CN101300273A

CN101300273A - TRAIL recipient 2 Polypeptides and antibodies

Info

Publication number: CN101300273A
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: 2008-11-05
Anticipated expiration: 2026-08-28
Also published as: ZA200802575B; CN101300273B; UA97096C2

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

This application claims the U.S. Provisional Application No. 60/713 submitted for 31st in August in 2005,433, and the U.S. Provisional Application No. 60/713,478 submitted for 31st in August in 2005 priority.U.S. Provisional Application No. 60/713,433 and 60/713,478 is herein by reference in its entirety for any purpose.

Invention field

There is provided polypeptide.There is provided the antibody or antigen-binding domains for combining such polypeptide.Additionally provide the method for obtaining the antibody for combining TNF (TNF) apoptosis induction ligand related (" TRAIL ") acceptor 2 (TR-2), methods described includes applying at least one such polypeptide to animal, and the antibody for combining TR-2 is obtained from animal.There is provided the antibody reacted with TR-2.Production and the cell of the TR-2 antibody reacted are additionally provided, comprising the pharmaceutical composition with the TR-2 antibody reacted, using the method with the TR-2 antibody reacted, and the kit with the TR-2 antibody reacted is included.Additionally provide by applying the method that such polypeptide reduces or prevented antibody to be combined with TR-2.

Background of invention

Interaction between TR-2 and its ligand/TRAIL is worked in the induction of apoptosis (see, e.g., Almasan et al., Cytokine ＆ Growth Factor Reviews 14：337-348(2003)).TRAIL is also referred to as Apo2L/TRAIL; it is 4 members (TRAIL acceptors (" TR ") 1-4) with TNF receptor superfamilies; and same several parts to related, soluble, opsteoprotegerin (suspected of osteoprotegerin, osteoprotegerin) (" OGP ") acceptor interaction.The combination of TRAIL and TR-1 or TR-2 on cell surface triggers the apoptosis of that cell.After TRAIL and TR-1 or TR-2 initial combinations, intracellular protein is recruited to the intracellular death domain of acceptor, forms Signaling complex.Some intracellular caspases are recruited to compound, and their own activates and activates other caspase successively and intracellular apoptosis cascade wherein.TR-3 and TR-4 and OPG lacks the intracellular domain for being responsible for apoptotic signal transmission.Therefore, TRAIL and TR-3, TR-4 or OPG combination do not trigger apoptosis.TR-3 and TR-4 are also known as " inveigling " acceptor, and their overexpression has shown and protected cell not by the apoptosis induction via TRAIL.TR-2 is expressed in various cells, including liver, brain, mammary gland, kidney, colon, lung, spleen, thymus gland, PBLC, prostate, testis, ovary, uterus and the various tissues along intestines and stomach.(see, e.g., Walczak et al., EMBO is J.16：5386-5397(1997)；Spierings et al., J.Histochem.Cytochem.52：821-831(2004)).Although TRAIL and TRAIL acceptors are wide expressions, they are most active in the apoptosis induction of transformed cells.(see, e.g., Daigle et al., Swiss Med.Wkly.131：231-237(2001)).

Summary of the invention

There is provided the isolated polypeptide for including at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a in certain embodiments：

Wherein CDR1a is glycine comprising amino acid sequence a b c d e f g h i j k l, wherein amino acid a, 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

Wherein CDR2a includes amino acid 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 tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；With

The polypeptide combination TRAIL acceptors 2 (TR-2) wherein combined with antibody light chain.

There is provided include the isolated polypeptide selected from least one following complementary determining regions (CDR) in certain embodiments：

SEQ ID NO：2 amino acid 26-35；

SEQ ID NO：2 amino acid 50-66；

SEQ ID NO：2 amino acid 99-110；

SEQ ID NO：4 amino acid 26-37；

SEQ ID NO：4 amino acid 52-67；

SEQ ID NO：4 amino acid/11 00-109；

SEQ ID NO：6 amino acid 26-37；

SEQ ID NO：6 amino acid 52-67；

SEQ ID NO：6 amino acid/11 00-109；

SEQ ID NO：8 amino acid 26-37；

SEQ ID NO：8 amino acid 52-67；

SEQ ID NO：8 amino acid/11 00-109；

SEQ ID NO：10 amino acid 26-35；

SEQ ID NO：10 amino acid 50-66；

SEQ ID NO：10 amino acid 99-110；

SEQ ID NO：12 amino acid 26-35；

SEQ ID NO：12 amino acid 50-66；

SEQ ID NO：12 amino acid 99-111；

SEQ ID NO：14 amino acid 26-35；

SEQ ID NO：14 amino acid 50-65；

SEQ ID NO：14 amino acid 98-111；

SEQ ID NO：16 amino acid 26-37；

SEQ ID NO：16 amino acid 52-67；

SEQ ID NO：16 amino acid/11 00-109；

SEQ ID NO：18 amino acid 26-35；

SEQ ID NO：18 amino acid 50-66；

SEQ ID NO：18 amino acid 99-105；

SEQ ID NO：20 amino acid 26-35；

SEQ ID NO：20 amino acid 50-66；

SEQ ID NO：20 amino acid 99-118；

SEQ ID NO：22 amino acid 26-35；

SEQ ID NO：22 amino acid 50-66；

SEQ ID NO：22 amino acid 99-118；

SEQ ID NO：24 amino acid 26-35；

SEQ ID NO：24 amino acid 50-65；

SEQ ID NO：24 amino acid 98-108；

SEQ ID NO：26 amino acid 26-35；

SEQ ID NO：26 amino acid 50-66；

SEQ ID NO：26 amino acid 99-110；

SEQ ID NO：28 amino acid 26-35；

SEQ ID NO：28 amino acid 50-66；

SEQ ID NO：28 amino acid 99-117；

SEQ ID NO：30 amino acid 26-37；

SEQ ID NO：30 amino acid 52-67；

SEQ ID NO：30 amino acid/11 00-111；

SEQ ID NO：32 amino acid 26-37；

SEQ ID NO：32 amino acid 52-67；

SEQ ID NO：32 amino acid/11 00-111；

SEQ ID NO：34 amino acid 26-37；

SEQ ID NO：34 amino acid 52-67；With

SEQ ID NO：34 amino acid/11 00-111；

The polypeptide combination TR-2 wherein combined with antibody light chain.

There is provided the isolated polypeptide for including at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b in certain embodiments：

Wherein CDR1b includes amino acid 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 lysine；Amino acid b1 is selected from threonine, alanine 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 valine；Amino acid g1 is selected from serine, leucine or arginine；Amino acid h1 is selected from threonine, serine, isoleucine, asparagine, arginine, histidine or tyrosine；Amino acid i1 is selected from tyrosine, arginine, tryptophan, aspartic acid or serine；J1 is selected from leucine, isoleucine, asparagine, tyrosine or serine；Amino acid k1 is selected from asparagine, glycine, valine, alanine or leucine；Amino acid l1 is selected from tyrosine, alanine or asparagine or is not present；Amino acid m1 is selected from asparagine or lysine or is not present；Amino acid n1 is selected from tyrosine, asparagine or isoleucine or is not present；Amino acid o1 is selected from leucine or tyrosine or is not present；Amino acid p1 is selected from aspartic acid or leucine or is not present；Valine, alanine or threonine are selected from amino acid q1 or are not present；

Wherein CDR2b includes amino acid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from alanine, aspartic acid, leucine, tryptophan, glycine or valine；Amino acid s1 is selected from threonine, valine, glycine or alanine；Amino acid t1 is serine；Amino acid u1 is selected from serine, asparagine or threonine；Amino acid v1 is selected from leucine, phenylalanine or arginine；Amino acid w1 is selected from glutamine, alanine or glutamic acid；Serine, arginine or threonine are selected from amino acid x1；

Wherein CDR3b is selected from glutamine, methionine, leucine or histidine comprising amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', wherein amino acid y1；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；With

The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

SEQ ID NO：36 amino acid 24-34；

SEQ ID NO：36 amino acid 50-56；

SEQ ID NO：36 amino acid 89-97；

SEQ ID NO：38 amino acid 24-34；

SEQ ID NO：38 amino acid 50-56；

SEQ ID NO：38 amino acid 89-97；

SEQ ID NO：40 amino acid 24-34；

SEQ ID NO：40 amino acid 50-56；

SEQ ID NO：40 amino acid 89-97；

SEQ ID NO：42 amino acid 24-34；

SEQ ID NO：42 amino acid 50-56；

SEQ ID NO：42 amino acid 89-97；

SEQ ID NO：44 amino acid 24-34；

SEQ ID NO：44 amino acid 50-56；

SEQ ID NO：44 amino acid 89-97；

SEQ ID NO：46 amino acid 24-34；

SEQ ID NO：46 amino acid 50-56；

SEQ ID NO：46 amino acid 89-97；

SEQ ID NO：48 amino acid 24-40；

SEQ ID NO：48 amino acid 56-62；

SEQ ID NO：48 amino acid 95-103；

SEQ ID NO：50 amino acid 24-39；

SEQ ID NO：50 amino acid 55-61；

SEQ ID NO：50 amino acid 94-102；

SEQ ID NO：52 amino acid 24-40；

SEQ ID NO：52 amino acid 56-62；

SEQ ID NO：52 amino acid 95-103；

SEQ ID NO：54 amino acid 24-34；

SEQ ID NO：54 amino acid 50-56；

SEQ ID NO：54 amino acid 89-97；

SEQ ID NO：56 amino acid 24-34,

SEQ ID NO：56 amino acid 50-56；

SEQ ID NO：56 amino acid 89-97；

SEQ ID NO：58 amino acid 24-40；

SEQ ID NO：58 amino acid 56-62；

SEQ ID NO：58 amino acid 95-103；

SEQ ID NO：60 amino acid 24-34；

SEQ ID NO：60 amino acid 50-56；

SEQ ID NO：60 amino acid 89-97；

SEQ ID NO：62 amino acid 24-34；

SEQ ID NO：62 amino acid 50-56；

SEQ ID NO：62 amino acid 89-97；

SEQ ID NO：64 amino acid 24-35；

SEQ ID NO：64 amino acid 51-57；

SEQ ID NO：64 amino acid 90-88；

SEQ ID NO：66 amino acid 24-34；

SEQ ID NO：66 amino acid 50-57；

SEQ ID NO：66 amino acid 89-97；

SEQ ID NO：68 amino acid 24-34；

SEQ ID NO：68 amino acid 50-56；With

SEQ ID NO：68 amino acid 89-97；

The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

In certain embodiments there is provided the separation polynucleotides of the sequence comprising coded polypeptide, the polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a：

Wherein CDR1a comprising amino acid sequence a b c d e f g h i j k l, wherein amino acid a be glycine, amino acid b be 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

Wherein CDR2a includes amino acid 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 tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；

Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；With

The polypeptide combination TR-2 wherein combined with antibody light chain.

In certain embodiments there is provided the separation polynucleotides of the sequence comprising coded polypeptide, the polypeptide includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b：

Wherein CDR2b includes amino acid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from alanine, aspartic acid, leucine, tryptophan, glycine or valine；Amino acid s1 is selected from threonine, valine, glycine or alanine；Amino acid t1 is serine；Amino acid u1 is selected from serine, asparagine or threonine；Amino acid v1 is selected from leucine, phenylalanine or arginine；Amino acid w1 is selected from glutamine, alanine or glutamic acid；Serine, arginine or threonine are selected from amino acid x1；Wherein CDR3 includes amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine, leucine or histidine；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；With

The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

In certain embodiments there is provided the anti-TR-2 antibody of the separation comprising variable region and constant region, wherein the antibody is included：

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

Wherein CDR1a includes amino acid 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

Wherein with antibody light chain with reference to described in the first polypeptide combination TR-2；With

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

Wherein CDR3b includes amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine, leucine or histidine；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；With

Second of polypeptide combination TR-2 wherein combined with heavy chain of antibody.

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

In certain embodiments there is provided cell, it is included：

(a) the first polynucleotides for the sequence for encoding the first polypeptide are included, the first described polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a, wherein CDR1a comprising amino acid sequence a b c d e f g h i j k l, wherein amino acid a be glycine, amino acid b be 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；Wherein with antibody light chain with reference to described in the first polypeptide combination TR-2；With

(b) second of polynucleotides of the sequence comprising second of polypeptide of coding, second of polypeptide includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b,

Wherein CDR3b includes amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine, leucine or histidine；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；Second of polypeptide combination TR-2 wherein combined with heavy chain of antibody.

In certain embodiments there is provided with epitope specificity combination separation antibody, the epitope is by selected from following at least one antibody specificity combinations：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.

There is provided comprising selected from SEQ ID NO in certain embodiments：94、SEQ ID NO：95 and SEQ ID NO：The polypeptide of 96 at least one amino acid sequence.

There is provided substantially by selected from SEQ ID NO in certain embodiments：94、SEQ IDNO：95 and SEQ ID NO：The polypeptide of 96 at least one amino acid sequence composition.

There is provided combine to be selected from SEQ ID NO in certain embodiments：94、SEQ ID NO：95 and SEQ ID NO：The antibody or antigen-binding domains of 96 at least one amino acid sequence.

In certain embodiments there is provided the method for obtaining the antibody for combining TR-2, methods described, which includes applying to animal, is selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：96 at least one polypeptide, and the antibody for combining TR-2 is obtained from animal.

In certain embodiments there is provided the method for reducing or preventing antibody to be combined with TR-2, methods described is by applying comprising selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ IDNO：The polypeptide of 96 at least one amino acid sequence.

In certain embodiments there is provided the method for reducing or preventing antibody to be combined with TR-2, methods described is by applying by selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ IDNO：The polypeptide of 96 at least one amino acid sequence composition.

Brief description

Fig. 1 shows the immunity inoculation timetable used in the transgenic mice of expression human immunoglobulin gene in embodiment 1 for TR-2-His constructs, via footpad inoculation (

group

1,2 and 3) or via intraperitoneal injection (group 4 and 5).

Fig. 2 shows the work according to embodiment 1, and some blood samples for measuring the selected mouse described in Fig. 1 are directed to antigen TR-2 reactive ELISA determination method results.

Fig. 3 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody As：1) with light chain (SEQ ID NO：35) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：2) with light chain (SEQ ID NO：36) amino acid sequence of variable region.

Fig. 4 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody B：3) with light chain (SEQ ID NO：37) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：4) with light chain (SEQ ID NO：38) amino acid sequence of variable region.

Fig. 5 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody C：5) with light chain (SEQ ID NO：39) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：6) with light chain (SEQ ID NO：40) amino acid sequence of variable region.

