and can be changed and do not destroy active suitable molecular domains.In certain embodiments, can identify in similar polypeptide it is conservative residue and molecular moiety.In certain embodiments, even can such zone enforcement conservative amino acid replacement not being destroyed to biological activity maybe can not adversely affect polypeptide structure, and described zone may be important to biological activity, includes but not limited to, the CDRs of antibody may be maybe important to structure.

In addition, in certain embodiments, those skilled in the art can summarize structure function research and identify in similar polypeptide activity and/or the important residue of structure.Consider this type of relatively, in certain embodiments, can predict in corresponding similar polypeptide the importance of amino-acid residue in polypeptide to activity or the important amino-acid residue of structure.In certain embodiments, the technician can select chemically similarly amino-acid substitution this type of predicts important amino-acid residue.

In certain embodiments, the technician can also analyze three-dimensional structure and the aminoacid sequence with respect to that structure in similar polypeptide.Consider this type of information, the technician can predict the amino-acid residue comparison of the antibody with regard to its three-dimensional structure.In certain embodiments, the technician can select the amino-acid residue on protein surface to prediction not carry out radical change, because this type of residue may relate to the important interaction with other molecules.In addition, in certain embodiments, the technician can be created in the test variant that each amino acid needed residue place comprises the single amino acids displacement.In certain embodiments, can use subsequently activation measurement screening variant well known by persons skilled in the art.For example, in certain embodiments, the ability screening variant that can be combined with TR-2 with regard to it.In certain embodiments, this type of variant can be for collecting the information about suitable variant.For example, in certain embodiments, if find change for particular amino acid residue causes destroying, reduce or inappropriate activity unwelcomely, can avoid having so the variant of this type of change.In other words, the information based on being collected by this class normal experiment, the technician can easily determine wherein separately or with other sudden change combinations, the amino acid that further displacement should be avoided.

Many scientific publications have been devoted to the prediction of secondary structure.Referring to Moult J., Curr.Op.in Biotech., 7 (4): 422-427 (1996), the people such as Chou, Biochemistry, 13 (2): 222-245 (1974); The people such as Chou, Biochemistry, 113 (2): 211-222 (1974); The people such as Chou, Adv.Enzymol.Relat.Areas Mol.Biol., 47:45-148 (1978); The people such as Chou, Ann.Rev.Biochem., the people such as 47:251-276 and Chou, Biophys.J., 26:367-384 (1979).In addition, computer program can be used for the aid forecasting secondary structure at present.A kind of method of prediction secondary structure is based on the homology modeling.For example, 2 peptide species or the protein that have the sequence identity that is greater than 30% or be greater than 40% similarity have similar topology usually.The immediate development of protein structure database (PDB) provides the secondary structure predictability strengthened, comprises possible folding number in polypeptide or protein structure.Referring to people such as Holm, Nucl.Acid.Res., 27 (1): 244-247 (1999).Have been proposed in given polypeptide or protein that to have limited number folding, and, after the structure of closing number of keys solved, it is obviously more accurate that structure prediction will become.Referring to, for example, the people such as Brenner, Curr.Op.Struct.Biol., 7 (3): 369-376 (1997).

The other illustrative methods of prediction secondary structure includes but not limited to, " wearing silk (threading) " (Jones, D., Curr.Opin.Struct.Biol., 7 (3): 377-87 (1997); The people such as Sippl, Structure, 4 (1): 15-19 (1996)), " signature analysis " (people such as Bowie, Science, 253:164-170 (1991); The people such as Gribskov, Meth.Enzym., 183:146-159 (1990); The people such as Gribskov, Proc.Nat.Acad.Sci., 84 (13): 4355-4358 (1987)), and " contact of evolving " (referring to Holm, the same (1999), and Brenner, the same (1997) .)

In certain embodiments, can easily calculate identity and the similarity of related polypeptide by currently known methods.These class methods include but not limited to, those that describe in following reference: Computational Molecular Biology, Lesk, A.M., editor, OxfordUniversity Press, New York (1988); Biocomputing:Informatics and GenomeProjects, Smith, D.W., editor, Academic Press, New York (1993); ComputerAnalysis of Sequence Data, Part 1, Griffin, A.M. and Grimn, H.G, editor, Humana Press, New Jersey (1994); Sequence Analysis in MolecularBiology, von Heinje, G, Academic Press (1987); Sequence AnalysisPrimer, Gribskov, M. and Devereux, J., editor, M.Stockton Press, NewYork (1991); With the people such as Carillo, SIAM J.Applied Math., 48:1073 (1988).In certain embodiments, polypeptide has and about 90% or approximately 95% or approximately 96% or approximately 97% or approximately 98% or approximately 99% aminoacid sequence be equal to of the aminoacid sequence shown in Fig. 3-19.

In certain embodiments, the method for mensuration identity is designed to produce maximum coupling between the sequence of test.In certain embodiments, some method of measuring identity the public can with computer program in described.Some computer program of measuring the identity between 2 sequences includes but not limited to, the GCG routine package comprises GAP (people such as Devereux, Nucl.Acid.Res., 12:387 (1984); Genetics Computer Group, Universityof Wisconsin, Madison, WI, BLASTP, BLASTN and FASTA (people such as Altschul, J.Mol.Biol., 215:403-410 (1990)).The BLASTX program can be from the state-run bioinformation of U.S. center (National Center for Biotechnology Information) (NCBI) and other resources openly obtain (BLAST Manual, the people such as Altschul, NCB/NLM/NIH Bethesda, MD 20894; The people such as Altschul, the same (1990)).In certain embodiments, Smith Waterman algorithm known in the art also can be used for measuring identity.

Can only produce the short Region Matching of 2 sequences for some comparison scheme of comparing 2 aminoacid sequences, and this little comparison zone can have very high sequence identity, although not significantly association between 2 full length sequences.Therefore, in certain embodiments, selected Comparison Method (GAP program) will cause crossing over the amino acid whose comparison of at least 50 adjacency of target polypeptide.

For example, use computerized algorithm GAP (Genetics Computer Group, Universityof Wisconsin, Madison, WI), 2 polypeptide of sequence identity per-cent to be determined are compared for other amino acid whose optimum matching of its minute (" matched spans ", as measured by algorithm).In certain embodiments, (this is calculated as the average diagonal lines of 3X to the open point penalty of breach; " average diagonal lines " is the diagonal averages of the comparator matrix of use; " diagonal lines " is score or the number to each perfect amino acid coupling appointment by specific comparator matrix) and breach extends point penalty (this normally 1/10 takes advantage of breach opening point penalty) and comparator matrix, and for example PAM 250 or BLOSUM 62 are used together with algorithm.In certain embodiments, the standard comparator matrix is also used by algorithm.For PAM 250 comparator matrixs referring to, for example, the people such as Dayhoff, Atlasof Protein Sequence and Structure, 5 (3) (1978), for BLOSUM 62 comparator matrixs referring to people such as Henikoff, Proc.Natl.Acad.Sci USA, 89:10915-10919 (1992).

In certain embodiments, the parameter about the peptide sequence comparison comprises following:

Algorithm: the people such as Needleman, J.Mol.Biol., 48:443-453 (1970);

Comparator matrix: from people such as Henikoff, the BLOSUM 62 of the same (1992);

Breach point penalty: 12

Notch length point penalty: 4

Similarity threshold value: 0

In certain embodiments, the GAP program can be used together with above-mentioned parameter.In certain embodiments, above-mentioned parameter be use the GAP algorithm for polypeptide relatively (together with for the end breach without point penalty) default parameter.

According to some embodiment, amino-acid substitution be following those: reduce proteoclastic susceptibility (1), (2) reduce the susceptibility to oxidation, (3) change the binding affinity that protein complex is formed, (4) change binding affinity, and/or (4) give or modify physical chemistry or the functional performance for this type of polypeptide.According to some embodiment, can be in naturally occurring sequence (in certain embodiments, in polypeptide portion outside forming the structural domain of intermolecular contact) carry out single or multiple amino-acid substitutions (conservative amino acid replacement in certain embodiments).

In certain embodiments, the constitutional features that conservative amino acid replacement generally can not change parental array basically (for example, replace amino acid and should not be tending towards destroying the spiral existed in parental array, or the secondary structure of the other types of the sign parental array that breaks).Generally acknowledged polypeptide secondary and the example of tertiary structure are described in following reference: for example, Proteins, Structuresand Molecular Principles (Creighton, editor, W.H.Freeman and Company, New York (1984)); Introduction to Protein Structure (C.Branden and J.Tooze, editor, Garland Publishing, New York, N.Y. (1991)); With the people such as Thornton, Nature 354:105 (1991).

As used herein, term " polypeptide fragment " refers to have the polypeptide of N-terminal and/or C-terminal deletion.In certain embodiments, fragment length is 5-500 amino acid at least.Be to be understood that in certain embodiments, fragment length is at least 5,6,8,10,14,20,50,70,100,150,200,250,300,350,400,450 or 500 amino acid.

Peptide analogs is commonly used for to have and is similar to those the non-peptide medicine of characteristic of template peptide in pharmaceutical industries.The non-peptide compound of these types is called as " peptide mimics " or " plan peptide ".Fauchere, J.Adv.Drug Res.15:29 (1986); Veber and the 392nd page of Freidinger TINS (1985); With the people such as Evans, J.Med.Chem.30:1229 (1987).Usually by means of computerized this compounds of molecule modeling developing.Above the peptide mimics of useful peptide can be for generation of similar treatment or prophylactic effect structurally to be similar to treatment.Usually, intend peptide and structurally be similar to the example polypeptide (, polypeptide with biochemical characteristic or pharmacological activity) people's antibody for example, but have by method well-known in the art optionally by the one or more peptide bonds that are selected from following key and replace:--CH ₂nH--,--CH ₂s--,--CH ₂-CH ₂--,--CH=CH-(cis and trans),--COCH ₂--,--CH (OH) CH ₂-and--CH ₂sO--.In certain embodiments, can use one or more amino acid (for example D-Lys replacement 1B) peptide more stable with generation of the D-amino acid system displacement consensus sequence of same type.In addition, the constraint peptide that comprises consensus sequence or substantially the same consensus sequence variant generally can pass through methods known in the art (Rizo and Gierasch Ann.Rev.Biochem.61:387 (1992)) and produce; For example, and be not limited to, by adding the inside cysteine residues that can form the intramolecular disulfide bridge that makes the peptide cyclisation.

Term " specific-binding agent " refers to the natural or non-native molecules of being combined with target-specific.The example of specific-binding agent includes but not limited to, protein, peptide, nucleic acid, carbohydrate, lipid and micromolecular compound.In certain embodiments, specific-binding agent is antibody.In certain embodiments, specific-binding agent is the antigen binding domain territory.

Term " specific binding " refers to that avidity that specific-binding agent is combined with target is greater than the ability of itself and non-target.In certain embodiments, specific binding refers to that the avidity of being combined with target is compared at least 10,50,100,250,500 or 1000 times greatly of the avidity of non-target.In certain embodiments, avidity is measured by avidity ELISA assay method.In certain embodiments, avidity is measured by the BIAcore assay method.In certain embodiments, avidity is measured by kinetics methodology.In certain embodiments, avidity is measured by balance/dissolution method.

Term " for the specific-binding agent of TR-2 " refers to the specific-binding agent of any part of specific binding TR-2.In certain embodiments, the specific-binding agent for TR-2 is the antibody for TR-2.In certain embodiments, specific-binding agent is the antigen binding domain territory.

" antibody " or " antibody peptide " refers to complete antibody or its fragment.In certain embodiments, antibody fragment can be to compete the binding fragment of specific binding with complete antibody.Term " antibody " also comprises polyclonal antibody and monoclonal antibody.In certain embodiments, binding fragment produces by recombinant DNA technology.In certain embodiments, binding fragment produces by enzymatic or the chemical chop of complete antibody.In certain embodiments, does binding fragment produce by recombinant DNA technology? repeat.Binding fragment includes but not limited to, Fab, Fab ', F (ab ') 2, Fv and single-chain antibody.Non-Fab includes but not limited to, the Fc fragment.In certain embodiments, antibody is combined with epitope specificity, and described epi-position is by being selected from least one following antibodies specific combination: Ab A, Ab B, Ab C, Ab D, Ab E, Ab F, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, Ab P and Ab Q.Term " antibody " also comprises the anti-unique antibody with the variable region specific binding of another kind of antibody.In certain embodiments, the variable region specific binding of anti-unique antibody and anti-TR-2 antibody.In certain embodiments, anti-unique antibody can or be blocked the activity of anti-TR-2 antibody for detection of the existence of specific anti-TR-2 antibody in sample.

As used herein, term " anti-TR-2 antibody " means the antibody with the TR-2 specific binding.In certain embodiments, anti-TR-2 antibody is combined with the TR-2 epitope specificity, described TR-2 epi-position and at least one antibodies that is selected from Ab A to Q.In various embodiments, TR-2 can be the TR-2 of any kind, includes but not limited to people, cynomolgus monkey, mouse and rabbit.For some assay method of measuring antibodies specific, be that the technician is well-known, and include but not limited to, ELISA, ELISPOT, western blotting, BIAcore assay method, solution avidity binding assay, T cell co-stimulatory assay method and T cell migration assay method.

As used herein, term " separation antibody " means such antibody, its (1) is not containing its common found at least some protein therewith, (2) be substantially free of from identical source for example from other protein of identical type, (3) by from different types of cell expressing, or (4) do not exist at occurring in nature.

" polyclonal antibody " refers to the heterogeneous mixture from the antibody of the different epi-position combinations of same antigen to term.

" monoclonal antibody " refers to the antibody set by identical nucleic acid molecule encoding to term.In certain embodiments, monoclonal antibody is by single hybridoma or other clone or produce by transgene mammal.The identical epi-position of the general identification of monoclonal antibody.Term " mono-clonal " is not limited to any concrete grammar for the preparation of antibody.

Term " CDR grafted antibody " refers to that wherein the CDR from a kind of antibody is inserted into the antibody in the framework of another kind of antibody.In certain embodiments, CDR antibody and framework by it the derivative antibody derivative by it is different sorts.In certain embodiments, CDR antibody and framework by it the derivative antibody derivative by it is different isotypes.

Term " polyspecific " refers to wherein the antibody of 2 or more variable regions and different epi-position combinations.Epi-position can be on identical or different target.In certain embodiments, multi-specificity antibody is " bi-specific antibody " of 2 kinds of different epi-positions on the identical or different antigen of identification.

Term " catalytic antibody " refers to wherein adhere to the antibody of one or more catalysed partials.In certain embodiments, catalytic antibody is the cytotoxic antibody that comprises the cytotoxicity part.

Term " humanized antibody " refers to the wherein all or part derived from human of antibody framework region, but all or part in one or more CDR district is derived from another kind antibody of mouse for example.

Term " people's antibody " and " human antibody " are used interchangeably, and refer to that wherein CDR and framework comprise human sequence's antibody basically.In certain embodiments, human antibody produces in non-human mammal, includes but not limited to mouse, rat and Lagomorph.In certain embodiments, human antibody produces in hybridoma.In certain embodiments, the human antibody restructuring produces.

In certain embodiments, anti-TR-2 antibody comprises:

In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:2 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:36.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:4 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:38.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:6 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:40.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:8 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:42.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:10 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:44.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:12 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:46.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ IDNO:14 and the second polypeptide that comprises the CDRs as shown in SEQ IDNO:48.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:16 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:50.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:18 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:52.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:20 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:54.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:22 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:56.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:24 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:58.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:26 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:60.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:28 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:62.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:30 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:64.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:32 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:66.In certain embodiments, anti-TR-2 antibody comprises: or the first polypeptide that comprises the complementary determining region (CDRs) as shown in SEQ ID NO:34 and the second polypeptide that comprises the CDRs as shown in SEQ ID NO:68.In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide as shown in above paragraph [079] and the as above the second polypeptide as shown in paragraph [084].In certain embodiments, anti-TR-2 antibody comprises: the first polypeptide as shown in above paragraph [080] and the as above the second polypeptide as shown in paragraph [085].In certain embodiments, anti-TR-2 antibody is people's antibody.In certain embodiments, anti-TR-2 antibody comprises detectable label.In certain embodiments, anti-TR-2 antibody is chimeric antibody.

" chimeric antibody " refers to have the antibody variable region with another kind of molecule first kind that for example antibody constant region of other second kind merges.Referring to, for example U.S. Patent number 4,816, and 567 and the people such as Morrison, Proc Natl Acad Sci (USA), 81:6851-6855 (1985).In certain embodiments, first kind can be different from second kind.In certain embodiments, first kind can be identical with second kind.Partly prepared by the known array that in certain embodiments, chimeric antibody can transplant to mate anti-TR-2 antibody variable region by mutagenesis or CDR.CDR transplants and relates generally to the CDRs from having required specific antibody is transplanted on the framework region (FRs) of another kind of antibody.

In certain embodiments, generally to be interpreted as its each binding site be identical to the bivalent antibody except " polyspecific " or " multifunctionality " antibody.

When making amount with the acceptor of ligand binding, excessive antibody is reduced by least approximately 20%, 40%, 60%, 80%, 85% or when more (as competed in binding assay and measure in vitro), antibody suppresses adhering to of part and acceptor basically.

Term " epi-position " refers to can be by the molecular moiety of specific-binding agent combination.Exemplary epi-position can comprise can with any polypeptide determiner of immunoglobulin (Ig) and/or φt cell receptor specific binding.Exemplary epi-position determinant includes but not limited to, the chemically reactive surface grouping of molecule, such as but not limited to amino acid, sugared side chain, phosphoryl and alkylsulfonyl.In certain embodiments, the epi-position determinant can have specific Three Dimensions Structure, and/or specific charge characteristic.In certain embodiments, epi-position is the antigen zone by antibodies.Epi-position can be adjacency or non-adjacent.In certain embodiments, epi-position can be simulated, because they comprise such three-dimensional structure, it is similar to the epi-position for generation of antibody, but be not included in the amino-acid residue of finding in that epi-position for generation of antibody, or only be included in some amino-acid residues of finding in that epi-position for generation of antibody.

Term " suppress and/or neutralizing epitope " refer to when by specific-binding agent in conjunction with the time, cause the epi-position of interior, the external and/or active minimizing of biology in situ of body.In certain embodiments, neutralizing epitope be positioned on the biologic activity zone of target or with the biologic activity zone combination of target.

Term " activation epi-position " refer to when by specific-binding agent in conjunction with the time, the epi-position that causes interior, the external and/or active activation of biology in situ of body or maintain.In certain embodiments, the activation epi-position be positioned on the biologic activity zone of target or with the biologic activity zone combination of target.

In certain embodiments, epi-position is by being selected from least one following antibodies specific combination: Ab A, Ab B, Ab C, Ab D, Ab E, Ab F, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, Ab P and Ab Q.In some this type of embodiment, epi-position is purifying basically.In some this type of embodiment, epi-position concentration is 1nM at least.In some this type of embodiment, epi-position concentration is 1nM-5nM.In some this type of embodiment, epi-position concentration is 5nM-10nM.In some this type of embodiment, epi-position concentration is 10nM-15nM at least.

In certain embodiments, antibody is combined with epitope specificity, described epi-position is by being selected from least one following antibodies specific combination: Ab A, Ab B, Ab C, Ab D, Ab E, AbF, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, AbP and Ab Q, and be purifying basically.In some this type of embodiment, antibody concentration is 1nM at least.In some this type of embodiment, antibody concentration is 1nM-5nM.In some this type of embodiment, antibody concentration is 5nM-10nM.In some this type of embodiment, antibody concentration is 10nM-15nM at least.

In certain embodiments, amino acid/11-85 specific binding of antibody and ripe people TR-2, and be purifying basically.In some this type of embodiment, antibody concentration is 1nM at least.In some this type of embodiment, antibody concentration is 1nM-5nM.In some this type of embodiment, antibody concentration is 5nM-10nM.In some this type of embodiment, antibody concentration is 10nM-15nM at least.

In certain embodiments, antibody be selected from least one following antibody competition and be combined epi-position: Ab A, Ab B, Ab C, Ab D, Ab E, Ab F, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, Ab P and Ab Q.In certain embodiments, antibody is purifying basically.In some this type of embodiment, antibody concentration is 1nM at least.In some this type of embodiment, antibody concentration is 1nM-5nM.In some this type of embodiment, antibody concentration is 5nM-10nM.In some this type of embodiment, antibody concentration is 10nM-15nM at least.

In certain embodiments, antibody be selected from least one following antibody competition and be combined the amino acid/11 of ripe people TR-2-85:Ab A, Ab B, Ab C, Ab D, Ab E, Ab F, Ab G, Ab H, Ab I, Ab J, Ab K, Ab L, Ab M, Ab N, Ab O, Ab P and Ab Q.In certain embodiments, antibody is purifying basically.In some this type of embodiment, antibody concentration is 1nM at least.In some this type of embodiment, antibody concentration is 1nM-5nM.In some this type of embodiment, antibody concentration is 5nM-10nM.In some this type of embodiment, antibody concentration is 10nM-15nM at least.

Term " reagent " is in this article for mixture, the biomacromolecule that means chemical compound, chemical compound or the extract prepared by biomaterial.

As used herein, term " mark " refers to any molecule that can be detected.In certain embodiments, antibody can carry out mark by mixing radiolabeled amino acid.In certain embodiments, the biotin moiety that avidin that can be by mark (for example, comprising the fluorescent mark that can be detected by optics or colorimetry or the Streptavidin of enzymatic activity) is detected can adhere to antibody.In certain embodiments, mark can be mixed in another kind of reagent, or mark and another kind of reagent are adhered to, described another kind of reagent successively with the purpose antibodies.In certain embodiments, mark can be mixed in antibody, or mark and antibody are adhered to, described antibody is combined with the purpose antibodies specific successively.In certain embodiments, label or tag also can be treated.The whole bag of tricks of labeling polypeptide and glycoprotein is known in the art and can uses.The mark of some general classes includes but not limited to, enzymatic, fluorescence, chemoluminescence and radio-labeling.Some mark example about polypeptide includes but not limited to, following: radio isotope or radionuclide are (for example, ³h, ¹⁴c, ¹⁵n, ³⁵s, ⁹⁰y, ⁹⁹tc, ¹¹¹in, ¹²⁵i, ¹³¹i), fluorescent mark (fluorescein isothiocyanate (FITC) for example, rhodamine, the lanthanon phosphorescent substance, phycoerythrin (PE)), enzymatic labelling (for example, horseradish peroxidase, beta-galactosidase enzymes, luciferase, alkaline phosphatase, glucose oxidase, glucose-6-phosphate dehydrogenase, alcoholdehydrogenase, malate dehydrogenase (malic acid dehydrogenase), penicillinase, luciferase? repeat), chemiluminescent labeling, biotinyl, with predetermined polypeptide epi-position by the identification of secondary reporter molecule (for example, leucine zipper is to sequence, binding site about secondary antibodies, the metal combining structure territory, epitope tag).In certain embodiments, mark adheres to reduce potential steric hindrance by the spacerarm of all lengths.

As used herein, term " sample " includes but not limited to, from the material of any amount of being or former being.This type of being includes but not limited to, people, mouse, monkey, rat, rabbit and other animals.This type of material includes but not limited to, blood, serum, urine, cell, organ, tissue, bone, marrow, lymphoglandula and skin.

As used herein, term " pharmaceutical agent or medicine ", while referring to suitably be applied to the patient, can be induced chemical compound or the composition of required treatment effect.

As used herein, term " conditioning agent " is the compound that changes or change molecular activity or function.For example, and in the situation that do not have activity that conditioning agent observes or function magnitude relatively, conditioning agent can cause some activity of molecule or increase or the minimizing in the function magnitude.In certain embodiments, conditioning agent is to reduce at least one activity of molecule or the inhibitor of function magnitude.The exemplary activity of some of molecule and function include but not limited to, binding affinity, enzymatic activity and signal transduction.Some exemplary inhibitor comprises but is not limited to, protein, peptide, antibody, peptide body (peptibodies), carbohydrate and organic molecule.The exemplary peptides body is for example being described in WO 01/83525.

As used herein, " purifying basically " means the purpose kind is the dominant kind (that is, it is abundanter than any other the indivedual kinds in composition on mole foundation) existed.In certain embodiments, the part of purifying is the composition at least about 50% (on mole foundation) of all macromole kinds that wherein the purpose kind comprises existence basically.In certain embodiments, basically the composition of purifying will comprise all macromole kinds of existing in composition surpass approximately 80%, 85%, 90%, 95% or 99%.In certain embodiments, the purpose kind is purified to composition wherein basically by the basic homogeneity (by the conventional sense method, at composition, can not detect contamination class) of single macromole species composition.

Term " patient " comprises the humans and animals experimenter.

According to some embodiment, provide the clone of expressing anti-TR-2 antibody.

The chimeric antibody of the sequence that comprises at least part of human sequence and another kind is provided in certain embodiments.In certain embodiments, with the antibody of the antibody sequence that does not contain the host, compare, this type of chimeric antibody can produce the immunne response reduced in the host.For example, in some cases, the purpose animal can be as the model about the particular person disease.In order to study the impact of antibody on that disease in the animal host, can use from different types of antibody.But, in some cases, can cause the immunne response for the antibody in host animal self from this antibody-like of another kind, thereby hinder the assessment of these antibody.In certain embodiments, the magnitude that the anti-antibody of using the aminoacid sequence of replacing the anti-TR-2 antibody of part from the antibody aminoacid sequence of host animal can reduce host animal is replied.

In certain embodiments, chimeric antibody comprises heavy chain and light chain, and wherein the variable region of light chain and heavy chain is from first kind, and the constant region of light chain and heavy chain is from second kind.In certain embodiments, the heavy chain of antibody constant region is the heavy chain of antibody constant region of the kind except the people.In certain embodiments, antibody light chain constant region is the antibody light chain constant region of the kind except the people.In certain embodiments, the heavy chain of antibody constant region is human antibody heavy chain's constant region, and antibody heavy chain variable region is the antibody heavy chain variable region of the kind except the people.In certain embodiments, antibody light chain constant region is human antibody light chain constant region, and antibody chain variable region is the antibody chain variable region of the kind except the people.The exemplary antibodies constant region includes but not limited to, people's antibody constant region, cynomolgus monkey antibody constant region, mouse antibodies constant region and rabbit antibody constant region.The exemplary antibodies variable region includes but not limited to, people's antibody variable region, mouse antibodies variable region, pig antibody variable region, cavy antibody variable region, cynomolgus monkey antibody variable region and rabbit antibody variable region.In certain embodiments, in heavy chain and light chain, the framework region of variable region can be used the framework region replacement derived from other antibody sequences.

Some exemplary chimeric antibody can produce by the well-known method of those of ordinary skills.In certain embodiments, can make the polynucleotide of second kind of the polynucleotide of first kind of encoding heavy chain variable region and encoding heavy chain constant region merge.In certain embodiments, can make the polynucleotide of first kind of encoded light chain variable region merge with the nucleotide sequence of second kind of coding constant region of light chain.In certain embodiments, for example the nucleotides sequence of these fusions can be listed in, in single expression vector (, plasmid) or a plurality of expression vector and introduce in cell.In certain embodiments, can use the cell that comprises at least one expression vector to prepare polypeptide.In certain embodiments, the nucleotides sequence of these fusions can be listed in expression vector separately or single expression vector and introduce in cell.In certain embodiments, host cell expression heavy chain and light chain, described heavy chain and light chain combine to produce antibody.In certain embodiments, can use the cell Dispersal risk that comprises at least one expression vector.For the production of the illustrative methods with expressing antibody, discuss hereinafter.

In certain embodiments, for the conservative modification of weight and the light chain of anti-TR-2 antibody (with accordingly for the modification of coding nucleotide) by generation have be similar to original antibody those function and the antibody of chemical feature.By contrast, in certain embodiments, basic modification in the function of anti-TR-2 antibody and/or chemical feature can complete by the displacement in the aminoacid sequence of selecting heavy and light chain, described displacement is significantly different on the following impact maintained at it: (a) replacement areas for example as the molecular backbone chain structure in folding or helical conformation, (b) minute charge of the electron or the hydrophobicity at target site place, or (c) side chain volume.

Some amino acid needed displacement (that no matter guard or nonconservative) can the time be determined in this type of displacement of needs by those skilled in the art.In certain embodiments, amino-acid substitution can be for the identification of the important residue of anti-TR-2 antibody, for example can increase or reduce for those of the effector function of the avidity of the antibody of TR-2 or antibody.

In certain embodiments, the effect of anti-TR-2 antibody can be assessed by the minimizing in the amount of measuring disease symptoms.In certain embodiments, the purpose disease can be caused by pathogenic agent.In certain embodiments, disease can be set up by additive method in the animal host, and described method comprises material (for example carcinogens) introducing and genetic manipulation.In certain embodiments, effect is assessed by one or more adverse events that detect in the animal host.Term " adverse events " includes but not limited to, accepts the adverse effect in the animal host of antibody, described adverse effect be do not accept in the animal host of this antibody non-existent.In certain embodiments, adverse events includes but not limited to, fever, the immunne response for antibody, inflammation and/or animal host's death.

Various antibody to antigen-specific can produce in many ways.In certain embodiments, the antigen that comprises the purpose epi-position for example can be introduced, in animal host's (, mouse), thereby be produced the antibody special to that epi-position.In some cases, the antibody special to the purpose epi-position can obtain from biological sample, and described biological sample is taken from Natural Exposure in the host of epi-position.In some cases, human normal immunoglobulin (Ig) locus is introduced in the mouse that wherein endogenous Ig gene has been inactivated, the chance of acquisition human monoclonal antibodies (MAbs) is provided.

Naturally occurring antibody structure

Naturally occurring antibody structure unit generally comprises tetramer.Each this type of tetramer generally is comprised of 2 pairs of identical polypeptide chains, and every pair has a total length " gently " chain (about 25kDa in certain embodiments) and a total length " weight " chain (about 50-70kDa in certain embodiments).Term " heavy chain " comprises having enough variable region sequences to give the specific any polypeptide to specific antigen.The total length heavy chain comprises variable region structural domain V _h, and 3 constant region domain Cs _h1, C _h2 and C _h3.V _hstructural domain is positioned at the N-terminal of polypeptide, and C _h3 structural domains are positioned at C-terminal.As used herein, term " heavy chain " comprises full length antibody heavy chain and fragment thereof.

Term " light chain " comprises having enough variable region sequences to give the specific any polypeptide to specific antigen.Full-length light chains comprises variable region structural domain V _l, and the constant region domain C _l.As heavy chain, the variable region structural domain of light chain is positioned at the N-terminal of polypeptide.As used herein, term " light chain " comprises full length antibody light chain and fragment thereof.

The N-terminal of every chain partly comprises general be responsible for approximately 100-110 of antigen recognition or the variable region of amino acids (V in heavy chain more _hwith light chain in V _l).The general restriction of the C-terminal of every chain may be responsible for the constant region (C in heavy chain of effector function _hwith light chain in C _l).The effect of antibody mediated effect comprises activating complement and stimulates conditioning phagocytosis.People's light chain generally is classified as κ and lambda light chain.Heavy chain generally is classified as μ, δ, γ, α or ε, and the isotype of antibody is limited and is respectively IgM, IgD, IgG, IgA and IgE.IgG has several subclass, includes but not limited to IgG1, IgG2, IgG3 and IgG4.IgM has subclass and includes but not limited to, IgM1 and IgM2.IgA is subdivided into similarly subclass and includes but not limited to, IgA1 and IgA2.In total length in light and heavy chain, variable and constant region general by approximately 12 or more " J " district of amino acids be connected, and heavy chain also comprises approximately 10 or " D " district of amino acids more.Referring to, for example, Fundamental Immunology the 7th chapter (Paul, W., editor, the 2nd edition, Raven Press, N.Y. (1989)).The variable region that each light/heavy chain is right generally forms antigen binding site.

Variable region generally demonstrates the identical general structure of the relatively conservative framework region (FR) connected by 3 hypervariable regions, and described hypervariable region is also referred to as complementary determining region or CDRs.CDRs from every counterweight and light chain generally aims at by framework region, and this makes it possible to be combined with specificity epitope.From the N-terminal to the C-terminal, light and variable region of heavy chain generally comprises structural domain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.Amino acid about each structural domain distributes generally according to Kabat Sequences of Proteins of Immunological Interest (NationalInstitutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia and LeskJ.Mol.Biol.196:901-917 (1987); The people such as Chothia, the definition of Nature 342:878-883 (1989).

As discussed above, there is the antibody fragment of several types.The Fab fragment is by the C of a light chain and a heavy chain _h1 and variable region form.The heavy chain of Fab molecule can not form disulfide linkage with another heavy chain molecule.Fab ' fragment comprises a light chain and a heavy chain, and described heavy chain is included in C _h1 and C _hmore constant region between 2 structural domains can form interchain disulfide bond to form F (ab ') 2 molecules thereby make between 2 heavy chains.The Fab fragment is similar to F (ab ') 2 molecules, except the constant region in the molecule heavy chain extends to C _houtside 2 structural domain ends.The Fv district comprises to conduct oneself with dignity and the variable region of light chain, but lacks constant region.Single-chain antibody is such Fv molecule, wherein weighs and by nonrigid connector, is connected to form the wall scroll polypeptide chain with variable region of light chain, and described polypeptide chain forms the antigen binding domain territory.Exemplary single-chain antibody discusses in detail in for example WO 88/01649 and U.S. Patent number 4,946,778 and 5,260,203.The C that the Fc fragment comprises heavy chain _h2 and C _h3 structural domains, and be included in C _h1 and C _hbetween 2 structural domains, more constant region, can form interchain disulfide bond thereby make between 2 heavy chains.

In certain embodiments, functional domain C _h1, C _h2, C _h3 and insertion sequence can be reorganized to produce different antibody constant regions.For example, in certain embodiments, this type of hybrid constant region can be for the transformation period in serum, for the assembling of antibody tetramer and folding and/or be optimized for the effector function improved.In certain embodiments, the antibody constant region of modification can produce by following method: a single point sudden change is introduced in the aminoacid sequence of constant region, and the character of just improving is for example above enumerated one or more in those and tested resulting antibody.

In certain embodiments, change a kind of antibody of isotype is transformed into to different isotypes by isotype, and do not lose its specificity to certain target molecules.The method of isotype conversion includes but not limited in other, directly recombinant technology (referring to for example, U.S. Patent number 4,816,397) and cell-cell-fusion techniques (referring to for example U.S. Patent number 5,916,771).In certain embodiments, use above-described technology or other modes known in the art antibody can be transformed into to another subclass from a subclass, and do not lose its specificity to certain target molecules, and include but not limited to, be transformed into IgG1, IgG3 or IgG4 subclass from the IgG2 subclass.

Dual specific or bifunctional antibody

Dual specific or bifunctional antibody be generally have 2 different heavy/light chain is to the artificial hybrid antibody with 2 different binding sites.Bi-specific antibody can produce by the whole bag of tricks, includes but not limited to, hybridoma merges or Fab ' fragment connects.Referring to, for example, Songsivilai and Lachmann Clin.Exp.Immunol.79:315-321 (1990), the people such as Kostelny, J.Immunol.148:1547-1553 (1992).

Some preparation of antibody

In certain embodiments, antibody can be expressed in the clone except hybridoma cell line.In certain embodiments, the coding specific antibodies comprises that the sequence of chimeric antibody can be for transforming suitable mammalian host cell.According to some embodiment, conversion can be by for introducing polynucleotide any currently known methods in host cell, comprise, for example polynucleotide are packaged into (or in virus vector) in virus, and with virus transduction host cell or by using operation transfection carrier known in the art, as by U.S. Patent number 4,399,216; 4,912,040; 4,740,461; With 4,959,455 is illustrative.

In certain embodiments, expression vector comprises any polynucleotide sequence that this paper discusses.In certain embodiments, provide the method for preparing polypeptide, described method is included in the polynucleotide that are suitable for wherein comprising and expresses to produce under the condition of polypeptide, in comprising the cell of any above-mentioned expression vector, produces polypeptide.

In certain embodiments, expression vector comprises polynucleotide, the sequence that described polynucleotide comprise coded polypeptide, described polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1a, CDR2a and CDR3a, wherein CDR1a comprises aminoacid sequence a b c d e f g hi j k l, and wherein amino acid a is glycine; Amino acid b is selected from glycine, tyrosine or phenylalanine; Amino acid c is selected from Serine or Threonine; Amino acid d is selected from Isoleucine or phenylalanine; Amino acid e is selected from Serine, Threonine or l-asparagine; Amino acid f is selected from Serine, aspartic acid, tyrosine, l-asparagine, Threonine or glycine; Amino acid g is selected from glycine, aspartic acid or tyrosine; Amino acid h is selected from glycine, aspartic acid, tyrosine, l-asparagine or Serine; Amino acid i is selected from tyrosine, Isoleucine, Histidine, methionine(Met) or tryptophane; Amino acid j is selected from l-asparagine, tyrosine, Histidine, Serine or phenylalanine; Amino acid k is tryptophane or does not exist; Be Serine or do not exist with amino acid l; Wherein CDR2a comprises aminoacid sequence m n o p q r s t u v w x y z a ' b ' c ', and wherein amino acid m is selected from tryptophane, tyrosine, Histidine, α-amino-isovaleric acid, L-glutamic acid or Serine; Amino acid n is selected from methionine(Met) or Isoleucine; Amino acid o is selected from l-asparagine, tyrosine, Serine, tryptophane or Histidine; Amino acid p is selected from proline(Pro), tyrosine, Serine, arginine, Histidine or l-asparagine; Amino acid q is selected from l-asparagine, Serine or aspartic acid; Amino acid r is selected from Serine or glycine; Amino acid s is selected from aspartic acid, Serine, Threonine or arginine; Amino acid t is selected from l-asparagine, Threonine, L-Ala, Isoleucine or tyrosine; Amino acid u is selected from Threonine, tyrosine, leucine, Methionin, l-asparagine or Isoleucine; Amino acid v is selected from glycine, tyrosine, aspartic acid or halfcystine; Amino acid w is selected from tyrosine or l-asparagine; Amino acid x is selected from L-Ala or proline(Pro); Amino acid y is selected from glutamine, Serine or aspartic acid; Amino acid z is selected from Methionin, leucine or Serine; Amino acid a ' is selected from phenylalanine, Methionin or α-amino-isovaleric acid; Amino acid b ' is selected from glutamine, Serine or Methionin; Be glycine or do not exist with amino acid c '; Wherein CDR3a comprises aminoacid sequence d ' e ' f ' g ' h ' i ' j ' k ' l ' m ' n ' o ' p ' q ' r ' s ' t ' u ' v ' w ', and wherein amino acid d ' is selected from tryptophane, aspartic acid, glycine, Serine or L-glutamic acid; Amino acid e ' is selected from l-asparagine, aspartic acid, glycine, arginine, Serine, α-amino-isovaleric acid or leucine; Amino acid f ' is selected from Histidine, Serine, L-Ala, tyrosine, proline(Pro), l-asparagine, glycine or Threonine; Amino acid g ' is selected from tyrosine, Serine, L-Ala, arginine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid h ' is selected from glycine, L-Ala, Serine, l-asparagine, methionine(Met), tyrosine, tryptophane, halfcystine or aspartic acid; Amino acid i ' is selected from Serine, tryptophane, glycine, phenylalanine, aspartic acid, tyrosine or Threonine; Amino acid j ' is selected from glycine, Threonine, Serine, leucine, α-amino-isovaleric acid, l-asparagine, tryptophane or tyrosine; Amino acid k ' is selected from Serine, phenylalanine, aspartic acid, tryptophane, glycine or tyrosine or does not exist; Amino acid l ' is selected from Histidine, aspartic acid, L-Ala, tryptophane, tyrosine, Serine, phenylalanine, α-amino-isovaleric acid or glycine or does not exist; Amino acid m ' is selected from phenylalanine, tyrosine, L-glutamic acid, proline(Pro), aspartic acid, halfcystine, Isoleucine or methionine(Met) or does not exist; Amino acid n ' is selected from aspartic acid, phenylalanine, L-Ala, leucine or Serine or does not exist; Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline(Pro) or α-amino-isovaleric acid or does not exist; Amino acid p ' is selected from leucine, aspartic acid or tyrosine or does not exist; Amino acid q ' is selected from Serine or tyrosine or does not exist; Amino acid r ' is tyrosine or does not exist; Amino acid s ' is selected from glycine or tyrosine or does not exist; Amino acid t ' is selected from glycine or methionine(Met) or does not exist; Amino acid u ' is selected from methionine(Met) or aspartic acid or does not exist; Amino acid v ' is selected from aspartic acid or α-amino-isovaleric acid or does not exist; Be α-amino-isovaleric acid or do not exist with amino acid w '; The described polypeptide of wherein being combined with light chain of antibody is in conjunction with TR-2.In certain embodiments, provide the method for preparing polypeptide, described method is included in the polynucleotide that are suitable for wherein comprising and expresses to produce under the condition of polypeptide, in comprising the cell of above-mentioned expression vector, produces polypeptide.

In certain embodiments, expression vector comprises polynucleotide, the sequence that described polynucleotide comprise coded polypeptide, described polypeptide comprises at least one complementary determining region (CDR) that is selected from CDR1b, CDR2b and CDR3b, wherein CDR1b comprises aminoacid sequence a1 b1 c1 d1 e1f1 g1 h1 i1 j1 k1 l1 m1 n1 o1 p1 q1, and wherein amino acid a1 is selected from arginine or Methionin; Amino acid b1 is selected from Threonine, L-Ala or Serine; Amino acid c1 is Serine; Amino acid d1 is glutamine; Amino acid e1 is selected from Serine or glycine; Amino acid f1 is selected from Isoleucine, leucine or α-amino-isovaleric acid; Amino acid g1 is selected from Serine, leucine or arginine; Amino acid h1 is selected from Threonine, Serine, Isoleucine, l-asparagine, arginine, Histidine or tyrosine; Amino acid i1 is selected from tyrosine, arginine, tryptophane, aspartic acid or Serine; J1 is selected from leucine, Isoleucine, l-asparagine, tyrosine or Serine; Amino acid k1 is selected from l-asparagine, glycine, α-amino-isovaleric acid, L-Ala or leucine; Amino acid l1 is selected from tyrosine, L-Ala or l-asparagine or does not exist; Amino acid m1 is selected from l-asparagine or Methionin or does not exist; Amino acid n1 is selected from tyrosine, l-asparagine or Isoleucine or does not exist; Amino acid o1 is selected from leucine or tyrosine or does not exist; Amino acid p1 is selected from aspartic acid or leucine or does not exist; Be selected from α-amino-isovaleric acid, L-Ala or Threonine or do not exist with amino acid q1; Wherein CDR2b comprises aminoacid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from L-Ala, aspartic acid, leucine, tryptophane, glycine or α-amino-isovaleric acid; Amino acid s1 is selected from Threonine, α-amino-isovaleric acid, glycine or L-Ala; Amino acid t1 is Serine; Amino acid u1 is selected from Serine, l-asparagine or Threonine; Amino acid v1 is selected from leucine, phenylalanine or arginine; Amino acid w1 is selected from glutamine, L-Ala or L-glutamic acid; Be selected from Serine, arginine or Threonine with amino acid x1; Wherein CDR3b comprises aminoacid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' el ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine(Met), leucine or Histidine; Amino acid z1 is selected from glutamine or Methionin; Amino acid a1 ' is selected from Serine, Threonine, L-Ala, Histidine, tyrosine or phenylalanine; Amino acid b1 ' is selected from tyrosine, leucine, l-asparagine or glycine; Amino acid c1 ' is selected from Serine, glutamine, Isoleucine or Methionin; Amino acid d1 ' is selected from Threonine, phenylalanine, tyrosine, L-Ala or Serine; Amino acid e1 ' is proline(Pro); Amino acid f1 ' is selected from leucine, phenylalanine, tryptophane, Serine or arginine; Be selected from Threonine or Serine with amino acid g1 '; The described polypeptide of wherein being combined with heavy chain of antibody is in conjunction with TR-2.In certain embodiments, provide the method for preparing polypeptide, described method is included in the polynucleotide that are suitable for wherein comprising and expresses to produce under the condition of polypeptide, in comprising the cell of above-mentioned expression vector, produces polypeptide.In certain embodiments, provide the cell that comprises at least one above-mentioned expression vector.In certain embodiments, provide the method for preparing polypeptide, described method is included in the polynucleotide that are suitable for wherein comprising and expresses to produce under the condition of polypeptide, in comprising the cell of above-mentioned expression vector, produces polypeptide.

In certain embodiments, expression vector is expressed anti-TR-2 heavy chain of antibody.In certain embodiments, expression vector is expressed anti-TR-2 light chain of antibody.In certain embodiments, expression vector is expressed anti-TR-2 heavy chain of antibody and anti-TR-2 light chain of antibody.In certain embodiments, provide the method for anti-TR-2 antibody, described method is included in the polynucleotide that are suitable for wherein comprising and expresses to produce under the condition of antibody, in comprising the cell of at least one expression vector described herein, produces antibody.

In certain embodiments, the transfection of use operation can be depended on host to be transformed.Some method that is used for heterologous polynucleotide is introduced in mammalian cell is known in the art, and include but not limited to, transfection, protoplast fusion, the electroporation of the transfection of dextran mediation, calcium phosphate precipitation, poly-quaternary ammonium salt (polybrene) mediation, one or more polynucleotide are encapsulated in liposome, and by the direct microinjection of DNA in core.

Some mammal cell line that can be used as the host for expressing is known in the art, and include but not limited to, the immortalized cell line that can obtain from American type culture collection (ATCC), include but not limited to Chinese hamster ovary (CHO) cell, E5 cell, HeLa cell, young hamster kidney (BHK) cell, monkey-kidney cells (COS), human hepatocellular carcinoma cell line (for example Hep G2), NS0 cell, SP20 cell, Per C6 cell, 293 cells and many other clones.In certain embodiments, can be by determining which clone has the antibody that high expression level and generation have a composition antigenic binding property and selects clone.

In certain embodiments, can comprise control sequence by transfected carrier in host cell, described control sequence is operably connected with the polynucleotide of the anti-TR-2 antibody of coding.In certain embodiments, control sequence promotes the expression of the polynucleotide of connection, thereby causes the polypeptide production by the polynucleotide encoding connected.In certain embodiments, carrier also comprises the polynucleotide sequence that allows the karyomit(e) dependent/non-dependent in host cell to copy.Exemplary carrier includes but not limited to, plasmid (such as BlueScript, puc etc.), clay and YACS.

Some antibody purposes

According to some embodiment, antibody is for detection of the specific antigen in sample.In certain embodiments, this allows to identify the cell or tissue of producing protein.For example, in certain embodiments, anti-TR-2 antibody can exist for detection of the TR-2 in sample.In certain embodiments, comprise that for detection of the existence of the TR-2 in sample or non-existent method (a) makes anti-TR-2 antibody and sample combination; (b) antibody that makes to be combined with antigen separates with unconjugated antibody; (c) detect the existence of the antibody of being combined with antigen or do not exist.

Wherein antibody can include but not limited to for detection of existence or the non-existent assay method of antigen, ELISA and western blotting.In certain embodiments, anti-TR-2 antibody can carry out mark.In certain embodiments, anti-TR-2 antibody can be detected by the traget antibody with anti-TR-2 antibodies.In certain embodiments, provide for detection of the TR-2 in sample and existed or non-existent test kit.In certain embodiments, this test kit comprises anti-TR-2 antibody and for detection of the reagent of antibody.

In certain embodiments, antibody can be for the part of separation chemistry basically such as but not limited to protein.In certain embodiments, antibody adheres to " matrix ", and described matrix is for making the fixing support material of antibody.Matrix includes but not limited to, pipe, dull and stereotyped (that is, porous flat plate), pearl be microballon, strainer, ball and film for example.In certain embodiments, matrix can be by the preparation of water-insoluble material, such as but not limited to polycarbonate resin, silicone resin or nylon resin.Include but not limited to Mierocrystalline cellulose, agarose, polyacrylamide, dextran, polystyrene, polyvinyl alcohol and porous silicon for the exemplary substrates of using at affinity chromatography.Have the much chromatography matrix be obtained commercially, it includes but not limited to, Sepharose 2B, Sepharose4B, Sepharose 6B and other forms of Sepharose (Pharmacia); Bio-Gel (with various forms of Bio-Gel for example Biogel A, P or CM), Cellex (with various forms of Cellex for example Cellex AE or Cellex-CM), Chromagel A, Chromagel P and Enzafix (Wako Chemical Indus.).The use of antibody affinity column is known to persons of ordinary skill in the art.In certain embodiments, the method for separating of TR-2 comprises that (a) adheres to TR-2 antibody and matrix; (b) sample that makes to comprise TR-2 is exposed to the antibody of part (a); (c) separate TR-2.In certain embodiments, provide the test kit for separating of TR-2.In certain embodiments, this test kit comprises the anti-TR-2 antibody that adheres to matrix and for separating of the reagent of TR-2.

As used herein, term " affinity chromatography " mean by utilize material between interaction (for example, avidity) method of the purpose material in the isolated or purified sample, described material is to for example antigen and antibody, enzyme and substrate or acceptor and part.

In certain embodiments, be combined with specified protein and block and the interactional antibody of other binding compounds may have therepic use.In this application, when the purposes of anti-TR-2 Antybody therapy disease or symptom is discussed, this type of purposes can comprise anti-TR-2 antibody self; The composition that comprises anti-TR-2 antibody; And/or the purposes of the combination treatment that comprises anti-TR-2 antibody and one or more other activeconstituentss.When anti-TR-2 antibody is used for " treatment " disease or symptom, this type for the treatment of can comprise or not comprise the prevention of disease or symptom.In certain embodiments, anti-TR-2 antibody can be blocked the interaction of TR-2 acceptor and its ligand/TRAIL.In certain embodiments, anti-TR-2 antibody can activate the TR-2 acceptor.In certain embodiments, anti-TR-2 antibody can constitutively activate TR-2 acceptor.Because TR-2 is relevant to apoptosis, so in certain embodiments, anti-TR-2 antibody is wished necrocytosis or stops in the disease treatment of necrocytosis to have therepic use therein.This type of disease includes but not limited to, cancer, inflammation and the virus infection relevant to any tissue of expressing TR-2.

In certain embodiments, anti-TR-2 antibody is used separately.In certain embodiments, anti-TR-2 antibody at least one other therapeutical agent before using, use.In certain embodiments, other therapeutical agent of anti-TR-2 antibody and at least one is used simultaneously and is used.In certain embodiments, anti-TR-2 antibody at least one other therapeutical agent after using, use.The exemplary treatment agent includes but not limited to, at least one other cancer therapeutic agent.Exemplary cancer therapeutic agent includes but not limited to, radiotherapy and chemotherapy.

In certain embodiments, anti-TR-2 antibody pharmaceutical compositions can be used in combination treatment, with other agent combination.In certain embodiments, combination treatment comprises the anti-TR-2 antibody combined with at least one anti-angiogenic agent.Exemplary agents includes but not limited to, the chemical composition of external synthetic preparation, antibody, antigen binding domain territory, radionuclide and combination and conjugate.In certain embodiments, reagent can serve as agonist, antagonist, alllosteric (doubting as allosteric allosteric) conditioning agent or toxin.In certain embodiments, reagent can for example, for suppressing or stimulating its target (, acceptor or enzyme activate or suppress) thereby and promotion necrocytosis or cell growth inhibiting.

The treatment of exemplary chemotherapy includes but not limited to, antineoplastic agent includes but not limited to, alkylating agent includes but not limited to: mustargen includes but not limited to dichloromethyldiethylamine, endoxan, ifosfamide, melphalan and Chlorambucil; Nitrosourea, include but not limited to, carmustine BCNU, Luo Mositing, CCNU and semustine, Semustine; Temodal ^tM, temozolamide; Aziridine/methylmelamine, include but not limited to, triethylenemelamine (TEM), and three ethylenes, tespamin, thiophene is for group, hexamethylmelamine (HMM) and altretamine; Alkylsulfonate, include but not limited to, busulfan; Triazine, include but not limited to, dacarbazine (DTIC); Metabolic antagonist, include but not limited to, folacin is methotrexate and trimetrexate for example; Pyrimidine analogue, include but not limited to, 5 FU 5 fluorouracil (5FU); Floxuridine, gemcitabine, cytarabin (AraC, cytosine arabinoside), 5-azacytidine, and 2,2 '-the difluoro Deoxyribose cytidine; Purine analogue, include but not limited to, Ismipur, the 6-Tioguanine, imuran, 2 '-deoxycoformycin (pentoside), red hydroxyl nonyl VITAMIN B4 (EHNA), fludarabine phosphate, carat Qu Bin and 2-chlorodeoxyadenosine (2-CdA); Natural product includes but not limited to that anti-mitosis medicine is taxol for example; Vinca alkaloids, include but not limited to, vinealeucoblastine(VLB) (VLB), vincristine(VCR) and vinorelbine; Taxotere; Estramustine and estramustine phosphate; Ppipodophylotoxins (doubting as epipodophyllotoxins epipodophyllotoxin), include but not limited to Etoposide and teniposide; Microbiotic, include but not limited to, dactinomycin, daunomycin, Rubomycin C, Zorubicin, mitoxantrone, darubicin, bleomycin, Plicamycin, mithramycin, ametycin and actinomycin; Enzyme, include but not limited to, L-ASP; Biological response modifier, include but not limited to, interferon-' alpha ', IL-2, G-CSF and GM-CSF; Doxycyckine (doubting as doxycycline doxycycline); Irinotecan hydrochloride; Mix reagent, include but not limited to, platinium (doubting as platinum platinum) co-ordination complex is cis-platinum and carboplatin for example; Amerantrone; Include but not limited to mitoxantrone; Substitute urea, include but not limited to hydroxyurea; The methyl hydrazine derivative, include but not limited to, N-methyl hydrazine (MIH) and procarbazine; The adrenal cortex inhibitor, include but not limited to, mitotane (o, p '-DDD) and aminoglutethimide; Hormone and antagonist, include but not limited to, the adrenocortical steroid antagonist is prednisone and Equivalent for example, dexamethasone and aminoglutethimide; Gemzar ^tM, gemcitabine; Progesterone, include but not limited to, Hydroxyprogesterone caproate bp 98, Veramix and acetic acid megestrol; Oestrogenic hormon, include but not limited to, stilboestrol and Ethinylestradiol Equivalent; Estrogen antagonist, include but not limited to, Tamoxifen; Male sex hormone, include but not limited to, testosterone propionate and Fluoxymesterone/Equivalent; Androgen antagonist, include but not limited to, Drogenil, gonadotropin releasing hormone analogues and Leuprolide; With the non-steroid androgen antagonist, include but not limited to Drogenil.

The exemplary cancer therapy that can use together with anti-TR-2 antibody includes but not limited to, targeted therapy.The example of targeted therapy includes but not limited to, the use for the treatment of antibody.Exemplary treatment antibody includes but not limited to, mouse, mouse-people are chimeric, CDR transplanting, humanization and people's antibody and synthetic antibody, include but not limited to those that select by the screening antibodies library.Exemplary antibodies includes but not limited to, with cell surface protein Her2, CDC20, CDC33, mucin-like glycoprotein, with those of the EGF-R ELISA existed on tumour cell (EGFr) combination, and optionally induce the cell of the tumour cell to showing these protein to suppress and/or cytotoxic effect.Exemplary antibodies also includes but not limited to, HERCEPTIN ^tM, can be used for the treatment of the Herceptin of mammary cancer and other forms of cancer, RITUXAN ^tM, Rituximab, ZEVALIN ^tM, ibritumomab tiuxetan, and LYMPHOCIDE ^tM, can be used for the treatment of the epratuzumab of non_hodgkin lymphoma and other forms of cancer, GLEEVEC ^tM, can be used for the treatment of the imatinib mesylate of chronic myeloid leukemia and gastrointestinal stromal tumor and BEXXAR ^tM, can be used for the treatment of the iodine 131 tositumomab of non_hodgkin lymphoma.Some exemplary antibodies also comprises ERBITUX ^tM; IMC-C225; Lressa ^tM; Gefitinib; TARCEVA ^tM, ertinolib (doubting as erlotinib erlotinib); KDR (kinase domain acceptor) inhibitor; VEGF antibody and antagonist (for example, Avastin ^tMand VEGAF-TRAP); Anti-vegf receptor antibody and antigen binding domain territory; Anti-Ang-1 and Ang-2 antibody and antigen binding domain territory; Antibody and other Ang-1 and Ang-2 acceptor for Tie-2; The Tie-2 part; Antibody for the Tie-2 kinase inhibitor; With

alemtuzumab.In certain embodiments, cancer therapeutic agent is other polypeptide of selective induction apoptosis in tumour cell, includes but not limited to, the TNF-related polypeptide is TRAIL for example.

In certain embodiments, the apoptotic specific-binding agent that IGF-1R is expressed in the combination of antagonism ligand i GF-1 and/or IGF-2 and IGF-1R (" IGF-1R ") and promotion (includes but not limited to, anti-IGF-R1 antibody) thus with antagonism and promote to express apoptotic specific-binding agent (including but not limited to TRAIL and the anti-TR-2 antibody) formulated in combination of TRAIL-R2 or use.Exemplary anti-IGF-1 R antibodies is known in the art, and is disclosed in the WO 2006/069202 that for example on December 20th, 2005 submits to, and described patent is incorporated herein by reference for any purpose.

In certain embodiments, cancer therapeutic agent is to reduce the anti-angiogenic agent that blood vessel occurs.Some this type of reagent includes but not limited to, ERBITUX ^tM, IMC-C225; KDR (kinase domain acceptor) inhibitor (antibody of for example, being combined with the kinase domain receptor-specific and antigen binding domain territory); Anti-VEGF reagent (for example, the antibody of specific binding VEGF or antigen binding domain territory, or soluble VEGF-receptor or its ligand binding region) is AVASTIN for example ^tMor VEGF-TRAP ^tM; Anti-vegf receptor reagent antibody or the antigen binding domain territory of its specific binding (for example, with); EGFR inhibitor (for example, with antibody or the antigen binding domain territory of its specific binding) is ABX-EGF for example, Victibix, IRESSA ^tM, Gefitinib, TARCEVA ^tM, erlotinib, anti-Ang-1 and anti-Ang-2 reagent (for example, with itself or with its acceptor for example antibody or the antigen binding domain territory of Tie2/Tek specific binding); For example, with anti-Tie-2 kinase inhibitor antibody or the antigen binding domain territory of its specific binding (, with).In certain embodiments, one or more reagent that pharmaceutical composition can also comprise specific binding and suppress growth factor activity (for example, antibody, antigen binding domain territory or soluble receptors), pHGF (HGF for example, also referred to as dispersion factor) antagonist, and the antibody of its acceptor of specific binding " c-met " or antigen binding domain territory.

Exemplary anti-angiogenic agent includes but not limited to, Campath, IL-8, B-FGF, Tek antagonist (people such as Ceretti, U.S. Patent Application Publication No. 2003/0162712; U.S. Patent number 6,413,932); Anti-TWEAK reagent (for example, specific binding antibody or antigen binding domain territory, or soluble T WEAK receptor antagonist; Referring to, for example, Wiley, U.S. Patent number 6,727,225); The ADAM disintegrin structural domain of the combination of antagonism integrin and its part (people such as Fanslow, U.S. Patent Application Publication No. 2002/0042368); The anti-eph acceptor of specific binding and/or anti-ephrin antibody or antigen binding domain territory (U.S. Patent number 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124; And patent family member); Anti-PDGF-BB antagonist (for example, specific binding antibody or antigen binding domain territory) and antibody or the antigen binding domain territory of with the PDGF-BB ligand specificity, being combined, for example, with PDGFR kinase inhibitor antibody or the antigen binding domain territory of its specific binding (, with).

Exemplary angiogenesis inhibitor/antitumor agent includes but not limited to, SF-7784 (Pfizer, USA); Cilengitide (EPO 770622 for Merck KgaA, Germany); Piperazine Jia Tani eight sodium (pegaptanib octasodium) (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, U.S. Patent number 5,712,291); Ilomastat (Arriva, USA, U.S. Patent number 5,892,112); Emaxanib (Pfizer, USA, U.S. Patent number 5,792,783); Vatalanib (Novartis, Switzerland); Methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); Anecortave acetate (Alcon, USA); α-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); Anti-Vn Mab (Crucell, Netherlands); DAC: angiogenesis inhibitor (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (EP 970070 for Novartis, Switzerland); ARGENT technology (Ariad, USA); YIGSR-Strealth (Johnson & Johnson, USA); Fibrinogen-E fragment (BioActa, UK); Angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (EntreMed, USA); Angiogenesis inhibitor (Tripep, Sweden); Maspin (Sosei, Japan); Methoxyestradiol (Oncology SciencesCorporation, USA); ER-68203-00 (IVAX, USA); Benefin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (JP 02233610 for Fujisawa, Japan); Platelet factor 4 (EP 407122 for RepliGen, USA); Vegf antagonist (Borean, Denmark); Temsirolimus (CCI-779) (University of South Carolina, USA); RhuMAb-VEGF (pINN) (Genentech, USA); Angiogenesis inhibitor (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); Mab, α 5 β 3 integrins, Vitaxin and s-generation Vitaxin (Applied Molecular Evolution, USA and MedImmune USA);

gene therapy (Oxford BioMedica, UK); Hydrochloric acid enzastaurin (USAN) (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); Angiogenesis inhibitor (Alchemia, Australia); VEGF antagonist (Regeneron, USA); The angiogenesis inhibitor (XOMA, USA) that rBPI 21 and BPI are derivative; PI 88 (Progen, Australia); Cilengitide (pINN) (Merck KgaA, Germany; Munich TechnicalUniversity, Germany; Scripps Clinic and Research Foundation, USA); Cetuximab (INN) (Aventis, France); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292 (Telios, USA); Endostatin (Boston Children ' s Hospital, USA); Methoxyestradiol (Boston Childrens Hospital, USA); ZD 6474 (AstraZeneca, UK); ZD6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); Vatalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); Tissue factor pathway inhibitor (EntraMed, USA); Piperazine Jia Tani (Pinn) (GileadSciences, USA); Yellow ginger root alcohol (Yonsei University, South Korea); Vaccine, based on gene, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX 103 (San Diego, the Universityof California of USA); PX 478 (Pro1X, USA); Metastatin (EntreMed, USA); Troponin I (Harvard University, USA); SU 6668 (SUGEN, USA); OXI 4503 (OXiGENE, USA); O-guanidine (Dimensional Pharmaceuticals, USA); Motuporamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); Atiprimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); FGF2 cancer vaccine (EntreMed, USA); Urokinase plasminogen activator inhibitor (Dendreon, USA); Oglufanide (pINN) (Melmotte, USA); HIF-1 alpha inhibitor (Xenova, UK); CEP 5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); Angiocidin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korean Research Institute of ChemicalTechnology, South Korea); GW 2286 (GlaxoSmithKline, UK); EHT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); Drug delivery system, intraocular, methoxyestradiol (EntreMed, USA); Anginex (Maastricht University, Netherlands and Minnesota University, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); Tumor necrosis factor alpha inhibitors (National Institute on Aging, USA); SU 11248 (Pfizer, USA and SUGEN USA); ABT 518 (Abbott, USA); YH16 (YantaiRongchang, China); S-3APG (Boston Childrens Hospital, USA and EntreMed, USA); Mab, KDR (ImClone Systems, USA); Mab, α 5 β 1 (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK and Johnson& Johnson, USA); GFB 116 (South Florida University, USA and Yale University, USA); CS 706 (Sankyo, Japan); Combretastatin A4 prodrug (Arizona State University, USA); Chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan and TAP, USA); AG 13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (WayneState University, USA); CV 247 (Ivy Medical, UK); CKD 732 (ChongKun Dang, South Korea); Mab, vascular endothelial growth factor (Xenova, UK); Irsogladine (INN) (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); Squalamine (pINN) (Genaera, USA); RPI 4610 (Sirna, USA); Breast fucan vitriol (galacto fucansulphate) (Marinova, Australia); Heparanase inhibitors (InSight, Israel); KL 3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Shering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); Vegf receptor conditioning agent (Pharmacopeia, USA); VE-cadherin-2 antagonist (ImClone Systems, USA); Vasostatin (National Institutes of Health, USA); Vaccine, Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); TumStatin (BethIsrael Hospital, USA); The solubility FLT 1 of brachymemma (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 part (Regeneron, USA); With thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA).