Fig. 6 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody D：7) with light chain (SEQ ID NO：41) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：8) with light chain (SEQ ID NO：42) amino acid sequence of variable region.

Fig. 7 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody E：9) with light chain (SEQ ID NO：43) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：10) with light chain (SEQ ID NO：44) amino acid sequence of variable region.

Fig. 8 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody F：11) with light chain (SEQ ID NO：45) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：12) with light chain (SEQ ID NO：46) amino acid sequence of variable region.

Fig. 9 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody G：13) with light chain (SEQ ID NO：47) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：14) with light chain (SEQ ID NO：48) amino acid sequence of variable region.

Figure 10 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody H：15) with light chain (SEQ ID NO：49) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：16) with light chain (SEQ ID NO：50) amino acid sequence of variable region.

Figure 11 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody Is：17) with light chain (SEQ ID NO：51) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：18) with light chain (SEQ ID NO：52) amino acid sequence of variable region.

Figure 12 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody J：19) with light chain (SEQ ID NO：53) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：20) with light chain (SEQ ID NO：54) amino acid sequence of variable region.

Figure 13 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody K：21) with light chain (SEQ ID NO：55) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：22) with light chain (SEQ ID NO：56) amino acid sequence of variable region.

Figure 14 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody L：23) with light chain (SEQ ID NO：57) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：24) with light chain (SEQ ID NO：58) amino acid sequence of variable region.

Figure 15 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody M：25) with light chain (SEQ ID NO：59) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：26) with light chain (SEQ ID NO：60) amino acid sequence of variable region.

Figure 16 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody N：27) with light chain (SEQ ID NO：61) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：28) with light chain (SEQ ID NO：62) amino acid sequence of variable region.

Figure 17 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody O：29) with light chain (SEQ ID NO：63) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：30) with light chain (SEQ ID NO：64) amino acid sequence of variable region.

Figure 18 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody P：31) with light chain (SEQ ID NO：65) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：32) with light chain (SEQ ID NO：66) amino acid sequence of variable region.

Figure 19 shows heavy chain (the SEQ ID NO for encoding anti-TR-2 antibody Q：33) with light chain (SEQ ID NO：67) nucleotide sequence of variable region, and that antibody heavy chain (SEQID NO：34) with light chain (SEQ ID NO：68) amino acid sequence of variable region.

Figure 20 is on anti-TR-2 antibody As to Q (SEQ ID NOs：2nd, 4,6,8,10,12,14,16,18,20,22,24,26,28,30,32 and weight chain variable district 34) amino acid alignment.Show the framework region 1 to 3 (FR1, FR2 and FR3) and complementary determining region 1 to 3 (CDR1, CDR2 and CDR3) on each sequence.

Figure 21 is on anti-TR-2 antibody As to Q (SEQ ID NOs：36th, 38,40,42,44,46,48,50,52,54,56,58,60,62,64,66 and light chain variable district 68) amino acid alignment.Show the framework region 1 to 3 (FR1, FR2 and FR3) and complementary determining region 1 to 3 (CDR1, CDR2 and CDR3) on each sequence.

Figure 22 table shows the work according to embodiment 5, according to each with truncate and the anti-TR-2 antibody of some people of ability that chimeric N- avidin TR-2 protein is combined to 4 reactivity one of organize in classification.

Figure 23 shows the work according to embodiment 6, the people's N- avidins-TR-2 used in epitope mapping 13 kinds of truncate schematic diagrames.

Figure 24 bar chart shows the work according to embodiment 6, some anti-TR-2 antibody of people and the truncate combinations of N- avidins-TR-2.

Figure 25 shows the work according to embodiment 6, truncate and N- avidin-cyno/ people's TR-2 chimeras the schematic diagrames of the N- avidin-cyno TR-2 used in epitope mapping.

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

Figure 27 bar chart shows the work according to embodiment 6, the combination that some anti-TR-2 antibody of people and N- avidins-TR-2 are truncate, chimera and domain are replaced.

The detailed description of some embodiments

Division header used herein is only used for organizational goal and is not necessarily to be construed as the theme of limitation description.The All Files quoted in the application or the part of file, include but is not limited to, patent, patent application, article, book and paper, in order to which any purpose is especially herein by reference in its entirety.

Definition

Standard technique can be used for recombinant DNA, oligonucleotide synthesis and tissue cultures and conversion (for example, electroporation, lipofection).Progress that enzymatic reaction and purification technique can be generally completed according to the specification of manufacturer or such as this area or as described herein.Aforementioned techniques and operation typically can according to conventional method well-known in the art and as various with the progress described in more specific bibliography, the bibliography is cited and discusses from beginning to end in this specification.See, for example, Sambrook et al., Molecular Cloning：A LaboratoryManual (second edition, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y (1989)).It is specifically defined unless provided, the term being used in combination 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 are well-known in the art and those usually used.Standard technique can be used for chemical synthesis, chemical analysis, medicine preparation, preparation, delivering and patient treatment.

In this application, unless otherwise expressly specified, the use of odd number includes plural number.In this application, unless otherwise indicated, the use of "or" means "and/or".In addition, the use of term " comprising " is not restricted.Equally, unless otherwise expressly specified, term such as " element " or " component " includes element and component containing unit and containing the element and component for having more than subunit.

As used according to present disclosure, unless otherwise indicated, following terms should be understood with following implications：

It is as used herein, term " separation polynucleotides " should mean the polynucleotides of genome, cDNA or synthesis source or its some combination, because all or part of polynucleotides that its source " separation polynucleotides " (1) is not found with wherein " separating polynucleotides " in nature are combined, (2) it is connected with the polynucleotides that it is not attached thereto in nature, or the part of (3) not as larger sequence in nature occurs.

Term " polynucleotides " and " oligonucleotides " are used interchangeably, and as mentioned above mean length be at least ten base polymerized form nucleotides.In certain embodiments, base can include any type of nucleotides of at least one ribonucleotide, deoxyribonucleotide and modified forms.The term includes the DNA of single and double chain form.Term " polynucleotides " is also comprising such sequence, and it includes SEQ ID NOs：1st, the one or more in 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, polynucleotides have and the nucleotide sequence being equal of the nucleotide sequence shown in Fig. 3-19 about 90% or about 95% or about 96% or about 97% or about 98% or about 99%.There is provided the complementary polynucleotides of the specific polynucleotides with encoding some polypeptides as described herein in certain embodiments.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, the polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a, wherein CDR1a comprising amino acid sequence a b c d e f g h i j k l, wherein amino acid a be glycine, amino acid b be 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；Wherein CDR2a includes amino acid 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 tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；The polypeptide combination TR-2 wherein combined with antibody light chain.

In certain embodiments, polynucleotides include coding CDR2a sequence, wherein CDR2a includes amino acid 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 tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present.

In certain embodiments, polynucleotides include coding CDR3a sequence, the CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes at least two complementary determining region (CDRs) selected from CDR1a, CDR2a and CDR3a, wherein the polypeptide combination TR-2 combined with antibody light chain.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes CDR1a, CDR2a and CDR3a, wherein the polypeptide combination TR-2 combined with antibody light chain.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes heavy chain of antibody variable region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes human antibody heavy chain variable region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes heavy chain constant region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes people's heavy chain constant region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes such as SEQ IDNO：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：Amino acid sequence shown in 34.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes non-human heavy chain's constant region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes the heavy chain constant region of the species in addition to people.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide, which is included, is selected from least one following complementary determining regions (CDR)：SEQ ID NO：2 amino acid 26-35；SEQ ID NO：2 amino acid 50-66；SEQ ID NO：2 amino acid 99-110；SEQ ID NO：4 amino acid 26-37；SEQ ID NO：4 amino acid 52-67；SEQ ID NO：4 amino acid/11 00-109；SEQ ID NO：6 amino acid 26-37；SEQ ID NO：6 amino acid 52-67；SEQ ID NO：6 amino acid/11 00-109；SEQ ID NO：8 amino acid 26-37；SEQ ID NO：8 amino acid 52-67；SEQ ID NO：8 amino acid/11 00-109；SEQ ID NO：10 amino acid 26-35；SEQ ID NO：10 amino acid 50-66；SEQ ID NO：10 amino acid 99-110；SEQ ID NO：12 amino acid 26-35；SEQ ID NO：12 amino acid 50-66；SEQ ID NO：12 amino acid 99-111；SEQ ID NO：14 amino acid 26-35；SEQ ID NO：14 amino acid 50-65；SEQ ID NO：14 amino acid 98-111；SEQ ID NO：16 amino acid 26-37；SEQ ID NO：16 amino acid 52-67；SEQ ID NO：16 amino acid/11 00-109；SEQ ID NO：18 amino acid 26-35；SEQ ID NO：18 amino acid 50-66；SEQ ID NO：18 amino acid 99-105；SEQ ID NO：20 amino acid 26-35；SEQ ID NO：20 amino acid 50-66；SEQ ID NO：20 amino acid 99-118；SEQ ID NO：22 amino acid 26-35；SEQ ID NO：22 amino acid 50-66；SEQ ID NO：22 amino acid 99-118；SEQ ID NO：24 amino acid 26-35；SEQ ID NO：24 amino acid 50-65；SEQ ID NO：24 amino acid 98-108；SEQ ID NO：26 amino acid 26-35；SEQ ID NO：26 amino acid 50-66；SEQ ID NO：26 amino acid 99-110；SEQ ID NO：28 amino acid 26-35；SEQ ID NO：28 amino acid 50-66；SEQ ID NO：28 amino acid 99-117；SEQ ID NO：30 amino acid 26-37；SEQ ID NO：30 amino acid 52-67；SEQ ID NO：30 amino acid/11 00-111；SEQ ID NO：32 amino acid 26-37；SEQ ID NO：32 amino acid 52-67；SEQ ID NO：32 amino acid/11 00-111；SEQ ID NO：34 amino acid 26-37；SEQ ID NO：34 amino acid 52-67；With SEQ ID NO：34 amino acid/11 00-111, wherein the polypeptide combination TR-2 combined with antibody light chain.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes at least two 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, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes at least three 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, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：2 amino acid 26-35, SEQ ID NO：2 amino acid 50-66 and SEQ ID NO：2 amino acid 99-110.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：4 amino acid 26-37, SEQ ID NO：4 amino acid 52-67 and SEQ ID NO：4 amino acid/11 00-109.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQID NO：6 amino acid 26-37, SEQ ID NO：6 amino acid 52-67 and SEQ IDNO：6 amino acid/11 00-109.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：8 amino acid 26-37, SEQ ID NO：8 amino acid 52-67 and SEQ ID NO：8 amino acid/11 00-109.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：10 amino acid 26-35, SEQ ID NO：10 amino acid 50-66 and SEQ ID NO：10 amino acid 99-110.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：12 amino acid 26-35, SEQ ID NO：12 amino acid 50-66 and SEQ ID NO：12 amino acid 99-111.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：14 amino acid 26-35, SEQ ID NO：14 amino acid 50-65 and SEQ ID NO：14 amino acid 98-111.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：16 amino acid 26-37, SEQ ID NO：16 amino acid 52-67 and SEQ ID NO：16 amino acid/11 00-109.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：18 amino acid 26-35, SEQ ID NO：18 amino acid 50-66 and SEQ ID NO：18 amino acid 99-105.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：20 amino acid 26-35, SEQ ID NO：20 amino acid 50-66 and SEQ ID NO：20 amino acid 99-118.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：22 amino acid 26-35, SEQ ID NO：22 amino acid 50-66 and SEQ ID NO：22 amino acid 99-118.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：24 amino acid 26-35, SEQ ID NO：24 amino acid 50-65 and SEQ ID NO：24 amino acid 98-108.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：26 amino acid 26-35, SEQ ID NO：26 amino acid 50-66 and SEQ ID NO：26 amino acid 99-110.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：28 amino acid 26-35, SEQ ID NO：28 amino acid 50-66 and SEQ ID NO：28 amino acid 99-117.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：30 amino acid 26-37, SEQ ID NO：30 amino acid 52-67 and SEQ ID NO：30 amino acid/11 00-111.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：32 amino acid 26-37, SEQ ID NO：32 amino acid 52-67 and SEQ ID NO：32 amino acid/11 00-111.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：34 amino acid 26-37, SEQ ID NO：34 amino acid 52-67 and SEQ ID NO：34 amino acid/11 00-111.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, the polypeptide includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b, wherein CDR1b includes amino acid 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 lysine；Amino acid b1 is selected from threonine, alanine 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 valine；Amino acid g1 is selected from serine, leucine or arginine；Amino acid h1 is selected from threonine, serine, isoleucine, asparagine, arginine, histidine or tyrosine；Amino acid i1 is selected from tyrosine, arginine, tryptophan, aspartic acid or serine；J1 is selected from leucine, isoleucine, asparagine, tyrosine or serine；Amino acid k1 is selected from asparagine, glycine, valine, alanine or leucine；Amino acid l1 is selected from tyrosine, alanine or asparagine or is not present；Amino acid m1 is selected from asparagine or lysine or is not present；Amino acid n1 is selected from tyrosine, asparagine or isoleucine or is not present；Amino acid o1 is selected from leucine or tyrosine or is not present；Amino acid p1 is selected from aspartic acid or leucine or is not present；Valine, alanine or threonine are selected from amino acid q1 or are not present；Wherein CDR2b includes amino acid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from alanine, aspartic acid, leucine, tryptophan, glycine or valine；Amino acid s1 is selected from threonine, valine, glycine or alanine；Amino acid t1 is serine；Amino acid u1 is selected from serine, asparagine or threonine；Amino acid v1 is selected from leucine, phenylalanine or arginine；Amino acid w1 is selected from glutamine, alanine or glutamic acid；Serine, arginine or threonine are selected from amino acid x1；Wherein CDR3b is selected from glutamine, methionine, leucine or histidine comprising amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', wherein amino acid y1；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes at least two complementary determining region (CDRs) selected from CDR1b, CDR2b and CDR3b, wherein the polypeptide combination TR-2 combined with heavy chain of antibody.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes CDR1b, CDR2b and CDR3b, wherein the polypeptide combination TR-2 combined with heavy chain of antibody.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes antibody light chain variable region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes human antibody light chain variable region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes constant region of light chain.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes people's constant region of light chain.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes such as 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：Amino acid sequence shown in 68.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes non-human light chain's constant region.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes the constant region of light chain of the species in addition to people.

In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide, which is included, is selected from least one following complementary determining regions (CDR)：SEQ ID NO：36 amino acid 24-34；SEQ ID NO：36 amino acid 50-56；SEQ ID NO：36 amino acid 89-97；SEQ ID NO：38 amino acid 24-34；SEQ ID NO：38 amino acid 50-56；SEQ ID NO：38 amino acid 89-97；SEQ ID NO：40 amino acid 24-34；SEQ ID NO：40 amino acid 50-56；SEQ ID NO：40 amino acid 89-97；SEQ ID NO：42 amino acid 24-34；SEQ ID NO：42 amino acid 50-56；SEQ ID NO：42 amino acid 89-97；SEQ ID NO：44 amino acid 24-34；SEQ ID NO：44 amino acid 50-56；SEQ ID NO：44 amino acid 89-97；SEQ ID NO：46 amino acid 24-34；SEQ ID NO：46 amino acid 50-56；SEQ ID NO：46 amino acid 89-97；SEQ ID NO：48 amino acid 24-40；SEQ ID NO：48 amino acid 56-62；SEQ ID NO：48 amino acid 95-103；SEQ ID NO：50 amino acid 24-39；SEQ ID NO：50 amino acid 55-61；SEQ ID NO：50 amino acid 94-102；SEQ ID NO：52 amino acid 24-40；SEQ ID NO：52 amino acid 56-62；SEQ ID NO：52 amino acid 95-103；SEQ ID NO：54 amino acid 24-34；SEQ ID NO：54 amino acid 50-56；SEQ ID NO：54 amino acid 89-97；SEQ ID NO：56 amino acid 24-34；SEQ ID NO：56 amino acid 50-56；SEQ ID NO：56 amino acid 89-97；SEQ ID NO：58 amino acid 24-40；SEQ ID NO：58 amino acid 56-62；SEQ ID NO：58 amino acid 95-103；SEQ ID NO：60 amino acid 24-34；SEQ ID NO：60 amino acid 50-56；SEQ ID NO：60 amino acid 89-97；SEQ ID NO：62 amino acid 24-34；SEQ ID NO：62 amino acid 50-56；SEQ ID NO：62 amino acid 89-97；SEQ ID NO：64 amino acid 24-35；SEQ ID NO：64 amino acid 51-57；SEQ ID NO：64 amino acid 90-88；SEQ ID NO：66 amino acid 24-34；SEQ ID NO：66 amino acid 50-57；SEQ ID NO：66 amino acid 89-97；SEQ ID NO：68 amino acid 24-34；SEQ ID NO：68 amino acid 50-56；With SEQ ID NO：68 amino acid 89-97；The polypeptide combination TR-2 wherein combined with heavy chain of antibody.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes at least two CDRs in SEQ IDNOS.36,38,40,42,44,46,48,50,52,54,56,58,60,62,64 or 68.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes at least three 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, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：36 amino acid 24-34, SEQ ID NO：36 amino acid 50-56 and SEQ ID NO：36 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：38 amino acid 24-34, SEQ ID NO：38 amino acid 50-56 and SEQ ID NO：38 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQID NO：40 amino acid 24-34, SEQ ID NO：40 amino acid 50-56 and SEQID NO：40 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：42 amino acid 24-34, SEQ IDNO：42 amino acid 50-56 and SEQ ID NO：42 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ IDNO：44 amino acid 24-34, SEQ ID NO：44 amino acid 50-56 and SEQ IDNO：44 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：46 amino acid 24-34, SEQ ID NO：46 amino acid 50-56 and SEQ ID NO：46 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：48 amino acid 24-40, SEQ ID NO：48 amino acid 56-62 and SEQ ID NO：48 amino acid 95-103.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：50 amino acid 24-39, SEQ ID NO：50 amino acid 55-61 and SEQ ID NO：50 amino acid 94-102.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：52 amino acid 24-40, SEQ ID NO：52 amino acid 56-62 and SEQ ID NO：52 amino acid 95-103.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：54 24-34, SEQ ID NO：54 amino acid 50-56 and SEQ ID NO：54 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：56 amino acid 24-34, SEQ ID NO：56 amino acid 50-56 and SEQ ID NO：56 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：58 amino acid 24-40, SEQ ID NO：58 amino acid 56-62 and SEQ ID NO：58 amino acid 95-103.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：60 amino acid 24-34, SEQ ID NO：60 amino acid 50-56 and SEQ ID NO：60 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：62 amino acid 24-34, SEQ ID NO：62 amino acid 50-56 and SEQ ID NO：62 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：64 amino acid 24-35, SEQ ID NO：64 amino acid 51-57 and SEQ ID NO：64 amino acid 90-88.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：66 amino acid 24-34, SEQ ID NO：66 amino acid 50-57 and SEQ ID NO：66 amino acid 89-97.In certain embodiments, polynucleotides include the sequence of coded polypeptide, and the polypeptide includes SEQ ID NO：68 amino acid 24-34, SEQ ID NO：68 amino acid 50-56 and SEQ ID NO：68 amino acid 89-97.

In certain embodiments, application discusses encoding antibody weight and some polynucleotides of light chain.In certain embodiments, application discusses some polynucleotides of encoding antibody heavy variable region.In certain embodiments, application discusses some polynucleotides of encoding human antibody's weight chain variable district.In certain embodiments, application discusses some polynucleotides of encoding antibody light variable region.In certain embodiments, application discusses some polynucleotides of encoding human antibody's light chain variable district.In certain embodiments, application discusses some polynucleotides of encoding antibody heavy constant region.In certain embodiments, application discusses some polynucleotides of encoding human antibody's heavy chain constant region.In certain embodiments, some polynucleotides of the heavy chain constant region of the species application discusses coding in addition to people.In certain embodiments, application discusses some polynucleotides of encoding antibody light constant region.In certain embodiments, application discusses some polynucleotides of encoding human antibody's constant region of light chain.In certain embodiments, some polynucleotides of the antibody light chain constant region of the species application discusses coding in addition to people.In certain embodiments, application discusses some polynucleotides of coding single-chain antibody.

In certain embodiments, these antibody weight and light chain polynucleotides and polypeptide are human antibody weight and light chain polynucleotides and polypeptide.In certain embodiments, polynucleotides include such as SEQ IDNOS.SEQ ID NOs：1st, the nucleotide sequence shown in 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, polynucleotides include such nucleotide sequence, and it has one or more missings of one or more nucleotides of those sequences, adds and/or replace.In certain embodiments, polynucleotides include the nucleotide sequence of encoding amino acid sequence, and the amino acid sequence includes such as SEQID NOS：2nd, the amino acid sequence shown in 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.There is provided the variable region sequences for including complementary determining region (CDRs) such as CDR1 to CDR3 in certain embodiments.In certain embodiments, variable region polynucleotide and polypeptide are people's variable region polynucleotide and polypeptide.

Term " naturally occurring nucleotides " includes deoxyribonucleotide and ribonucleotide.Deoxyribonucleotide includes but is not limited to, adenosine, guanine, cytimidine and thymidine.Ribonucleotide includes but is not limited to, adenosine, cytimidine, thymidine and uracil.Term " nucleotides of modification " includes but is not limited to, the nucleotides of with modification or displacement glycosyl group etc..Term " polynucleotides key " includes but is not limited to, and polynucleotides key is such as thiophosphate, phosphorodithioate, phosphoroselenoate, two phosphoroselenoates, phosphoroanilothioate, phoshoraniladate, phosphoramidate.See, e.g., LaPlanche et al., Nucl.Acids Res.14：9081(1986)；Stec et al., J.Am.Chem.Soc.106：6077(1984)；Stein et al., Nucl.AcidsRes.16：3209(1988)；Zon et al., Anti-Cancer Drug Design 6：539(1991)；Zon et al., Oligonucleotides and Analogues：A Practical Approach, the 87-108 pages (F.Eckstein, editor, Oxford University Press, Oxford England (1991))；Stec et al., U.S. Patent number 5,151,510；Uhlmann and Peyman Chemical Reviews90：543(1990).In certain embodiments, polynucleotides can include the mark for detecting.

Term " isolated polypeptide " refers to any polypeptide, its (1) is without it generally by least some of protein being found therewith, (2) substantially free of other protein from identical source for example from identical type, (3) by being not present from the expression of different types of cell, or (4) in nature.

Term " polypeptide ", " peptide " and " protein " is used interchangeably herein, and refers to the polymer of 2 or more the amino acid interconnected by peptide bond or the peptide bond of modification, i.e. peptide isostere.The term application is in the amino acid polymer for including naturally occurring amino acid, and wherein one or more amino acid residues are the amino acid polymers of the chemical analog of non-naturally occurring amino acid or naturally occurring amino acid.Amino acid polymer can include the one or more amino acid residues modified by one or more natural processes such as post translational processing, and/or the one or more amino acid residues modified by one or more chemical modification technologies known in the art.

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

" variant " of reference polypeptide refers to relative to reference polypeptide, the polypeptide with one or more amino acid replacements, deletion or insertion.In certain embodiments, the variant of reference polypeptide has the posttranslational modification site (that is, glycosylation site) changed.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 is not limited to, glycosylation variants.Glycosylation variants include being compared with reference polypeptide, the variant that the number and/or type of wherein glycosylation site have been changed.In certain embodiments, the glycosylation variants of reference polypeptide include the N linked glycosylation sites of numbers more more or less than reference polypeptide.In certain embodiments, N linked glycosylation sites are characterised by sequence Asn-X-Ser or Asn-X-Thr, wherein the amino acid residue for being appointed as X can be any amino acid residue in addition to proline.In certain embodiments, the glycosylation variants of reference polypeptide join the rearrangement of carbohydrate chain comprising N, and wherein one or more N linked glycosylation sites (be usually naturally occurring those) are eliminated, and one or more new N connection sites are generated.

The variant of reference polypeptide includes but is not limited to, cysteine variants.In certain embodiments, cysteine variants include such variant, and wherein one or more cysteine residues of reference polypeptide are replaced by one or more non-cysteine residues；And/or one or more non-cysteine residues of reference polypeptide are replaced by one or more cysteine residues.In certain embodiments, cysteine variants are probably useful, when the necessary refolding of particular polypeptide is into biologically active conformation, for example, after the separation of insoluble occlusion body.In certain embodiments, the cysteine variants of reference polypeptide have the cysteine residues less than reference polypeptide.In certain embodiments, the cysteine variants of reference polypeptide have the cysteine of even number, are preferably minimized so as to be interacted as caused by azygous cysteine.In certain embodiments, cysteine variants have cysteine residues more more than native protein.

" derivative " of reference polypeptide refers to：Polypeptide：(1) one or more modifications of one or more amino acid residues with reference polypeptide；And/or (2) wherein one or more peptidyl keys are replaced with one or more non-peptidyl linker keys；And/or (3) wherein N-terminal and/or C-terminal has been modified.Some example sex modifications include but is not limited to, acetylation, it is acylated, ADP- ribosylation, amidatioon, biotinylation, the covalent attachment of riboflavin, the covalent attachment of heme moiety, the covalent attachment of nucleotides or nucleotide derivative, the covalent attachment of lipid or lipid derivate, the covalent attachment of phosphatidylinositols, crosslinking, cyclisation, disulfide formation, demethylation, the formation of covalent cross-linking, the formation of cystine, the formation of pyroglutamate, formyl, gamma-carboxylation, glycosylation, GPI anchors into, hydroxylating, iodate, methylate, myristoylation, oxidation, proteolysis are processed, phosphorylation, isoprenylation, racemization, selenizing (selenoylation), sulphation, transfer RNA mediation amino acid to protein addition such as arginyl and ubiquitination.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 is not limited to, the amino acid sequence modified by natural process such as post translational processing or by chemical modification technology well-known in the art.In certain embodiments, modification can occur from anywhere in polypeptide, including peptide backbone, amino acid side chain and amino or carboxyl terminal.In some such embodiments, modification can be present at several sites of given polypeptide with identical or different degree.In certain embodiments, the modification that polypeptide includes many types, deletion, addition and/or the displacement of one or more amino acid of such as native sequences are given.In certain embodiments, polypeptide can be branch and/or ring-type.Ring-type, branch and the ring type polypeptide of branch can be after translating natural process (include but is not limited to, ubiquitination) produce, or can be prepared by synthetic method.As described below, term " polypeptide " is also comprising such sequence, the amino acid sequence of its heavy chain comprising antibody and/or light chain, the 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：2nd, 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 includes the sequence of one or more deletions, addition and/or the displacement of one or more amino acid with those sequences.In certain embodiments, some peptide sequences include at least one complementary determining region (CDR).

In certain embodiments, polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a, wherein CDR1a includes amino acid 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；Wherein CDR2a includes amino acid 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 tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；The polypeptide combination TR-2 wherein combined with antibody light chain.

In certain embodiments, polypeptide includes at least two complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a, wherein the polypeptide combination TR-2 combined with antibody light chain.In certain embodiments, polypeptide includes CDR1a, CDR2a and CDR3a, wherein the polypeptide combination TR-2 combined with antibody light chain.

In certain embodiments, polypeptide includes heavy chain of antibody variable region.In certain embodiments, polypeptide includes human antibody heavy chain variable region.In certain embodiments, polypeptide includes heavy chain constant region.In certain embodiments, polypeptide includes people's heavy chain constant region.In certain embodiments, polypeptide includes such as 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：Amino acid sequence shown in 34.In certain embodiments, polypeptide includes non-human heavy chain's constant region.In certain embodiments, polypeptide includes the heavy chain constant region of the species in addition to people.