Some cancer therapeutic agent includes but not limited to: Thalidomide and thalidomide analogs (N-(2,6-dioxo-3-piperidyl) phthalimide); Tecogalan sodium (sulfated polysaccharides peptide glycan);

Bortezomib; Rapamycin; TAN 1120 (8-acetyl group-7,8,9,10-tetrahydrochysene-6,8,11-trihydroxy-1-methoxyl group-10-[[octahydro-5-hydroxyl-2-2-(hydroxypropyl)]-4,10-dimethyl pyrans is [3,4-d]-1 also, 3,6-dioxa azepine Xin Ying-8-yl] the oxygen base)-5,12-aphthacene diketone); Suradista (7,7 '-[carbonyl two [imino group (1-methyl isophthalic acid H-pyrroles-4,2-bis-bases) carbonyl imino group (1-methyl isophthalic acid H-pyrroles-4,2-bis-bases) carbonyl imino group]] two-1,3-naphthalenedisulfonic acid tetrasodium salt); SU 302; SU 301; SU 1498 ((E)-2-cyano group-3-[4-hydroxyl-3, two (1-Methylethyl) phenyl of 5-]-N-(3-phenyl propyl)-2-acrylamide); SU 1433 (4-(6,7-dimethyl-2-quinoxalinyl)-1,2-Benzenediol); ST 1514; SR 25989; Soluble T ie-2; The SERM derivative, Pharmos; Semaxanib (pINN) (3-[(3,5-dimethyl-1H-pyrroles-2-yl) methylene]-1,3-dihydro-2H-indol-2-one); S 836; RG 8803; RESTIN; R 440 (3-(1-Methyl-1H-indole-3-yl)-4-(1-methyl-6-nitro-1H-indol-3-yl)-1H-pyrroles-2,5-diketone); R 123942 (1-[6-(1,2,4-thiadiazoles-5-yl)-3-pyridazinyl]-N-[3-(trifluoromethyl) phenyl]-the 4-piperidinamine); Prolyl hydroxylase inhibitors; Progressive raising (progression elevated) gene; Prinomastat (INN) ((S)-2,2-dimethyl-4-[[p-(4-pyridine oxygen base) phenyl] sulfonyl]-3-thiomorpholine first hydroximic acid); NV 1030; NM 3 (8-hydroxyl-6-methoxyl group-Alpha-Methyl-1-oxo-1H-2-chromene-3-acetic acid); NF 681; NF 050; MIG; METH 2; METH 1;(α-[1-[4-[5-[4-[2-(3 for manassantinB, the 4-Dimethoxyphenyl)-2-hydroxyl-1-methyl ethoxy]-the 3-methoxyphenyl] tetrahydrochysene-3,4-dimethyl-2-furyl]-the 2-methoxyphenoxy] ethyl]-1,3-benzo dioxole-5-methyl alcohol); The KDR monoclonal antibody; α 5 β 3 integrin monoclonal antibodies; LY 290293 (2-amino-4-(3-pyridine radicals)-4H-naphtho-[1,2-b] pyrans-3-formonitrile HCN); KP0201448; KM 2550; The integrin specific peptide; INGN 401; GYKI 66475; GYKI 66462; Greenstatin (101-354-plasminogen (people)); The gene therapy of rheumatoid arthritis, prostate cancer, oophoroma, glioma, endostatin, colorectal cancer, ATF BTPI, anti-angiogenic generation gene, AI or Angiogenesis; The gelatinase inhibitor, FR 111142 (4,5-dihydroxy-2-hexenoic acid 5-methoxyl group-4-[2-methyl-3-(3-methyl-2-butene base) Oxyranyle]-1-oxaspiro [2.5] suffering-6-base ester); Forfenimex (pINN) (S)-alpha-amido-3-hydroxyl-4-(hydroxymethyl) phenylacetic acid); Fibronectin antagonist (1-acetyl group-L-prolyl-L-histidyl--L-seryl-L-cysteinyl-altheine); The fibroblast growth factor acceptor inhibitor; The fibroblast growth factor antagonist; FCE 27164 (7,7 '-[carbonyl two [imino group (1-methyl isophthalic acid H-pyrroles-4,2-bis-bases) carbonyl imino group (1-methyl isophthalic acid H-pyrroles-4,2-bis-bases) carbonyl imino group]] two-1,3,5-naphthalene trisulfonic acid six sodium salts); FCE 26752 (8,8 '-[carbonyl two [imino group (1-methyl isophthalic acid H-pyrroles-4,2-bis-bases) carbonyl imino group (1-methyl isophthalic acid H-pyrroles-4,2-bis-bases) carbonyl imino group]] two-1,3, the 6-naphthalene trisulfonic acid); Endothelial mononuclear cell activating polypeptide II; The VEGFR ASON; Anti-angiogenesis and ANFs; The ANCHOR angiostatin; Endostatin; The Del-1 angiogenic proteins; CT 3577; Contortrostatin; CM 101; Chondroitinase AC; CDP 845; CanStatin; BST 2002; BST 2001; BLS 0597; BIBF 1000; ARRESTIN; Apomigren (1304-1388-type XV collagen (people's gene COL15A1 α 1-chain precursor)); Angiostatin (angioinhibin); AaATIII; A 36;9 α-fluorine Medroxyprogesterone acetate (the fluoro-6-methyl of (6-α)-17-(acetoxyl group)-9--pregnant-4-alkene-3,20-diketone); 2-methyl-2-phthaloyl imino glutaric acid (2-(1,3-dihydro-1-oxo-2H-iso-indoles-2-yl)-2-methylglutaric acid); The monoclonal antibody BC-1 of Y90 mark; Semaxanib (3-(4,5-dimethyl pyrrole-2-methylene) Indolin-2-one) (C15H14N2O); PI 88 (phosphoric acid sweet dew pentose sulfate); Alvocidib (the 4H-1-benzopyran-4-one, 2-(2-chlorphenyl)-5,7-dihydroxy-8-(3-hydroxyl-1-methyl-4-piperidyl)-cis-(-)-) (C21H20ClNO5); E 7820; SU11248 (the fluoro-2-oxo-1 of 5-[3-, 2-indylidene-(3Z)-ylmethyl]-2,4-dimethyl-1H-pyrroles-3-formic acid (2-diethylamino ethyl) acid amides) (C22H27FN4O2); Squalamine (cholestane-7,24-glycol, 3-[[3-[(4-aminobutyl) aminopropyl] amino]-, 24-(hydrogen sulfate), (3. β, 5. α, 7. α)-] (C34H65N3O5S); Eriochrome Black T; AGM 1470 (carbamic acid, (chloracetyl)-, 5-methoxyl group-4-[2-methyl-3-(3-methyl-2-butene base) Oxyranyle]-1-oxaspiro [2,5] suffering-6-base ester, [3R-[3 α, 4 α (2R, 3R), 5 β, 6 β]]) (C19H28ClNO6); AZD 9935; BIBF 1000; AZD 2171; ABT 828; The KS-proleulzin; Uteroglobin; A6; NSC 639366 (1-[3-(diethylamino)-2-hydroxypropyl amino]-4-(oxirane-2-ylmethyl amino) anthraquinone fumarate) (C24H29N3O4.C4H4O4); ISV 616; Anti-ED-B fusion; HUI 77; Troponin 1; The BC-1 monoclonal antibody; SPV 5.2; ER 68203; (3-(3 for CKD 731,4, the 5-trimethoxyphenyl)-2 (E)-acrylic acid (3R, 4S, 5S, 6R)-4-[2 (R)-methyl-3 (R)-3 (R)-(3-methyl-2-butene base) oxirane-2-yl]-5-methoxyl group-1-oxaspiro [2.5] suffering-6-base ester) (C28H38O8); IMC-1C11; AaATIII; SC 7; CM 101; Angiocol; Kringle 5; CKD 732 (3-[4-[2-(dimethylamino) ethyoxyl] phenyl]-2 (E)-acrylic acid) (C29H41NO6); U 995;Canstatin; SQ 885; CT 2584 (1-[11-(dodecyl amino)-10-hydroxyl undecyl-3,7-dimethylxanthine] (C30H55N5O3); Salmosin; EMAP II; TX 1920 (1-(4-methyl piperazine)-2-(2-nitro-1H-1-imidazole radicals)-1-ethane ketone) (C10H15N5O3); α-v β-x inhibitor; CHIR 11509 (two (4-methoxyphenyl) formamides of N-(1-propinyl) glycyl-[N-(2-naphthyl)] glycyl-[N-(carbamyl ylmethyl)] glycine) (C36H37N5O6); BST2002; BST 2001; B 0829; FR 111142; 4,5-dihydroxy-2 (E)-hexenoic acid (3R, 4S, 5S, 6R)-4-[1 (R), 2 (R)-epoxies-1,5-dimethyl-4-hexenyl]-5-methoxyl group-1-oxaspiro [2.5] octane-6-base ester (C22H34O7); And inhibitors of kinases, include but not limited to N-(4-chlorphenyl)-4-(4-pyridylmethyl)-1-phthalazines amine (phthalazinamine); The chloro-3-of 4-[4-[[[[4-(trifluoromethyl) phenyl] amino] carbonyl] amino] phenoxy group]-N-methyl-2-pyridine carboxamide; N-[2-(diethylamino) ethyl]-5-[(5-is fluoro-1,2-dihydro-2-oxo--3H-indylidene-3-yl) methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide; 3-[(4-is bromo-2, the 6-difluorophenyl) methoxyl group]-5-[[[[4-(1-pyrrolidinyl) butyl] amino] carbonyl] amino]-the 4-isothiazole formamide; N-(the bromo-2-fluorophenyl of 4-)-6-methoxyl group-7-[(1-methyl-4-piperidines) methoxyl group]-4-quinazoline amine; 3-[5,6,7,13-tetrahydrochysene-9-[(1-methyl ethoxy) methyl]-5-oxo-12H-indeno [2,1-a] pyrrolo-[3,4-c] carbazole-12-yl] the propyl diester DMG; N-[5-[[[5-(1,1-dimethyl ethyl)-2-oxazolyl] methyl] sulfenyl]-the 2-thiazolyl]-the 4-piperidine formamide; The chloro-4-[(3-fluorophenyl of N-[3-) methoxyl group] phenyl]-6-[5-[[[2-(methyl sulphonyl) ethyl] amino] methyl]-the 2-furyl]-4-quinazoline amine; 4-[(4-methyl isophthalic acid-piperazinyl) methyl]-N-[4-methyl-3-[[4-(3-pyridine radicals)-2-pyrimidine radicals] amino]-phenyl] benzamide; N-(the chloro-4-fluorophenyl of 3-)-7-methoxyl group-6-[3-(4-morpholinyl) propoxyl group]-4-quinazoline amine; N-(3-ethynyl phenyl)-6, two (2-the methoxy ethoxy)-4-quinazoline amine of 7-;N-(3-((((2R)-1-methyl-2-pyrrolidinyl) methyl) oxygen base)-5-(trifluoromethyl) phenyl)-2-((3-(1,3-oxazole-5-yl) phenyl) amino)-Niacinamide; 2-(((4-fluorophenyl) methyl) amino)-N-(3-((((2R)-1-methyl-2-pyrrolidinyl) methyl) oxygen base)-5-(trifluoromethyl) phenyl)-Niacinamide; N-[3-(azetidine-3-ylmethoxy)-5-trifluoromethyl-phenyl]-2-(the fluoro-benzylamino of 4-)-niacinamide; The fluoro-N-of 6-(4-(1-Methylethyl) phenyl)-2-((4-pyridylmethyl) amino)-Niacinamide; 2-((4-pyridylmethyl) amino)-N-(3-(((2S)-2-pyrrolidinyl methyl) oxygen base)-5-(trifluoromethyl) phenyl)-Niacinamide; N-(3-(1,1-dimethyl ethyl)-1H-pyrazoles-5-yl)-2-((4-pyridylmethyl) amino)-Niacinamide; N-(3,3-dimethyl-2,3-dihydro-1-benzofuran-6-yl)-2-((4-pyridylmethyl) amino)-Niacinamide; N-(3-((((2S)-1-methyl-2-pyrrolidinyl) methyl) oxygen base)-5-(trifluoromethyl) phenyl)-2-((4-pyridylmethyl) amino)-Niacinamide; 2-((4-pyridylmethyl) amino)-N-(3-((2-(1-pyrrolidinyl) ethyl) oxygen base)-4-(trifluoromethyl) phenyl)-Niacinamide; N-(3,3-dimethyl-2,3-dihydro-1H-indoles-6-yl)-2-((4-pyridylmethyl) amino)-Niacinamide; N-(4-(pentafluoroethyl group)-3-(((2S)-2-pyrrolidinyl methyl) oxygen base) phenyl)-2-((4-pyridylmethyl) amino)-Niacinamide; N-(3-((3-azetidine ylmethyl) oxygen base)-5-(trifluoromethyl) phenyl)-2-((4-pyridylmethyl) amino)-Niacinamide; N-(3-(4-piperidyl oxygen base)-5-(trifluoromethyl) phenyl)-2-((2-(3-pyridine radicals) ethyl) amino)-Niacinamide; N-(4,4-dimethyl-1,2,3,4-tetrahydro-isoquinoline-7-yl)-2-(1H-indazole-6-base amino)-niacinamide; 2-(1H-indazole-6-base amino)-N-[3-(1-methylpyrroline-2-ylmethoxy)-5-trifluoromethyl-phenyl]-niacinamide; N-[1-(2-dimethylamino-acetyl group)-3,3-dimethyl-2,3-dihydro-1H-indoles-6-yl]-2-(1H-indazole-6-base amino)-niacinamide; 2-(1H-indazole-6-base amino)-N-[3-(pyrrolin-2-ylmethoxy)-5-trifluoromethyl-phenyl]-niacinamide;N-(1-acetyl group-3,3-dimethyl-2,3-dihydro-1H-indoles-6-yl)-2-(1H-indazole-6-base amino)-niacinamide; N-(4,4-dimethyl-1-oxo-1,2,3,4-tetrahydro-isoquinoline-7-yl)-2-(1H-indazole-6-base amino)-niacinamide; N-[4-(tert-butyl group)-3-(3-piperidyl propyl group) phenyl] [2-(1H-indazole-6-base amino) (3-pyridine radicals)] formamide; N-[5-(tert-butyl group) isoxazole-3-base] [2-(1H-indazole-6-base amino) (3-pyridine radicals)] formamide; And N-[4-(tert-butyl group) phenyl] [2-(1H-indazole-6-base amino) (3-pyridine radicals)] formamide, and be disclosed in the inhibitors of kinases in following patent: U.S. Patent number 6,258,812; 6,235,764; 6,630,500; 6,515,004; 6,713,485; 5,521,184; 5,770,599; 5,747,498; 5,990,141; U.S. Patent Application Publication No. US2003/0105091; With Patent Cooperation Treaty publication number WO01/37820; WO01/32651; WO02/68406; WO02/66470; WO02/55501; WO04/05279; WO04/07481; WO04/07458; WO04/09784; WO02/59110; WO99/45009; WO98/35958; WO00/59509; WO99/61422; WO00/12089; And WO00/02871, described publication is hereby incorporated by for any purpose separately.

TR-2 is at various cells, comprises liver, brain, kidney, colon, mammary gland, lung, spleen, thymus gland, peripheral blood lymphocyte, pancreas, prostate gland, testis, ovary, uterus and along GI various tissues.Example T R-2 associated cancer includes but not limited to, liver cancer, the cancer of the brain, kidney, mammary cancer, carcinoma of the pancreas, colorectal carcinoma, lung cancer (small cell lung cancer and nonsmall-cell lung cancer), spleen cancer, thymus gland or hemocyte cancer (that is, leukemia), prostate cancer, carcinoma of testis, ovarian cancer, uterus carcinoma, cancer of the stomach, H&N squamous cell carcinoma, melanoma and lymphoma.

In certain embodiments, anti-TR-2 antibody can be used separately or together with at least one the other therapeutical agent that is used for the treatment of cancer.In certain embodiments, anti-TR-2 antibody is combined with the other therapeutical agent for the treatment of significant quantity.The exemplary treatment agent that can use together with anti-TR-2 antibody includes but not limited to, the antibiotic geldanamycin family member of anisamycin (doubting as anisomycin Anisomycin); Pro-HGF; NK2; The c-Met inhibitor peptides; The antagonist of Grb2Src homologue 2; The Gab1 conditioning agent; Dominant Src; The von-Hippel-Landau inhibitor, include but not limited to, wortmannin; The P13 kinase inhibitor, other anti-autogenic therapies, anti-EGFr, cox 2 inhibitor, Celebrex of being subject to ^tM, celecoxib, Vioxx ^tM, rofecoxib; Vascular endothelial growth factor (VEGF), VEGF conditioning agent, fibroblast growth factor (FGF), FGF conditioning agent, Urogastron (EGF); The EGF conditioning agent; Keratinocyte growth factor (KGF), KGF associated molecule, KGF conditioning agent; And matrix metalloproteinase (MMP) conditioning agent.

In certain embodiments, anti-TR-2 antibody is used to treat various cancers together with particular therapeutic agent.In certain embodiments, consider symptom and required treatment level, can use 2 kinds, 3 kinds or more kinds of reagent.When compound is used together with one or more other components, other combinations of compound and one or more can together with, separately or in turn (for example,, with medicament forms) use.In certain embodiments, this type of reagent can provide together by being included in same preparation.In certain embodiments, this type of reagent can provide with anti-TR-2 antibody together with being included in same preparation.In certain embodiments, this type of reagent can separately be prepared and provide together by being included in the treatment test kit.In certain embodiments, this type of reagent can separately be prepared with anti-TR-2 antibody and provide together with being included in the treatment test kit.In certain embodiments, this type of reagent can separately provide.

In certain embodiments, when using by gene therapy, the gene of coded protein reagent and/or anti-TR-2 antibody can be included in same vehicle.In certain embodiments, the gene of coded protein reagent and/or anti-TR-2 antibody can be under the control in identical promoters district.In certain embodiments, the gene of coded protein reagent and/or anti-TR-2 antibody can be in the carrier separated.

In certain embodiments, anti-TR-2 antibody can be used for the treatment of the non-human animal, such as pet (dog, pig, bird, primates etc.), and raise and train farm-animals (horse?, ox, sheep, pig, bird etc.).In some this type of situation, suitable dosage can be determined according to the body weight of animal.For example, in certain embodiments, can use the dosage of 0.2-1mg/kg.In certain embodiments, dosage can determine according to the surface-area of animal, and exemplary dose is 0.1-20mg/in ², or 5-12mg/m ².For animalcule, for example dog or cat, in certain embodiments, suitable dosage is 0.4mg/kg.In certain embodiments, anti-TR-2 antibody can be used weekly one or many by injection or other suitable pathways, until the symptom of animal improves, or it can be used indefinitely.

Be to be understood that single patient can be different to replying of aforementioned medicine or combination treatment, and the suitable effective combination of medicine can be decided by his or her doctor for each patient.

Cynomolgus monkey provides the useful model about some disease.Exemplary disease includes but not limited to, transplant rejection syndrome and inflammatory bowel sample disease.When the effect of test person MAb in cynomolgus monkey human disease model, in certain embodiments, determine whether anti-TR-2MAb is useful with comparable horizontal integration TR-2 in people and cynomolgus monkey.

In certain embodiments, anti-TR-2 antibody can be the part of the conjugate molecules that comprises the anti-TR-2 antibody of all or part and cytotoxic agent.Term " cytotoxic agent " refers to suppress or stop the cell function and/or cause necrocytosis or the material of destruction.This term includes but not limited to, radio isotope (for example, I ¹³¹, I ¹²⁵, Y ⁹⁰and Re ¹⁸⁶), chemotherapeutic, and the toxin enzymatic activity toxin of bacterium, fungi, plant or animal-origin for example, or its fragment.The agent of exemplary cells toxin includes but not limited to, Doxorubicin, Zorubicin, 5 FU 5 fluorouracil, cytarabin (" AraC "), endoxan, thiophene is for group, taxotere (docetaxel), busulfan, Cytoxin, taxol, methotrexate, cis-platinum, melphalan, vinealeucoblastine(VLB), bleomycin, Etoposide, ifosfamide, ametycin, mitoxantrone, vincristine(VCR), vinorelbine, carboplatin, teniposide, daunomycin, carminomycin, aminopterin, gengshengmeisu, mitomycin, the Ai Sibo mycin, melphalan and other relevant mustargen.

In certain embodiments, anti-TR-2 antibody can be the part of the conjugate molecules that comprises the anti-TR-2 antibody of all or part and prodrug.In certain embodiments, term " prodrug " refers to precursor or the derivative form of pharmaceutically active substances.In certain embodiments, with parent drug, compare, prodrug is less to the cytotoxicity of cell, and can be activated or be transformed into more active cytotoxicity parent form by enzymatic.The exemplary precursors medicine includes but not limited to, the phosphate-containing prodrug, containing sulfo-phosphate precursor medicine, the containing sulfate prodrug, containing the peptide prodrug, the prodrug that D-is amino acid modified, the glycosylation prodrug, containing the beta-lactam prodrug, containing the optional phenoxy acetamide prodrug replaced with containing the optional phenylacetamide prodrug replaced, can be transformed into 5-flurocytosine and other 5-FUD prodrugs of more active cytotoxicity free drug.The example that can be derivatized to the cytotoxic drug of prodrug includes but not limited to, above-described those cytotoxic agents.Referring to, for example, U.S. Patent number 6,702,705.

In certain embodiments, the specific cell precursor in the prodrug part target patient of the antibody moiety targeted cells toxicity part of antibody conjugates by making molecule or molecule works.In the situation that anti-TR-2 antibody, this type of conjugate molecules can for, for example, in certain embodiments, destroy the cell of abnormality proliferation, for example cancer cells.

In certain embodiments, provide treatment patient's method, described method comprises the anti-TR-2 antibody of administering therapeutic significant quantity.In certain embodiments, provide treatment patient's method, described method comprises the antibody conjugates of administering therapeutic significant quantity.In certain embodiments, as mentioned above, antibody is combined with at least one other therapeutical agent for the treatment of significant quantity.

As mentioned above, in certain embodiments, anti-TR-2 antibody can be used with one or more other drugs that are applied to same patient simultaneously, and every kind of medicine is used according to the scheme that is suitable for that medicine.This type for the treatment of comprises pretreat, treats simultaneously, treats in turn and alternate scheme.The other example of this type of medicine includes but not limited to, antiviral agent, microbiotic, pain killer, reflunomide, inflammatory cytokine antagonist, DMARDs, non-steroidal anti-inflammatory drugs, chemotherapy, angiogenesis inhibitor and vasculogenesis stimulator.

In certain embodiments, various medical conditions are used with the anti-TR-2 antibody of another kind of apoptotic stimulus thing combination and are treated.For example, in certain embodiments, anti-TR-2 antibody can be used in composition, and described composition also comprises stimulates one or more apoptotic compounds.In certain embodiments, anti-TR-2 antibody and apoptotic stimulus thing can be used as composition separately and are used, and these can be used by identical or different approach.

In certain embodiments, provide pharmaceutical composition, described pharmaceutical composition comprise treat significant quantity antibody together with pharmaceutically acceptable thinner, carrier, solubilizing agent, emulsifying agent, sanitas and/or adjuvant.

In certain embodiments, pharmaceutical composition is provided, described pharmaceutical composition comprises the antibody for the treatment of significant quantity and at least one other therapeutical agent for the treatment of significant quantity, together with pharmaceutically acceptable thinner, carrier, solubilizing agent, emulsifying agent, sanitas and/or adjuvant.

In certain embodiments, acceptable preparation material is preferably nontoxic to acceptor on used dosage and concentration.In certain embodiments, antibody of the present invention provides in the preparation without buffer reagent as in being disclosed in PCT/US06/22599, and described patent was submitted on June 8th, 2006, for any purpose is incorporated herein by reference.

In certain embodiments, pharmaceutical composition can comprise for modifying, maintain or preserve for example preparation material of pH, perviousness, viscosity, transparency, color, isotonicity, smell, sterility, stability, dissolving or rate of release, absorption or the infiltration of composition.In certain embodiments, suitable preparation material includes but not limited to, amino acid (for example glycine, glutamine, l-asparagine, arginine or Methionin); Biocide; Antioxidant (for example xitix, S-WAT or sodium bisulfite); Buffer reagent (for example, borate, supercarbonate, Tris-HCl, Citrate trianion, phosphoric acid salt or other organic acids); Swelling agent (for example N.F,USP MANNITOL or glycine); Sequestrant (for example ethylenediamine tetraacetic acid (EDTA) (EDTA)); Complexing agent (for example caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin); Filling agent; Monose; Disaccharides; For example, with other carbohydrate (glucose, seminose or dextrin); Protein (for example serum albumin, gelatin or immunoglobulin (Ig)); Tinting material, seasonings and thinner; Emulsifying agent; Hydrophilic polymer (for example polyvinylpyrrolidone); Low molecular weight polypeptide; Salify counter ion (for example sodium); Sanitas (for example Zephiran chloride, phenylformic acid, Whitfield's ointment, Thiomersalate, phenylethyl alcohol, methyl p-hydroxybenzoate, propylparaben, chlorhexidine, Sorbic Acid or hydrogen peroxide); Solvent (for example glycerine, propylene glycol or polyoxyethylene glycol); Sugar alcohol (for example N.F,USP MANNITOL or Sorbitol Powder); Suspension agent; Tensio-active agent or wetting agent (polyethers for example, EG, sorbitan ester, polysorbate is polysorbate20, polysorbate80 for example, trotyl, trometamol, Yelkin TTS, cholesterol, tyloxapal (tyloxapol)); Stability enhancer (for example sucrose or Sorbitol Powder); Tension-elevating agent (basic metal halogenide for example, preferably sodium-chlor or potassium, N.F,USP MANNITOL?, Sorbitol Powder); Delivery vehicle; Thinner; Vehicle and/or medicine adjuvant.(Remington ' s Pharmaceutical Sciences, the 18th edition, A.R.Gennaro, editor, Mack Publishing Company (1990).

In certain embodiments, antibody and/or other treatment molecule are connected with Increased Plasma Half-life vehicle known in the art.This type of vehicle includes but not limited to, Fc structural domain, polyoxyethylene glycol and dextran.This type of vehicle is described in for example U.S. Patent number 6,660,843 and disclosed PCT application number WO 99/25044.

In certain embodiments, the optimal drug composition will by those skilled in the art depend on for example expect route of administration, send mode and required dosage is determined.Referring to, for example, Remington ' s Pharmaceutical Sciences, the same.In certain embodiments, such composition may affect the interior rate of release of stability, body and the interior clearance rate of body of physical state, antibody.

In certain embodiments, the main media thing in pharmaceutical composition or carrier can be water or non-water in nature.For example, in certain embodiments, suitable vehicle or carrier can be water for injection, normal saline solution or artificial cerebrospinal fluid, may be supplemented with the other materials common for the composition of parenteral administration.In certain embodiments, neutral buffered saline or the salt solution that mixes with serum albumin are further exemplary vehicle.In certain embodiments, the Tris damping fluid that pharmaceutical composition comprises about pH 7.0-8.5, or the acetate buffer of about pH4.0-5.5, it may further include Sorbitol Powder or its suitable substituent.In certain embodiments, pharmaceutical composition is the acetate buffer that comprises about pH 4.0-5.5, polyol (polyvalent alcohol) and optionally water or the liquid preparation of tensio-active agent, wherein said composition does not comprise salt, for example, sodium-chlor, and wherein said composition is oozed for patient etc.Exemplary polyol includes but not limited to, sucrose, glucose, Sorbitol Powder and N.F,USP MANNITOL.Exemplary surfactants includes but not limited to, polysorbate.In certain embodiments, pharmaceutical composition is water or the liquid preparation of the acetate buffer, Sorbitol Powder and the polysorbate that comprise about pH 5.0, and wherein said composition does not comprise salt, for example, sodium-chlor, and wherein said composition is oozed for patient etc.Some exemplary composition is for example finding in U.S. Patent number 6,171,586.Other pharmaceutical carrier includes but not limited to, oil comprises oil, animal oil, vegetables oil, peanut oil, soybean oil, mineral oil, sesame wet goods.In certain embodiments, glucose and aqueous glycerin solution also can be used as liquid vehicle, particularly for Injectable solution.In certain embodiments, to there is the selected composition of required purity and the optional preparaton (Remington ' sPharmaceutical Sciences of freeze-drying agglomerate or aqueous solution form, the same) mix, can prepare comprise antibody, together with or not together with at least one the composition of other therapeutical agent for storing.In addition, in certain embodiments, use suitable excipient solution (for example, sucrose) as thinner can by comprise antibody, together with or together with at least one, the composition of other therapeutical agent is not formulated as lyophilized products.

In certain embodiments, anti-TR-2 antibody is used with the form of the acceptable composition of physiology, the recombinant protein that the acceptable composition of described physiology comprises the purifying of being combined with physiology acceptable carrier, vehicle or thinner.In certain embodiments, examples of such carriers is nontoxic to acceptor on used dosage and concentration.In certain embodiments, the preparation of such composition can relate to makes anti-TR-2 antibody and following substances combination: buffer reagent, antioxidant be xitix for example, low molecular weight polypeptide (for example, have be less than 10 amino acid whose those), protein, amino acid, carbohydrate is glucose, sucrose or dextrin for example, sequestrant is EDTA for example, gsh and/or other stablizers and vehicle.In certain embodiments, suitable dosage is determined, and can be become according to selected route of administration in the standard medicine-feeding test.In certain embodiments, according to suitable industrial standards, can also add sanitas, this includes but not limited to, phenylcarbinol.In certain embodiments, the amount of using and frequency can be based on this type of factor as the character of disease to be treated and severity, requiredly reply, age of patient and condition etc. determine.

In certain embodiments, pharmaceutical composition can be selected to send for parenteral.The preparation of some this type of pharmaceutically acceptable composition is in art technology.

In certain embodiments, formulation component exists to use the acceptable concentration in site.In certain embodiments, use damping fluid so that composition is maintained at physiology pH or slightly lower pH, be generally approximately 8 pH of about 5-.

In certain embodiments, when expection during parenteral administration, pharmaceutical composition can be in acceptable vehicle pharmaceutically, comprise required antibody, together with or not together with pyrogen-free, the acceptable aqueous solution form of parenteral of other therapeutical agent.In certain embodiments, for the vehicle of parenteral injection, be sterile distilled water, wherein antibody, together with or together with other therapeutical agent, be not configured to that suitable rot-resistant is aseptic, isotonic solution.In certain embodiments, preparation can relate to that for example particle, polymerizable compound (for example poly(lactic acid) or polyglycolic acid), pearl or liposome formulation desired molecule are separated in injectable microsphere, biological erosion with reagent, described reagent may provide control or the sustained release of product, and described product can be sent via depot injection subsequently.In certain embodiments, hyaluronic acid also can be used, and may have the effect that promotes the time length in circulation.In certain embodiments, can use implantable drug delivery device to introduce desired molecule.

In certain embodiments, pharmaceutical composition can be prepared for sucking.In certain embodiments, antibody, together with or the other therapeutical agent together with at least one not, can be configured to dry powder for sucking.In certain embodiments, comprise antibody, together with or together with at least one, the inhalation solution of other therapeutical agent can be with propelling agent preparation for aerosol delivery.In certain embodiments, solution can be vaporific.Pulmonary administration further describes in PCT publication number WO94/20069, and described patent has been described the pulmonary delivery of the protein of chemically modified.

In certain embodiments, the expection preparation can be used by oral area.In certain embodiments, the antibody of using by this way, together with or the other therapeutical agent together with at least one not, can together with or not together with solid dosage for example Tablet and Capsula coordinate in those carriers commonly used prepare.In certain embodiments, capsule can be designed as in gi tract when bioavailability maximize and system before the active part of delivery formulations on some during minimum degradation.In certain embodiments, can comprise the absorption of at least one other reagent with enhancing antibody and/or any other therapeutical agent.In certain embodiments, can also use thinner, seasonings, low melt wax, vegetables oil, lubricant, suspension agent, tablet disintegrant and/or tackiness agent.

In certain embodiments, pharmaceutical composition can relate to the antibody of significant quantity, together with or the other therapeutical agent together with at least one not, and be suitable for non-toxic excipients prepared by tablet and mix.In certain embodiments, by making tablet dissolved in sterilized water or another kind of suitable vehicle, can prepare the solution of unit dosage.Suitable vehicle includes but not limited to, inert diluent, for example calcium carbonate, sodium carbonate or sodium bicarbonate, lactose or calcium phosphate; With tackiness agent for example starch, gelatin and gum arabic; With lubricant for example Magnesium Stearate, stearic acid and talcum.

Other pharmaceutical composition will be apparent for those skilled in the art, be included in and continue or control in delivery formulation to relate to antibody, together with or the not preparation of other therapeutical agent together with at least one.Some exemplary continue or control delivery formulations in include but not limited to, particulate, porous bead and depot injection are separated in liposome vectors, biological erosion.Some example technique for the preparation of some preparation is well known by persons skilled in the art.Referring to for example, PCT publication number WO93/15722, the control that described patent has been described for the porous polymerizing particulate of delivering drugs composition discharges.In certain embodiments, extended release preparation can comprise for example semipermeable polymers matrix of film or microencapsulation form of formed article form.Sustained release matrix includes but not limited to, polyester, hydrogel, polylactide (U.S. Patent number 3,773,919 and EP 058,481), Pidolidone and the γ ethyl-Pidolidone salt copolymer (people such as Sidman, Biopolymers, 22:547-556 (1983)), poly-(2-hydroxyethyl-methacrylate) (people such as Langer, J.Biomed.Mater.Res., 15:167-277 (1981) and Langer, Chem.Tech., 12:98-105 (1982)), the ethylene vinyl acetate (people such as Langer, the same) and poly-D (-)-3-hydroxybutyric acid (EP 133,988).In certain embodiments, sustained-release composition can also comprise liposome, and in certain embodiments, described liposome can be by any preparation the in several method known in the art.Referring to for example, the people such as Eppstein, Proc.Natl.Acad.Sci.USA, 82:3688-3692 (1985); EP 036,676; EP 088,046 and EP 143,949.

In certain embodiments, for the pharmaceutical composition of using in body, be aseptic.In certain embodiments, for the pharmaceutical composition used in body by making aseptic via aseptic membrane filtration.In certain embodiments, when composition is lyophilized, with the sterilizing of aseptic filter membrane, can before or after freeze-drying and reconstruct, carry out.In certain embodiments, for the composition of parenteral administration, can store with lyophilized form or at solution.In certain embodiments, parenteral composition generally is placed in the container with sterile access port, and described container for example has can be by intravenous solution bag or the phial of the stopper of subcutaneous injection needle-penetration.

In certain embodiments, after pharmaceutical composition is prepared, it can be used as solution, suspension, gel, milk sap, solid, or is stored in aseptic phial as dehydration or lyophilized powder.In certain embodiments, this type of preparation can be for example, to store by form or the form (, lyophilized form) that is reconstructed before using.

In certain embodiments, provide the test kit for the production of the single dose unit of using.In certain embodiments, this test kit can each self-contained the second container that has the first container of drying protein and have water preparation.In certain embodiments, the test kit that has comprised the pre-filled syringe (for example fluid injector and lyosyringes) that comprises list and/or multicompartmented.

In certain embodiments, the significant quantity of the pharmaceutical composition used in treatment will depend on for example treats background and purpose, and described pharmaceutical composition comprises antibody, together with or the other therapeutical agent together with at least one not.Those skilled in the art are to be understood that according to some embodiment, therefore the appropriate dose level be used for the treatment of complies with by part the molecule of sending, antibody, together with or together with at least one, other therapeutical agent will be for its indication, route of administration, and patient size (body weight, body surface or organ size) and/or condition (age and general health) and become.In certain embodiments, the clinician can progressively increase dosage and revise route of administration to obtain optimum curative effect.In certain embodiments, depend on above-mentioned factor, general dosage can be for about 0.1 μ g/kg-up to about 100mg/kg or more.In certain embodiments, dosage can be for 0.1 μ g/kg-up to about 100mg/kg; Or 1 μ g/kg-up to about 100mg/kg; Or 5 μ g/kg-up to about 100mg/kg; Or 0.1mg/kg-is up to about 100mg/kg.

The pharmacokinetic parameter of the antibody in the preparation that in certain embodiments, administration frequency will be considered to use and/or any other therapeutical agent.In certain embodiments, the clinician will use composition until reach the dosage that obtains required effect.In certain embodiments, therefore said composition can be used as single dose, or along with past time as 2 times or more times dosage (this can comprise or not comprise the desired molecule of same amount), or use as continuous infusion via implanted device or conduit.Further improve some method of suitable dosage in art technology.In certain embodiments, suitable dosage can be determined by the dose response data with suitable.

In certain embodiments, the route of administration of pharmaceutical composition is according to currently known methods, oral area for example, by approach in (in essence), Intraventricular, intramuscular, intraocular, intra-arterial, portal vein in intravenously, intraperitoneal, brain or in damage via injection; By sustained release system or pass through implanted device.In certain embodiments, said composition can be by injecting or continuous infusion or use by implanted device.

As mentioned above, in certain embodiments, any effective route of administration can be for using anti-TR-2 antibody.If injection, so in certain embodiments, anti-TR-2 antibody can be for example via in intraarticular, intravenously, intramuscular, damage, in intraperitoneal, encephalic, nose, suck or by injecting or using by the subcutaneous route of continuous infusion.Exemplary application process includes but not limited to, from the sustained release of implant, aerosol sucks, eye drops, the oral area preparation, comprise pill, syrup, lozenge and chewing gum, and local formulation example is as lotion, gelifying agent, sprays, ointment and other suitable technology.

In certain embodiments, when the treatment disease relevant with the lung illness, it is favourable by suction, using.In certain embodiments, the culturing cell that anti-TR-2 antibody can be expressed antibody by implantation is used.In certain embodiments, by use, encode in one or more carrier bodies of anti-TR-2 antibody or self cell of producing the patient is induced in vitro transfection.In certain embodiments, by injection encode naked DNA or the liposome encapsulated dna of anti-TR-2 antibody, or, by other transfection methods, this carrier can be introduced in patient's cell.When other biologically active cpds combined administration of anti-TR-2 antibody and one or more, in certain embodiments, these can be by identical or use by different approaches, and can be together, separately or use in turn.

In certain embodiments, said composition can be carried out topical application via implanting the film, sponge or the another kind of suitable material that have absorbed or encapsulated desired molecule on it.In certain embodiments, when using implanted device, this device can be implanted in any suitable tissue or organ, and sending of desired molecule can be via diffusion, time controlled released bolus or continuous administration.

In certain embodiments, may wish to use and comprise antibody in vitro mode, together with or the pharmaceutical composition of other therapeutical agent together with at least one not.In this type of embodiment, make the cell, tissue and/or the organ that have taken out from the patient be exposed to pharmaceutical composition, described pharmaceutical composition comprises antibody, together with or the other therapeutical agent together with at least one not, after this, cell, tissue and/or organ are planted and are got back in the patient subsequently.

In certain embodiments, antibody and any other therapeutical agent can be sent by implanting some cell, described cell using method for example described herein those carry out genetic modification, to express and secrete polypeptide.In certain embodiments, this type of cell can be animal or human's cell, and can be autologous homology, allos or xenogenesis.In certain embodiments, this cell can be immortalization.In certain embodiments, in order to reduce the probability of immunne response, this cell can be encapsulated to avoid the infiltration of peripheral organization.In certain embodiments, packaged material is generally biocompatible, semipermeability polymeric shell or film, and it allows one or more proteins to discharge but prevents that cell from destroying by patient's immunity system or from other injurious factors of peripheral organization.

Embodiment

Embodiment 1

the production of some human monoclonal antibodies

The anti-TR-2 antibody of people is produced with one of 2 kinds of methods.Make to express human immunoglobulin gene's transgenic mice

be exposed to people TR-2.Use hybridoma technology to produce the anti-TR-2 monoclonal antibody of some people by these mouse.Use the XenoMax technology to produce some other the anti-TR-2 monoclonal antibody of people by those mouse, described XenoMax technology merged selection lymphocyte antibody method (" SLAM ") technology (referring to, for example, U.S. Patent number 5,627,052; With the people such as Babcook, Proc.Natl.Acad.Sci.USA 93:7843-7848 (1996)).

The method of producing the anti-TR-2 monoclonal antibody of people for the transgenic mice expressing the human immunoglobulin gene is as follows.As shown in fig. 1,5 groups of recombinant human TR-2 with C-terminal six histidine marks (TR-2-His) for mouse (ripe aminoacid sequence ALITQQDLAPQQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWND LLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMV KVGDCTPWSDIECVHKESGTKHSGEAPAVEETVTSSPGTPASRSGSSHHHHHH (SEQ ID NO:140)) (Genbank reference number NM-003842) carry out immunity.Group 1, organize 3, organize 4 and organize the antibody (Fig. 2) that mouse in 5 is transformed to produce the IgG2 isotype.Mouse in group 2 is transformed to produce the antibody (Fig. 2) of IgG4 isotype.Group 1 comprises 7 mouse, organizes 2 and comprises 8 mouse, organizes 3 and comprises 8 mouse, organizes 4 and comprises 10 mouse, and organize 5 and comprise 5 mouse.Group 1, organize 2 and organize mouse in 3 and carry out immunity (10 μ g/ injection) by TR-2-His being expelled in palmula, and organize 4 and group 5 in mouse carry out intraperitoneal immunity (10 μ g/ injection) with TR-2-His.In the time of the 0th day, by described approach, use 10 μ g antigens.When appointed interval, to mouse, use booster shots.Organize 1 little, mouse and there are 9 booster shots, in the time of the 5th, 11,14,18,24,28,34,42 and 46 days.Group 2 and group 3 mouse have 7 booster shots; About organize 2 those the 3rd, 7,10,14,17,24 and 27 days the time, and about organize 3 those the 5th, 8,15,21,26,30 and 33 days the time.Group 4 and group 5 mouse have 5 booster shots, in the time of the 14th, 28,42,56 and 72 days.Each comprises 10 μ g TR-2-His and adjuvants with each booster shots first, and described adjuvant is Titermax Gold (organizing 1,2 and 3), alum gel (organizing 1,2 and 3), complete Freund's adjuvant (CFA) (organizing 4 and 5), incomplete Freund's adjuvant (IFA) (organizing 4 and 5) or DulbeccoShi phosphate buffered saline (PBS) (D-PBS) (organizing 1,2,3,4 and 5) (referring to Fig. 1).Inject rear (organizing 4 and 5) at 3 times, at 4 times, inject rear (organizing 1,2 and 3), at 6 times, inject rear (organizing 1 and 2) and inject rear (organizing 1) at 10 times mouse is got to blood.As shown in Figure 2, get blood is assessed by ELISA the reactivity of TR-2-His at every turn.

The ELISA assay method is carried out as follows.By the coated porous flat plate of soluble T R-2-His (0.5 μ g/mL) for Passive intake that spends the night in 4.Coated hole is washed and seals 30 minutes with breast.Make the combination of every kind of mice serum of 10 μ L and 40 μ L breast, and in different dull and stereotyped holes incubation 1 hour, 1.25 hours or 2 hours.Flat board washes with water 5 times.Make subsequently dull and stereotyped and incubation 1 hour at room temperature together with the mountain of final concentration 1 μ g/mL goat anti-human igg Fc specificity horseradish peroxidase is puted together antibody (Pierce).Flat board washes with water 5 times.Make dull and stereotyped and incubation together with K indigo plant substrate (Neogen) 30 minutes.Negative control comprises the blank well that lacks TR-2-His, the hole of incubation together with the natural G2 serum that comprises TR-2-His but lack anti-TR-2 antibody with expection.

Method for the production of the anti-TR-2 monoclonal antibody of people is as follows.For the XenoMax technology, separation of C D19+B cell from high immune transgenic mouse, results when after immunization is initial the 37th day of described high immune transgenic mouse (mouse M712-7 is from group 3) or the 76th day (mouse M564-1 is from group 4, and mouse M564-3, M564-5 and M563-5 are from group 5).The B cell cultures also is divided into to plasmocyte subsequently to allow it to increase in 1 week.The supernatant liquor that comprises secretory antibody is preserved for further analysis, and be frozen in the substratum that comprises 10%DMSO and 90%FCS-80celcius of the plasmocyte in each hole degree (doubt as celsius, degree centigrade).For hybridoma technology, as shown in fig. 1, from the cell of the high immune transgenic mouse of residue the 31st, 37 or 46 days time results for further analyzing.

For the XenoMax technology, by the ELISA supernatant liquor from the B cell culture for the existence screening of the antibody of TR-2 just.Use as follows anti-human IgG antibody test reagent to detect anti-TR-2 antibody by assessment and the combination of immobilization TR-2-His.By coated dull and stereotyped with soluble T R-2-His (0.5 μ g/mL) in 4 ℃ of Passive intakes that spend the night.Water after 30 minutes, makes 10 μ L cell conditioned medium liquid and the combination of 40 μ L breast from each single hybridoma by the sealing of the hole in flat board by flat board washing 5 times and with breast, and in the holes of different flat boards incubation 1 hour, 1.25 hours or 2 hours.Flat board washes with water 5 times, and with incubation 1 hour at room temperature together with the mountain goat anti-human igg Fc specificity horseradish peroxidase (Pierce) of final concentration 1 μ g/mL is puted together antibody.Flat board washes with water 5 times.Make dull and stereotyped and incubation together with K indigo plant substrate (Neogen) 30 minutes.Negative control comprises the blank well that lacks TR-2-His, and uses expection to lack the hole of the natural G2 serum of anti-TR-2 antibody.Screen positive for TR-2-His for the second time by ELISA, to confirm to produce the cell identity of the special antibody to TR-2.

The antibody reacted with TR-2 that uses the apoptosis assay method to induce the ability screening of WM-266 melanoma cells (ATCC catalog number (Cat.No.) CRL-1676) apoptosis above to identify with regard to it.As the density overnight incubation in the normal cell substratum with 4500 cells/well in microtiter plate of the WM-266 cell by the ATCC suggestion.For the B cell cultures, in 180 μ L apoptosis substratum mixtures (cell culture medium that comprises 1 μ g/mL cycloheximide (CHX) and 0.5% foetal calf serum (" FCS ")), add 20 μ L antigen-specific b cells culture supernatant or contrast B cell culture supernatant.Removal is from the substratum of WM-266 cell and add antibody-apoptosis substratum mixture, each 1 row in cell.Cell incubation together with antibody-apoptosis substratum is occurred with the permission apoptosis in 20 hours.Final concentration with 0.5 μ g/mL and 2.5 μ g/mL adds DNA binding fluorescent dyes propidium iodide (Sigma) and Hoechst 33342 (Molecular Probes) in each hole respectively.Hoechst 33342 can see through film, and therefore mark is lived and dead cell; Propidium iodide can not see through film, and therefore can only the mark dead cell.After 1 hour, catch image and analysis of cells overall number (by the amount of assessment Hoechst mark) and the dead cell overall number (by assessing the amount of propidium iodide mark) in each hole in 37 ℃.The apoptosis percentage test is (propidium iodide-positive cell/Hoechst-positive cell) * 100.

For the XenoMax technology, use haemolysis plaque measurement method to select to be used for rescuing from the antibody in the several holes that show best apoptosis induction.Make the TR-2-His biotinylation and be coated on the coated sheep red blood cell (SRBC) of Streptavidin.The plasmocyte in corresponding antigen-specific hole is thawed, and strengthen under the existence of serum incubation together with antigen coated red corpuscle at the anti-human IgG of complement and cavy.Production causes the sheep red blood cell (SRBC) cracking around them for the plasmocyte of the antibody of TR-2-His, and therefore allows to identify the antigen-specific plasmocyte in mixture.Those plasmocyte separate from mixture by single celled micrurgy.

After separating required single plasmocyte, from those cells, extract mRNA.Make the mRNAs of coding weight and light chain variable sequence be transformed into cDNA, and increased by reverse transcriptase PCR, use homing sequence and the special merger antisense primer of constant region to human IgG2 and people κ mRNA.Primer sequence provides in following table 2:

Primer locates to introduce above-mentioned restriction site at 5 ' end (BglII) and the 3 ' end (Xba1) of heavy chain cDNA.Similarly, primer locates to introduce above-mentioned restriction site at 5 ' end (BglII) and the 3 ' end (NheI) of κ chain cDNA.

Variable heavy chain cDNA amplicon uses the restriction enzyme suitable to restriction site to be digested, and described restriction site adds between the reaction period at PCR.By the product cloning of that digestion to having in each of clone's IgG1, the IgG2 of compatible overhang and IgG4 expression vector.IgG2 and IgG4 expression vector are digested with BamHI and XbaI, to produce the compatible overhang for subclone.The IgG1 expression construct is digested with BamHI and NheI, to produce the compatible overhang for subclone.Be cloned into interior these carriers that produce of multiple clone site of carrier pcDNA3.1+/Hygro (Invitrogen) by the constant domain by human IgG1, IgG2 and IgG4.

Variable light chain cdna amplicon uses the restriction enzyme suitable to restriction site to be digested equally, and described restriction site adds between the reaction period at PCR.By the product cloning of that digestion, in the IgK expression vector, described IgK expression vector is digested with BamHI and NheI, to produce the compatible overhang for subclone.Be cloned into that carrier of the interior generation of multiple clone site of carrier pcDNA4.1+/Neo (Invitrogen) by the constant domain by people IgK gene.

Subsequently by the 60mm ware of heavy chain and light chain expression vector people's tire kidney 293 cell (ATCC catalog number (Cat.No.) CRL-1573) that lipofection to 70% converges altogether.For 24 hours, allow the transfectional cell secretion to there is the specific recombinant antibodies identical with Plasmablast.Results are from the supernatant liquor (3mL) of HEK 293 cells, and confirm that with sandwich ELISA the secretion of complete antibody is with the specific detection human IgG.With dull and stereotyped the same for combination, contrast is dull and stereotyped to be coated with 2mg/mL mountain goat anti-human igg H+L O/N.Flat board washes with water 5 times.For 7 holes from undiluted lipofection supernatant liquor, recombinant antibodies carries out titration in 1: 2.Dull and stereotyped with dH2O washing 5 times.For secretion and 2 kinds of binding assays, add the mountain goat anti-human igg Fc specificity HRP of final concentration 1 μ g/mL to put together antibody in room temperature 1 hour.Dull and stereotyped with dH2O washing 5 times.By adding tetramethyl benzidine (TMB) within 30 minutes, to make dull and stereotyped colour developing, and end ELISA by adding 1M phosphoric acid.

Except above-described XenoMax method, use hybridoma technology to obtain some antibody.Put to death immune mouse by cervical dislocation, and gather in the crops from per share draining lymph node and merged.Separate lymphoidocyte by grinding with release cells from tissue in Dulbecco improvement Eagle substratum (" DMEM ").To reclaim cell is resuspended in DMEM.Counting cells, and add 0.9mL DMEM/100 1,000,000 lymphocytes in cell mass, so that the gently but complete resuspension of cell.Make cell and the 100 μ L CD90 of resuspension ⁺magnetic bead/10,000 ten thousand cells one arise from 4 ℃ of incubations 15 minutes.The cell suspending liquid of magnetic mark (is comprised up to 10 ⁸positive cell is (or up to 2 * 10 ⁹total cell)) be loaded into LS ⁺on post.This post is washed with DMEM.Collect total effluent liquid as CD90 ^-negative part, this expection mainly comprises the B cell.

Will be from enrichment B cell and the non-secretion myelomatosis P3X63Ag of washing above by the ratio with 1: 1 ^*.653 (ATCC (CRL 1580, referring to for example, the people such as Kearney, J.Immunol.123,1979,1548-1550)) mixes and is merged cell.Cell mixture is by the centrifugal agglomerate that forms lightly under 800xg.Remove supernatant liquor fully from cell after, by 2-4ml PRONASE A solution (CalBiochem for cell; Be dissolved in the 0.5mg/ml of phosphate buffered saline (PBS) (" PBS ")) process and to be no more than 2 minutes.Add 3-5mL foetal calf serum (" FBS ") to end enzymic activity, and make electricity consumption urge cytogamy solution (" ECFS ") (0.3M sucrose, 0.1mM magnesium acetate, 0.1mM lime acetate) suspension is adjusted to the 40mL cumulative volume.Centrifugal rear removal supernatant liquor and cell is resuspended in 40mL ECFS.Repeat this washing step and again cell be resuspended in 40mL ECFS to 2 * 10 ⁶the concentration of cell/mL.

The short cytogamy of electricity is used fusion producer (model ECM2001, Genetronic, Inc.) to carry out.The fusion chamber size of using is 2.0mL, uses following instrument to set: comparison condition: 50V, 50 seconds; Film breaks under 3000V, 30 microseconds; Hold-time after merging: 3 seconds.

After the short cytogamy of electricity occurs, take out carefully cell suspending liquid and transfer in the sterile tube of the hybridoma substratum that comprises equal volume under aseptic condition from merge chamber, described hybridoma substratum comprises DMEM, (JRH Biosciences), 15%FBS (Hyclone), is supplemented with L-glutaminate, penicillin/streptomycin, OPI (oxaloacetate, pyruvate salt, Sigma I8405) and IL-6 (Boehringer Mannheim).Make cell in 37 ℃ of incubation 15-30 minute, and descend centrifugal 5 minutes at 400xg (1000rpm) subsequently.The hybridoma that cell is resuspended to lightly to small volume is selected in substratum (being supplemented with the hybridoma substratum of 0.5x hyaluronic acid (Sigma)).Based on altogether 5 * 10 ⁶the final bed board volume in B cell/96 holes flat boards and 200 μ L/ holes, select substratum suitably to adjust cumulative volume with more hybridomas.Cell is mixed gently, be drawn in 96 hole flat boards, and allow growth.The 7th or 10 days the time, remove half substratum, and select substratum to resupply cell with fresh hybridoma.

Cultivate after 14 days, by ELISA, with regard to the TR-2 monoclonal antibody specific, screen the hybridoma supernatant liquor.In first screening, at coated damping fluid, (the 0.1M carbonate buffer solution, pH 9.6, NaHCO with 50 μ L/ hole TR-2 protein (2 μ g/mL) for ELISA dull and stereotyped (Fisher catalog number (Cat.No.) 12-565-136) ₃8.4g/L) in be coated with, in 4 ℃, be incubated overnight subsequently.After incubation, flat board washs 1 time with lavation buffer solution (being dissolved in the 0.05%Tween 20 of PBS).Add 200 μ L/ holes sealings damping fluids (being dissolved in the 0.5%BSA of 1xPBS, 0.1%Tween 20,0.01% Thiomersalates), and make at room temperature incubation 1 hour of flat board.After incubation, dull and stereotyped with lavation buffer solution washing 1 time.Add the aliquots containig (50 μ L/ hole) of hybridoma supernatant liquor and positive and negative control, and make at room temperature incubation 2 hours of flat board.The positive control used from start to finish is the serum from the immune XenoMouse animal of height, and negative control is the serum from the XenoMouse animal of KLH-immunity.After incubation, dull and stereotyped with lavation buffer solution washing 3 times.Add 100 μ L/ holes to detect the anti-huIgGFc-HRP of antibody goat (Caltag Inc. catalog number (Cat.No.) H10507, working concentration is 1: 2000 extent of dilution), and make at room temperature incubation 1 hour of flat board.After incubation, dull and stereotyped with lavation buffer solution washing 3 times.Add 100 μ L/ hole TMB (BioFX Lab. catalog number (Cat.No.) TMSK-0100-01), and allow dull and stereotyped colour developing approximately 10 minutes (until negative control hole has just started Show Color).Add subsequently 50 μ L/ holes to end solution (TMB Stop Solution (BioFX Lab. catalog number (Cat.No.) STPR-0100-01), and 450nm wavelength place reads dull and stereotyped on the ELISA flat bed reader.

Antibody by hybridoma production is used above-described identical apoptosis assay method to be analyzed.The WM-266 cell with the density of 4500 cells/well in normal substratum in microtiter plate overnight incubation.Use not containing FCS and comprise that in addition the cell culture medium of 1.8 μ g/mL cycloheximides and 0.9%FCS prepares 2x apoptosis substratum mixture.Use the negative control anti-KLH antibody parallel titration hybridoma supernatant liquor 1: 2 (in 2x apoptosis substratum mixture) of microtiter plate separately by the isotype coupling.Remove substratum from the WM-266 cell, and add 100 μ L antibody-apoptosis substratum mixture, each 1 row in celliferous each hole of bag.The microtiter plate incubation is occurred with the permission apoptosis in 20 hours.Final concentration with 0.5 μ g/mL and 2.5 μ g/mL adds DNA binding fluorescent dyes propidium iodide (Sigma) and Hoechst33342 (Molecular Probes) in each hole respectively., catch the fluoroscopic image in each hole and analyze dead cell overall number (PI) and total cellular score order (Hoechst) after 1 hour in 37 ℃.The apoptosis percentage test is (PI-positive cell/Hoechst-positive cell) * 100.

Use XenoMax or hybridoma method to obtain 17 kinds of different anti-TR-2 antibody (antibody A-Q).All antibody is checked order, and identified the sequence (referring to Fig. 3-19) of heavy and variable region of light chain.The heavy chain of 17 kinds of antibody and light chain comparison are shown in Figure 20 and 21.

Use and above-described a kind of similar apoptosis assay method some antibody of performance testing apoptosis-induced in cell with regard to it.The WM-266 melanoma cells in microtiter plate with the density of 4500 cells/well overnight incubation in normal substratum.In a separate microtiter plate, a titration of recombinant antibodies to be tested, the appropriate positive control (M413, mouse IgG1 anti-TR-2 antibody having a heavy chain variable sequence: MEVQLVESGGGLVQPGGSLKLSCAASGFTFSTYGMSWVRQTPDKRLELVALINSQGGSTYNSDSVKGRFTISRDNARNTLYLQMSSLKSEDTAMYYCARRDYESLDSWGQGTSVTVSSG (SEQ ID NO: 141) and a light chain variable sequence: DIVLTQSPASLPVSLGQRATISCRASESVEYSGTSLIQWYRQKPGQPPKLLIYAASNVDSEVPARFSGSGSGTDFSLYIHPVEEDDIAMYFCQQSRKVPWTFGGGTKLEIKRTDAAPGLEAA (SEQ ID NO: 142)) and the isotype-matched negative control antibody (anti potential not show activity TR-2 antibody), such that the final concentration of the antibody will contain 0.0001μg/mL- range 5μg/mL's.Mixed antibody in the apoptosis substratum that comprises final concentration 0.9 μ g/mL CHX and 0.45%FCS.Remove substratum from the WM-266 cell, and add antibody-apoptosis substratum mixture in cell.Cultivate after 20 hours, with propidium iodide (Sigma) and Hoechst 33342 (Molecular Probes) staining cell., catch the image in each hole and analyze dead cell overall number (PI) and total cellular score order (Hoechst) after 1 hour in 37 ℃.The apoptosis percentage test is (PI-positive cell/Hoechst-positive cell) * 100.With in M413 or the cell by above-described some anti-TR-2 antibody treatment, observing significant necrocytosis.

Embodiment 2

the dynamic analysis that anti-TR-2 antibody is combined with TR-2

Use

the kinetics that the anti-TR-2 antibody A of instrumental analysis is combined with TR-2 to Q.Use conventional amine to be coupled at

the preparation anti-human antibody of highdensity goat surface on chip.The anti-TR-2 antibody of every kind of purifying is diluted to approximately 1 μ g/ml in the HBS-P running buffer that comprises 100 μ g/ml BSA.Every kind of anti-TR-2 antibody is used 2 minute contact time and washing in 5 minutes to be caught on the surface separated, to stablize the anti-TR-2 antibody surface on chip.

In order to analyze the kinetics of TR-2 and every kind of single anti-TR-2 antibodies, in 25 ℃ by 226nM recombinant human TR-2-His (in embodiment 1 describe) Dynamic Injection on every kind of anti-TR-2 surface upper 1 minute (using kinject), be to dissociate the phase in 5 minutes subsequently.From on other surfaces, observe separately in conjunction with in deduct baseline shift, described baseline shift results from anti-TR-2 antibody surface the damping fluid injection that lacks TR-2.In addition, carry out stdn about the data of TR-2 and anti-TR-2 antibodies for the amount of the monoclonal antibody of catching on every kind of surface.Each data set to the overall matching of the interaction model of 1: 1 to determine binding kinetics.The k obtained for every kind of antibody _a, k _dand K _dvalue is shown in table 3.

Table 3: in the kinetics of 25 ℃ of TR-2 and anti-TR-2 antibodies

Antibody	k _a(M ^-1s ^-1)	k _d(s ^-1)	K _d(nM)
				A	5.3×10 ⁵	3.7×10 ^-3	6.9
B	5.7×10 ⁵	1.1×10 ^-2	19
				C	6.8×10 ⁵	2.6×10 ^-3	3.9
D	6.2×10 ⁵	2.7×10 ^-3	4.5
				E	8.7×10 ⁵	1.8×10 ^-3	2.1
F	3.8×10 ⁵	5.0×10 ^-3	13
				G	6.0×10 ⁵	1.9×10 ^-2	31
H	8.6×10 ⁵	8.4×10 ^-3	9.8
				I	2.9×10 ⁵	1.3×10 ^-3	4.4
J	5.7×10 ⁵	7.1×10 ^-3	12
				K	6.8×10 ⁵	1.2×10 ^-2	18
L	6.0×10 ⁵	1.1×10 ^-2	18
				M	3.4×10 ⁵	1.2×10 ^-2	37
N	8.1×10 ⁵	5.5×10 ^-2	68 ^*
				O	4.4×10 ⁵	8.4×10 ^-3	19
P	8.1×10 ⁵	2.7×10 ^-2	33 ^*
				Q	1.2×10 ⁶	1.6×10 ^-2	13 ^*

^*data presentation about that sample is heterogeneous and very poor to model-fitting in 1: 1.