In certain embodiments, polypeptide, which is included, is selected from least one following complementary determining regions (CDR)：SEQ ID NO：2 amino acid 26-35；SEQ ID NO：2 amino acid 50-66；SEQ ID NO：2 amino acid 99-110；SEQ ID NO：4 amino acid 26-37；SEQ ID NO：4 amino acid 52-67；SEQ ID NO：4 amino acid/11 00-109；SEQ ID NO：6 amino acid 26-37；SEQ ID NO：6 amino acid 52-67；SEQ ID NO：6 amino acid/11 00-109；SEQ ID NO：8 amino acid 26-37；SEQ ID NO：8 amino acid 52-67；SEQ ID NO：8 amino acid/11 00-109；SEQ ID NO：10 amino acid 26-35；SEQ ID NO：10 amino acid 50-66；SEQ ID NO：10 amino acid 99-110；SEQ ID NO：12 amino acid 26-35；SEQ ID NO：12 amino acid 50-66；SEQ ID NO：12 amino acid 99-111；SEQ ID NO：14 amino acid 26-35；SEQ ID NO：14 amino acid 50-65；SEQ ID NO：14 amino acid 98-111；SEQ ID NO：16 amino acid 26-37；SEQ ID NO：16 amino acid 52-67；SEQ ID NO：16 amino acid/11 00-109；SEQ ID NO：18 amino acid 26-35；SEQ ID NO：18 amino acid 50-66；SEQ ID NO：18 amino acid 99-105；SEQ ID NO：20 amino acid 26-35；SEQ ID NO：20 amino acid 50-66；SEQ ID NO：20 amino acid 99-118；SEQ ID NO：22 amino acid 26-35；SEQ ID NO：22 amino acid 50-66；SEQ ID NO：22 amino acid 99-118；SEQ ID NO：24 amino acid 26-35；SEQ ID NO：24 amino acid 50-65；SEQ ID NO：24 amino acid 98-108；SEQ ID NO：26 amino acid 26-35；SEQ ID NO：26 amino acid 50-66；SEQ ID NO：26 amino acid 99-110；SEQ ID NO：28 amino acid 26-35；SEQ ID NO：28 amino acid 50-66；SEQ ID NO：28 amino acid 99-117；SEQ ID NO：30 amino acid 26-37；SEQ ID NO：30 amino acid 52-67；SEQ ID NO：30 amino acid/11 00-111；SEQ ID NO：32 amino acid 26-37；SEQ ID NO：32 amino acid 52-67；SEQ ID NO：32 amino acid/11 00-111；SEQ ID NO：34 amino acid 26-37；SEQ ID NO：34 amino acid 52-67；With SEQ ID NO：34 amino acid/11 00-111；The polypeptide combination TR-2 wherein combined with antibody light chain.In certain embodiments, polypeptide includes at least two 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 includes at least three 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, polypeptide includes SEQ ID NO：2 amino acid 26-35, SEQ ID NO：2 amino acid 50-66 and SEQ ID NO：2 amino acid 99-110.In certain embodiments, polypeptide includes SEQ ID NO：4 amino acid 26-37, SEQ IDNO：4 amino acid 52-67 and SEQ ID NO：4 amino acid/11 00-109.In certain embodiments, polypeptide includes SEQ ID NO：6 amino acid 26-37, SEQ ID NO：6 amino acid 52-67 and SEQ ID NO：6 amino acid/11 00-109.In certain embodiments, polypeptide includes SEQ ID NO：8 amino acid 26-37, SEQ ID NO：8 amino acid 52-67 and SEQ ID NO：8 amino acid/11 00-109.In certain embodiments, polypeptide includes SEQ ID NO：10 amino acid 26-35, SEQ ID NO：10 amino acid 50-66 and SEQ ID NO：10 amino acid 99-110.In certain embodiments, polypeptide includes SEQ ID NO：12 amino acid 26-35, SEQ ID NO：12 amino acid 50-66 and SEQ ID NO：12 amino acid 99-111.In certain embodiments, polypeptide includes SEQ ID NO：14 amino acid 26-35, SEQ ID NO：14 amino acid 50-65 and SEQ ID NO：14 amino acid 98-111.In certain embodiments, polypeptide includes SEQ ID NO：16 amino acid 26-37, SEQ ID NO：16 amino acid 52-67 and SEQ ID NO：16 amino acid/11 00-109.In certain embodiments, polypeptide includes SEQ ID NO：18 amino acid 26-35, SEQ ID NO：18 amino acid 50-66 and SEQ ID NO：18 amino acid 99-105.In certain embodiments, polypeptide includes SEQ ID NO：20 amino acid 26-35, SEQ ID NO：20 amino acid 50-66 and SEQ ID NO：20 amino acid 99-118.In certain embodiments, polypeptide includes SEQ ID NO：22 amino acid 26-35, SEQ ID NO：22 amino acid 50-66 and SEQ ID NO：22 amino acid 99-118.In certain embodiments, polypeptide includes SEQ ID NO：24 amino acid 26-35, SEQ ID NO：24 amino acid 50-65 and SEQ ID NO：24 amino acid 98-108.In certain embodiments, polypeptide includes SEQ ID NO：26 amino acid 26-35, SEQ ID NO：26 amino acid 50-66 and SEQ ID NO：26 amino acid 99-110.In certain embodiments, polypeptide includes SEQ ID NO：28 amino acid 26-35, SEQ ID NO：28 amino acid 50-66 and SEQ ID NO：28 amino acid 99-117.In certain embodiments, polypeptide includes SEQ ID NO：30 amino acid 26-37, SEQ ID NO：30 amino acid 52-67 and SEQ ID NO：30 amino acid/11 00-111.In certain embodiments, polypeptide includes SEQ ID NO：32 amino acid 26-37, SEQ ID NO：32 amino acid 52-67 and SEQ ID NO：32 amino acid/11 00-111.In certain embodiments, polypeptide includes SEQ ID NO：34 amino acid 26-37, SEQ ID NO：34 amino acid 52-67 and SEQ ID NO：34 amino acid/11 00-111.

In certain embodiments, polypeptide includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b, wherein CDR1b includes amino acid 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 lysine；Amino acid b1 is selected from threonine, alanine 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 valine；Amino acid g1 is selected from serine, leucine or arginine；Amino acid h1 is selected from threonine, serine, isoleucine, asparagine, arginine, histidine or tyrosine；Amino acid i1 is selected from tyrosine, arginine, tryptophan, aspartic acid or serine；J1 is selected from leucine, isoleucine, asparagine, tyrosine or serine；Amino acid k1 is selected from asparagine, glycine, valine, alanine or leucine；Amino acid l1 is selected from tyrosine, alanine or asparagine or is not present；Amino acid m1 is selected from asparagine or lysine or is not present；Amino acid n1 is selected from tyrosine, asparagine or isoleucine or is not present；Amino acid o1 is selected from leucine or tyrosine or is not present；Amino acid p1 is selected from aspartic acid or leucine or is not present；Valine, alanine or threonine are selected from amino acid q1 or are not present；Wherein CDR2b includes amino acid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from alanine, aspartic acid, leucine, tryptophan, glycine or valine；Amino acid s1 is selected from threonine, valine, glycine or alanine；Amino acid t1 is serine；Amino acid u1 is selected from serine, asparagine or threonine；Amino acid v1 is selected from leucine, phenylalanine or arginine；Amino acid w1 is selected from glutamine, alanine or glutamic acid；Serine, arginine or threonine are selected from amino acid x1；Wherein CDR3b is selected from glutamine, methionine, leucine or histidine comprising amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', wherein amino acid y1；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

In certain embodiments, polypeptide includes at least two complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b, wherein the polypeptide combination TR-2 combined with heavy chain of antibody.In certain embodiments, polypeptide includes CDR1b, CDR2b and CDR3b, wherein the polypeptide combination TR-2 combined with heavy chain of antibody.

In certain embodiments, polypeptide includes antibody light chain variable region.In certain embodiments, polypeptide includes human antibody light chain variable region.In certain embodiments, polypeptide includes constant region of light chain.In certain embodiments, antibody includes people's constant region of light chain.In certain embodiments, polypeptide includes such as 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：Amino acid sequence shown in 68.In certain embodiments, polypeptide includes non-human light chain's constant region.In certain embodiments, polypeptide includes the constant region of light chain of the species in addition to people.

In certain embodiments, polypeptide, which is included, is selected from least one following complementary determining regions (CDR)：SEQ ID NO：36 amino acid 24-34；SEQ ID NO：36 amino acid 50-56；SEQ ID NO：36 amino acid 89-97；SEQ ID NO：38 amino acid 24-34；SEQ ID NO：38 amino acid 50-56；SEQ ID NO：38 amino acid 89-97；SEQ ID NO：40 amino acid 24-34；SEQ ID NO：40 amino acid 50-56；SEQ ID NO：40 amino acid 89-97；SEQ ID NO：42 amino acid 24-34；SEQ ID NO：42 amino acid 50-56；SEQ ID NO：42 amino acid 89-97；SEQ ID NO：44 amino acid 24-34；SEQ ID NO：44 amino acid 50-56；SEQ ID NO：44 amino acid 89-97；SEQ ID NO：46 amino acid 24-34；SEQ ID NO：46 amino acid 50-56；SEQ ID NO：46 amino acid 89-97；SEQ ID NO：48 amino acid 24-40；SEQ ID NO：48 amino acid 56-62；SEQ ID NO：48 amino acid 95-103；SEQ ID NO：50 amino acid 24-39；SEQ ID NO：50 amino acid 55-61；SEQ ID NO：50 amino acid 94-102；SEQ ID NO：52 amino acid 24-40；SEQ ID NO：52 amino acid 56-62；SEQ ID NO：52 amino acid 95-103；SEQ ID NO：54 amino acid 24-34；SEQ ID NO：54 amino acid 50-56；SEQ ID NO：54 amino acid 89-97；SEQ ID NO：56 amino acid 24-34, SEQ ID NO：56 amino acid 50-56；SEQ ID NO：56 amino acid 89-97；SEQ ID NO：58 amino acid 24-40；SEQ ID NO：58 amino acid 56-62；SEQ ID NO：58 amino acid 95-103；SEQ ID NO：60 amino acid 24-34；SEQ ID NO：60 amino acid 50-56；SEQ ID NO：60 amino acid 89-97；SEQ ID NO：62 amino acid 24-34；SEQ ID NO：62 amino acid 50-56；SEQ ID NO：62 amino acid 89-97；SEQ ID NO：64 amino acid 24-35；SEQ ID NO：64 amino acid 51-57；SEQ ID NO：64 amino acid 90-88；SEQ ID NO：66 amino acid 24-34；SEQ ID NO：66 amino acid 50-57；SEQ ID NO：66 amino acid 89-97；SEQ ID NO：68 amino acid 24-34；SEQ ID NO：68 amino acid 50-56；With SEQ ID NO：68 amino acid 89-97；The polypeptide combination TR-2 wherein combined with heavy chain of antibody.In certain embodiments, polypeptide includes at least two 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 includes at least three 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 includes SEQ ID NO：36 amino acid 24-34, SEQ ID NO：36 amino acid 50-56 and SEQ ID NO：36 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：38 amino acid 24-34, SEQID NO：38 amino acid 50-56 and SEQ ID NO：38 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：40 amino acid 24-34, SEQ IDNO：40 amino acid 50-56 and SEQ ID NO：40 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：42 amino acid 24-34, SEQ ID NO：42 amino acid 50-56 and SEQ ID NO：42 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：44 amino acid 24-34, SEQ ID NO：44 amino acid 50-56 and SEQ ID NO：44 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：46 amino acid 24-34, SEQ ID NO：46 amino acid 50-56 and SEQ ID NO：46 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：48 amino acid 24-40, SEQ ID NO：48 amino acid 56-62 and SEQ ID NO：48 amino acid 95-103.In certain embodiments, polypeptide includes SEQ ID NO：50 amino acid 24-39, SEQ ID NO：50 amino acid 55-61 and SEQ ID NO：50 amino acid 94-102.In certain embodiments, polypeptide includes SEQ ID NO：52 amino acid 24-40, SEQ ID NO：52 amino acid 56-62 and SEQ ID NO：52 amino acid 95-103.In certain embodiments, polypeptide includes SEQ ID NO：54 amino acid 24-34, SEQ ID NO：54 amino acid 50-56 and SEQ ID NO：54 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：56 amino acid 24-34, SEQ ID NO：56 amino acid 50-56 and SEQ ID NO：56 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：58 amino acid 24-40, SEQ ID NO：58 amino acid 56-62 and SEQ ID NO：58 amino acid 95-103.In certain embodiments, polypeptide includes SEQ ID NO：60 amino acid 24-34, SEQ ID NO：60 amino acid 50-56 and SEQ ID NO：60 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：62 amino acid 24-34, SEQ ID NO：62 amino acid 50-56 and SEQ ID NO：62 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：64 amino acid 24-35, SEQ ID NO：64 amino acid 51-57 and SEQ ID NO：64 amino acid 90-88.In certain embodiments, polypeptide includes SEQ ID NO：66 amino acid 24-34, SEQ ID NO：66 amino acid 50-57 and SEQ ID NO：66 amino acid 89-97.In certain embodiments, polypeptide includes SEQ ID NO：68 amino acid 24-34, SEQ ID NO：68 amino acid 50-56 and SEQ ID NO：68 amino acid 89-97.

Term " naturally occurring " means the object that can be found in nature when applied to object.For example, what can be separated from the source in nature is present in biological (including virus), and by the polypeptide or polynucleotides manually modified intentionally it is not in the lab or otherwise naturally occurring.

As it is used herein, term " being operably connected " refers to the component of the relation that they are worked in its expection mode in permission.For example, in the background of polynucleotide sequence, when control sequence and coded sequence are combined with each other in this way so that the expression of coded sequence under conditions of compatible with the function of coded sequence when reaching, control sequence " can be operably connected " with coded sequence.

Term " control sequence " refers to the polynucleotide sequence of expression and the processing for the coded sequence that them can be influenceed in combination.The property of such control sequence can be different according to host organism.Some exemplary control sequences on prokaryotes include but is not limited to, promoter, ribosome bind site and transcription terminator.Include but is not limited on Eukaryotic some exemplary control sequences, promoter, enhancer and transcription terminator.In certain embodiments, " control sequence " can include homing sequence and/or fusion partner sequence.

In certain embodiments, when the first and second of polynucleotide encoding sequence are transcribed into single adjacent mRNA, the single adjacent mRNA can be translated into single adjacent polypeptide, and the first polynucleotide encoding sequence is operably connected with second of polynucleotide encoding sequence.

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

As it is used herein, when two or more polypeptides are by being translated into single adjacent peptide sequence or being attached by the way that they are synthesized into single adjacent peptide sequence, two or more polypeptides are " fusions ".In certain embodiments, two or more fused polypeptides can be translated by two or more polynucleotide encoding sequences that are operably connected in vivo.In certain embodiments, two or more fused polypeptides can be translated by two or more polynucleotide encoding sequences that are operably connected in vitro.

If as it is used herein, the polypeptide of every kind of connection can be worked in its expection mode, then two or more polypeptides " operationally merging ".

In certain embodiments, the first polypeptide comprising 2 or more different polypeptide units is considered as being connected with second of polypeptide, as long as the different polypeptide units of at least one of the first polypeptide are connected with second of polypeptide.As non-limitative example, in certain embodiments, it is all it is following in the case of antibody be considered as being connected with second of polypeptide：(a) one of heavy chain polypeptide of second of polypeptide and antibody is connected；(b) one of light chain polypeptide of second of polypeptide and antibody is connected；(c) one of heavy chain polypeptide of first molecule of second of polypeptide and antibody is connected, and second of second molecule and antibody the connection of one of light chain polypeptide；First and second molecule and first of antibody and second heavy chain polypeptide connection of second polypeptide, and three of second polypeptide and four molecule and first of antibody and second light chain polypeptide connection (d).