Embodiment 3

cell kill determination method

Carry out cell kill determination method with the anti-TR-2 antibody of some people of describing in embodiment 2, to determine every kind of antibody, trigger the degree of apoptosis and necrocytosis.The anti-TR-2 antibody of some people and mouse anti-TR-2 antibody M412 and M413 are fixed in the difference hole of 96 porin matter G coated dull and stereotyped (reactin conjugated protein G is coated dull and stereotyped, Pierce catalog number (Cat.No.) 15131).M412 is that the anti-TR-2 antibody of mouse IgG 1 has the weight chain variable sequence: KVQLQQSGTELVKPGASVKLSCKASGYTFTEYIIHWVKQRSGQGLEWIGWFYPGSG YIKYNEKFKDKATMTADKSSSTVYMELSRLTSEDSAVYFCTRHEEDGYYAAYWGQG TLVTVSA, (SEQ ID NO:143) and light chain variable sequence: DIVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWYQQKPGQSPKLLIYWASTRHT GVPDRFTGSGSGTDYTLTISSVQAEDLALYYCQQHYSTPYTFGGGTKLEIKR, (SEQ ID NO:144).

M412 is that the anti-TR-2 antibody of mouse IgG 1 has the weight chain variable sequence: KVQLQQSGTELVKPGASVKLSCKASGYTFTEYIIHWVKQRSGQGLEWIGWFYPGSG YIKYNEKFKDKATMTADKSSSTVYMELSRLTSEDSAVYFCTRHEEDGYYAAYWGQG TLVTVSA, (SEQ ID NO:143) and light chain variable sequence: DIVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWYQQKPGQSPKLLIYWASTRHT GVPDRFTGSGSGTDYTLTISSVQAEDLALYYCQQHYSTPYTFGGGTKLEIKR, (SEQ ID NO:144).M413 is the anti-TR-2 antibody of mouse IgG 1 as described in Example 1.Every kind of antibody adds in first hole with the concentration of 50 μ g/ml, and carries out 1 in each of 7 other holes: the 3x serial dilution.Every kind of antibody dilution carries out in triplicate.Flat board is before use in 4 ℃ of incubations 24 hours.Each is for hole after substratum (RPMI adds 10%FBS) washing, and by one of 4 kinds of different clones bed board, to every kind of sessile antibody, density is 50,000 cells/well in the cumulative volume of 200 μ L.Test cell system is COLO 205 cells (human colon adenocarcinoma), MDA-231 cell (human breast carcinoma), WM35 cell (human melanoma) and WM793 cell (human melanoma).Cell is at 37 ℃/6%CO ₂lower incubation 24 hours, subsequently with ³the H-thymidine is incubation 6 hours together.Survivaling cell per-cent is processed and is mixed in cell by mensuration ³h-thymidine level is with respect to mixing in untreated cell ³h-thymidine level is assessed.By measuring with respect to untreated cell, make the survival of processing cell reduce by 50% antibody concentration, obtained the ED of every kind of antibody by the cell survival titration curve ₅₀.People's antibody is for the ED of COLO 205 cells ₅₀scope is 0 μ g/ml-3.25 μ g/ml.Mouse antibodies M412 and M413 have respectively the ED of 1.85 μ g/ml and 0.07 μ g/ml for those cells ₅₀s.People's antibody is for the ED of MDA-231 cell ₅₀scope is 0.05 μ g/ml-0.5 μ g/ml.Mouse antibodies M412 and M413 have respectively the ED of 0.6 μ g/ml and 0.07 μ g/ml for those cells ₅₀s.People's antibody is for the ED of WM35 cell ₅₀scope is 0.1 μ g/ml-0.6 μ g/ml.Mouse antibodies M412 and M413 have respectively the ED of 1.85 μ g/ml and 0.07 μ g/ml for those cells ₅₀s.People's antibody is for the ED of WM793 cell ₅₀scope is 0.2 μ g/ml-0.2 μ g/ml.Mouse antibodies M412 and M413 have respectively the ED of 1.85 μ g/ml and 0.05 μ g/ml for those cells ₅₀s.

Embodiment 4

people TR-2 in tumor cell line expresses

Expression screening human tumor cell line with regard to TR-2.The clone of using comprises from mammary gland, central nervous system, colon, liver, lung, uterine cervix, uterus, ovary, pancreas, prostate gland and kidney, and leukemia and melanomatous those.

The array of use based on cell determines that the TR-2 on human tumor cells expresses.In brief, will be in 100Ml CBA damping fluid (PBS, 3%FBS, 0.02% trinitride) 4 * 10 ⁵cell is assigned in each hole of 96 hole flat boards at the bottom of 20 V-arrangements.To adding CBA damping fluid (150 μ L) in each hole and dull and stereotyped centrifugal so that cell being rotated down.Discard substratum and, to the antibody-solutions (antibody A is to one of Q) that adds 100 μ L 10 μ g/ml in cell mass, described cell mass is resuspended in the PBS (" mensuration damping fluid ") that comprises 2%PBS.At incubation on ice after 25 minutes, cell washing 1 time in measuring damping fluid.Xiang Kongzhong adds 100 μ L time level mountain goat anti-human igg Fc specificity horseradish peroxidase (HRP, Pierce), and makes dull and stereotyped incubation on ice 20 minutes.Dull and stereotyped with washing in the mensuration damping fluid 2 times, and at room temperature add 100 μ L tmb substrate (ZYMED) 10 minutes.Make dull and stereotyped centrifugal and every kind of supernatant liquor of 50 μ L is transferred to and comprised in the clean flat board that 50 μ L end solution (BioFX Laboratories).Use SpectraMax/plus reader (Molecular Devices) to carry out the optical density(OD) reading at the 450nm place.Make data normalization by deducting the optical density value that derives from the isotype control antibodies.

Several clone has the OD that is greater than 0.1 in this assay method ₄₅₀, comprise breast cancer cell line HS 578.T (OD 0.122) and T-47D (OD 0.112), colon carcinoma cell line TE 671 (u) (OD 0.109), HT-29 (OD 0.193), SW-948 (OD 0.122), KM-12 (OD 0.354) and HCC-2998 (OD 0.133), hepatoma cell line NCI-N87 (OD 0.154) and NCI-SNU-5 (OD 0.137), Leukemia Cell Lines HL-60 (OD 0.233) and hPBMC (OD 0.131), Lines JY (OD 0.118), CCRF-CEM (OD0.106), NCI-H2126 (OD 0.108) and NCI-H460 (OD 0.122), melanoma cell series SK-mel-5 (OD 0.131), LOX IMVI (OD 0.102), RPMR 7951 (OD0.101) and UACC-62 (OD 0.127), pancreatic carcinoma HPAF II (OD 0.117) and CAPAN-1 (OD 0.101), prostate cancer cell line LNCaP (OD 0.174), with renal carcinoma cell line Caki-1 (OD 0.148) and UO-31 (OD 0.104).TR-2 maximum in the tumor cell line of research expresses in colon carcinoma cell line KM-12 and HT-29, and finds in Leukemia Cell Lines HL-60.In central nervous system, minicell liver, uterine cervix, uterus or the ovarian cancer cell line of research, none has the OD450 that is greater than background.

For the TR-2 expression characteristic of determining that human tumor cells is fastened, with the anti-TR-2 antibody of mouse, M412 measures above-mentioned human tumor cell line.The TR-2 that the assay method of use based on cell measured on human tumor cells expresses.In brief, will be in 100Ml CBA damping fluid (PBS, 3%FBS, 0.02% trinitride) 4 * 10 ⁵cell is assigned in each hole of 96 hole flat boards at the bottom of 20 V-arrangements.To adding CBA damping fluid (150 μ L) in each hole and dull and stereotyped centrifugal so that cell being rotated down.Discard substratum and, to the anti-TR-2 monoclonal antibody of the mouse M412 that adds 100 μ L 10 μ g/ml in cell mass, described cell mass is resuspended in the PBS (" mensuration damping fluid ") that comprises 2%PBS.At incubation on ice after 25 minutes, cell washing 1 time in measuring damping fluid.Xiang Kongzhong adds 100 μ L time level goat anti-mouse IgG Fc specificity horseradish peroxidase (HRP, Pierce), and makes dull and stereotyped incubation on ice 20 minutes.Dull and stereotyped with washing in the mensuration damping fluid 2 times, and at room temperature add 100 μ g tmb substrate (ZYMED) 10 minutes.Make dull and stereotyped centrifugal and every kind of supernatant liquor of 50 μ L is transferred to and comprised in the clean flat board that 50 μ L end solution (BioFX Laboratories).Use SpectraMax/pIus reader (Molecular Devices) to carry out the optical density(OD) reading at the 450nm place.Make data normalization by deducting the optical density value that derives from the isotype control antibodies.

Much clone has TR-2 and expresses.The highest expresser (have the OD450nm that is greater than 0.3 those) comprises breast cancer cell line HS 578.T (OD 0.403), MDA-MB-231 (OD 0.408) and T-47D (OD 0.366), CNS cancerous cell line SF-295 (OD 0.354) and U251 (OD 0.323), colon carcinoma cell line HCT-116 (OD 0.41), HT-29 (OD0.869), SW-707 (OD 0.323), SW-948 (OD 0.423), KM-12 (OD 0.77) and HCC-2998 (OD 0.635), hepatoma cell line NCI-SNU-1 (OD 0.354), Leukemia Cell Lines A 673 (OD 0.347), Lines HOP-62 (OD 0.313), HOP-62 (OD 0.47), NCI-H2126 (OD 0.501), NCI-H460 (OD 0.326), small cell lung cancer? clone A549 (OD 0.381), melanoma cell series LOX IMVI (OD0.573), RPMI 7951 (OD 0.322) and UACC-62 (OD 0.319), ovarian cancer cell line IGROV1 (OD 0.312), prostate cancer cell line DU 145 (OD 0.372), 22Rv1 (OD 0.301) and LNCaP (OD 0.63), and renal carcinoma cell line Caki-1 (OD 0.93), Caki-2 (OD 0.443), SN12C (OD 0.313) and UO-31 (OD 0.331).TR-2 maximum in the tumor cell line of the anti-TR-2 antibody treatment with mouse expresses in renal carcinoma cell line Caki-1, and finds in Colon cancer cell line HT-29 and KM-12.

Embodiment 5

antibody cross reaction

As people such as Jia, described in J.Immunol.Methods 288:91-98 (2004), assess the ability that anti-other antibody of TR-2 antibody blocking of some people are combined with TR-2.Use and directly take from Luminex 100User ' s Manual, the coupling operation of Version 1.7 is puted together pearl and anti-human IgG antibody.After integument activates, according to the specification sheets of manufacturers, make they and the anti-hIgG mAb of Pharmingen mouse coupling.Carry out 2 experiments.In experiment for the first time, make at room temperature incubation 2 hours of coated pearl.In experiment for the second time, coated pearl is incubated overnight in 4 ℃.When incubation finishes, make coated bead seal close and use subsequently the Coulter cell counter to be counted.Puting together pearl uses immediately or is stored in 4 ℃ of dark for using in the future.

Carry out the anti-TR-2 antibody classification based on the epi-position cross reactivity by following step.At first, make to arise from 4 ℃ with reference antibody (" reference antibody ") respectively from the anti-hIgG mixture of every group of pearl-mouse above is incubated overnight on turner.Reference antibody is selected from above-described anti-TR-2 antibody A-Q.After antibody capture, 2000 anti-hIgG of every kind of pearl-mouse are merged in 1 pipe together with reference to the Ab mixture, and tightly add subsequently in each holes of 96 hole flat boards and aspirated.To adding TR-2 (50ng) and incubation 1 hour at room temperature in each hole.After washing hole, to adding the another kind of anti-TR-2 antibody of people (" detection antibody ") of 100-500ng/mL and incubation 2 hours at room temperature in each hole.After the washing of hole, use 1 μ g/ml biotinylation form reference antibody is caught and used the anti-hIgG of identical mono-clonal mouse to detect the detection antibody of combination.After incubation and washing hole, add 0.5 μ g/ml SA-PE.Make at room temperature incubation 30 minutes of mixture, and use subsequently Luminex 100 to detect the phycoerythrin signals.The hole of using another group to lack antigen as negative control to help data analysis.

Data are analyzed in 2 step processes.At first, use the negative control value to make data normalization.Secondly, the ability that anti-TR-2 antibody hinders one or more other anti-TR-2 antibodies according to it is carried out cluster.For cluster analysis, by standardized intensity matrix, produce dissimilarity matrix.The value of antibody based in average dissimilarity degree matrix carried out cluster, uses SPLUS 2000 to assemble nested hierarchical cluster sub-routine and Manhattan tolerance, uses the input dissimilarity matrix of actual average dissimilarity matrix.

Based on this discovery, antibody is put into to 4 different epi-position groups.In any one group, one of group membership and TR-2 in conjunction with blocking-up another member on the same group and the combination of TR-2 mutually.Yet, for example, organize one of 1 member and TR-2 in conjunction with not blocking the combination of one of

group

2,3 or 4 members with TR-2.Those groups are shown in Figure 22.

Embodiment 6

epitope mapping

In order to identify the TR-2 specific regions important with some described anti-TR-2 antibodies, carry out epitope mapping research.N-avidin-TR-2 construct is prepared by following method: pcr amplification come the self-template resource about mature T R-2 (MacFarlane, 1997) encoding sequence, and with such direction, be cloned in the pCEP4 carrier (Invitrogen) that comprises chicken avidin sequence by it, make after the HindIII site is inserted, the TR-2 sequence connects at the C-terminal place of avidin sequence.Forward primer about the encoding sequence of mature T R-2 is GTAAGCAAGCTTGGCTC TGATCACCCAACAAGA (SEQID NO:145), and reverse primer is GATTAGGGATCCAGAGGCAGGAGTCCCTGG (SEQ ID NO:146).The aminoacid sequence of resulting avidin-TR-2 fusion rotein is LSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGTYTTAVTATSNEIK ESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRNGKEVLKTMWLLRS SVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAPQQRAAPQQKRSSP SEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTT RNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHS GEAPAVEETVTSSPGTPAS (SEQ ID NO:69).

Comprise 12 kinds of truncate as described below being synthesized of molecule of N-avidin and people TR-2.3 kinds of C-terminal truncate (TR-2-1 to TR-2-3) that molecule only has people TR-2, and 9 kinds of molecules have the N of people TR-2 and truncate (TR-2-4 to TR-2-13) (the schematically showing in Figure 23) at C-terminal place.The truncate polynucleotide of encoding human TR-2 use primer described below to prepare by pcr amplification.In order to form each in 12 kinds of molecules, the people TR-2 of the brachymemma that will be produced by amplification inserts in the pCEP4 carrier (Invitrogen) that comprises chicken avidin sequence as above.Use the polynucleotide of the amino acid/11-43 of forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer TAGTTGGGATCCTCAGGAGATGCAATCTCT ACCGT (SEQ ID NO:147) amplification coding mature T R-2.The aminoacid sequence of TR-2-1 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCPPGHHISEDGRDCIS (SEQ ID NO:70).

Use the polynucleotide of the amino acid/11-85 of forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer GGTAGTGGATCCTCACTGACACACTGTGTTTCTGG (SEQ ID NO:148) amplification coding mature T R-2.The aminoacid sequence of TR-2-2 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQ (SEQ ID NO:71).

Use the polynucleotide of the amino acid/11-126 of forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer GTAATGGGATCCTCAGACACATTCGATGTCACTCC (SEQ ID NO:149) amplification coding mature T R-2.The aminoacid sequence of TR-2-3 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSD IECV (SEQ ID NO:72).

Use the polynucleotide of the amino acid/11 6-43 of forward primer GTAATGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO:150) and reverse primer TAGTTGGGATCCTCAGGAGATGCAATCTCTACCGT (SEQ ID NO:147) amplification coding mature T R-2.The aminoacid sequence of TR-2-4 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCIS (SEQID NO:73).

Use the polynucleotide of the amino acid/11 6-85 of forward primer GTAATGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO:150) and reverse primer GGTAGTGGATCCTCACTGACACACTGTGTTTCTGG (SEQ ID NO:148) amplification coding mature T R-2.The aminoacid sequence of TR-2-5 is MVHATSPLLLLLLLSLALVAPGLSARKCSITGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRN TVCQ (SEQ ID NO:74).

Use the polynucleotide of the amino acid/11 6-126 of forward primer GTAATGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO:150) and reverse primer GTAATGGGATCCTCAGACACATTCGATGTCACTCC (SEQ ID NO:149) amplification coding mature T R-2.The aminoacid sequence of TR-2-6 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRN TVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECV (SEQ ID NO:75).

Use the polynucleotide of the amino acid 42-85 of forward primer GATTGAAAGCTTGATCTCCTGCAAATATGGACAG (SEQ ID NO:151) and reverse primer GGTAGTGGATCCTCACTGACACACTGTGTTTCTGG (SEQ ID NO:148) amplification coding mature T R-2.The aminoacid sequence of TR-2-7 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLISCKYGQDYS THWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQ (SEQ ID NO:76).

Use the polynucleotide of the amino acid 42-126 of forward primer GATTGAAAGCTTGATCTCCTGCAAATATGGACAG (SEQ ID NO:151) and reverse primer GTAATGGGATCCTCAGACACATTCGATGTCACTCC (SEQ ID NO:149) amplification coding mature T R-2.The aminoacid sequence of TR-2-9 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLISCKYGQDYS THWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGC PRGMVKVGDCTPWSDIECV (SEQ ID NO:77).

Use the polynucleotide of the amino acid 85-154 of forward primer GTAATGAAGCTTGCAGTGCGAAGAAGGCACCT (SEQ ID NO:152) and reverse primer GATTAGGGATCCAGAGGCAGGAGTCCCTGG (SEQ ID NO:146) amplification coding mature T R-2.The aminoacid sequence of TR-2-10 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLQCEEGTFREE DSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHSGEAPAVEETVTSSPG TPAS (SEQ ID NO:78).

Use the polynucleotide of the amino acid 42-154 of forward primer GATTGAAAGCTTGATCTCCTGCAAATATGGACAG (SEQ ID NO:151) and reverse primer GATTAGGGATCCAGAGGCAGGAGTCCCTGG (SEQ ID NO:146) amplification coding mature T R-2.The aminoacid sequence of TR-2-11 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLISCKYGQDYS THWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGC PRGMVKVGDCTPWSDIECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQ ID NO:79).

Use the polynucleotide of the amino acid/11 6-66 of forward primer TGATTGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO:150) and reverse primer GATGGAGGATCCTCAACACCTGGTGCAGCGCAAG (SEQ ID NO:153) amplification coding mature T R-2.The aminoacid sequence of TR-2-12 is MVHATSPLLLLLLLSLALVAPGLSARKCSITGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISYKYGQDYSTHWNDLLFCLRCTRC (SEQ ID NO:80).

Use the polynucleotide of the amino acid/11 6-74 of forward primer TGATTGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO:150) and reverse primer GTAAGTGGATCCTCAGCAGGGACTTAGCTCCACT (SEQ ID NO:154) amplification coding mature T R-2.The aminoacid sequence of TR-2-13 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELS (SEQ ID NO:81).Comprise the N-avidin and truncate 4 kinds of as described below being synthesized of molecule from the TR-2 of cynomolgus monkey.Use the polynucleotide of the amino acid/11-132 of the ripe cyno TR-2 of forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQID NO:155) and reverse primer GTTGATGGATCCTTCTTTGTGGACACTCGAT (SEQ ID NO:156) amplification coding.Cyno TR-2, the aminoacid sequence of (short) is PLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGTYTTAVT ATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRNGKEVLK TMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDPQRRAAP QQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKCDSGEVE VSSCTTTRNTVCQCEEGTFREEDSPEICRKCRTGCPRGMVKVKDCTPWSDIECPQR RIQT, (SEQ ID NO:82).

Use the polynucleotide of the amino acid/11-154 of the ripe cynoTR-2 of forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQID NO:155) and reverse primer GTAGTTGGATCCTCAAGAAGCAGGAGTCCCAGGG (SEQ ID NO:157) amplification coding.The aminoacid sequence of cyno TR-2 (length) is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVS SCTTTRNTVCQCEEGTFREEDSPEICRKCRTGCPRGMVKVKDCTPWSDIECVHKES GTKHTGEVPAVEKTVTTSPGTPAS (SEQ ID NO:83).

Use the polynucleotide of the amino acid/11-85 of the ripe cynoTR-2 of forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQID NO:155) and reverse primer GTATGAGGGATCCTCACTGACACACCGTGTTTCTGG (SEQ ID NO:158) amplification coding.The aminoacid sequence of cyno TR-2 (length) is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVSSCTTTRNTVCQ (SEQ ID NO:84).

Use the polynucleotide of the amino acid/11 6-85 of forward primer GTATGGAAGCTTGCCACAACAAAAGAGATCCAGC (SEQ ID NO:159) and reverse primer GTATGAGGGATCCTCACTGACACACCGTGTTTCTGG (SEQ ID NO:158) the ripe cyno TR-2 that increases.The aminoacid sequence of cyno 16-85 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPIE GLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKCDSGEVEVSSCTTTRN TVCQ (SEQ ID NO:85).

As shown in Figure 25,4 kinds of N-avidin-fusion mosaics of the same preparation of the different piece of end user TR-2 and cyno TR-2.2 kinds of PCR products that have overlapping end by preparation build every kind of construct, and described PCR product is used identical 5 ' to increase together with 3 ' primer subsequently.In order to form every kind of mosaic, subsequently the polynucleotide of amplification are subcloned in the pCEP4 carrier (Invitrogen) that comprises chicken avidin sequence as above.The comparison of people, cyno (short) and mouse TR-2 sequence is shown in Figure 26.

Cyno/ people's mosaic #1 is prepared by following: use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQ ID NO:155) and reverse primer GGACCTCTTTTGTTGTGGAGCCGCTCTTCGCTGG (SEQ IDNO:159), the increase ripe cyno TR-2 zone of corresponding amino acid/11-16, and use forward primer CAGCGAAGAGCGGCTCCACAACAAAAG AGGTCCAG (SEQ ID NO:160) and reverse primer GGTAGTGGATCCTCACTGACACACTGTGTTTCTGG (SEQ ID NO:148), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11 7-85.Use is about the forward primer of cyno TR-2 amino acid/11-16 fragment, above (SEQ ID NO:155), with the reverse primer about people TR-2 amino acid/11 7-85 fragment, above (SEQ ID NO:148), carry out the overlapping PCR of cyno and people TR-2 fragment.Aminoacid sequence about cyno/ people's mosaic #1 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQ (SEQ ID NO:86).

Cyno/ people's mosaic #2 is prepared by following: use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQ ID NO:155) and reverse primer GGACCTCTTTTGTTGTGGAGCCGCTCTTCGCTGG (SEQ IDNO:159), the increase ripe cyno TR-2 zone of corresponding amino acid/11-16, and use forward primer CAGCGAAGAGCGGCTCCACAACAAAA GAGGTCCAG (SEQ ID NO:160) and reverse primer GATTAGGGATCCTCAAGAGGCAGGAGTCCCTGG (SEQ ID NO:146), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11 7-154.Use is about the forward primer of cyno TR-2 amino acid/11-16 fragment, above (SEQ ID NO:155), with the reverse primer about people TR-2 amino acid/11 7-154 fragment, above (SEQ ID NO:146), carry out the overlapping PCR of cyno and people TR-2 fragment.Aminoacid sequence about cyno/ people's mosaic #2 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPSEGLCPPGHHISEDGRDYISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSD IECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQID NO:87).

Cyno/ people's mosaic #3 is prepared by following: use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer GGATCTCTTTTGTTGTGGGGCCGCTCTCTGCTGG G (SEQ ID NO:161), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11-16, and use forward primer CAGCAGAGAGCGGCCCCACAACAAAAGAGATCCAGC (SEQ ID NO:162) and reverse primer GTATGAGGGATCCTCACTGACACACCGTGTTTCTGG (SEQ ID NO:158), the increase ripe cyno TR-2 zone of corresponding amino acid/11 7-85.Use is about the forward primer of people TR-2 amino acid/11-16 fragment, above (SEQ ID NO:145), with the reverse primer about cyno TR-2 amino acid/11 7-85 fragment, above (SEQ ID NO:158), carry out the overlapping PCR of cyno and people TR-2 fragment.Aminoacid sequence about cyno/ people's mosaic #3 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVSSCTTTRNTVCQ (SEQ ID NO:88).

Cyno/ people's mosaic #4 is prepared by following: use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer GGATCTCTTTTGTTGTGGGGCCGCTCTCTGCTGG G (SEQ ID NO:161), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11-16, and use forward primer CAGCAGAGAGCGGCCCCACAACAAAAGAGATCCAGC (SEQ ID NO:162) and reverse primer GTAGTTGGATCCTCAAGAAGCAGGAGTCCCAGGG (SEQ ID NO:157), the increase ripe cyno TR-2 zone of corresponding amino acid/11 7-154.Use is about the forward primer of people TR-2 amino acid/11-16 fragment, above (SEQ ID NO:145), with the reverse primer about cyno TR-2 amino acid/11 7-154 fragment, above (SEQ ID NO:157), carry out the overlapping PCR of cyno and people TR-2 fragment.Aminoacid sequence about cyno/ people's mosaic #4 is MVHATSPLLLLLLLS LALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGTYTTAVTATSNEIKES PLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRNGKEVLKTMWLLRSSV NDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAPQQRAAPQQKRSSPTE GLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKCDSGEVEVSSCTTTRN TVCQCEEGTFREEDSPEICRKCRTGCPRGMVKVKDCTPWSDIECVHKESGTKHTGE VPAVEKTVTTSPGTPAS (SEQ ID NO:89).Under the background merged at the N-avidin, build the TR-2 protein of 4 kinds of other modifications by the short zone of replacing people TR-2 by corresponding mouse TR-2 sequence.People/mouse TR-2#1 comprises from the mouse TR-2 sequence of amino acid/11-22 with from the people TR-2 sequence of amino acid 23-150.People/mouse TR-2#2 comprises from the people TR-2 sequence of amino acid/11-28 with from the mouse TR-2 sequence of amino acid 29-34.People/mouse TR-2#3 comprises from the people TR-2 sequence of amino acid/11-53 with from the mouse TR-2 sequence of amino acid 54-59.People/mouse TR-2#4 comprises from the people TR-2 sequence of amino acid/11-66 with from the mouse TR-2 sequence of amino acid 67-75.In order to form the protein of every kind of modification, subsequently the polynucleotide of amplification are subcloned in the pCEP4 carrier (Invitrogen) that comprises chicken avidin sequence as above.

People/mouse TR-2#1 is prepared by following: use forward primer CAGCGGCCGGAGGAGAGCCCCTCAGAGGGATTGT (SEQ ID NO:163) and reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO:164), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid 23-150, and use forward primer TGAATGAAGCTTGGTTCCAGTAACAGCTAACCCA (SEQ ID NO:165) and reverse primer TCCCTCTGAGGGGCTCTCCTCCGGCCGCTGTAG (SEQ ID NO:166), the increase ripe mouse TR-2 zone of corresponding amino acid/11-22.Use is about the forward primer of mouse TR-2 amino acid/11-22 fragment, above (SEQ ID NO:165), with the reverse primer about people TR-2 amino acid 23-150 fragment, above (SEQ ID NO:164), carry out the overlapping PCR of people and mouse TR-2 fragment.Aminoacid sequence about people/mouse TR-2#1 is MVHATSPLLLLLLLSLALVAPGLSARKCSITGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLVPVTANPAHN RPAGLQRPEESPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCD SGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDI ECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQ ID NO:90).

People/mouse TR-2#2 is prepared by following: use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer CAGGTACTGGCCTGCTAGACACAATCCCTCTGAGGGG (SEQ ID NO:167), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11-28, use forward primer CTAGCAGGCCAGTACCTGTCAGAAGACGGTAGAGATTGC (SEQ ID NO:168) and reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO:164), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid 35-150, and use forward primer CAGGTACTGGCCTGCTAGACACAATCCCTCTGAGGGG (SEQ IDNO:169) and reverse primer CTAGCAGGCCAGTACCTGTCAGAAGACGGTAGAGATTGC (SEQ ID NO:170), the increase ripe mouse TR-2 zone of corresponding amino acid 29-34.Use is about the forward primer of people TR-2 amino acid/11-28 fragment, above (SEQ ID NO:145), with the reverse primer about people TR-2 amino acid 35-150 fragment, above (SEQ ID NO:170), carry out the overlapping PCR of people and mouse TR-2 fragment.Aminoacid sequence about people/mouse TR-2#2 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCLAGQYLSEDGRDCISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSD IECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQ ID NO:91).

People/mouse TR-2#3 is prepared by following: use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer TGAATCCAGAGAATGGTTGGAGTGAGTGCTATAGTCCTG TC (SEQID NO:171), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11-53, and use forward primer TCCAACCATTCTCTGGATTCA TGCTTGCGCTGCACCAGG (SEQ ID NO:172) and reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO:173), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid 60-154.Above-mentioned primer comprises the Nucleotide of the mouse TR-2 of the corresponding amino acid 54-59 that encodes.Use is about the forward primer of people TR-2 amino acid/11-53 fragment, above (SEQ ID NO:145), with the reverse primer about people TR-2 amino acid 60-154 fragment, above (SEQ ID NO:173), carry out the overlapping PCR of people and mouse TR-2 fragment.Aminoacid sequence about people/mouse TR-2#3 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSD IECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQ IDNO:92).

People/mouse TR-2#4 is prepared by following: use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO:145) and reverse primer TCGGGTTTCTACGACTTTATCTTCCTTACACCTGGTGCAGCGCAAG (SEQ ID NO:174), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid/11-66, and use forward primer AAGGAAGATAAAGTCGTAGAAACCCGATGCACCACGACCAGAAAC AC (SEQID NO:175) and reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO:176), the increase one-tenth acquaintance TR-2 zone of corresponding amino acid 76-154.Above-mentioned primer comprises the Nucleotide of the mouse TR-2 of the corresponding amino acid 67-75 that encodes.Use is about the forward primer of people TR-2 amino acid/11-66 fragment, above (SEQID NO:145), with the reverse primer about people TR-2 amino acid 76-154 fragment, above (SEQ ID NO:176), carry out the overlapping PCR of people and mouse TR-2 fragment.Aminoacid sequence about people/mouse TR-2#4 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRS SVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAPQQRAAPQQKRSSP SEGLCPPGHHISEDGRDCISCKYGQDYSTHSNHSLDSCLRCTRCDSGEVELSPCTT TRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKH SGEAPAVEETVTSSPGTPAS (SEQ ID NO:93).

Carry out the expression of avidin fusion rotein by transient transfection people 293T adherent cell in ventilation T75 tissue culture flasks.Cell in the DMEM of the FBS that contains 10% dialysis and 1x pen-step-glutamine at 37 ℃ and 5%CO ₂lower growth also maintains.For transfection is had to preparation, by approximately 3 * 10 ⁶the 293T cell is inoculated in each of clean T75 flask of a series of 15ml of comprising growth mediums, and all flask grow overnight approximately 20 hours.As follows pCEP4-Avidin (N)-TR-2 construct is transfected in different cells separately.Under the existence of Opti-MEM substratum (Invitrogen), 15 μ g DNA are mixed, to form the DNA-Lipofectamine mixture with 75 μ L Lipofectamine2000 (Invitrogen).Make the mixture incubation 20 minutes.Between incubation period, sucking-off growth medium replacing with 15mLOpti-MEM from the T75 flask.After incubation, every kind of transfection composite is inoculated in different flasks, and in 37 ℃ of incubation 4-5 hour.When incubation period finishes, the Opti-MEM substratum in each flask is replaced with to fresh growth medium.After transfection approximately 48 hours, the results conditioned medium also was transferred to 50ml pipe (Falcon).Make pipe under 2000xg in 4 ℃ centrifugal 10 minutes to remove cell and fragment, and be transferred to subsequently clean 50mL pipe.Prepare equally the control flask that lacks transfection DNA according to same approach, produce for the negative control conditioned medium in conjunction with experiment.

The assay method of use based on quantitative FACS measured the concentration of every kind of N-avidin-TR-2 fusion rotein.Catch the avidin fusion rotein on 6.7 μ m vitamin H polystyrene beads (Spherotech, Inc.).Prepare 2 kinds of samples for every kind of fusion rotein: 5 μ L (approximately 3.5 * 10 ⁵) pearl suspension adds 20 μ L 1x conditioned mediums, and 5 μ L pearl suspension add 200 μ L1x conditioned mediums.All samples is accompanying rotation incubation 1 hour at room temperature.By centrifugal and remove conditioned medium with the PBS that comprises 0.5%BSA (BPBS) washing from every kind of sample.Anti-anti-avidin antibody (Vector Labs, Burlingame, CA) solution-dyed avidin pearl with the 200 μ L 0.5 μ g/mL goat FITC marks that are dissolved in BPBS.Allow reaction at room temperature to carry out 45 minutes, wherein reaction tubes is covered by paper tinsel.After incubation, again pass through centrifugal and collect pearl with the BPBS washing, and being resuspended in 0.5ml BPBS for analyzing.Use FACScan (Becton Dickinson Bioscience) to detect FITC fluorescence.Use is transformed into protein quality by the derivative typical curve of restructuring avidin by signal.

Assess 2 kinds of anti-TR-2 antibody of people and people TR-2 is truncate, people TR-2 and from the TR-2 combination separately of cynomolgus monkey.Binding assay carries out as follows.Vitamin H pearl as above is loaded one of about 100ng N-avidin TR-2 fusion rotein/3.5 * 10 ⁵pearl and reach volume with growth medium.The anti-TR-2 monoclonal antibody of people that pearl is puted together with 1 μ g FITC is mixed in 0.2mLBPBS.At room temperature incubation, after 1 hour, adds 3mL BPBS and passes through to collect in centrifugal 5 minutes under 750xg antibody-pearl mixture.Agglomerate washs in 3mL BPBS.By the antibody that facs analysis detects with avidin-the pearl mixture is combined.For every kind of sample record average fluorescent strength.Use those antibody of being combined with the conditioned medium that lacks TR-2 as negative control (" Neg CM ").The results are shown in Figure 24.

The binding pattern that 2 kinds of antibody is observed is similar.The strongest combination of observing is about positive control people TR-2, and wherein average fluorescent strength is 7349.The antibody of observing (as with fluorescence intensity measurement) is 6561-6693 with the combination of truncate TR-2-2, with truncate TR-2-3 and TR-2-5 in conjunction with being 3158-3866, with truncate TR-2-6 in conjunction with being 1959-2202, and with truncate TR-2-1 in conjunction with being 662-759.This antibody with from the total length TR-2 of cynomolgus monkey, in conjunction with (as with fluorescence intensity measurement), be 666-764.Truely as the background classes by conjunction with about experiment determine, this antibody not with mouse or rat TR-2, or with truncate TR-2-4, TR-2-7, TR-2-9, TR-2-10, TR-2-11, TR-2-12 or TR-2-13, be not combined.

TR-2-1 is that the C-terminal of TR-2 after amino acid 43 is truncate, and TR-2-2 ,-3 ,-5 and-6 all comprises at least amino acid/11 6-85.When the whole zone from amino acid/11-85 exists (referring to the result about TR-2-2), in conjunction with occurring.The interpolation of amino acid 86-126 makes in conjunction with reducing by approximately 2 times (comparing the result about TR-2-2 and TR-2-3).Not existing of amino acid/11-15 from the TR-2N end in TR-2-2 makes in conjunction with reducing by approximately 2 times (comparing the result about TR-2-2 and TR-2-5).Not existing with the interpolation of amino acid 86-126 of simultaneous amino acid/11-15 makes in conjunction with reducing by approximately 3 times (comparing the result about TR-2-2 and TR-2-6).The elimination of residue 44-85 (TR-2-1) make in conjunction be reduced to for TR-2-2, observe that approximately 11%.Those results are pointed out amino acid/11-15 (SEQ ID NO:94; ALITQQDLAPQQRAA) and 44-85 (SEQ ID NO:95; CKYGQDYSTHWNDLL FCLRCTRCDSGEVE LSPCTTTRNTVCQ) the one or more residues in zone are important for the combination of the anti-TR-2 antibody of those 2 kinds of people and people TR-2.

The anti-TR-2 antibody of evaluator and cyno TR-2 is truncate, people/cyno mosaic and the people TR-2 combination separately that comprises some mouse TR-2 structural domain also.The strong combination of anti-TR-2 antibody and total length people TR-2 (fluorescence intensity (" FI ") 5681).The combination of the total length cynoTR-2 of anti-TR-2 antibody and microscler formula is compared to approximately 5 times of that minimizings (FI 1573) of total length people TR-2.For the total length cyno TR-2 (FI 209) of short-form and the combination of background level only observe to(for) the truncate 17-154 of cyno TR-2 (FI51), cyno 1-85 (FI 11) and cyno 17-85 (FI 8).

The anti-TR-2 antibody of some people and the chimeric combination of cyno/ people TR-2 (referring to Figure 27) have also been assessed.The antibody of observing and 4 kinds of chimeric combinations (FI) are as follows: cyno/ people's mosaic #1:FI 5977; Cyno/ people's mosaic #2:FI 47; Cyno/ people's mosaic #3:FI 12; Cyno/ people's mosaic #4:FI 1507.As above, the combination of the antibody of observing and total length people TR-2 is 5681, and the combination of antibody and total length cyno TR-2 is 1573 (microscler formulas) and 209 (short-form).

Because antibody and cyno/ people's mosaic #1 in conjunction with being similar to that with truncate TR-2-5, so, under the background of human amino acid 17-85, with corresponding cyno sequence replacement amino acid/11-16, obviously can not affect antibodies.Yet, under the background of total length people TR-2 (cyno/ people #2), replace amino acid/11-16 by corresponding cyno sequence and obviously cancel combination, confirm the part at least one the Amino acid profile epi-position from the 1-16 zone.With cyno/ people's mosaic #3 and #4 in conjunction with than that significantly the weakening with total length people TR-2, hint human sequence's amino acid/11 7-85 is in conjunction with being important.In a word, human sequence 1-85 zone (SEQ ID NO:96; ALITQQDLAPQQRAAPQQKRS SPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCT TTRNTVCQ) one or more amino acid in relate to the epi-position combination.Similarly, the various human sequences that replace in amino acid/11-85 zone by corresponding mouse sequence significantly weaken antibodies (referring to Figure 27), further confirm that one or more amino acid in that zone relate to the epi-position combination.

Sequence table

<110>AMGEN?INC.

<120 > polypeptide and antibody

<130>06843.0109-00304

<140>

<141>

<150>60/713,433

<151>2005-08-31

<150>60/713,478

<151>2005-08-31

<160>204

<170>PatentIn?Ver.3.3

<210>1

<211>363

<212>DNA

<213 > homo sapiens

<400>1

caggtgcagc?tggtgcagtc?tggggctgag?gtgaagaagc?ctggggcctc?agtgaaggtc?60

tcctgcaagg?cttctggata?caccttcacc?agttatgata?tcaactgggt?gcgacaggcc?120

actggacaag?ggcttgagtg?gatgggatgg?atgaacccta?acagtgataa?cacaggctat?180

gcacagaagt?tccagggcag?agtcaccatg?accaggaaca?cctccataag?cacagcctac?240

atggagttga?gcagcctgag?atctgaggac?acggccgtgt?attactgtgc?gagatggaat?300

cactatggtt?cggggagtca?ttttgactac?tggggccagg?gaaccctggt?caccgtctcc?360

tca 363

<210>2

<211>121

<212>PRT

<213 > homo sapiens

<400>2

Gln?Val?Gln?Leu?Val?Gln?Ser?Gly?Ala?Glu?Val?Lys?Lys?Pro?Gly?Ala

1 5 10 15

Ser?Val?Lys?Val?Ser?Cys?Lys?Ala?Ser?Gly?Tyr?Thr?Phe?Thr?Ser?Tyr

20 25 30

Asp?Ile?Asn?Trp?Val?Arg?Gln?Ala?Thr?Gly?Gln?Gly?Leu?Glu?Trp?Met

35 40 45

Gly?Trp?Met?Asn?Pro?Asn?Ser?Asp?Asn?Thr?Gly?Tyr?Ala?Gln?Lys?Phe

50 55 60

Gln?Gly?Arg?Val?Thr?Met?Thr?Arg?Asn?Thr?Ser?Ile?Ser?Thr?Ala?Tyr

65 70 75 80

Met?Glu?Leu?Ser?Ser?Leu?Arg?Ser?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Trp?Asn?His?Tyr?Gly?Ser?Gly?Ser?His?Phe?Asp?Tyr?Trp?Gly

100 105 110

Gln?Gly?Thr?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>3

<211>360

<212>DNA

<213 > homo sapiens

<400>3

caggtgcagc?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagc?agtggtggtc?actactggag?ctggatccgc?120

cagcacccag?ggaagggcct?ggagtggatt?gggtacatct?attacagtgg?gagcacctac?180

tacaacccgt?ccctcaagag?tcgagttacc?atatcagtag?acacgtctaa?gaaccagttc?240

tccctgaagc?tgagctctgt?gactgccgcg?gacacggccg?tgtattattg?tgcgagagat?300

gacagcagtg?gctggggttt?tgactactgg?ggccagggaa?tcctggtcac?cgtctcctca?360

<210>4

<211>120

<212>PRT

<213 > homo sapiens

<400>4

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?His?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asp?Asp?Ser?Ser?Gly?Trp?Gly?Phe?Asp?Tyr?Trp?Gly?Gln

100 105 110

Gly?Ile?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>5

<211>360

<212>DNA

<213 > homo sapiens

<400>5

caggtgcagc?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagc?agtggtggtc?actactggag?ctggatccgc?120

cagcacccag?ggaagggcct?ggagtggatt?gggtacatct?attacagtgg?gagcgcctac?180

tacaacccgt?ccctcaagag?tcgagttacc?atatcagtag?acacgtctaa?gaaccagttc?240

tccctgaagc?tgagctctgt?gactgccgcg?gacacggccg?tgtattactg?tgcgagagat?300

gacagcagtg?gctggggttt?tgactactgg?ggccagggaa?tcctggtcac?cgtctcctca?360

<210>6

<211>120

<212>PRT

<213 > homo sapiens

<400>6

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Sar?Ser?Gly

20 25 30

Gly?His?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Ala?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asp?Asp?Ser?Ser?Gly?Trp?Gly?Phe?Asp?Tyr?Trp?Gly?Gln

100 105 110

Gly?Ile?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>7

<211>360

<212>DNA

<213 > homo sapiens

<400>7

caggtgcagc?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagc?agtggtggtc?actactggag?ctggatccgc?120

cagcacccag?ggaagggcct?ggagtggatt?gggtacatct?attacagtgg?gagcgcctac?180

tacaacccgt?ccctcaagag?tcgagttacc?atatcagtag?acacgtctaa?gaaccagttc?240

tccctgaagc?tgagctctgt?gactgccgcg?gacacggccg?tgtattactg?tgcgagagat?300

gacagcagtg?gctggggttt?tgactactgg?ggccagggaa?tcctggtcac?cgtctcctca?360

<210>8

<211>120

<212>PRT

<213 > homo sapiens

<400>8

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?His?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Ala?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asp?Asp?Ser?Ser?Gly?Trp?Gly?Phe?Asp?Tyr?Trp?Gly?Gln

100 105 110

Gly?Ile?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>9

<211>363

<212>DNA

<213 > homo sapiens

<400>9

caggtgcagc?tggtggagtc?tgggggaggc?ttggtcaagc?ctggagggtc?cctgagactc?60

tcctgtgcag?cctctggatt?caccttcagt?gactactaca?tgaactggat?ccgccaggct?120

ccagggaagg?gactggagtg?ggtttcacac?attagtagta?gtggtagtat?cttagactac?180

gcagactctg?tgaagggccg?attcaccatc?tccagggaca?acgccaagaa?ctcactgtat?240

ctgcaaatga?acagcctgag?agtcgaggac?acggccgtgt?attactgtgc?gagagatggg?300

gctgcagctg?gtacggatgc?ttttgatctc?tggggccaag?ggacaatggt?caccgtctct?360

tca 363

<210>10

<211>121

<212>PRT

<213 > homo sapiens

<400>10

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Lys?Pro?Gly?Gly

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Asp?Tyr

20 25 30

Tyr?Met?Asn?Trp?Ile?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ser?His?Ile?Ser?Ser?Ser?Gly?Ser?Ile?Leu?Asp?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Lys?Asn?Ser?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Val?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Asp?Gly?Ala?Ala?Ala?Gly?Thr?Asp?Ala?Phe?Asp?Leu?Trp?Gly

100 105 110

Gln?Gly?Thr?Met?Val?Thr?Val?Ser?Ser

115 120

<210>11

<211>366

<212>DNA

<213 > homo sapiens

<400>11

caggtgcagc?tggtggagtc?tgggggaggc?gtggtccagc?ctgggaggtc?cctgagactc?60

tcctgtgcag?cgtctggatt?caccttcagt?tactatggca?tacactgggt?ccgccaggct?120

ccaggcaagg?ggctggagtg?ggtggcagtt?atatggtatg?atggaagtaa?taaatactat?180

gcagactccg?tgaagggccg?attcaccatc?tccagagaca?attccaagaa?cacgctgtat?240

ctgcaaatga?acagcctgag?agccgaggac?acggctgtgt?attactgtgc?gagagggagg?300

tatagcagct?cgtcctggtg?gtacttcgat?ctctggggcc?gtggcaccct?ggtcactgtc?360

tcctca 366

<210>12

<211>122

<212>PRT

<213 > homo sapiens

<400>12

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Tyr?Tyr

20 25 30

Gly?Ile?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ala?Val?Ile?Trp?Tyr?Asp?Gly?Ser?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Ash?Ser?Lys?Asn?Thr?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Gly?Arg?Tyr?Ser?Ser?Ser?Ser?Trp?Trp?Tyr?Phe?Asp?Leu?Trp

100 105 110

Gly?Arg?Gly?Thr?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>13

<211>366

<212>DNA

<213 > homo sapiens

<400>13

caggtgcagg?ctgagcagtc?gggcccagga?ctggtgaagc?cttcggagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagt?aattactact?ggagctggat?ccggcagccc?120

ccagggaagg?gactggagtg?gattgggtat?atctattaca?gtgggagcac?caagtacaac?180

ccctccctca?agagtcgagt?caccatatca?gtagacacgt?ccaagaacca?gttctccctg?240

aagctaacct?ctgtgaccac?tgcggacacg?gccgtgtatt?actgtgcgag?agactcccct?300

cgtggattta?gtggctacga?ggcttttgac?tcctggggcc?agggaaccct?ggtcaccgtc?360

tcctca 366

<210>14

<211>122

<212>PRT

<213 > homo sapiens

<400>14

Gln?Val?Gln?Ala?Glu?Gln?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Glu

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Asn?Tyr

20 25 30

Tyr?Trp?Ser?Trp?Ile?Arg?Gln?Pro?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Ile

35 40 45

Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Lys?Tyr?Asn?Pro?Ser?Leu?Lys

50 55 60

Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe?Ser?Leu

65 70 75 80

Lys?Leu?Thr?Ser?Val?Thr?Thr?Ala?Asp?Thr?Ala?Val?Tyr?Tyr?Cys?Ala

85 90 95

Arg?Asp?Ser?Pro?Arg?Gly?Phe?Ser?Gly?Tyr?Glu?Ala?Phe?Asp?Ser?Trp

100 105 110

Gly?Gln?Gly?Thr?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>15

<211>360

<212>DNA

<213 > homo sapiens

<400>15

caggtgcagc?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagc?agtgataatt?actactggag?ctggatccgc?120

cagcacccag?ggaagggcct?ggagtggatt?gggtacatct?attacagtgg?gagcacctac?180

tacaacccgt?ccctcaagag?tcgagttacc?atatcagtag?acacgtctaa?gaaccagttc?240

tccctgaagc?tgagctctgt?gactgccgcg?gacacggccg?tgtattactg?tgcgagagga?300

gttaactgga?actttctttt?tgatatctgg?ggccaaggga?caatggtcac?cgtctcttca?360

<210>16

<211>120

<212>PRT

<213 > homo sapiens

<400>16

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Asp

20 25 30

Asn?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Gly?Val?Asn?Trp?Asn?Phe?Leu?Phe?Asp?Ile?Trp?Gly?Gln

100 105 110

Gly?Thr?Met?Val?Thr?Val?Ser?Ser

115 120

<210>17

<211>348

<212>DNA

<213 > homo sapiens

<400>17

caggtgcagc?tggtggagtc?tgggggaggc?ttggtcaagc?ctggagggtc?cctgagactc?60

tcctgtgcag?cctctggatt?caccttcagt?gactactaca?tgagctggat?ccgccaggct?120

ccagggaagg?ggctggagtg?ggtttcatac?attagtagaa?gtggtagtac?catatactac?180

gcagactctg?tgaagggccg?attcaccatc?tccagggaca?acgccaagaa?ctcactgtat?240

ctgcaaatga?acagcctgag?agccgaggac?acggccgtgt?attactgtgc?gagatcttta?300

ggcggtatgg?acgtctgggg?ccaagggacc?acggtcaccg?tctcctca 348

<210>18

<211>116

<212>PRT

<213 > homo sapiens

<400>18

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Lys?Pro?Gly?Gly

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Asp?Tyr

20 25 30

Tyr?Met?Ser?Trp?Ile?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ser?Tyr?Ile?Ser?Arg?Ser?Gly?Ser?Thr?Ile?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Lys?Asn?Ser?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Ser?Leu?Gly?Gly?Met?Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr?Val

100 105 110

Thr?Val?Ser?Set

115

<210>19

<211>387

<212>DNA

<213 > homo sapiens

<400>19

caggtgcagc?tggtggagtc?tgggggaggc?gtggtccagc?ctgggaggtc?cctgagactc?60

tcctgtgcag?cgtctggatt?caccttcaat?aactatggca?tgcactgggt?ccgccaggct?120

ccaggcaagg?ggctggagtg?ggtggcagtt?atatggtatg?atggaagtaa?taaatactat?180

gcagactccg?tgaagggccg?attcaccatc?tccagagaca?attccaagaa?cacgctgtat?240

ctgcaaatga?acagcctgag?agccgaggac?acggctgtgt?attactgtgc?gagagatagg?300

accgtatata?gcaactcgtc?acccttttac?tactactact?acggtatgga?cgtctggggc?360

caagggacca?cggtcaccgt?ctcctca 387

<210>20

<211>129

<212>PRT

<213 > homo sapiens

<400>20

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Asn?Asn?Tyr

20 25 30

Gly?Met?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ala?Val?Ile?Trp?Tyr?Asp?Gly?Ser?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ser?Lys?Asn?Thr?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Asp?Arg?Thr?Val?Tyr?Ser?Asn?Ser?Ser?Pro?Phe?Tyr?Tyr?Tyr

100 105 110

Tyr?Tyr?Gly?Met?Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser

115 120 125

Ser

<210>21

<211>387

<212>DNA

<213 > homo sapiens

<400>21

caggtgcagc?tggtggagtc?tgggggaggc?gtggtccagc?ctgggaggtc?cctgagactc?60

tcctgtgcag?cgtctggatt?caccttcagt?acctatggca?tgcactgggt?ccgccaggct?120

ccaggcaagg?ggctggagtg?ggtggcagtt?atatggtatg?atggaagtaa?taaatactat?180

gcagactccg?tgaagggccg?attcaccatc?tccagagaca?attccaagaa?cacgctgtat?240

ctgcaaatga?acagcctgag?agccgaggac?acggctgtgt?attattgtgc?gagagatagg?300

accgtatata?gcagctcgtc?acccttttac?tactactact?acggtatgga?cgtctggggc?360

caagggacca?cggtcaccgt?ctcctca 387

<210>22

<211>129

<212>PRT

<213 > homo sapiens

<400>22

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Thr?Tyr

20 25 30

Gly?Met?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ala?Val?Ile?Trp?Tyr?Asp?Gly?Ser?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ser?Lys?Asn?Thr?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Asp?Arg?Thr?Val?Tyr?Ser?Ser?Ser?Ser?Pro?Phe?Tyr?Tyr?Tyr

100 105 110

Tyr?Tyr?Gly?Met?Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser

115 120 125

Ser

<210>23

<211>357

<212>DNA

<213 > homo sapiens

<400>23

caggtgcagc?tacagcagtg?gggcgcacga?ctgttgaagc?cttcggagac?cctgtccctc?60

acctgcgctg?tctatggtgg?gtccttcagt?ggttactact?ggagctggat?ccgccagccc?120

ccagggaagg?ggctggagtg?gattggggaa?atcaatcata?gtggaagcac?caactacaac?180

ccgtccctca?agagtcgagt?caccatatca?gtagacacgt?ccaagaacca?gttctccctg?240

aagctgaggt?ctgtgaccgc?cgcggacacg?gctgtgtatt?actgtgcgag?agggggaagc?300

agtggctact?ggtacttcga?tctctggggc?cgtggcaccc?tggtcactgt?ctcctca 357

<210>24

<211>119

<212>PRT

<213 > homo sapiens

<400>24

Gln?Val?Gln?Leu?Gln?Gln?Trp?Gly?Ala?Arg?Leu?Leu?Lys?Pro?Ser?Glu

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Ala?Val?Tyr?Gly?Gly?Ser?Phe?Ser?Gly?Tyr

20 25 30

Tyr?Trp?Ser?Trp?Ile?Arg?Gln?Pro?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Ile

35 40 45

Gly?Glu?Ile?Asn?His?Ser?Gly?Ser?Thr?Asn?Tyr?Asn?Pro?Ser?Leu?Lys

50 55 60

Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe?Ser?Leu

65 70 75 80

Lys?Leu?Arg?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr?Cys?Ala

85 90 95

Arg?Gly?Gly?Ser?Ser?Gly?Tyr?Trp?Tyr?Phe?Asp?Leu?Trp?Gly?Arg?Gly

100 105 110

Thr?Leu?Val?Thr?Val?Ser?Ser

115

<210>25

<211>363

<212>DNA

<213 > homo sapiens

<400>25

gaggtgcagg?tggtggagtc?tgggggaggc?ctggtcaagc?ctggggggtc?cctgagactc?60

tcctgtgcag?cctctggatt?caccttcagt?agctatagca?tgaactgggt?ccgccaggct?120

ccagggaagg?ggctggagtg?ggtctcatcc?attagtagta?gtagtagtta?catatactac?180

gcagactcag?tgaagggccg?attcaccatc?tccagagaca?acgccaagaa?ctcactgtat?240

ctgcaaatga?acagcctgag?agccgaggac?acggctgtgt?attactgtgc?gagggggggc?300

agcagctggt?acggggactg?gttcgacccc?tggggccagg?gaaccctggt?caccgtctcc?360

tca 363

<210>26

<211>121

<212>PRT

<213 > homo sapiens

<400>26

Glu?Val?Gln?Val?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Lys?Pro?Gly?Gly

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Ser?Tyr

20 25 30

Ser?Met?Asn?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ser?Ser?Ile?Ser?Ser?Ser?Ser?Ser?Tyr?Ile?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Lys?Asn?Ser?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Gly?Gly?Ser?Ser?Trp?Tyr?Gly?Asp?Trp?Phe?Asp?Pro?Trp?Gly

100 105 110

Gln?Gly?Thr?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>27

<211>378

<212>DNA

<213 > homo sapiens

<400>27

cagctggtgg?agtctggggg?aggcgtggtc?cagcctggga?ggtccctgag?actctcctgt?60

gcagcgtctg?gattcacctt?cagtagctat?ggcatgcact?gggtccgcca?ggctccaggc?120

aaggggctgg?agtgggtggc?agttatatgg?tatgatggaa?gaaataaata?ctatgcagac?180

tccgtgaagg?gccgattcac?catctccaga?gacaattcca?agaacacgct?gtatctgcaa?240

atgaacagcc?tgagagccga?ggacacggct?gtgtattact?gtgcgagaga?agtgggatat?300

tgtactaatg?gtgtatgctc?ctactactac?tacggtatgg?acgtctgggg?ccaagggacc?360

acggtcaccg?tctcctca 378

<210>28

<211>126

<212>PRT

<213 > homo sapiens

<400>28

Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg?Ser?Leu

1 5 10 15

Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Ser?Tyr?Gly?Met

20 25 30

His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val?Ala?Val

35 40 45

Ile?Trp?Tyr?Asp?Gly?Arg?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val?Lys?Gly

50 55 60

Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ser?Lys?Asn?Thr?Leu?Tyr?Leu?Gln

65 70 75 80

Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys?Ala?Arg

85 90 95

Glu?Val?Gly?Tyr?Cys?Thr?Asn?Gly?Val?Cys?Ser?Tyr?Tyr?Tyr?Tyr?Gly

100 105 110

Met?Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser?Ser

115 120 125

<210>29

<211>366

<212>DNA

<213 > homo sapiens

<400>29

caggtgcagc?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagc?agtggtgatt?acttctggag?ctggatccgc?120

cagctcccag?ggaagggcct?ggagtgcatt?gggcacatcc?ataacagtgg?gaccacctac?180

tacaatccgt?ccctcaagag?tcgagt?acc?atatcagtag?acacgtctaa?gaagcagttc?240

tccctgaggc?tgagttctgt?gactgccgcg?gacacggccg?tatattactg?tgcgagagat?300

cgagggggtg?actactacta?tggtatggac?gtctggggcc?aagggaccac?ggtcaccgtc?360

tcctca 366

<210>30

<211>122

<212>PRT

<213 > homo sapiens

<400>30

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Asp?Tyr?Phe?Trp?Ser?Trp?Ile?Arg?Gln?Leu?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Cys?Ile?Gly?His?Ile?His?Asn?Ser?Gly?Thr?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Lys?Gln?Phe

65 70 75 80

Ser?Leu?Arg?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asp?Arg?Gly?Gly?Asp?Tyr?Tyr?Tyr?Gly?Met?Asp?Val?Trp

100 105 110

Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser?Ser

115 120

<210>31

<211>366

<212>DNA

<213 > homo sapiens

<400>31

caggtgcagc?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcagtg?tctctggtgg?ctccatcagc?agtggtggtt?actactggag?ctggatccgc?120

cagcacccag?ggaagggcct?ggagtggatt?gggtacatct?attacagtgg?gagcacctac?180

tgcaacccgt?ccctcaagag?tcgagttacc?atatcagtcg?acacgtctaa?gaaccagttc?240

tccctgaagc?tgagctctgt?gactgccgcg?gacacggccg?tgtattactg?tgcgagagac?300

aatggttcgg?ggagttatga?ctggttcgac?ccctggggcc?agggaatcct?ggtcaccgtc?360

tcctca 366

<210>32

<211>122

<212>PRT

<213 > homo sapiens

<400>32

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Ser?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Cys?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asp?Asn?Gly?Ser?Gly?Ser?Tyr?Asp?Trp?Phe?Asp?Pro?Trp

100 105 110

Gly?Gln?Gly?Ile?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>33

<211>366

<212>DNA

<213 > homo sapiens

<400>33

caggtgcaga?tgcaggagtc?gggcccagga?ctggtgaagc?cttcacagac?cctgtccctc?60

acctgcactg?tctctggtgg?ctccatcagc?agtggtgatt?actactggag?ctggatccgc?120

cagcacccag?ggaagaacct?ggagtggatt?gggtacatct?attacagtgg?gagcacctac?180

tacaacccgt?ccctcaagag?tcgagttacc?atatcagtag?acacgtctaa?gaaccagttc?240

tccctgaagc?tgagctctgt?gactgccgcg?gacacggccg?tgtattactg?tgcgagagac?300

aatggttcgg?ggagttatga?ctggttcgac?ccctggggcc?agggaaccct?ggtcaccgtc?360

tcctca 366

<210>34

<211>122

<212>PRT

<213 > homo sapiens

<400>34

Gln?Val?Gln?Met?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Asp?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Asn?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asp?Asn?Gly?Ser?Gly?Ser?Tyr?Asp?Trp?Phe?Asp?Pro?Trp

100 105 110

Gly?Gln?Gly?Thr?Leu?Val?Thr?Val?Ser?Ser

115 120

<210>35

<211>321

<212>DNA

<213 > homo sapiens

<400>35

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgcc?gggcaagtca?gagcattagc?atttatttaa?attggtatca?gcagaaacca?120

gggaaagccc?ctaagctcct?gatctatgct?gcatccagtt?tgcaaagtgg?ggtcccatta?180

aggttcagtg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?tctgcaacct?240

gaagatattg?caacttacta?ctgtcaacag?agttacaaaa?ccccgctcac?tttcggcgga?300

gggaccaagg?tggagatcaa?a 321

<210>36

<211>107

<212>PRT

<213 > homo sapiens

<400>36

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Ser?Ile?Ser?Ile?Tyr

20 25 30

Leu?Asn?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Leu?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Ile?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ser?Tyr?Lys?Thr?Pro?Leu

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>37

<211>321

<212>DNA

<213 > homo sapiens

<400>37

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgttggaga?cagagtcacc?60

atcacttgcc?gggcaagtca?gggccttaga?aatgatttag?gctggtttca?gcagaaacca?120

gggaaagtca?ctaagcgcct?gatctatgct?gcatccagtt?tgcaaagagg?ggtcccatca?180

aggttcagcg?gcagtggatc?tgggacagaa?ttcactctca?caatcagcag?cctgcagcct?240

gaagattttg?caacttatta?ctgtctacag?cattatagtt?tcccgtggac?gttcggccaa?300

gggaccaagg?tggagatcaa?a 321

<210>38

<211>107

<212>PRT

<213 > homo sapiens

<400>38

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Leu?Arg?Asn?Asp

20 25 30

Leu?Gly?Trp?Phe?Gln?Gln?Lys?Pro?Gly?Lys?Val?Thr?Lys?Arg?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Arg?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Glu?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Leu?Gln?His?Tyr?Ser?Phe?Pro?Trp