In certain embodiments, word " the first polypeptide is connected with second of polypeptide " includes following situations：(a) the only one molecule of the first polypeptide is connected with the only one molecule of second of polypeptide；(b) the only one molecule of the first polypeptide is connected with more than one molecule of second of polypeptide；(c) the first polypeptide is connected more than a molecule with the only one molecule of second of polypeptide；The first polypeptide more than a molecule with more than one molecule of second polypeptide be connected (d).In certain embodiments, when connection molecule comprising the first polypeptide more than a molecule and second polypeptide only one molecule when, the first polypeptide all or fewer than all molecules can be with second of polypeptid covalence or non-covalent linking.In certain embodiments, when connection molecule comprising the first polypeptide more than a molecule when, other molecule covalents or non-covalent linking that one or more molecules of the first polypeptide can with the first polypeptide.

As it is used herein, " nonrigid connector " refers to according to its chemical constitution, any connector in three dimensions is not fixed in prediction.Whether it is elastic that those skilled in the art can predict that specific connector is expected in background at it.In certain embodiments, the peptide connector comprising 3 or more amino acid is nonrigid connector.

As it is used herein, 20 kinds of conventional amino acids and its abbreviation follow common usage.Referring to Immunology--A Synthesis (second edition, E.S.Golub and D.R.Gren, editor, Sinauer Associates, Sunderland, Mass. (1991)).In certain embodiments, one or more unconventional amino acids can be introduced in polypeptide.Term " unconventional amino acid " refer to be not one of 20 kinds of conventional amino acids any amino acid.Term " non-naturally occurring amino acid " refers to the amino acid that can not find in nature.Non-naturally occurring amino acid is the subgroup of unconventional amino acid.Unconventional amino acid includes but is not limited to, the stereoisomer of 20 kinds of conventional amino acids is (for example, D- amino acid), alpha-non-natural amino acid such as α-, α-disubstituted amino acid, N- alkyl amino acids, lactic acid, homoserine, same cysteine, 4-Hydroxyproline, Gla, ε-N, N, N- trimethyl lysines, ε-N-acetyllysine, O- phosphoserines, N- acetyl serines, N-formylmethionine, 3-Methyl histidine, 5- hydroxylysines, σ-N- methylarginines, and other similar amino acid known in the art and imino acid are (for example, 4-Hydroxyproline).In polypeptide symbol used herein, according to normal usage and convention, left direction is amino terminal direction and right direction is carboxyl terminal direction.

In certain embodiments, conservative amino acid replacement includes the displacement with one or more unconventional amino acid residues.In certain embodiments, unconventional amino acid residue is introduced by chemical peptide symthesis rather than by synthesis in biosystem.

Term " acidic residues " refers to when being introduced between 2 other identical or different amino acid residues in polypeptide, the amino acid residue of the D- or L- forms comprising at least one acidic-group.In certain embodiments, acidic residues include the side chain containing at least one acidic-group.Illustrative acid residue includes but is not limited to, aspartic acid (D) and glutamic acid (E).In certain embodiments, acidic residues can be unconventional amino acid.

Term " aromatic residue " refers to the amino acid residue of the D- or L- forms comprising at least one aromatic group.In certain embodiments, aromatic residue includes the side chain containing at least one aromatic group.Exemplary aromatic race residue includes but is not limited to, phenylalanine (F), tyrosine (Y) and tryptophan (W).In certain embodiments, aromatic residue can be unconventional amino acid.

Term " alkaline residue " refers to when being introduced close to identical or different one or more amino acid residues in polypeptide, can include the amino acid residue of the F- or L- forms of at least one basic group.In certain embodiments, alkaline residue includes the side chain containing at least one basic group.Exemplary basic residue includes but is not limited to, histidine (H), lysine (K) and arginine (R).In certain embodiments, alkaline residue can be unconventional amino acid.

Term " Neutral hydrophilic residue " refers to when being introduced close to identical or different one or more amino acid residues in polypeptide, the amino acid residue of the D- or L- forms comprising at least one hydrophily and/or polar group but not comprising acid or basic group.In certain embodiments, Neutral hydrophilic residue includes the side chain containing at least one Neutral hydrophilic group.Exemplary neutral hydrophilic residue includes but is not limited to, alanine (A), cysteine (C), serine (S), threonine (T), asparagine (N) and glutamine (Q).In certain embodiments, Neutral hydrophilic residue can be unconventional amino acid.

Term " lipophilic residue " and " Laa " refer to the amino acid residue of the D- or L- forms with least one uncharged, aliphatic and/or aromatic group.In certain embodiments, lipophilic residue includes the side chain containing at least one uncharged, aliphatic and/or aromatic group.Exemplary lipophilic residue includes but is not limited to, alanine (A), phenylalanine (F), isoleucine (I), leucine (L), nor-leucine (Nile), methionine (M), valine (V), tryptophan (W) and tyrosine (Y).In certain embodiments, lipophilic residue can be unconventional amino acid.

Term " amphipathic residue " refers to the amino acid residue for turning into hydrophily or D- the or L- forms of lipophilic residue.Exemplary amphipathic residue includes but is not limited to, alanine (A).In certain embodiments, amphipathic residue can be unconventional amino acid.

Term " non-functional residue " refers to when being introduced close to identical or different one or more amino acid residues in polypeptide, lacks the amino acid residue of acid, alkalescence and D- the or L- forms of aromatic group.In certain embodiments, non-functional residue includes the side chain containing at least one non-functional group.Exemplary non-functional residue includes but is not limited to, methionine (M), glycine (G), alanine (A), valine (V), isoleucine (I), leucine (L) and nor-leucine (Nle).In certain embodiments, non-functional residue can be unconventional amino acid.

In certain embodiments, glycine (G) and proline (P) are can to influence the considered amino acid residue of polypeptide chain direction.

In certain embodiments, conservative substitution can be related to replaces a kind of member of residue type with the member of identical residue type.As non-limitative example, in certain embodiments, conservative substitution can be related to replaces acidic residues such as D with different acidic residues such as E.In certain embodiments, non-conservative displacement can be related to replaces a kind of member of residue type with the member of different residue types.As non-limitative example, in certain embodiments, non-conservative displacement can be related to replaces acidic residues such as D with alkaline residue such as K.In certain embodiments, another radical amino acid replacement of cysteine residues, to prevent the disulfide formation with that position in polypeptide.

In the preparation of conservative or non-conservative displacement, according to some embodiments, it may be considered that the hydrophilic index of amino acid.The hydrophilic index of every kind of amino acid can be specified based on its hydrophobicity and charge characteristic.The hydrophilic index of 20 kinds of naturally occurring amino acid is：Isoleucine (+4.5)；Valine (+4.2)；Leucine (+3.8)；Phenylalanine (+2.8)；Cysteine/cystine (+2.5)；Methionine (+1.9)；Alanine (+1.8)；Glycine (- 0.4)；Threonine (- 0.7)；Serine (- 0.8)；Tryptophan (- 0.9)；Tyrosine (- 1.3)；Proline (- 1.6)；Histidine (- 3.2)；Glutamic acid (- 3.5)；Glutamine (- 3.5)；Aspartic acid (- 3.5)；Asparagine (- 3.5)；Lysine (- 3.9)；With arginine (- 4.5).

Importance of the hydropathic amino acid index in interactive biological function is given to protein is known in the art.Kyte et al., J.Mol.Biol., 157：105-131(1982).It is known to have similar hydrophilic index or other amino acid of score with some amino acid substitutions in some cases, and still retain similar bioactivity.In the change carried out based on hydrophilic index, in certain embodiments, include amino acid replacement of its hydrophilic index in ± 2.In certain embodiments, it is included in those in ± 1, and in certain embodiments, is included in those in ± 0.5.

This area should also be understood that the displacement of Similar amino acids can effectively be carried out based on hydrophily, particularly when resulting biological function protein or peptide are intended in immunological embodiments in use, as in this case.In certain embodiments, it is as by it, adjacent to being determined the hydrophily of amino acid, the maximum local average hydrophilicity of protein is related to its immunogenicity and antigenicity, i.e., related to the biological property of polypeptide.

Following hydrophilicity values are specified to these amino acid residues：Arginine (+3.0)；Lysine (+3.0)；Aspartic acid (+3.0 ± 1)；Glutamic acid (+3.0 ± 1)；Serine (+0.3)；Asparagine (+0.2)；Glutamine (+0.2)；Glycine (0)；Threonine (- 0.4)；Proline (- 0.5 ± 1)；Alanine (- 0.5)；Histidine (- 0.5)；Cysteine (- 1.0)；Methionine (- 1.3)；Valine (- 1.5)；Leucine (- 1.8)；Isoleucine (- 1.8)；Tyrosine (- 2.3)；Phenylalanine (- 2.5) and tryptophan (- 3.4).In the change based on similar hydrophilicity value progress, in certain embodiments, include amino acid replacement of its hydrophilicity value in ± 2.In certain embodiments, it is included in those in ± 1, and in certain embodiments, is included in those in ± 0.5.In some cases, it is also based on epitope of the hydrophily identification from primary amino acid sequence.These regions are also referred to as " epitopic core regions ".

Exemplary amino acid displacement is listed in table 1.

Table 1：Amino acid replacement

Original Residue	Exemplary permutation	More specifically exemplary permutation
Original Residue	Exemplary permutation	More specifically exemplary permutation	Ala	Val, Leu, Ile	Val
Arg	Lys, Gln, Asn	Lys	Ala	Val, Leu, Ile	Val
Arg	Lys, Gln, Asn	Lys	Asn	Gln	Gln
Asp	Glu	Glu	Asn	Gln	Gln
Asp	Glu	Glu	Cys	Ser, Ala	Ser
Gln	Asn	Asn	Cys	Ser, Ala	Ser
Gln	Asn	Asn	Glu	Asp	Asp
Gly	Pro, Ala	Ala	Glu	Asp	Asp
Gly	Pro, Ala	Ala	His	Asn, Gln, Lys, Arg	Arg
Ile	Leu, Val, Met, Ala, Phe, nor-leucine	Leu	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 permutation	More specifically exemplary permutation
Original Residue	Exemplary permutation	More specifically exemplary permutation	Lys	Arg, Isosorbide-5-Nitrae diaminourea-butyric acid, Gln, Asn	Arg
Met	Leu, Phe, Ile	Leu	Lys	Arg, Isosorbide-5-Nitrae diaminourea-butyric acid, Gln, Asn	Arg
Met	Leu, Phe, Ile	Leu	Phe	Leu, Val, Ile, Ala, Tyr	Leu
Pro	Ala	Gly	Phe	Leu, Val, Ile, Ala, Tyr	Leu
Pro	Ala	Gly	Ser	Thr, Ala, Cys	Thr
Thr	Ser	Ser	Ser	Thr, Ala, Cys	Thr
Thr	Ser	Ser	Trp	Tyr, Phe	Tyr
Tyr	Trp, Phe, Thr, Ser	Phe	Trp	Tyr, Phe	Tyr
Tyr	Trp, Phe, Thr, Ser	Phe	Val	Ile, Met, Leu, Phe, Ala, nor-leucine	Leu

Similarly, as it is used herein, unless otherwise indicated, the left distal end of single stranded polynucleotide sequence is 5 ' ends；The left direction of double-stranded polynucleotide sequence is referred to as 5 ' directions.The 5 ' of nascent RNA transcript to 3 ' addition directions are referred to herein as transcriptional orientation；Have on DNA and RNA identicals sequence and be that the sequence area of 5 ' to 5 ' ends of RNA transcript is referred to herein as " upstream sequence "；Have on DNA and RNA identicals sequence and be that the sequence area of 3 ' to 3 ' ends of RNA transcript is referred to herein as " downstream sequence ".

In certain embodiments, conservative amino acid replacement includes non-naturally occurring amino acid residue, and it is introduced typically by chemical peptide symthesis rather than by synthesis in biosystem.Those non-naturally occurring amino acid residues include but is not limited to, the amino acid moiety of peptidomimetic and other reverses or reverse form.

Technical staff is possible to determine the suitable displacement variant of reference polypeptide as described herein using widely-known technique.In certain embodiments, it is considered as by targeting to active unessential target region, those skilled in the art can identify the suitable molecular domains that can be changed without destroying activity.In certain embodiments, it can identify in similar polypeptide it is conservative residue and molecular moiety.In certain embodiments, conservative amino acid replacement can even be implemented on such region without destroying bioactivity or polypeptide structure can not adversely be influenceed, the region may be important to bioactivity, include but is not limited to, the CDRs of antibody, or may be important to structure.

In addition, in certain embodiments, those skilled in the art can be summarized in structure function research identification similar polypeptide to activity and/or the important residue of structure.In view of such comparison, in certain embodiments, the importance to the amino acid residue of activity or the important amino acid residue of structure in polypeptide in correspondence similar polypeptide can be predicted.In certain embodiments, technical staff can select the important amino acid residue of the such prediction of amino acid replacement being chemically similar.

In certain embodiments, technical staff can also analyze the three-dimensional structure and amino acid sequence relative to that structure in similar polypeptide.In view of this type of information, technical staff can predict that the amino acid residue of the antibody for its three-dimensional structure is compared.In certain embodiments, technical staff can be selected not to predicting that the amino acid residue on protein surface carries out radical change, because such residue may relate to the important interaction with other molecules.In addition, in certain embodiments, technical staff can occur in the test variant replaced at each required amino acid residue comprising single amino acids.In certain embodiments, activation measurement screening variant well known by persons skilled in the art can then be used.For example, in certain embodiments, the ability screening variant that can be combined with regard to it with TR-2.In certain embodiments, such variant can be used for collecting the information on suitable variant.For example, in certain embodiments, if it find that the change for particular amino acid residue causes destruction, unwelcomely reduce or inappropriate activity, then the variant with such change can be avoided.In other words, based on the information collected by this class normal experiment, technical staff can readily determine that wherein individually or with other mutation combinations, further replace the amino acid that should be avoided by.

Many scientific publications have been working on the prediction of secondary structure.Referring to Moult J., Curr.Op.in Biotech., 7 (4)：422-427 (1996), Chou et al., Biochemistry, 13 (2)：222-245(1974)；Chou et al., Biochemistry, 113 (2)：211-222(1974)；Chou et al., Adv.Enzymol.Relat.Areas Mol.Biol., 47：45-148(1978)；Chou et al., Ann.Rev.Biochem., 47：251-276 and Chou et al., Biophys.J, 26：367-384(1979).In addition, computer program is presently available for aid forecasting secondary structure.Predict that a kind of method of secondary structure is based on homology modeled.For example, 2 kinds of polypeptides or protein with the sequence identity more than 30% or the similitude more than 40% generally have similar topology.The recent development of Protein structure databases (PDB) has been provided for enhanced secondary structure predictability, including possible folding number in polypeptide or protein structure.Referring to Holm et al., Nucl.Acid.Res., 27 (1)：244-247(1999).Have been proposed in and there is a limited number of folding in given polypeptide or protein, and critical number structure solved after, structure prediction will be apparent more accurate.See, e.g., Brenner et al., Curr.Op.Struct.Biol., 7 (3)：369-376(1997).