85 90 95

Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>39

<211>321

<212>DNA

<213 > homo sapiens

<400>39

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgttggaga?cagagtcacc?60

atcacttgcc?gggcaagtca?gggccttaga?aatgatttag?gctggtttca?gcagaaacca?120

gggaaagccc?ctaagcgcct?gatctatgct?gcatccagtt?tgcaaagagg?ggtcccatca?180

aggttcagcg?gcagtggatc?tgggacagaa?ttcactctca?caatcagcag?cctgcagcct?240

gaagatttta?caacttattt?ctgtctacag?cataatagtt?tcccgtggac?gttcggccaa?300

gggaccaagg?tggaaatcaa?a 321

<210>40

<211>107

<212>PRT

<213 > homo sapiens

<400>40

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Leu?Arg?Asn?Asp

20 25 30

Leu?Gly?Trp?Phe?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Arg?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Arg?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Glu?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Thr?Thr?Tyr?Phe?Cys?Leu?Gln?His?Asn?Ser?Phe?Pro?Trp

85 90 95

Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>41

<211>321

<212>DNA

<213 > homo sapiens

<400>41

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgttggaga?cagagtcacc?60

atcacttgcc?gggcaagtca?gggccttaga?aatgatttag?gctggtttca?gcagaaacca?120

gggaaagccc?ctaagcgcct?gatctatgct?gcatccagtt?tgcaaagagg?ggtcccatca?180

aggttcagcg?gcagtggatc?tgggacagaa?ttcactctca?caatcagcag?cctgcagcct?240

gaagatttta?caacttattt?ctgtctacag?cataatagtt?tcccgtggac?gttcggccaa?300

gggaccaagg?tggaaatcaa?a 321

<210>42

<211>107

<212>PRT

<213 > homo sapiens

<400>42

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Leu?Arg?Asn?Asp

20 25 30

Leu?Gly?Trp?Phe?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Arg?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Arg?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Glu?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Thr?Thr?Tyr?Phe?Cys?Leu?Gln?His?Asn?Ser?Phe?Pro?Trp

85 90 95

Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>43

<211>321

<212>DNA

<213 > homo sapiens

<400>43

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgcc?ggtcaagtca?gagcattagt?aactatataa?attggtatca?acagagacca?120

gggaaagccc?cgaacctcct?gatccatgat?gtatccagtt?tccaaagtgc?ggtcccatca?180

aggttcagtc?gcagtggatc?tgggacagtt?ttcactctca?ccatcagcag?tctgcaacct?240

gaagattttg?caacttactt?ctgtcaacag?acttacatta?ccccattcac?tttcggccct?300

gggaccaaag?tggatatcaa?a 321

<210>44

<211>107

<212>PRT

<213 > homo sapiens

<400>44

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ser?Ser?Gln?Ser?Ile?Ser?Asn?Tyr

20 25 30

Ile?Asn?Trp?Tyr?Gln?Gln?Arg?Pro?Gly?Lys?Ala?Pro?Asn?Leu?Leu?Ile

35 40 45

His?Asp?Val?Ser?Ser?Phe?Gln?Ser?Ala?Val?Pro?Ser?Arg?Phe?Ser?Arg

50 55 60

Ser?Gly?Ser?Gly?Thr?Val?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Phe?Cys?Gln?Gln?Thr?Tyr?Ile?Thr?Pro?Phe

85 90 95

Thr?Phe?Gly?Pro?Gly?Thr?Lys?Val?Asp?Ile?Lys

100 105

<210>45

<211>321

<212>DNA

<213 > homo sapiens

<400>45

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgcc?gggcgagtca?gggcattagc?aattatttag?cctggtatca?gcagaaacca?120

gggaaagttc?ctaagctcct?gatctatgct?gcatccactt?tgcaatcagg?ggtcccatct?180

cggttcagtg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?cctgcagcct?240

gaagatgttg?caacttatta?ctgtcaaaag?tataacagtg?ccccgctcac?tttcggcgga?300

gggaccaagg?tggagatcaa?a 321

<210>46

<211>107

<212>PRT

<213 > homo sapiens

<400>46

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Asn?Tyr

20 25 30

Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Val?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Thr?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Val?Ala?Thr?Tyr?Tyr?Cys?Gln?Lys?Tyr?Asn?Ser?Ala?Pro?Leu

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>47

<211>339

<212>DNA

<213 > homo sapiens

<400>47

gacatcgtga?tgacccagtc?tccagactcc?ctggctgtgt?ctctgggcga?gagggccacc?60

atcaactgca?agtccagcca?gagtgtttta?tacaggtcca?acaataagat?ctacttagct?120

tggtaccagc?agaaaccagg?acagcctcct?aagctgctca?tttactgggc?atcgacccgg?180

gaatccgggg?tccctgaccg?attcagtggc?agcgggtctg?ggacagattt?cactctcacc?240

atcagcagcc?tgctggctga?agatgtggca?gtttattact?gtcagcaata?ttatagtact?300

ccattcactt?tcggccctgg?gaccaaagtg?gatatcaaa 339

<210>48

<211>113

<212>PRT

<213 > homo sapiens

<400>48

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Asp?Ser?Leu?Ala?Val?Ser?Leu?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Ile?Asn?Cys?Lys?Ser?Ser?Gln?Ser?Val?Leu?Tyr?Arg

20 25 30

Ser?Asn?Asn?Lys?Ile?Tyr?Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln

35 40 45

Pro?Pro?Lys?Leu?Leu?Ile?Tyr?Trp?Ala?Ser?Thr?Arg?Glu?Ser?Gly?Val

50 55 60

Pro?Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr

65 70 75 80

Ile?Ser?Ser?Leu?Leu?Ala?Glu?Asp?Val?Ala?Val?Tyr?Tyr?Cys?Gln?Gln

85 90 95

Tyr?Tyr?Ser?Thr?Pro?Phe?Thr?Phe?Gly?Pro?Gly?Thr?Lys?Val?Asp?Ile

100 105 110

Lys

<210>49

<211>336

<212>DNA

<213 > homo sapiens

<400>49

gatattgtga?tgactcagtc?tccactctcc?ctgcccgtca?cccctggaga?gccggcctcc?60

atctcctgca?ggtctagtca?gagcctcctg?cgtcgtaatg?gatacaacta?tttggattgg?120

tacctgcaga?agccagggca?gtctccacaa?ctcctgatct?atttgggttc?taatcgggcc?180

tccggggtcc?cagacaggtt?cagtggcagt?ggatcaggca?cagattttac?actgaaaatc?240

agcagagtgg?aggctgagga?tgttggggtt?tattactgca?tgcaagctct?acaaactccg?300

ctcactttcg?gcggagggac?cgaggtggag?atcaaa 336

<210>50

<211>112

<212>PRT

<213 > homo sapiens

<400>50

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Leu?Ser?Leu?Pro?Val?Thr?Pro?Gly

1 5 10 15

Glu?Pro?Ala?Ser?Ile?Ser?Cys?Arg?Ser?Ser?Gln?Ser?Leu?Leu?Arg?Arg

20 25 30

Asn?Gly?Tyr?Asn?Tyr?Leu?Asp?Trp?Tyr?Leu?Gln?Lys?Pro?Gly?Gln?Ser

35 40 45

Pro?Gln?Leu?Leu?Ile?Tyr?Leu?Gly?Ser?Asn?Arg?Ala?Ser?Gly?Val?Pro

50 55 60

Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Lys?Ile

65 70 75 80

Ser?Arg?Val?Glu?Ala?Glu?Asp?Val?Gly?Val?Tyr?Tyr?Cys?Met?Gln?Ala

85 90 95

Leu?Gln?Thr?Pro?Leu?Thr?Phe?Gly?Gly?Gly?Thr?Glu?Val?Glu?Ile?Lys

100 105 110

<210>51

<211>339

<212>DNA

<213 > homo sapiens

<400>51

gacatcgtga?tgacccagtt?tccagactcc?ctggctgtgt?ctctgggcga?gagggccacc?60

atcaactgca?agtccagcca?gagtgtttta?cacagctcca?acaataagaa?ctacttaact?120

tggtaccagc?tgaaaccagg?acagcctcct?aagttgctca?tttactgggc?atctacccgg?180

gaatccgggg?tccctgaccg?attcagtggc?agcgggtctg?ggacagattt?cactctcacc?240

atcagcagcc?tgcaggctga?agatgtggca?gtttattact?gtcaccaata?ttatagtact?300

ccgtccagtt?ttggccaggg?gaccaagctg?gagatcaaa 339

<210>52

<211>113

<212>PRT

<213 > homo sapiens

<400>52

Asp?Ile?Val?Met?Thr?Gln?Phe?Pro?Asp?Ser?Leu?Ala?Val?Ser?Leu?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Ile?Asn?Cys?Lys?Ser?Ser?Gln?Ser?Val?Leu?His?Ser

20 25 30

Ser?Asn?Asn?Lys?Asn?Tyr?Leu?Thr?Trp?Tyr?Gln?Leu?Lys?Pro?Gly?Gln

35 40 45

Pro?Pro?Lys?Leu?Leu?Ile?Tyr?Trp?Ala?Ser?Thr?Arg?Glu?Ser?Gly?Val

50 55 60

Pro?Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr

65 70 75 80

Ile?Ser?Ser?Leu?Gln?Ala?Glu?Asp?Val?Ala?Val?Tyr?Tyr?Cys?His?Gln

85 90 95

Tyr?Tyr?Ser?Thr?Pro?Ser?Ser?Phe?Gly?Gln?Gly?Thr?Lys?Leu?Glu?Ile

100 105 110

Lys

<210>53

<211>321

<212>DNA

<213 > homo sapiens

<400>53

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgttggaga?cagagtcacc?60

atcacttgcc?ggacaagtca?gagcattagc?acctatttaa?attggtatca?gcagaaacca?120

gggaaagccc?ctaagctcct?gatctctgct?acatccagtt?tgcaaagtgg?ggtcccatca?180

aggttcagtg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?tctgcaacct?240

gaagattttg?caacttacta?ctgtcaacag?agttacagta?ccccgctcac?tttcggcgga?300

gggaccaagg?tggagatcaa?a 321

<210>54

<211>107

<212>PRT

<213 > homo sapiens

<400>54

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Thr?Ser?Gln?Ser?Ile?Ser?Thr?Tyr

20 25 30

Leu?Asn?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Ser?Ala?Thr?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ser?Tyr?Ser?Thr?Pro?Leu

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>55

<211>321

<212>DNA

<213 > homo sapiens

<400>55

gacatccaga?tgacccagtc?tccatcctcc?ctgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgcc?gggcaagtca?gagcattagc?agctatttaa?attggtatca?gcagaaacca?120

gggaaagccc?ctaagctcct?gatctctgct?acatccagtt?ttcaaagtgg?ggtcccatca?180

aggttcagtg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?tctgcaacct?240

gaagattttg?cagcttacta?ctgtcaacag?agttacagta?ccccgctcac?tttcggcgga?300

gggaccaagg?tggagatcaa?a 321

<210>56

<211>107

<212>PRT

<213 > homo sapiens

<400>56

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Ser?Ile?Ser?Ser?Tyr

20 25 30

Leu?Asn?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Ser?Ala?Thr?Ser?Ser?Phe?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Ala?Tyr?Tyr?Cys?Gln?Gln?Ser?Tyr?Ser?Thr?Pro?Leu

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>57

<211>339

<212>DNA

<213 > homo sapiens

<400>57

gacatcgtga?tgacccagtc?tccagactcc?ctggctgtgt?ctctgggcga?gagggccacc?60

atcaactgca?agtccagcca?gagtgtttta?cacagctcca?acaataagaa?ttatttagtt?120

tggtaccagc?agaaaccagg?acagcctcct?aagctgctca?tttactgggc?atctacccgg?180

gaatccgggg?tccctgaccg?attcagtggc?agcgggtctg?ggacagattt?cactctcacc?240

atcagcagcc?tgcaggctga?agatgtggca?gtttattact?gtcagcaata?ttatagtact?300

cctctcactt?tcggcggagg?gaccaaggtg?gagatcaaa 339

<210>58

<211>113

<212>PRT

<213 > homo sapiens

<400>58

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Asp?Ser?Leu?Ala?Val?Ser?Leu?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Ile?Asn?Cys?Lys?Ser?Ser?Gln?Ser?Val?Leu?His?Ser

20 25 30

Ser?Asn?Asn?Lys?Asn?Tyr?Leu?Val?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln

35 40 45

Pro?Pro?Lys?Leu?Leu?Ile?Tyr?Trp?Ala?Ser?Thr?Arg?Glu?Ser?Gly?Val

50 55 60

Pro?Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr

65 70 75 80

Ile?Ser?Ser?Leu?Gln?Ala?Glu?Asp?Val?Ala?Val?Tyr?Tyr?Cys?Gln?Gln

85 90 95

Tyr?Tyr?Ser?Thr?Pro?Leu?Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile

100 105 110

Lys

<210>59

<211>321

<212>DNA

<213 > homo sapiens

<400>59

gacatccaga?tgacccagtc?tccatcttcc?gtgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgtc?gggcgagtca?gggtattagc?agctggttag?tctggtatca?gcagaaacca?120

gggaaagccc?ctaagctcct?gatctatgct?gcatccagtt?tgcaaagtgg?ggtcccatca?180

aggttcagcg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?cctgcagcct?240

gaagattttg?caacttacta?ttgtcagcag?gctaacagtt?tccctttcac?tttcggcgga?300

gggaccaagg?tggagatcaa?a 321

<210>60

<211>107

<212>PRT

<213 > homo sapiens

<400>60

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Val?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Ser?Trp

20 25 30

Leu?Val?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ala?Asn?Ser?Phe?Pro?Phe

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>61

<211>321

<212>DNA

<213 > homo sapiens

<400>61

gacatccaga?tgacccagtc?tccatcctca?ctgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgtc?gggcgagtca?gggcattagc?aattatttag?cctggtttca?gcagaaacca?120

gggaaagccc?ctaagtccct?gatctatgct?gcatccagtt?tgcaaagtgg?ggtcccatca?180

aaattcagcg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?cctgcagcct?240

gaagattttg?caacttatta?ctgccaacag?tataatagtt?accctctcac?tttcggcgga?300

gggaccaagg?tggagatcaa?a 321

<210>62

<211>107

<212>PRT

<213 > homo sapiens

<400>62

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Asn?Tyr

20 25 30

Leu?Ala?Trp?Phe?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Ser?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Lys?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Tyr?Asn?Ser?Tyr?Pro?Leu

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>63

<211>324

<212>DNA

<213 > homo sapiens

<400>63

gaaattgtgt?tgacgcagtc?tccaggcacc?ctgtctttgt?ctccagggga?aagagccacc?60

ctctcctgca?gggccagtca?gggtattagt?agaagctact?tagcctggta?ccagcagaaa?120

cctggccagg?ctcccagcct?cctcatctat?ggtgcatcca?gcagggccac?tggcatccca?180

gacaggttca?gtggcagtgg?gtctgggaca?gacttcactc?tcaccatcag?cagactggag?240

cctgaagatt?ttgcagtgta?ttactgtcaa?caatttggta?gttcaccgtg?gacgttcggc?300

caagggacca?aggtggaaat?caaa 324

<210>64

<211>108

<212>PRT

<213 > homo sapiens

<400>64

Glu?Ile?Val?Leu?Thr?Gln?Ser?Pro?Gly?Thr?Leu?Ser?Leu?Ser?Pro?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Leu?Ser?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Arg?Ser

20 25 30

Tyr?Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln?Ala?Pro?Ser?Leu?Leu

35 40 45

Ile?Tyr?Gly?Ala?Ser?Ser?Arg?Ala?Thr?Gly?Ile?Pro?Asp?Arg?Phe?Ser

50 55 60

Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Arg?Leu?Glu

65 70 75 80

Pro?Glu?Asp?Phe?Ala?Val?Tyr?Tyr?Cys?Gln?Gln?Phe?Gly?Ser?Ser?Pro

85 90 95

Trp?Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>65

<211>321

<212>DNA

<213 > homo sapiens

<400>65

gacattcaga?tgacccagtc?tccatcctcc?gtgtctgcat?ctgtaggaga?cagagtcacc?60

atcacttgtc?gggcgagtca?gggtattagc?agctggttag?cctggtatca?gcagaaacca?120

gggaaagccc?caaagttcct?gatctttgtt?gcatccagtt?tccaaagtgg?ggtcccatca?180

aggttcagcg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?cctgcagcct?240

gaagattttg?caacttacta?ttgtcaacag?gctaacagtt?tccctcggac?gttcggccaa?300

gggaccaagg?tggaaatcaa?a 321

<210>66

<211>107

<212>PRT

<213 > homo sapiens

<400>66

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Val?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Ser?Trp

20 25 30

Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Phe?Leu?Ile

35 40 45

Phe?Val?Ala?Ser?Ser?Phe?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ala?Asn?Ser?Phe?Pro?Arg

85 90 95

Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>67

<211>321

<212>DNA

<213 > homo sapiens

<400>67

gacatccaga?tgacccagtc?tccatcttcc?gtgtctgcat?ctgttggaga?cagagtcacc?60

atcacttgtc?gggcgagtca?gggtattagc?agctggttag?cctggtatca?gcagaaacca?120

gggaaagccc?ctaagttcct?gatctttgtt?gcatccagtt?tgcaaagtgg?ggtcccatca?180

aggttcagcg?gcagtggatc?tgggacagat?ttcactctca?ccatcagcag?cctgcagcct?240

gaagattttg?caacttacta?ttgtcaacag?gctaacagtt?tccctcggac?gttcggccaa?300

gggaccaagg?tggaaatcaa?a 321

<210>68

<211>107

<212>PRT

<213 > homo sapiens

<400>68

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Val?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Ser?Trp

20 25 30

Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Phe?Leu?Ile

35 40 45

Phe?Val?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ala?Asn?Ser?Phe?Pro?Arg

85 90 95

Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>69

<211>312

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic

<400>69

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser

275 280 285

Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val?Thr

290 295 300

Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>70

<211>201

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic

<400>70

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser

195 200

<210>71

<211>243

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>71

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln

<210>72

<211>284

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>72

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val

275 280

<210>73

<211>186

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>73

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Pro?Gln

145 150 155 160

Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His

165 170 175

Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser

180 185

<210>74

<211>228

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>74

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phs

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Pro?Gln

145 150 155 160

Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His

165 170 175

Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp

180 185 190

Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg

195 200 205

Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn

210 215 220

Thr?Val?Cys?Gln

225

<210>75

<211>269

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>75

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Pro?Gln

145 150 155 160

Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His

165 170 175

Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp

180 185 190

Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg

195 200 205

Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn

210 215 220

Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro

225 230 235 240

Glu?Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys

245 250 255

Val?Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val

260 265

<210>76

<211>202

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>76

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

5?0 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ile?Ser

145 150 155 160

Cys?Lys?Tyr?Gly?Gln?Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe

165 170 175

Cys?Leu?Arg?Cys?Thr?Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro

180 185 190

Cys?Thr?Thr?Thr?Arg?Asn?Thr?Val?Cys?Gln

195 200

<210>77

<211>243

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>77

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ile?Ser

145 150 155 160

Cys?Lys?Tyr?Gly?Gln?Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe

165 170 175

Cys?Leu?Arg?Cys?Thr?Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro

180 185 190

Cys?Thr?Thr?Thr?Arg?Asn?Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe

195 200 205

Arg?Glu?Glu?Asp?Ser?Pro?Glu?Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys

210 215 220

Pro?Arg?Gly?Met?Val?Lys?Val?Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile

225 230 235 240

Glu?Cys?Val

<210>78

<211>228

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>78

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Gln?Cys

145 150 155 160

Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu?Met?Cys?Arg?Lys

165 170 175

Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val?Gly?Asp?Cys?Thr

180 185 190

Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser?Gly?Thr?Lys?His

195 200 205

Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val?Thr?Ser?Ser?Pro?Gly

210 215 220

Thr?Pro?Ala?Ser

225

<210>79

<211>271

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>79

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ile?Ser

145 150 155 160

Cys?Lys?Tyr?Gly?Gln?Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe

165 170 175

Cys?Leu?Arg?Cys?Thr?Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro

180 185 190

Cys?Thr?Thr?Thr?Arg?Asn?Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe

195 200 205

Arg?Glu?Glu?Asp?Ser?Pro?Glu?Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys

210 215 220

Pro?Arg?Gly?Met?Val?Lys?Val?Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile

225 230 235 240

Glu?Cys?Val?His?Lys?Glu?Ser?Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro

245 250 255

Ala?Val?Glu?Glu?Thr?Val?Thr?Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

260 265 270

<210>80

<211>209

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>80

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Pro?Gln

145 150 155 160

Gln Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His

165 170 175

Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Tyr?Lys?Tyr?Gly?Gln?Asp

180 185 190

Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg

195 200 205

Cys

<210>81

<211>217

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>81

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Pro?Gln

145 150 155 160

Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His

165 170 175

Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp

180 185 190

Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg

195 200 205

Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser

210 215

<210>82

<211>290

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>82

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Pro

145 150 155 160

Ile?Thr?Arg?Gln?Ser?Leu?Asp?Pro?Gln?Arg?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Thr?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Lys?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Ile?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Lys?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Pro?Gln?Arg?Arg?Ile

275 280 285

Gln?Thr

290

<210>83

<211>312

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>83

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Pro

145 150 155 160

Ile?Thr?Arg?Gln?Ser?Leu?Asp?Pro?Gln?Arg?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Thr?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Lys?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Ile?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Lys?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser

275 280 285

Gly?Thr?Lys?His?Thr?Gly?Glu?Val?Pro?Ala?Val?Glu?Lys?Thr?Val?Thr

290 295 300

Thr?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>84

<211>243

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>84

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Pro

145 150 155 160

Ile?Thr?Arg?Gln?Ser?Leu?Asp?Pro?Gln?Arg?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Thr?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Lys?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln

<210>85

<211>228

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>85

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Pro?Gln

145 150 155 160

Gln?Lys?Arg?Ser?Ser?Pro?Ile?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His

165 170 175

Ile?Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp

180 185 190

Tyr?Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Lys

195 200 205

Cys?Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg?Asn

210 215 220

Thr?Val?Cys?Gln

225

<210>86

<211>243

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>86

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

5er?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Pro

145 150 155 160

Ile?Thr?Arg?Gln?Ser?Leu?Asp?Pro?Gln?Arg?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln

<210>87

<211>312

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>87

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Pro

145 150 155 160

Ile?Thr?Arg?Gln?Ser?Leu?Asp?Pro?Gln?Arg?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Tyr?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser

275 280 285

Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val?Thr

290 295 300

Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>88

<211>243

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>88

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Thr?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Lys?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln

<210>89

<211>312

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>89

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Thr?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Lys?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Ile?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Lys?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser

275 280 285

Gly?Thr?Lys?His?Thr?Gly?Glu?Val?Pro?Ala?Val?Glu?Lys?Thr?Val?Thr

290 295 300

Thr?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>90

<211>311

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>90

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Val?Pro

145 150 155 160

Val?Thr?Ala?Asn?Pro?Ala?His?Asn?Arg?Pro?Ala?Gly?Leu?Gln?Arg?Pro

165 170 175

Glu?Glu?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile?Ser

180 185 190

Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr?Ser

195 200 205

Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys?Asp

210 215 220

Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr?Val

225 230 235 240

Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu?Met

245 250 255

Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val?Gly

260 265 270

Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser?Gly

275 280 285

Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val?Thr?Ser

290 295 300

Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>91

<211>312

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>91

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Leu?Ala?Gly?Gln?Tyr?Leu

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 255 270

Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser

275 280 285

Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val?Thr

290 295 300

Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>92

<211>312

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>92

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr?Arg?Cys

210 215 220

Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn?Thr

225 230 235 240

Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro?Glu

245 250 255

Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys?Val

260 265 270

Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu?Ser

275 280 285

Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val?Thr

290 295 300

Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>93

<211>313

<212>PRT

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic peptide sequence

<400>93

Met?Val?His?Ala?Thr?Ser?Pro?Leu?Leu?Leu?Leu?Leu?Leu?Leu?Ser?Leu

1 5 10 15

Ala?Leu?Val?Ala?Pro?Gly?Leu?Ser?Ala?Arg?Lys?Cys?Ser?Leu?Thr?Gly

20 25 30

Lys?Trp?Thr?Asn?Asp?Leu?Gly?Ser?Asn?Met?Thr?Ile?Gly?Ala?Val?Asn

35 40 45

Ser?Lys?Gly?Glu?Phe?Thr?Gly?Thr?Tyr?Thr?Thr?Ala?Val?Thr?Ala?Thr

50 55 60

Ser?Asn?Glu?Ile?Lys?Glu?Ser?Pro?Leu?His?Gly?Thr?Gln?Asn?Thr?Ile

65 70 75 80

Asn?Lys?Arg?Thr?Gln?Pro?Thr?Phe?Gly?Phe?Thr?Val?Asn?Trp?Lys?Phe

85 90 95

Ser?Glu?Ser?Thr?Thr?Val?Phe?Thr?Gly?Gln?Cys?Phe?Ile?Asp?Arg?Asn

100 105 110

Gly?Lys?Glu?Val?Leu?Lys?Thr?Met?Trp?Leu?Leu?Arg?Ser?Ser?Val?Asn

115 120 125

Asp?Ile?Gly?Asp?Asp?Trp?Lys?Ala?Thr?Arg?Val?Gly?Ile?Asn?Ile?Phe

130 135 140

Thr?Arg?Leu?Arg?Thr?Gln?Lys?Glu?Gln?Leu?Leu?Ala?Ser?Leu?Ala?Leu

145 150 155 160

Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro?Gln?Gln

165 170 175

Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His?His?Ile

180 185 190

Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln?Asp?Tyr

195 200 205

Ser?Thr?His?Ser?Asn?His?Ser?Leu?Asp?Ser?Cys?Leu?Arg?Cys?Thr?Arg

210 215 220

Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg?Asn

225 230 235 240

Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser?Pro

245 250 255

Glu?Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val?Lys

260 265 270

Val?Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys?Glu

275 280 285

Ser?Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr?Val

290 295 300

Thr?Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

305 310

<210>94

<211>15

<212>PRT

<213 > homo sapiens

<400>94

Ala?Leu?Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala

1 5 10 15

<210>95

<211>42

<212>PRT

<213 > homo sapiens

<400>95

Cys?Lys?Tyr?Gly?Gln?Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe

1 5 10 15

Cys?Leu?Arg?Cys?Thr?Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro

20 25 30

Cys?Thr?Thr?Thr?Arg?Asn?Thr?Val?Cys?Gln

35 40

<210>96

<211>85

<212>PRT

<213 > homo sapiens

<400>96

Ala?Leu?Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro

1 5 10 15

Gln?Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His

20 25 30

His?Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln

35 40 45

Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr

50 55 60

Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg

65 70 75 80

Asn?Thr?Val?Cys?Gln

85

<210>97

<211>14

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>97

gtaggtgctg?tcct 14

<210>98

<211>14

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>98

tgagttccac?gaca 14

<210>99

<211>13

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>99

cttccaagcc?act 13

<210>100

<211>12

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>100

cargcactgt?ca 12

<210>101

<211>18

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>101

gtaggtgctg?tccttgct 18

<210>102

<211>21

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>102

ctctgtgaca?ctctcctggg?a 21

<210>103

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>103

gctctagatt?ggagggcgtt?atccaccttc?cact 34

<210>104

<211>39

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>104

aactagctag?cagttccaga?tttcaactgc?tcatcagat 39

<210>105

<211>18

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>105

gctcccgggt?agaagtca 18

<210>106

<211>22

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>106

acyagtgtgg?ccttgttggc?tt 22

<210>107

<211>26

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>107

gctctagagg?gygggaacag?agtgac 26

<210>108

<211>18

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>108

acgacaccgt?caccggtt 18

<210>109

<211>25

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>109

aagtagtcct?tgaccaggca?gccca 25

<210>110

<211>30

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>110

gctctagagg?gtgccagggg?gaagaccgat 30

<210>111

<211>28

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>111

gctctagagc?agggcgccag?ggggaaga 28

<210>112

<211>20

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>112

atgaggstcc?cygctcagct 20

<210>113

<211>21

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>113

atggaarccc?cagckcagct?t 21

<210>114

<211>22

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>114

atggtgttgc?agacccaggt?ct 22

<210>115

<211>35

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>115

gaagatctca?ccatgaggst?cccygctcag?ctyct 35

<210>116

<211>38

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>116

gaagatctca?ccatggaarc?cccagckcag?cttctctt 38

<210>117

<211>38

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>117

gaagatctca?ccatggtgtt?gcagacccag?gtcttcat 38

<210>118

<211>18

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>118

crtcwccacc?atgrcmwg 18

<210>119

<211>19

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>119

caccatgrcc?wgstyccct 19

<210>120

<211>20

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>120

accatggcct?ggrctcykct 20

<210>121

<211>19

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>121

caccatggcm?tggrycvyt 19

<210>122

<211>21

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>122

caccatggcy?tggryccmay?t 21

<210>123

<211>29

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>123

gaagatctca?ccatgrccwg?styccctct 29

<210>124

<211>32

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>124

gaagatctca?ccatggcctg?grctcykcts?yt 32

<210>125

<211>30

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>125

gaagatctca?ccatggcmtg?grycvy?tctc 30

<210>126

<211>30

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>126

gaagatctca?ccatggcytg?gryccmaytc 30

<210>127

<211>25

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<220>

<221 > modified base

<222>(23)

<223 > a, c, g or t

<400>127

caccatggas?tggacctgga?gvntc 25

<210>128

<211>28

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>128

caccatggac?atactttgyt?ccacgctc 28

<210>129

<211>23

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>129

caccatggar?ttkggrctbh?gct 23

<210>130

<211>30

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>130

caccatgaar?cayctgtggt?tcttcctyct 30

<210>131

<211>24

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>131

caccatgggg?tcaaccgyca?tcct 24

<210>132

<211>28

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>132

caccatgtct?gtctccttcc?tcatcttc 28

<210>133

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<220>

<221 > modified base

<222>(30)

<223 > a, c, g or t

<400>133

gaagatctca?ccatggastg?gacctggagv?ntcc 34

<210>134

<211>37

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>134

gaagatctca?ccatggacat?actttgytcc?acgctcc 37

<210>135

<211>41

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>135

gaagatctca?ccatggartt?kggrctbhgc?tggvttttyc?t?41

<210>136

<211>39

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>136

gaagatctca?ccatgaarca?yctgtggttc?ttcctyctc 39

<210>137

<211>32

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>137

gaagatctca?ccatggggtc?aaccgycatc?ct 32

<210>138

<211>37

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>138

gaagatctca?ccatgtctgt?ctccttcctc?atcttct 37

<210>139

<211>44

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>139

gaagatctca?ccatggactg?gacctggagg?atcctcttct?tggt 44

<210>140

<211>165

<212>PRT

<213 > homo sapiens

<400>140

Ala?Leu?Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro

1 5 10 15

Gln?Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His

20 25 30

His?Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln

35 40 45

Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr

50 55 60

Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg

65 70 75 80

Asn?Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser

85 90 95

Pro?Glu?Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val

100 105 110

Lys?Val?Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys

115 120 125

Glu?Ser?Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr

130 135 140

Val?Thr?Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser?Arg?Ser?Gly?Ser?Ser?His

145 150 155 160

His?His?His?His?His

165

<210>141

<211>119

<212>PRT

<213 > mouse kind

<400>141

Met?Glu?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Gln?Pro?Gly

1 5 10 15

Gly?Ser?Leu?Lys?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Thr

20 25 30

Tyr?Gly?Met?Ser?Trp?Val?Arg?Gln?Thr?Pro?Asp?Lys?Arg?Leu?Glu?Leu

35 40 45

Val?Ala?Leu?Ile?Asn?Ser?Gln?Gly?Gly?Ser?Thr?Tyr?Asn?Ser?Asp?Ser

50 55 60

Val?Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Arg?Asn?Thr?Leu

65 70 75 80

Tyr?Leu?Gln?Met?Ser?Ser?Leu?Lys?Ser?Glu?Asp?Thr?Ala?Met?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Arg?Asp?Tyr?Glu?Ser?Leu?Asp?Ser?Trp?Gly?Gln?Gly?Thr