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

In certain embodiments, the homogeneity and similitude of related polypeptide can be easily computed by known method.Such method includes but is not limited to, those described in following bibliography：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 Griffin, 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 Carillo et al., SIAM J.Applied Math., 48：1073(1988).In certain embodiments, polypeptide has and the amino acid sequence being equal of the amino acid sequence shown in Fig. 3-19 about 90% or about 95% or about 96% or about 97% or about 98% or about 99%.

In certain embodiments, the method for determining homogeneity is designed to produce the matching of maximum between the sequence of test.In certain embodiments, some methods for determining homogeneity are described in publicly available computer program.The some computer programs for determining the homogeneity between 2 sequences include but is not limited to, GCG program bags, including GAP (Devereux et al., Nucl.Acid.Res., 12：387(1984)；Genetics Computer Group, Universityof Wisconsin, Madison, WI, BLASTP, BLASTN and FASTA (Altschul et al., J.Mol.Biol., 215：403-410(1990)).BLASTX programs can disclose acquisition (BLAST Manual from US National Bioinformatics Institute (National Center for Biotechnology Information) (NCBI) and other resources, Altschul et al., NCB/NLM/NIH Bethesda, MD 20894；Altschul et al., ibid (1990)).In certain embodiments, Smith Waterman algorithms known in the art can also be used for determining homogeneity.

Some alignment schemes for comparing 2 amino acid sequences can only produce the short Region Matching of 2 sequences, and this small comparison area can have very high sequence identity, although not associated significantly between 2 full length sequences.Therefore, in certain embodiments, selected Comparison Method (GAP programs) will cause the comparison across at least 50 adjacent amino acid of target polypeptide.

For example, use computerized algorithm GAP (Genetics Computer Group, Universityof Wisconsin, Madison, WI), 2 polypeptides of Percentage of sequence identity to be determined are compared for the best match (" matched spans ", as determined by algorithm) of the amino acid of its difference.In certain embodiments, (this is calculated as 3X average diagonal lines to gap open penalty；" average diagonal line " is the diagonal averages of the comparator matrix used；" diagonal " is the score or number specified by specific comparator matrix to each perfect amino acid match) it is used together with gap extension penalty (this be typically 1/10 multiply gap open penalty) and comparator matrix such as PAM 250 or BLOSUM 62 with algorithm.In certain embodiments, standard comparing matrix is also used by algorithm.For the comparator matrixs of PAM 250 see, e.g., Dayhoff et al., Atlasof Protein Sequence and Structure, 5 (3) (1978), for the comparator matrixs of BLOSUM 62 referring to Henikoff et al., Proc.Natl.Acad.Sci USA, 89：10915-10919(1992).

In certain embodiments, include on the parameter that peptide sequence compares following：

Algorithm：Needleman et al., J.Mol.Biol., 48：443-453(1970)；

Comparator matrix：BLOSUM 62 from Henikoff et al., ibid (1992)；

Gap Penalty：12

Gap Length Penalty：4

Similarity threshold：0

In certain embodiments, GAP programs can be used together with above-mentioned parameter.In certain embodiments, above-mentioned parameter is to be used for the default parameter that polypeptide compares (together with for no penalty for end gaps) using GAP algorithms.

According to some embodiments, amino acid replacement be it is following those：(1) sensitiveness to proteolysis is reduced, (2) sensitiveness to oxidation is reduced, (3) binding affinity to protein complex formation is changed, (4) binding affinity is changed, and/or the physical chemistry or functional characteristic for such polypeptide are given or modified in (4).According to some embodiments, can be in naturally occurring sequence (in certain embodiments, in the overseas polypeptide portion of the structure for forming intermolecular contacts) carry out single or multiple amino acid replacements (in certain embodiments, conservative amino acid replacement).

In certain embodiments, conservative amino acid replacement typically can not substantially change the architectural feature of parental array (for example, spiral present in destruction parental array should not be tended to by replacing amino acid, or rupture characterizes the other kinds of secondary structure of parental array).Two grades of generally acknowledged polypeptide and the example of tertiary structure are described in following bibliography：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 Thornton et al., Nature 354：105(1991).

As it is used herein, term " polypeptide fragment " refers to the polypeptide with amino terminal and/or carboxyl terminal deletion.In certain embodiments, fragment length is at least 5-500 amino acid.It should 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 analogues are commonly used in pharmaceutical industries with the non-peptide drugs for being similar to the template peptide characteristic of those.The non-peptide compound of these types is referred to as " peptide mimics " or " peptidomimetic ".Fauchere, J.Adv.Drug Res.15：29(1986)；Veber and Freidinger TINS pages 392 (1985)；With Evans et al., J.Med.Chem.30：1229(1987).Molecule modeling generally by means of computerization develops such compound.The peptide mimics for being structurally similar to the upper useful peptide for the treatment of can be used for producing similar treatment or prevention effect.Usually, peptidomimetic is structurally similar to paradigm polypeptide (i.e., polypeptide with biochemical characteristic or pharmacological activity) such as human antibody, but with one or more peptide bonds by method well-known in the art optionally by being replaced selected from following keys：--CH₂NH--、--CH₂S--、--CH₂-CH₂--, -- CH=CH- (cis and trans), -- COCH₂--、--CH(OH)CH₂- and -- CH₂SO--.In certain embodiments, the D- amino acid system of same type can be used to replace one or more amino acid (for example D-Lys replace 1B) of consensus sequence to produce more stable peptide.In addition, the constraint peptide comprising consensus sequence or substantially the same consensus sequence variant can typically pass through methods known in the art (Rizo and Gierasch Ann.Rev.Biochem.61：387 (1992)) produce；For example, and be not limited to, by addition can be formed make peptide be cyclized intramolecular disulfide bridge internal cysteine residues.

Term " specific-binding agent " refers to the natural or non-native molecules combined with target-specific.The example of specific-binding agent includes but is 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 antigen binding regions.

Term " specific binding " refers to the affinity that specific-binding agent combined with target and is more than its ability with non-target.In certain embodiments, specific binding refer to the affinity that is combined with target than the affinity for non-target greatly at least 10,50,100,250,500 or 1000 times.In certain embodiments, affinity is determined by affinity ELISA determination methods.In certain embodiments, affinity is determined by BIAcore determination methods.In certain embodiments, affinity is determined by dynamic method.In certain embodiments, affinity is determined by balance/dissolution method.

Term " specific-binding agent for being directed to TR-2 " refers to specific binding TR-2 any portion of specific-binding agent.In certain embodiments, the specific-binding agent for TR-2 is the antibody for TR-2.In certain embodiments, specific-binding agent is antigen binding regions.

" antibody " or " antibody peptide " refers to complete antibody or its fragment.In certain embodiments, antibody fragment can be the binding fragment that specific binding is competed with complete antibody.Term " antibody " also includes polyclonal antibody and monoclonal antibody.In certain embodiments, binding fragment is produced by recombinant DNA technology.In certain embodiments, binding fragment is produced by the enzymatic or chemical cleavage of complete antibody.In certain embodiments, binding fragment is produced by recombinant DNA technologyRepeat.Binding fragment includes but is not limited to, Fab, Fab ', F (ab ') 2, Fv and single-chain antibody.Non- antigen-binding fragment includes but is not limited to, Fc fragments.In certain embodiments, antibody is combined with epitope specificity, and the epitope is by selected from following at least one antibody specificity combinations：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 includes the anti-unique antibody specifically bound with the variable region of another antibody.In certain embodiments, the variable region of anti-unique antibody and anti-TR-2 antibody is specifically bound.In certain embodiments, anti-unique antibody can be used for the activity of the presence for detecting specific anti-TR-2 antibody in sample or the anti-TR-2 antibody of blocking.

As it is used herein, term " anti-TR-2 antibody " means the antibody specifically bound with TR-2.In certain embodiments, anti-TR-2 antibody is combined with TR-2 epitope specificities, the TR-2 epitopes and at least one antibody binding selected from Ab A to Q.In various embodiments, TR-2 can be any kind of TR-2, include but is not limited to, people, machin, mouse and rabbit.Some determination methods for determining antibody specificity are that technical staff is well-known, and include but is not limited to, ELISA, ELISPOT, Western blotting, BIAcore determination methods, solution affinity binding assay, T cell costimulation determination method and T cell migration assay.

It is as used herein, term " separation antibody " means such antibody, its (1) is without it generally by least some of protein being found therewith, (2) substantially free of other protein from identical source for example from identical type, (3) by being not present from the expression of different types of cell, or (4) in nature.

" polyclonal antibody " refers to the heterogeneous mixture of the antibody combined with the different epitopes of same antigen to term.

" monoclonal antibody " refers to by the collection of antibodies of identical nucleic acid molecule encoding term.In certain embodiments, monoclonal antibody is produced by single hybridoma or other cell lines or by transgene mammal.Monoclonal antibody typically recognizes same epitope.Term " monoclonal " is not limited to use in any specific method for preparing antibody.

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

Term " polyspecific " refers to the antibody that wherein 2 or more variable regions are combined with different epitopes.Epitope can be on identical or different target.In certain embodiments, multi-specificity antibody is " bispecific antibody " of 2 kinds of different epitopes on the identical or different antigen of identification.

Term " catalytic antibody " refers to the antibody for wherein adhering to one or more catalysed partials.In certain embodiments, catalytic antibody is the cytotoxic antibody for including cytotoxic moieties.

Term " humanized antibody " refers to all or part of derived from human of wherein antibody framework, but all or part of antibody derived from another species such as mouse of one or more CDR regions.

Term " human antibody " and " human antibody " are used interchangeably, and refer to wherein CDR and framework basically comprises the antibody of human sequence.In certain embodiments, human antibody is produced in non-human mammal, is included but is not limited to, mouse, rat and Lagomorph.In certain embodiments, human antibody is produced in hybridoma.In certain embodiments, human antibody restructuring is produced.

In certain embodiments, anti-TR-2 antibody is included：

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

" chimeric antibody " refers to the antibody variable region with first species merged with the antibody constant region of for example other second species of another molecule.See, e.g. U.S. Patent number 4,816,567 and Morrison et al., Proc Natl Acad Sci (USA), 81：6851-6855(1985).In certain embodiments, first species can be differently configured from second species.In certain embodiments, first species can be identical with second species.In certain embodiments, chimeric antibody can be prepared by mutagenesis or CDR transplanting with matching the known array part of anti-TR-2 antibody variable regions.CDR transplanting relates generally to be transplanted on the framework region of another antibody (FRs) from the CDRs with required specific antibody.

In certain embodiments, the bivalent antibody in addition to " polyspecific " or " multifunctionality " antibody is generally understood that each of which binding site is identical.

When excessive antibodies make with the amount of the acceptor of ligand binding reduce at least about 20%, 40%, 60%, 80%, 85% or more (as measured in competition binding determination method in vitro) when, antibody substantially suppresses the attachment of part and acceptor.

Term " epitope " refers to the molecular moiety combined by specific-binding agent.Exemplary epitope can include any polypeptide determinant that can be specifically bound with immunoglobulin and/or φt cell receptor.Exemplary Epitopic determinants include but is not limited to, the chemically reactive surface packet of molecule, such as, but not limited to amino acid, sugared side chain, phosphoryl and sulfonyl.In certain embodiments, Epitopic determinants can have specific Three Dimensions Structure, and/or specific charge characteristic.In certain embodiments, epitope is the antigenic domains by antibody binding.Epitope can be adjacent or non-adjacent.In certain embodiments, epitope can be simulation, because they include such three-dimensional structure, it is similar to the epitope for being used for producing antibody, but be not included in for produce antibody that epitope in the amino acid residue that finds, or be contained only in for produce antibody that epitope in some amino acid residues for finding.

Term " suppressing and/or neutralizing epitope " refers to when being combined by specific-binding agent, causes the epitope of the active reduction of internal, external and/or biology in situ.In certain embodiments, neutralizing epitope is located on the biological activity region of target or combined with the biological activity region of target.

Term " activation epitope " refers to when being combined by specific-binding agent, causes the epitope that internal, external and/or biology in situ is activated or maintains.In certain embodiments, activation epitope is located on the biological activity region of target or combined with the biological activity region of target.

In certain embodiments, epitope is by selected from following at least one antibody specificity combinations：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 such embodiments, epitope is substantially purified.In some such embodiments, epitope concentration is at least 1nM.In some such embodiments, epitope concentration is 1nM-5nM.In some such embodiments, epitope concentration is 5nM-10nM.In some such embodiments, epitope concentration is at least 10nM-15nM.

In certain embodiments, antibody is combined with epitope specificity, and the epitope is by selected from following at least one antibody specificity combinations：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 substantially purify.In some such embodiments, antibody concentration is at least 1nM.In some such embodiments, antibody concentration is 1nM-5nM.In some such embodiments, antibody concentration is 5nM-10nM.In some such embodiments, antibody concentration is at least 10nM-15nM.

In certain embodiments, the amino acid/11-85 of antibody and ripe people TR-2 is specifically bound, and is substantially purified.In some such embodiments, antibody concentration is at least 1nM.In some such embodiments, antibody concentration is 1nM-5nM.In some such embodiments, antibody concentration is 5nM-10nM.In some such embodiments, antibody concentration is at least 10nM-15nM.

In certain embodiments, antibody is with being selected from following at least one antibody competition combination epitopes：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 substantially purified.In some such embodiments, antibody concentration is at least 1nM.In some such embodiments, antibody concentration is 1nM-5nM.In some such embodiments, antibody concentration is 5nM-10nM.In some such embodiments, antibody concentration is at least 10nM-15nM.

In certain embodiments, people TR-2 of the antibody with being combined maturation selected from following at least one antibody competitions amino acid/11-85：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 substantially purified.In some such embodiments, antibody concentration is at least 1nM.In some such embodiments, antibody concentration is 1nM-5nM.In some such embodiments, antibody concentration is 5nM-10nM.In some such embodiments, antibody concentration is at least 10nM-15nM.

Term " reagent " is used herein to mean that chemical compound, the mixture of chemical compound, large biological molecule or the extract prepared by biomaterial.

As it is used herein, term " mark " refers to any molecule that can be detected.In certain embodiments, antibody can be marked by mixing radiolabeled amino acid.In certain embodiments, the biotin moiety that can be detected by the avidin (for example, including the fluorescence labeling that can be detected by optics or colorimetric method or the Streptavidin of enzymatic activity) of mark can adhere to antibody.In certain embodiments, can will be in another reagent of mark incorporation, or mark is adhered to another reagent, another reagent successively with purpose antibody binding.In certain embodiments, mark can be mixed in antibody, or mark is adhered to antibody, the antibody is combined with purpose antibody specificity successively.In certain embodiments, label or tag can also be treatment.The various methods of labeling polypeptide and glycoprotein are known in the art and can used.The mark of some general classes includes but is not limited to, enzymatic, fluorescence, chemiluminescence and radioactive label.Some mark examples on polypeptide include but is not limited to, following：Radio isotope or radionuclide (for example,³H、¹⁴C、¹⁵N、³⁵S、⁹⁰Y、⁹⁹Tc、¹¹¹In、¹²⁵I、¹³¹I), fluorescence labeling (such as fluorescein isothiocynate (FITC), rhodamine, lanthanide series phosphor, phycoerythrin (PE)), enzymatic labelling are (for example, horseradish peroxidase, beta galactosidase, luciferase, alkaline phosphatase, glucose oxidase, glucose-6-phosphate dehydrogenase, alcohol dehydrogenase, malic dehydrogenase, penicillase, luciferaseRepeat), chemiluminescent labeling, biotinyl, and the predetermined polypeptide epitope (for example, leucine zipper pair sequences, binding site, metal binding domain, epitope tag on secondary antibodies) recognized by secondary reporter molecule.In certain embodiments, mark adheres to reduce potential steric hindrance by the spacerarm of various length.

As it is used herein, term " sample " includes but is not limited to, from being or before being any amount material.Such being includes but is not limited to, people, mouse, monkey, rat, rabbit and other animals.Such material includes but is not limited to, blood, serum, urine, cell, organ, tissue, bone, marrow, lymph node and skin.

As it is used herein, term " pharmaceutical agent or medicine " refers to when being suitably applied to patient, the chemical compound or composition of response to treatment needed for can inducing.

As it is used herein, term " conditioning agent " is transformation or the compound for changing molecular activity or function.For example, being compared with the activity or function magnitude observed in the case of in the absence of conditioning agent, conditioning agent can cause increasing or decreasing in some activity or function magnitude of molecule.In certain embodiments, conditioning agent is the inhibitor of at least one activity or function magnitude that reduce molecule.The some Exemplary actives and function of molecule include but is not limited to, binding affinity, enzymatic activity and signal transduction.Some exemplary inhibitor include but is not limited to, protein, peptide, antibody, peptibody (peptibodies), carbohydrate and organic molecule.Exemplary peptibody is described in such as WO 01/83525.

As it is used herein, " substantially purifying " means that purpose species is present dominant species (that is, it is more more rich than any other individual species in composition on a molar basis).In certain embodiments, the part substantially purified is the composition of wherein at least about 50% (on a molar basis) of the purpose species comprising all macromolecular species existed.In certain embodiments, the composition substantially purified will be more than about 80%, 85%, 90%, 95% or 99% comprising all macromolecular species present in composition.In certain embodiments, purpose species is purified to the basic homogeney (can not detect contamination class in composition by common detection methods) that wherein composition is substantially made up of single macromolecular species.

Term " patient " includes humans and animals subject.

According to some embodiments, there is provided the cell line for expressing anti-TR-2 antibody.

There is provided the chimeric antibody comprising at least part human sequence He the sequence of another species in certain embodiments.In certain embodiments, the antibody with the antibody sequence without host is compared, and such chimeric antibody can produce reduced immune response in host.For example, in some cases, purpose animal may be used as the model on specific human disease.In order to study influence of the antibody to that disease in animal reservoir, it can use from different types of antibody.But, in some cases, this antibody-like from another species can trigger for the immune response of itself of the antibody in host animal, so as to hinder the assessment of these antibody.In certain embodiments, the amino acid sequence for replacing the anti-TR-2 antibody in part with the antibody amino acids sequence from host animal can reduce the magnitude of the antiantibody response of host animal.

In certain embodiments, chimeric antibody includes heavy chain and light chain, and wherein the variable region of light chain and heavy chain is from first species, and the constant region of light chain and heavy chain comes from second species.In certain embodiments, heavy chain constant region is the heavy chain constant region of the species in addition to people.In certain embodiments, antibody light chain constant region is the antibody light chain constant region of the species in addition to people.In certain embodiments, heavy chain constant region is human antibody heavy chain's constant region, and heavy chain of antibody variable region is the heavy chain of antibody variable region of the species in addition to people.In certain embodiments, antibody light chain constant region is human antibody light chain constant region, and antibody light chain variable region is the antibody light chain variable region of the species in addition to people.Exemplary antibodies constant region includes but is not limited to, human antibody constant region, cynomolgus monkey constant region, mouse antibodies constant region and rabbit antibody constant region.Exemplary antibodies variable region includes but is not limited to, human antibody variable region, mouse antibody variable region, pig antibody variable region, guinea pig antibodies variable region, cynomolgus monkey variable region and rabbit antibody variable region.In certain embodiments, the framework region of variable region can use the framework region derived from other antibody sequences to replace in heavy chain and light chain.

Some exemplary chimeric antibody can be produced by the well-known method of those of ordinary skill in the art.In certain embodiments, can merge the polynucleotides of first species of encoding heavy chain variable region and the polynucleotides of second species of encoding heavy chain constant.In certain embodiments, the nucleotide sequence fusion of the polynucleotides of first species of coding light chain variable region and second species of coding constant region of light chain can be made.In certain embodiments, the nucleotides sequence of these fusions can be listed in single expression vector (for example, plasmid) or multiple expression vectors and introduces intracellular.In certain embodiments, the cell comprising at least one expression vector can be used to prepare polypeptide.In certain embodiments, the nucleotides sequence of these fusions can be listed in separated expression vector or single expression vector and introduces intracellular.In certain embodiments, host cell expression heavy chain and light chain, the heavy chain and light chain are combined to produce antibody.In certain embodiments, the cell comprising at least one expression vector can be used to prepare antibody.Illustrative methods for producing and expressing antibody are discussed below.

In certain embodiments, for anti-TR-2 antibody the conservative modification (and corresponding modification for being directed to coding nucleotide) of weight and light chain will be produced with the function of those and the antibody of chemical feature similar to original antibodies.By contrast, in certain embodiments, the displacement in the amino acid sequence that basic modification in the function and/or chemical feature of anti-TR-2 antibody can be by selecting weight and light chain be completed, and the displacement is dramatically different in its influence to following maintenances：(a) molecule charge or hydrophobicity at the molecular backbone structure in replacement areas such as such as folding or helical conformation, (b) target site, or (c) side-chain bulk.

Amino acid replacement needed for some (either guarding or non-conservative) can be determined by those skilled in the art when needing such displacement.In certain embodiments, amino acid replacement can be used for the important residue for identifying anti-TR-2 antibody, can for example increase or decrease for TR-2 antibody affinity or antibody effector function those.

In certain embodiments, reduction in the amount that the effect of anti-TR-2 antibody can be by measuring disease symptomses is assessed.In certain embodiments, purpose disease can be caused by pathogen.In certain embodiments, disease can be set up by other method in animal reservoir, and methods described includes material (such as carcinogenic substance) and introduced and genetic manipulation.In certain embodiments, effect is assessed by detecting one or more adverse events in animal reservoir.Term " adverse events " includes but is not limited to, and receives the adverse effect in the animal reservoir of antibody, and the adverse effect is non-existent in the animal reservoir for do not receive the antibody.In certain embodiments, adverse events include but is not limited to, have a fever, for antibody immune response, inflammation and/or animal reservoir death.

The various antibody of antigen-specific can be produced in many ways.In certain embodiments, the antigen comprising purpose epitope can be introduced in animal reservoir (for example, mouse), so as to produce the antibody special to that epitope.In some cases, the antibody special to purpose epitope can be obtained from biological sample, and the biological sample is derived from Natural Exposure in the host of epitope.In some cases, by there is provided the chance for obtaining human monoclonal antibodies (MAbs) in human immunoglobulin(HIg) (Ig) locus introducing mouse that wherein endogenous Ig gene has been inactivated.

Naturally occurring antibody structure

Naturally occurring antibody structural unit generally comprises tetramer.Each such tetramer is typically made up of 2 pairs of identical polypeptide chains, and each pair has total length " light " chain (in certain embodiments, about 25kDa) and total length " weight " chain (in certain embodiments, about 50-70kDa).Term " heavy chain " includes that there are enough variable region sequences to give specific any polypeptide to specific antigen.Total length heavy chain includes Variable domain V_H, and 3 constant region domain C _H1、C _H2 and C_H3。V_HDomain is located at the amino terminal of polypeptide, and C _H3 domains are located at carboxyl terminal.As it is used herein, term " heavy chain " includes full length antibody heavy chain and its fragment.

Term " light chain " includes that there are enough variable region sequences to give specific any polypeptide to specific antigen.Full-length light chains include Variable domain V_L, and constant region domain C_L.Such as heavy chain, the Variable domain of light chain is located at the amino terminal of polypeptide.As it is used herein, term " light chain " includes full length antibody light chain and its fragment.

The amino terminus portion of every chain includes being generally responsible for the variable region (V in heavy chain of the about 100-110 of antigen recognizing or more amino acid_HAnd light chain in V_L).The carboxyl terminal of every chain, which is typically limited, may be responsible for the constant region (C in heavy chain of effector function_HAnd light chain in C_L).Antibody mediated effect effect includes activating complement and stimulates conditioning phagocytosis.People's light chain is generally classified as κ and lambda light chain.Heavy chain is generally classified as μ, δ, γ, α or ε, and it is respectively IgM, IgD, IgG, IgA and IgE that the isotype of antibody, which is limited,.IgG has several subclass, includes but is not limited to, IgG1, IgG2, IgG3 and IgG4.There is IgM subclass to include but is not limited to, IgM1 and IgM2.IgA is similarly subdivided into subclass and included but is not limited to, IgA1 and IgA2.In total length gently and in heavy chain, general " J " area by about 12 or more amino acid of variable and constant region is connected, and heavy chain also includes " D " area of about 10 or more amino acid.See, e.g., the chapters of Fundamental Immunology the 7th (Paul, W., editor, second edition, Raven Press, N.Y (1989)).The variable region of each light/heavy chain pair generally forms antigen binding site.

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

As discussed above, there is the antibody fragment of several types.Fab fragments are by a light chain and the C of a heavy chain _H1 and variable region composition.The heavy chain of Fab molecules can not be with another heavy chain molecule formation disulfide bond.Fab ' fragments include a light chain and a heavy chain, and the heavy chain is included in C _H1 and C_HMore constant regions between 2 domains, so that interchain disulfide bond can be formed between 2 heavy chains to form the molecules of F (ab ') 2.Fab fragments are similar to the molecules of F (ab ') 2, except the constant region in molecule heavy chain extends to C_HOutside 2 domain ends.Fv areas include the variable region to conduct oneself with dignity with light chain, but lack constant region.Single-chain antibody is such Fv molecules, wherein heavy and light chain variable district is attached to form wall scroll polypeptide chain, the polypeptide chain formation antigen binding regions by nonrigid connector.Exemplary single-chain antibody is discussed in detail in such as WO 88/01649 and U.S. Patent number 4,946,778 and 5,260,203.Fc fragments include the C of heavy chain _H2 and C _H3 domains, and included in C _H1 and C_HMore constant regions between 2 domains, so that interchain disulfide bond can be formed between 2 heavy chains.

In certain embodiments, functional domain C _H1、C _H2、C _H3 and insetion sequence can be reorganized to produce different antibody constant regions.For example, in certain embodiments, assembling and folding and/or optimized for improved effector function that such hybrid constant region can be for the half-life period in serum, for antibody tetramer.In certain embodiments, the antibody constant region of modification can be produced by following methods：In the amino acid sequence that single point mutation is introduced to constant region, and the antibody obtained by one or more tests in those is for example listed above in the property just improved.

In certain embodiments, a kind of antibody of isotype is transformed into by different isotypes by isotype conversion, without losing its specificity to certain target molecules.The method of isotype conversion includes but is not limited in other, direct recombinant technique (see, for example, U.S. Patent number 4,816,397) and cell-cell fusion techniques (see, for example, U.S. Patent number 5,916,771).In certain embodiments, antibody from a subclass can be transformed into another subclass using technique described above or other modes known in the art, without losing its specificity to certain target molecules, include but is not limited to, IgG1, IgG3 or IgG4 subclass are transformed into from IgG2 subclass.

Bispecific or bifunctional antibody

Bispecific or bifunctional antibody are usually the artificial hybrid antibody with 2 different weight/light chains pair binding site different with 2.Bispecific antibody can be produced by various methods, be included but is not limited to, hybridoma fusion or the connection of Fab ' fragments.See, e.g., Songsivilai and Lachmann Clin.Exp.Immunol.79：315-321 (1990), Kostelny et al., J.Immunol.148：1547-1553(1992).

Some preparations of antibody

In certain embodiments, antibody can be expressed in the cell line in addition to hybridoma cell line.In certain embodiments, sequence of the encoding particular antibodies including chimeric antibody can be used for converting suitable mammalian host cell.According to some embodiments, conversion can pass through any known method for polynucleotides to be introduced in host cell, including, for example polynucleotides are packaged into virus (or in viral vector), and with viral transduction host cell or by using operation transfection carrier known in the art, such as by U.S. Patent number 4,399,216；4,912,040；4,740,461；Illustrated with 4,959,455.

In certain embodiments, expression vector includes any polynucleotide sequence being discussed herein.In certain embodiments there is provided the method for preparing polypeptide, methods described, which is included in, to be suitable for the polynucleotides that wherein include and expresses that under conditions of producing polypeptide, polypeptide is produced in the cell comprising any of above expression vector.

In certain embodiments, expression vector includes polynucleotides, the polynucleotides include the sequence of coded polypeptide, the polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a, wherein CDR1a includes amino acid 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；Wherein CDR2a includes amino acid 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 tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；The polypeptide combination TR-2 wherein combined with antibody light chain.In certain embodiments there is provided the method for preparing polypeptide, methods described, which is included in, to be suitable for the polynucleotides that wherein include and expresses that under conditions of producing polypeptide, polypeptide is produced in the cell comprising above-mentioned expression vector.