100 105 110

Ser?Val?Thr?Val?Ser?Ser?Gly

115

<210>142

<211>122

<212>PRT

<213 > mouse kind

<400>142

Asp?Ile?Val?Leu?Thr?Gln?Ser?Pro?Ala?Ser?Leu?Pro?Val?Ser?Leu?Gly

1 5 10 15

Gln?Arg?Ala?Thr?Ile?Ser?Cys?Arg?Ala?Ser?Glu?Ser?Val?Glu?Tyr?Ser

20 25 30

Gly?Thr?Ser?Leu?Ile?Gln?Trp?Tyr?Arg?Gln?Lys?Pro?Gly?Gln?Pro?Pro

35 40 45

Lys?Leu?Leu?Ile?Tyr?Ala?Ala?Ser?Asn?Val?Asp?Ser?Glu?Val?Pro?Ala

50 55 60

Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Ser?Leu?Tyr?Ile?His

65 70 75 80

Pro?Val?Glu?Glu?Asp?Asp?Ile?Ala?Met?Tyr?Phe?Cys?Gln?Gln?Ser?Arg

85 90 95

Lys?Val?Pro?Trp?Thr?Phe?Gly?Gly?Gly?Thr?Lys?Leu?Glu?Ile?Lys?Arg

100 105 110

Thr?Asp?Ala?Ala?Pro?Gly?Leu?Glu?Ala?Ala

115 120

<210>143

<211>119

<212>PRT

<213 > mouse kind

<400>143

Lys?Val?Gln?Leu?Gln?Gln?Ser?Gly?Thr?Glu?Leu?Val?Lys?Pro?Gly?Ala

1 5 10 15

Ser?Val?Lys?Leu?Ser?Cys?Lys?Ala?Ser?Gly?Tyr?Thr?Phe?Thr?Glu?Tyr

20 25 30

Ile?Ile?His?Trp?Val?Lys?Gln?Arg?Ser?Gly?Gln?Gly?Leu?Glu?Trp?Ile

35 40 45

Gly?Trp?Phe?Tyr?Pro?Gly?Ser?Gly?Tyr?Ile?Lys?Tyr?Asn?Glu?Lys?Phe

50 55 60

Lys?Asp?Lys?Ala?Thr?Met?Thr?Ala?Asp?Lys?Ser?Ser?Ser?Thr?Val?Tyr

65 70 75 80

Met?Glu?Leu?Ser?Arg?Leu?Thr?Ser?Glu?Asp?Ser?Ala?Val?Tyr?Phe?Cys

85 90 95

Thr?Arg?His?Glu?Glu?Asp?Gly?Tyr?Tyr?Ala?Ala?Tyr?Trp?Gly?Gln?Gly

100 105 110

Thr?Leu?Val?Thr?Val?Ser?Ala

115

<210>144

<211>108

<212>PRT

<213 > mouse kind

<400>144

Asp?Ile?Val?Met?Thr?Gln?Ser?His?Lys?Phe?Met?Ser?Thr?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Ser?Ile?Thr?Cys?Lys?Ala?Ser?Gln?Asp?Val?Ser?Ser?Ala

20 25 30

Val?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln?Ser?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Trp?Ala?Ser?Thr?Arg?His?Thr?Gly?Val?Pro?Asp?Arg?Phe?Thr?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Tyr?Thr?Leu?Thr?Ile?Ser?Ser?Val?Gln?Ala

65 70 75 80

Glu?Asp?Leu?Ala?Leu?Tyr?Tyr?Cys?Gln?Gln?His?Tyr?Ser?Thr?Pro?Tyr

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Leu?Glu?Ile?Lys?Arg

100 105

<210>145

<211>33

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>145

gtaagcaagc?ttggctctga?tcacccaaca?aga 33

<210>146

<211>30

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>146

gattagggat?ccagaggcag?gagtccctgg 30

<210>147

<211>35

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>147

tagttgggat?cctcaggaga?tgcaatctct?accgt 35

<210>148

<211>35

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>148

ggtagtggat?cctcactgac?acactgtgtt?tctgg 35

<210>149

<211>35

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>149

gtaatgggat?cctcagacac?attcgatgtc?actcc 35

<210>150

<211>33

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>150

gtaatgaagc?ttgccacaac?aaaagaggtc?cag 33

<210>151

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>151

gattgaaagc?ttgatctcct?gcaaatatgg?acag 34

<210>152

<211>32

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>152

gtaatgaagc?ttgcagtgcg?aagaaggcac?ct 32

<210>153

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>153

gatggaggat?cctcaacacc?tggtgcagcg?caag 34

<210>154

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>154

gtaagtggat?cctcagcagg?gacttagctc?cact 34

<210>155

<211>29

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>155

gttagtaagc?ttggctccaa?tcacccgac 29

<210>156

<211>31

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>156

gttgatggat?ccttctttgt?ggacactcga?t 31

<210>157

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>157

gtagttggat?cctcaagaag?caggagtccc?aggg 34

<210>158

<211>36

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>158

gtatgaggga?tcctcactga?cacaccgtgt?ttctgg 36

<210>159

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>159

gtatggaagc?ttgccacaac?aaaagagatc?cagc 34

<210>160

<211>35

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>160

cagcgaagag?cggctccaca?acaaaagagg?tccag 35

<210>161

<211>35

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>161

ggatctcttt?tgttgtgggg?ccgctctctg?ctggg 35

<210>162

<211>36

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>162

cagcagagag?cggccccaca?acaaaagaga?tccagc 36

<210>163

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>163

cagcggccgg?aggagagccc?ctcagaggga?ttgt 34

<210>164

<211>33

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>164

gattgaggat?ccctaagagg?caggagtccc?tgg 33

<210>165

<211>34

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>165

tgaatgaagc?ttggttccag?taacagctaa?ccca 34

<210>166

<211>33

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>166

tccctctgag?gggctctcct?ccggccgctg?tag 33

<210>167

<211>37

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>167

caggtactgg?cctgctagac?acaatccctc?tgagggg 37

<210>168

<211>39

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>168

ctagcaggcc?agtacctgtc?agaagacggt?agagattgc 39

<210>169

<211>37

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>169

caggtactgg?cctgctagac?acaatccctc?tgagggg 37

<210>170

<211>39

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>170

ctagcaggcc?agtacctgtc?agaagacggt?agagattgc 39

<210>171

<211>41

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>171

tgaatccaga?gaatggt?tgg?agtgagtgct?atagtcctgt?c?41

<210>172

<211>39

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>172

tccaaccatt?ctctggattc?atgcttgcgc?tgcaccagg 39

<210>173

<211>33

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>173

gattgaggat?ccctaagagg?caggagtccc?tgg 33

<210>174

<211>46

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>174

tcgggtttct?acgactttat?cttccttaca?cctggtgcag?cgcaag 46

<210>175

<211>47

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>175

aaggaagata?aagtcgtaga?aacccgatgc?accacgacca?gaaacac?47

<210>176

<211>33

<212>DNA

<213 > artificial sequence

<220>

<223 > description of artificial sequence: synthetic primer

<400>176

gattgaggat?ccctaagagg?caggagtccc?tgg 33

<210>177

<211>118

<212>PRT

<213 > homo sapiens

<400>177

Gln?Val?Gln?Leu?Val?Gln?Ser?Gly?Ala?Glu?Val?Lys?Lys?Pro?Gly?Ala

1 5 10 15

Ser?Val?Lys?Val?Ser?Cys?Lys?Ala?Ser?Gly?Tyr?Thr?Phe?Thr?Ser?Tyr

20 25 30

Asp?Ile?Asn?Trp?Val?Arg?Gln?Ala?Thr?Gly?Gln?Gly?Leu?Glu?Trp?Met

35 40 45

Gly?Trp?Met?Asn?Pro?Asn?Ser?Gly?Asn?Thr?Gly?Tyr?Ala?Gln?Lys?Phe

50 55 60

Gln?Gly?Arg?Val?Thr?Met?Thr?Arg?Asn?Thr?Ser?Ile?Ser?Thr?Ala?Tyr

65 70 75 80

Met?Glu?Leu?Ser?Ser?Leu?Arg?Ser?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Tyr?Gly?Ser?Gly?Ser?Phe?Asp?Tyr?Trp?Gly?Gln?Gly?Thr?Leu

100 105 110

Val?Thr?Val?Ser?Ser?Ala

115

<210>178

<211>118

<212>PRT

<213 > homo sapiens

<400>178

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Ser?Ser?Gly?Trp?Phe?Asp?Tyr?Trp?Gly?Gln?Gly?Thr?Leu

100 105 110

Val?Thr?Val?Ser?Ser?Ala

115

<210>179

<211>117

<212>PRT

<213 > homo sapiens

<400>179

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Lys?Pro?Gly?Gly

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Asp?Tyr

20 25 30

Tyr?Met?Ser?Trp?Ile?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ser?Tyr?Ile?Ser?Ser?Ser?Gly?Ser?Thr?Ile?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Lys?Asn?Ser?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Ala?Ala?Gly?Ala?Phe?Asp?Ile?Trp?Gly?Gln?Gly?Thr?Met?Val

100 105 110

Thr?Val?Ser?Ser?Ala

115

<210>180

<211>119

<212>PRT

<213 > homo sapiens

<400>180

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Ser?Tyr

20 25 30

Gly?Met?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ala?Val?Ile?Trp?Tyr?Asp?Gly?Ser?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ser?Lys?Asn?Thr?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Tyr?Ser?Ser?Ser?Trp?Tyr?Phe?Asp?Leu?Trp?Gly?Arg?Gly?Thr

100 105 110

Leu?Val?Thr?Val?Ser?Ser?Ala

115

<210>181

<211>117

<212>PRT

<213 > homo sapiens

<400>181

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Glu

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Tyr

20 25 30

Tyr?Trp?Ser?Trp?Ile?Arg?Gln?Pro?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Ile

35 40 45

Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Asn?Tyr?Asn?Pro?Ser?Leu?Lys

50 55 60

Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe?Ser?Leu

65 70 75 80

Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr?Cys?Ala

85 90 95

Arg?Gly?Tyr?Ser?Gly?Tyr?Phe?Asp?Tyr?Trp?Gly?Gln?Gly?Thr?Leu?Val

100 105 110

Thr?Val?Ser?Ser?Ala

115

<210>182

<211>117

<212>PRT

<213 > homo sapiens

<400>182

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Asn?Trp?Asn?Phe?Asp?Ile?Trp?Gly?Gln?Gly?Thr?Met?Val

100 105 110

Thr?Val?Ser?Ser?Ala

115

<210>183

<211>114

<212>PRT

<213 > homo sapiens

<400>183

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Lys?Pro?Gly?Gly

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Asp?Tyr

20 25 30

Tyr?Met?Ser?Trp?Ile?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ser?Tyr?Ile?Ser?Ser?Ser?Gly?Ser?Thr?Ile?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Lys?Asn?Ser?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Gly?Met?Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser

100 105 110

Ser?Ala

<210>184

<211>123

<212>PRT

<213 > homo sapiens

<400>184

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Ser?Tyr

20 25 30

Gly?Met?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ala?Val?Ile?Trp?Tyr?Asp?Gly?Ser?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ser?Lys?Asn?Thr?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Tyr?Ser?Ser?Ser?Tyr?Tyr?Tyr?Tyr?Tyr?Gly?Met?Asp?Val?Trp

100 105 110

Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser?Ser?Ala

115 120

<210>185

<211>118

<212>PRT

<213 > homo sapiens

<400>185

Gln?Val?Gln?Leu?Gln?Gln?Trp?Gly?Ala?Gly?Leu?Leu?Lys?Pro?Ser?Glu

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Ala?Val?Tyr?Gly?Gly?Ser?Phe?Ser?Gly?Tyr

20 25 30

Tyr?Trp?Ser?Trp?Ile?Arg?Gln?Pro?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Ile

35 40 45

Gly?Glu?Ile?Asn?His?Ser?Gly?Ser?Thr?Asn?Tyr?Asn?Pro?Ser?Leu?Lys

50 55 60

Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe?Ser?Leu

65 70 75 80

Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr?Cys?Ala

85 90 95

Arg?Ser?Ser?Gly?Tyr?Trp?Tyr?Phe?Asp?Leu?Trp?Gly?Arg?Gly?Thr?Leu

100 105 110

Val?Thr?Val?Ser?Ser?Ala

115

<210>186

<211>117

<212>PRT

<213 > homo sapiens

<400>186

Glu?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Leu?Val?Lys?Pro?Gly?Gly

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Ser?Tyr

20 25 30

Ser?Met?Asn?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ser?Ser?Ile?Ser?Ser?Ser?Ser?Ser?Tyr?Ile?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ala?Lys?Asn?Ser?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Ser?Ser?Trp?Tyr?Trp?Phe?Asp?Pro?Trp?Gly?Gln?Gly?Thr?Leu?Val

100 105 110

Thr?Val?Ser?Ser?Ala

115

<210>187

<211>126

<212>PRT

<213 > homo sapiens

<400>187

Gln?Val?Gln?Leu?Val?Glu?Ser?Gly?Gly?Gly?Val?Val?Gln?Pro?Gly?Arg

1 5 10 15

Ser?Leu?Arg?Leu?Ser?Cys?Ala?Ala?Ser?Gly?Phe?Thr?Phe?Ser?Ser?Tyr

20 25 30

Gly?Met?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Lys?Gly?Leu?Glu?Trp?Val

35 40 45

Ala?Val?Ile?Trp?Tyr?Asp?Gly?Ser?Asn?Lys?Tyr?Tyr?Ala?Asp?Ser?Val

50 55 60

Lys?Gly?Arg?Phe?Thr?Ile?Ser?Arg?Asp?Asn?Ser?Lys?Ash?Thr?Leu?Tyr

65 70 75 80

Leu?Gln?Met?Asn?Ser?Leu?Arg?Ala?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys

85 90 95

Ala?Arg?Gly?Tyr?Cys?Thr?Asn?Gly?Val?Cys?Tyr?Tyr?Tyr?Tyr?Gly?Met

100 105 110

Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr?Val?Thr?Val?Ser?Ser?Ala

115 120 125

<210>188

<211>118

<212>PRT

<213 > homo sapiens

<400>188

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Tyr?Tyr?Tyr?Gly?Met?Asp?Val?Trp?Gly?Gln?Gly?Thr?Thr

100 105 110

Val?Thr?Val?Ser?Ser?Ala

115

<210>189

<211>120

<212>PRT

<213 > homo sapiens

<400>189

Gln?Val?Gln?Leu?Gln?Glu?Ser?Gly?Pro?Gly?Leu?Val?Lys?Pro?Ser?Gln

1 5 10 15

Thr?Leu?Ser?Leu?Thr?Cys?Thr?Val?Ser?Gly?Gly?Ser?Ile?Ser?Ser?Gly

20 25 30

Gly?Tyr?Tyr?Trp?Ser?Trp?Ile?Arg?Gln?His?Pro?Gly?Lys?Gly?Leu?Glu

35 40 45

Trp?Ile?Gly?Tyr?Ile?Tyr?Tyr?Ser?Gly?Ser?Thr?Tyr?Tyr?Asn?Pro?Ser

50 55 60

Leu?Lys?Ser?Arg?Val?Thr?Ile?Ser?Val?Asp?Thr?Ser?Lys?Asn?Gln?Phe

65 70 75 80

Ser?Leu?Lys?Leu?Ser?Ser?Val?Thr?Ala?Ala?Asp?Thr?Ala?Val?Tyr?Tyr

85 90 95

Cys?Ala?Arg?Gly?Ser?Gly?Ser?Tyr?Trp?Phe?Asp?Pro?Trp?Gly?Gln?Gly

100 105 110

Thr?Leu?Val?Thr?Val?Ser?Ser?Ala

115 120

<210>190

<211>106

<212>PRT

<213 > homo sapiens

<400>190

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Ser?Ile?Ser?Ser?Tyr

20 25 30

Leu?Asn?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ser?Tyr?Ser?Thr?Thr?Phe

85 90 95

Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105

<210>191

<211>108

<212>PRT

<213 > homo sapiens

<400>191

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Ser?Ile?Ser?Ser?Tyr

20 25 30

Leu?Asn?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ser?Tyr?Ser?Thr?Pro?Phe

85 90 95

Thr?Phe?Gly?Pro?Gly?Thr?Lys?Val?Asp?Ile?Lys?Arg

100 105

<210>192

<211>111

<212>PRT

<213 > homo sapiens

<400>192

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Leu?Ser?Leu?Pro?Val?Thr?Pro?Gly

1 5 10 15

Glu?Pro?Ala?Ser?Ile?Ser?Cys?Arg?Ser?Ser?Gln?Ser?Leu?Leu?His?Ser

20 25 30

Asn?Gly?Tyr?Asn?Tyr?Leu?Asp?Trp?Tyr?Leu?Gln?Lys?Pro?Gly?Gln?Ser

35 40 45

Pro?Gln?Leu?Leu?Ile?Tyr?Leu?Gly?Ser?Asn?Arg?Ala?Ser?Gly?Val?Pro

50 55 60

Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Lys?Ile

65 70 75 80

Ser?Arg?Val?Glu?Ala?Glu?Asp?Val?Gly?Val?Tyr?Tyr?Cys?Met?Gln?Ala

85 90 95

Leu?Gln?Thr?Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105 110

<210>193

<211>106

<212>PRT

<213 > homo sapiens

<400>193

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Val?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Ser?Trp

20 25 30

Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ala?Asn?Ser?Phe?Pro?Thr

85 90 95

Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys

100 105

<210>194

<211>106

<212>PRT

<213 > homo sapiens

<400>194

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Arg?Asn?Asp

20 25 30

Leu?Gly?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Arg?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Glu?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Leu?Gln?His?Asn?Ser?Trp?Thr?Phe

85 90 95

Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105

<210>195

<211>108

<212>PRT

<213 > homo sapiens

<400>195

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Asn?Tyr

20 25 30

Leu?Ala?Trp?Phe?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Ser?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Tyr?Asn?Ser?Tyr?Pro?Leu

85 90 95

Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105

<210>196

<211>114

<212>PRT

<213 > homo sapiens

<400>196

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Asp?Ser?Leu?Ala?Val?Ser?Leu?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Ile?Asn?Cys?Lys?Ser?Ser?Gln?Ser?Val?Leu?Tyr?Ser

20 25 30

Ser?Asn?Asn?Lys?Asn?Tyr?Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln

35 40 45

Pro?Pro?Lys?Leu?Leu?Ile?Tyr?Trp?Ala?Ser?Thr?Arg?Glu?Ser?Gly?Val

50 55 60

Pro?Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr

65 70 75 80

Ile?Ser?Ser?Leu?Gln?Ala?Glu?Asp?Val?Ala?Val?Tyr?Tyr?Cys?Gln?Gln

85 90 95

Tyr?Tyr?Ser?Thr?Pro?Leu?Thr?Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile

100 105 110

Lys?Arg

<210>197

<211>114

<212>PRT

<213 > homo sapiens

<400>197

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Asp?Ser?Leu?Ala?Val?Ser?Leu?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Ile?Asn?Cys?Lys?Ser?Ser?Gln?Ser?Val?Leu?Tyr?Ser

20 25 30

Ser?Asn?Asn?Lys?Asn?Tyr?Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln

35 40 45

Pro?Pro?Lys?Leu?Leu?Ile?Tyr?Trp?Ala?Ser?Thr?Arg?Glu?Ser?Gly?Val

50 55 60

Pro?Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr

65 70 75 80

Ile?Ser?Ser?Leu?Gln?Ala?Glu?Asp?Val?Ala?Val?Tyr?Tyr?Cys?Gln?Gln

85 90 95

Tyr?Tyr?Ser?Thr?Pro?Phe?Thr?Phe?Gly?Pro?Gly?Thr?Lys?Val?Asp?Ile

100 105 110

Lys?Arg

<210>198

<211>106

<212>PRT

<213 > homo sapiens

<400>198

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Asn?Tyr

20 25 30

Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Val?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Thr?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Val?Ala?Thr?Tyr?Tyr?Cys?Gln?Lys?Tyr?Asn?Ser?Ala?Thr?Phe

85 90 95

Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105

<210>199

<211>111

<212>PRT

<213 > homo sapiens

<400>199

Asp?Ile?Val?Met?Thr?Gln?Ser?Pro?Asp?Ser?Leu?Ala?Val?Ser?Leu?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Ile?Asn?Cys?Lys?Ser?Ser?Gln?Ser?Val?Leu?Tyr?Ser

20 25 30

Ser?Asn?Asn?Lys?Asn?Tyr?Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln

35 40 45

Pro?Pro?Lys?Leu?Leu?Ile?Tyr?Trp?Ala?Ser?Thr?Arg?Glu?Ser?Gly?Val

50 55 60

Pro?Asp?Arg?Phe?Ser?Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr

65 70 75 80

Ile?Ser?Ser?Leu?Gln?Ala?Glu?Asp?Val?Ala?Val?Tyr?Tyr?Cys?Gln?Gln

85 90 95

Tyr?Tyr?Ser?Thr?Phe?Gly?Gln?Gly?Thr?Lys?Leu?Glu?Ile?Lys?Arg

100 105 110

<210>200

<211>109

<212>PRT

<213 > homo sapiens

<400>200

Glu?Ile?Val?Leu?Thr?Gln?Ser?Pro?Gly?Thr?Leu?Ser?Leu?Ser?Pro?Gly

1 5 10 15

Glu?Arg?Ala?Thr?Leu?Ser?Cys?Arg?Ala?Ser?Gln?Ser?Val?Ser?Ser?Ser

20 25 30

Tyr?Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Gln?Ala?Pro?Arg?Leu?Leu

35 40 45

Ile?Tyr?Gly?Ala?Ser?Ser?Arg?Ala?Thr?Gly?Ile?Pro?Asp?Arg?Phe?Ser

50 55 60

Gly?Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Arg?Leu?Glu

65 70 75 80

Pro?Glu?Asp?Phe?Ala?Val?Tyr?Tyr?Cys?Gln?Gln?Tyr?Gly?Ser?Ser?Pro

85 90 95

Trp?Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105

<210>201

<211>108

<212>PRT

<213 > homo sapiens

<400>201

Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Val?Ser?Ala?Ser?Val?Gly

1 5 10 15

Asp?Arg?Val?Thr?Ile?Thr?Cys?Arg?Ala?Ser?Gln?Gly?Ile?Ser?Ser?Trp

20 25 30

Leu?Ala?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Leu?Leu?Ile

35 40 45

Tyr?Ala?Ala?Ser?Ser?Leu?Gln?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly

50 55 60

Ser?Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro

65 70 75 80

Glu?Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Ala?Asn?Ser?Phe?Pro?Arg

85 90 95

Thr?Phe?Gly?Gln?Gly?Thr?Lys?Val?Glu?Ile?Lys?Arg

100 105

<210>202

<211>129

<212>PRT

<213 > cynomolgus monkey

<400>202

Ala?Pro?Ile?Thr?Arg?Gln?Ser?Leu?Asp?Pro?Gln?Arg?Arg?Ala?Ala?Pro

1 5 10 15

Gln?Gln?Lys?Arg?Ser?Ser?Pro?Thr?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His

20 25 30

His?Ile?Ser?Glu?Asp?Ser?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln

35 40 45

Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Phe?Leu?Phe?Cys?Leu?Arg?Cys?Thr

50 55 60

Lys?Cys?Asp?Ser?Gly?Glu?Val?Glu?Val?Ser?Ser?Cys?Thr?Thr?Thr?Arg

65 70 75 80

Asn?Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser

85 90 95

Pro?Glu?Ile?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val

100 105 110

Lys?Val?Lys?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys

115 120 125

Glu

<210>203

<211>154

<212>PRT

<213 > homo sapiens

<400>203

Ala?Leu?Ile?Thr?Gln?Gln?Asp?Leu?Ala?Pro?Gln?Gln?Arg?Ala?Ala?Pro

1 5 10 15

Gln?Gln?Lys?Arg?Ser?Ser?Pro?Ser?Glu?Gly?Leu?Cys?Pro?Pro?Gly?His

20 25 30

His?Ile?Ser?Glu?Asp?Gly?Arg?Asp?Cys?Ile?Ser?Cys?Lys?Tyr?Gly?Gln

35 40 45

Asp?Tyr?Ser?Thr?His?Trp?Asn?Asp?Leu?Leu?Phe?Cys?Leu?Arg?Cys?Thr

50 55 60

Arg?Cys?Asp?Ser?Gly?Glu?Val?Glu?Leu?Ser?Pro?Cys?Thr?Thr?Thr?Arg

65 70 75 80

Asn?Thr?Val?Cys?Gln?Cys?Glu?Glu?Gly?Thr?Phe?Arg?Glu?Glu?Asp?Ser

85 90 95

Pro?Glu?Met?Cys?Arg?Lys?Cys?Arg?Thr?Gly?Cys?Pro?Arg?Gly?Met?Val

100 105 110

Lys?Val?Gly?Asp?Cys?Thr?Pro?Trp?Ser?Asp?Ile?Glu?Cys?Val?His?Lys

115 120 125

Glu?Ser?Gly?Thr?Lys?His?Ser?Gly?Glu?Ala?Pro?Ala?Val?Glu?Glu?Thr

130 135 140

Val?Thr?Ser?Ser?Pro?Gly?Thr?Pro?Ala?Ser

145 150

<210>204

<211>133

<212>PRT

<213 > mouse kind

<400>204

Pro?Val?Thr?Ala?Asn?Pro?Ala?His?Asn?Arg?Pro?Ala?Gly?Leu?Gln?Arg

1 5 10 15

Pro?Glu?Glu?Ser?Pro?Ser?Arg?Gly?Pro?Cys?Leu?Ala?Gly?Gln?Tyr?Leu

20 25 30

Ser?Glu?Gly?Asn?Cys?Lys?Pro?Cys?Arg?Glu?Gly?Ile?Asp?Tyr?Thr?Ser

35 40 45

His?Ser?Asn?His?Ser?Leu?Asp?Ser?Cys?Ile?Leu?Cys?Thr?Val?Cys?Lys

50 55 60

Glu?Asp?Lys?Val?Val?Glu?Thr?Arg?Cys?Asn?Ile?Thr?Thr?Asn?Thr?Val

65 70 75 80

Cys?Arg?Cys?Lys?Pro?Gly?Thr?Phe?Glu?Asp?Lys?Asp?Ser?Pro?Glu?Ile

85 90 95

Cys?Gln?Ser?Cys?Ser?Asn?Cys?Thr?Asp?Gly?Glu?Glu?Glu?Leu?Thr?Ser

100 105 110

Cys?Thr?Pro?Arg?Glu?Asn?Arg?Lys?Cys?Val?Ser?Lys?Thr?Ala?Trp?Ala

115 120 125

Ser?Trp?His?Lys?Leu

130

Claims

1. a separation antibody that comprises heavy chain and light chain, the CDR3 of the amino acid/11 00-111 that the CDR2 of the amino acid 52-67 that CDR1, the sequence that wherein said heavy chain comprises the amino acid 26-37 that sequence is SEQ ID NO:30 is SEQ ID NO:30 and sequence are SEQ ID NO:30, the CDR3 of the amino acid 90-98 that the CDR2 of the amino acid 51-57 that CDR1, the sequence that wherein said light chain comprises the amino acid 24-35 that sequence is SEQ ID NO:64 is SEQ ID NO:64 and sequence are SEQ ID NO:64, and wherein said antibodies TRAIL acceptor-2 (TR-2).

2. a separation antibody that comprises heavy chain and light chain, the aminoacid sequence that wherein said heavy chain comprises SEQ ID NO:30, the aminoacid sequence that wherein said light chain comprises SEQ ID NO:64, and wherein said antibodies TRAIL acceptor-2 (TR-2).

3. claim 1 or 2 separation antibody, wherein said antibody is fully human antibodies.

4. a pharmaceutical composition, the separation antibody that comprises any one in claim 1-3 and pharmaceutical acceptable carrier.

5. the polynucleotide of encoding antibody heavy chain and light chain, the CDR3 of the amino acid/11 00-111 that the CDR2 of the amino acid 52-67 that CDR1, the sequence that wherein said heavy chain comprises the amino acid 26-37 that sequence is SEQ ID NO:30 is SEQ ID NO:30 and sequence are SEQ ID NO:30, and the CDR2 of the wherein said light chain CDR1, the sequence that comprise the amino acid 24-35 that sequence the is SEQ ID NO:64 amino acid 51-57 that is SEQ ID NO:64 and the CDR3 of the amino acid 90-98 that sequence is SEQ ID NO:64.

6. the polynucleotide of encoding antibody heavy chain and light chain, the aminoacid sequence that wherein said heavy chain comprises SEQ ID NO:30, and the wherein said light chain aminoacid sequence that comprises SEQ ID NO:64.

7. claim 5 or 6 polynucleotide, its integral part that is expression vector.

8. a host cell that comprises the first polynucleotide and the second polynucleotide, the heavy chain of wherein said the first polynucleotide encoding antibody, the light chain of the second polynucleotide encoding antibody, the CDR1 that wherein said heavy chain comprises the amino acid 26-37 that sequence is SEQ ID NO:30, the CDR3 of the amino acid/11 00-111 that the CDR2 of the amino acid 52-67 that sequence is SEQ ID NO:30 and sequence are SEQ ID NO:30, the CDR1 that wherein said light chain comprises the amino acid 24-35 that sequence is SEQ ID NO:64, the CDR3 of the amino acid 90-98 that the CDR2 of the amino acid 51-57 that sequence is SEQ ID NO:64 and sequence are SEQ ID NO:64, and wherein said antibodies TRAIL acceptor-2 (TR-2).

9. the host cell of claim 8, the aminoacid sequence that wherein said heavy chain comprises SEQ ID NO:30.

10. the host cell of claim 9, the aminoacid sequence that wherein said light chain comprises SEQ ID NO:64.

11. the host cell of claim 8, the aminoacid sequence that wherein said light chain comprises SEQ ID NO:64.

12. the host cell of any one in claim 8-11, wherein said the first polynucleotide and the second polynucleotide are integral parts of expression vector.

13. the host cell of any one in claim 8-11, wherein said the first polynucleotide are integral parts of the first expression vector, and described the second polynucleotide are integral parts of the second expression vector.

14. the method for a Dispersal risk, be included under the condition that is suitable for described the first polynucleotide of host cell expression and described the second polynucleotide the host cell of any one in incubation claim 8-11, to produce described antibody.

15. the method for a Dispersal risk, be included in the host cell of incubation claim 12 under the condition that is suitable for described the first polynucleotide of host cell expression and described the second polynucleotide, to produce described antibody.

16. the method for a Dispersal risk, be included in the host cell of incubation claim 13 under the condition that is suitable for described the first polynucleotide of host cell expression and described the second polynucleotide, to produce described antibody.

17. the purposes of the antibody of any one in the medicine of preparation treatment patient's cancer in the claim 1-3 for the treatment of significant quantity.

18. the purposes of claim 17, wherein said cancer is selected from following at least one: liver cancer, the cancer of the brain, kidney, colorectal carcinoma, lung cancer, spleen cancer, thymic carcinoma, leukemia, prostate cancer, carcinoma of testis, ovarian cancer, uterus carcinoma, mammary cancer, carcinoma of the pancreas, cancer of the stomach, squamous cell carcinoma of the head and neck and lymphatic cancer.