In certain embodiments, expression vector includes polynucleotides, the polynucleotides include the sequence of coded polypeptide, the polypeptide includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b, wherein CDR1b includes amino acid 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 lysine；Amino acid b1 is selected from threonine, alanine 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 valine；Amino acid g1 is selected from serine, leucine or arginine；Amino acid h1 is selected from threonine, serine, isoleucine, asparagine, arginine, histidine or tyrosine；Amino acid i1 is selected from tyrosine, arginine, tryptophan, aspartic acid or serine；J1 is selected from leucine, isoleucine, asparagine, tyrosine or serine；Amino acid k1 is selected from asparagine, glycine, valine, alanine or leucine；Amino acid l1 is selected from tyrosine, alanine or asparagine or is not present；Amino acid m1 is selected from asparagine or lysine or is not present；Amino acid n1 is selected from tyrosine, asparagine or isoleucine or is not present；Amino acid o1 is selected from leucine or tyrosine or is not present；Amino acid p1 is selected from aspartic acid or leucine or is not present；Valine, alanine or threonine are selected from amino acid q1 or are not present；Wherein CDR2b includes amino acid sequence r1 s1 t1 u1 v1 w1 x1, and wherein amino acid r1 is selected from alanine, aspartic acid, leucine, tryptophan, glycine or valine；Amino acid s1 is selected from threonine, valine, glycine or alanine；Amino acid t1 is serine；Amino acid u1 is selected from serine, asparagine or threonine；Amino acid v1 is selected from leucine, phenylalanine or arginine；Amino acid w1 is selected from glutamine, alanine or glutamic acid；Serine, arginine or threonine are selected from amino acid x1；Wherein CDR3b includes amino acid sequence y1 z1 a1 ' b1 ' c1 ' d1 ' e1 ' f1 ' g1 ', and wherein amino acid y1 is selected from glutamine, methionine, leucine or histidine；Amino acid z1 is selected from glutamine or lysine；Amino acid a1 ' is selected from serine, threonine, alanine, histidine, tyrosine or phenylalanine；Amino acid b1 ' is selected from tyrosine, leucine, asparagine or glycine；Amino acid c1 ' is selected from serine, glutamine, isoleucine or lysine；Amino acid d1 ' is selected from threonine, phenylalanine, tyrosine, alanine or serine；Amino acid e1 ' is proline；Amino acid f1 ' is selected from leucine, phenylalanine, tryptophan, serine or arginine；Threonine or serine are selected from amino acid g1 '；The polypeptide combination TR-2 wherein combined with heavy chain of antibody.In certain embodiments there is provided the method for preparing polypeptide, methods described, which is included in, to be suitable for the polynucleotides that wherein include and expresses that under conditions of producing polypeptide, polypeptide is produced in the cell comprising above-mentioned expression vector.There is provided the cell for including at least one above-mentioned expression vector in certain embodiments.In certain embodiments there is provided the method for preparing polypeptide, methods described, which is included in, to be suitable for the polynucleotides that wherein include and expresses that under conditions of producing polypeptide, polypeptide is produced in the cell comprising above-mentioned expression vector.

In certain embodiments, expression vector expresses anti-TR-2 heavy chain of antibody.In certain embodiments, expression vector expresses anti-TR-2 antibody light chains.In certain embodiments, expression vector expresses anti-TR-2 heavy chain of antibody and anti-TR-2 antibody light chains.In certain embodiments there is provided the method for anti-TR-2 antibody, methods described, which is included in, to be suitable for the polynucleotides that wherein include and expresses that under conditions of producing antibody, antibody is produced in the cell comprising at least one expression vector described herein.

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

It is known in the art for can be used as some mammal cell lines of the host for expression, and include but is not limited to, the immortalized cell line that can be obtained from American type culture collection (ATCC), including but not limited to, Chinese hamster ovary (CHO) cell, E5 cells, HeLa cell, baby hamster kidney (BHK) cell, MK cells (COS), human hepatocellular carcinoma cell line (such as Hep G2), NS0 cells, SP20 cells, Per C6 cells, 293 cells and many other cell lines.In certain embodiments, there can be high expression level by determining which cell line and produce the antibody with composition antigenic binding property to select cell line.

In certain embodiments, the carrier that can be transfected in host cell includes control sequence, and the control sequence is operably connected with encoding the polynucleotides of anti-TR-2 antibody.In certain embodiments, control sequence promotes the expression of the polynucleotides of connection, so as to cause the polypeptide production of the polynucleotide encoding by connecting.In certain embodiments, the polynucleotide sequence that carrier is also replicated comprising the chromosome dependent/non-dependent allowed in host cell.Exemplary carrier includes but is not limited to, plasmid (such as BlueScript, puc), clay and YACS.

Some antibody purposes

According to some embodiments, antibody is used to detect the specific antigen in sample.In certain embodiments, this allows the cell or tissue of identification production protein.For example, in certain embodiments, anti-TR-2 antibody can be used for detecting that the TR-2 in sample is present.In certain embodiments, anti-TR-2 antibody and sample combination are made for detecting that the present or absent methods of the TR-2 in sample include (a)；(b) separate antibody and uncombined antibody with antigen binding；Detection and the existence or non-existence of the antibody of antigen binding (c).

Wherein antibody can be used for detecting that the present or absent determination method of antigen includes but is not limited to, ELISA and Western blotting.In certain embodiments, anti-TR-2 antibody can be marked.In certain embodiments, anti-TR-2 antibody can be detected by the labelled antibody with anti-TR-2 antibody bindings.There is provided for detecting the present or absent kits of the TR-2 in sample in certain embodiments.In certain embodiments, the kit includes anti-TR-2 antibody and the reagent for detecting antibody.

In certain embodiments, antibody can be used for substantially separate chemical part such as, but not limited to protein.In certain embodiments, antibody adheres to " matrix ", and the matrix is the support material for fixing antibody.Matrix includes but is not limited to, pipe, flat board (that is, porous flat plate), pearl such as microballon, filter, ball and film.In certain embodiments, matrix can be prepared by water-insoluble material, such as, but not limited to polycarbonate resin, organic siliconresin or nylon resin.Exemplary substrates for being used in affinity chromatography include but is not limited to, cellulose, agarose, polyacrylamide, glucan, polystyrene, polyvinyl alcohol and porous silicon.The chromatography matrix being obtained commercially in the presence of many, it includes but is not limited to, the Sepharose (Pharmacia) of Sepharose 2B, Sepharose4B, Sepharose 6B and other forms；Bio-Gel (and various forms of Bio-Gel such as Biogel A, P or CM), Cellex (and various forms of Cellex such as 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, including (a) for separating TR-2 method adheres to TR-2 antibody and matrix；(b) sample comprising TR-2 is made exposed to the antibody of part (a)；Separate TR-2 (c).There is provided the kit for separating TR-2 in certain embodiments.In certain embodiments, the kit is included with the anti-TR-2 antibody of matrix attachment and the reagent for separating TR-2.

It is as used herein, term " affinity chromatography " means by using the interaction between material pair (for example, affinity) purpose material in isolated or purified sample method, the material is to such as antigen and antibody, enzyme and substrate or acceptor and part.

In certain embodiments, combined with specified protein and block the antibody with the interaction of other binding compounds to have therapeutical uses.In this application, when discussing the purposes of anti-TR-2 Antybody therapies disease or symptom, such purposes can include anti-TR-2 antibody itself；Include the composition of anti-TR-2 antibody；And/or the purposes of the combination treatment comprising anti-TR-2 antibody and one or more other active components.When anti-TR-2 antibody is used for " treatment " disease or symptom, such treatment can include or not include the prevention of disease or symptom.In certain embodiments, anti-TR-2 antibody can block the interaction of TR-2 acceptors and its ligand/TRAIL.In certain embodiments, anti-TR-2 antibody can activate TR-2 acceptors.In certain embodiments, anti-TR-2 antibody can be with constitutively activated TR-2 acceptors.Because TR-2 is related to apoptosis, in certain embodiments, anti-TR-2 antibody may have therapeutical uses in the disease treatment for wherein wishing cell death or prevention cell death.Such disease includes but is not limited to, and cancer, inflammation and the virus related to any tissue for expressing TR-2 infects.

In certain embodiments, anti-TR-2 antibody is administered alone.In certain embodiments, anti-TR-2 antibody is applied before at least one other therapeutic agents are applied.In certain embodiments, anti-TR-2 antibody is applied with least one other therapeutic agents and is administered simultaneously.In certain embodiments, anti-TR-2 antibody is applied after at least one other therapeutic agents are applied.Exemplary treatment agent includes but is not limited to, at least one others cancer therapeutic agent.Exemplary cancers therapeutic agent includes but is not limited to, radiotherapy and chemotherapy.

In certain embodiments, anti-TR-2 antibody pharmaceutical compositions can be applied in combination treatment, i.e., with other agent combinations.In certain embodiments, combination treatment includes the anti-TR-2 antibody combined with least one anti-angiogenic agent.Exemplary agents include but is not limited to, external synthetically prepared Chemical composition that, antibody, antigen binding regions, radionuclide and combinations thereof and conjugate.In certain embodiments, reagent can serve as activator, antagonist, alllosteric (suspected of allosteric, allosteric) conditioning agents or toxin.In certain embodiments, reagent can be used for suppressing or stimulate its target (for example, acceptor or enzyme activition or suppression) and so as to promote cell death or suppress cell growth.

Exemplary chemical therapy for treating includes but is not limited to, and antitumor agent includes but is not limited to, and alkylating agent includes but is not limited to：Mustargen, includes but is not limited to, mechlorethamine, endoxan, ifosfamide, melphalan and Chlorambucil；Nitroso ureas, includes but is not limited to, BCNU BCNU, Luo Mositing, CCNU and Semustine, Semustine；Temodal^TM, temozolamide；Aziridine/methylmelamine, includes but is not limited to, triethylenemelamine (TEM), three ethylenes, tespamin, thiotepa, hexamethylmelamine (HMM) and hemel；Alkylsulfonate, includes but is not limited to, busulfan；Triazine, includes but is not limited to, dacarbazine (DTIC)；Antimetabolite, includes but is not limited to, folacin such as amethopterin and Trimetrexate；Pyrimidine analogue, includes but is not limited to, 5 FU 5 fluorouracil (5FU)；Floxuridine, gemcitabine, cytarabin (AraC, cytarabine), 5-azacitidine, and 2,2 '-difluoro deoxycytidine；Purine analogue, includes but is not limited to, Ismipur, 6- thioguanines, imuran, 2 '-deoxycoformycin (pentoside), red hydroxyl nonyl adenine (EHNA), fludarabine phosphate, carat Qu Bin and 2-chlorodeoxyadenosine (2-CdA)；Natural products, includes but is not limited to, anti-mitosis medicine such as taxol；Vinca alkaloids, includes but is not limited to, vincaleukoblastinum (VLB), vincristine and vinorelbine；Taxotere；Estramustine and estramustine phosphate；Ppipodophylotoxins (suspected of epipodophyllotoxins, epipodophyllotoxin), includes but is not limited to, Etoposide and Teniposide；Antibiotic, includes but is not limited to, actinomycin D, daunomycin, rubidomycin, adriamycin, mitoxantrone, darubicin, bleomycin, mithramycin, mithramycin, mitomycin C and D actinomycin D；Enzyme, includes but is not limited to, L-ASP；Biological response modifier, includes but is not limited to, interferon-' alpha ', IL-2, G-CSF and GM-CSF；Doxycyckine (suspected of doxycycline, doxycycline)；Irinotecan hydrochloride；Mix reagent, include but is not limited to, platinium (suspected of platinum, platinum) co-ordination complexs such as cis-platinum and carboplatin；Amerantrone；Including but not limited to, mitoxantrone；Urea is substituted, is included but is not limited to, hydroxycarbamide；Methyl hydrazine derivatives, include but is not limited to, N- methyl hydrazines (MIH) and procarbazine；Adrenal cortex inhibitor, includes but is not limited to, mitotane (o, p '-DDD) and aminoglutethimide；Hormone and antagonist, include but is not limited to, adrenocorticotro antagonist such as prednisone and equivalent, dexamethasone and aminoglutethimide；Gemzar^TM, gemcitabine；Progesterone, includes but is not limited to, hydroxyprogesterone caproate, Medroxyprogesterone Acetate and megestrol acetate；Estrogen, includes but is not limited to, diethylstilbestrol and ethinylestradiol equivalent；Antiestrogenic, includes but is not limited to, tamosifen；Androgen, includes but is not limited to, testosterone propionate and Fluoxymesterone/equivalent；Antiandrogen, includes but is not limited to, Drogenil, gonadotropin releasing hormone analogues and Leuprorelin；With non-steroid antiandrogen, include but is not limited to, Drogenil.

The exemplary cancers treatment that can be applied together with anti-TR-2 antibody includes but is not limited to, targeted therapy.The example of targeted therapy includes but is not limited to, the use for the treatment of antibody.Exemplary treatment antibody includes but is not limited to, mouse, mouse-human chimeric, CDR transplanting, humanization and human antibody and synthetic antibody, includes but is not limited to, those selected by screening antibodies library.Exemplary antibodies include but is not limited to, with cell surface protein Her2, CDC20, CDC33, mucin-like glycoprotein, those combined with EGF-R ELISA present on tumour cell (EGFr), and optionally induction is to the Carbazole alkaloid and/or cytotoxic effect of the tumour cell for showing these protein.Exemplary antibodies also include but is not limited to, HERCEPTIN^TM, can be used for the Herceptin for treating the cancer of breast cancer and other forms, RITUXAN^TM, Rituximab, ZEVALIN^TM, ibritumomab tiuxetan, and LYMPHOCIDE^TM, can be used for the epratuzumab for treating the cancer of non_hodgkin lymphoma and other forms, GLEEVEC^TM, can be used for the imatinib mesylate for treating chronic myeloid leukemia and gastrointestinal stromal tumor, and BEXXAR^TM, can be used for the iodine 131 tositumomab for treating non_hodgkin lymphoma.Some exemplary antibodies also include ERBITUX^TM；IMC-C225；lressa^TM；Gefitinib；TARCEVA^TM, ertinolib (suspected of erlotinib, Erlotinib)；KDR (kinase domain receptor) inhibitor；Anti-VEGF antibody and antagonist are (for example, Avastin^TMAnd VEGAF-TRAP)；Anti-vegf receptor antibody and antigen binding regions；Anti- Ang-1 and Ang-2 antibody and antigen binding regions；For Tie-2 antibody and other Ang-1 and Ang-2 acceptors；Tie-2 parts；For the antibody of Tie-2 kinase inhibitors；With

Alemtuzumab.In certain embodiments, cancer therapeutic agent is other polypeptides of the selective induction apoptosis in tumour cell, is included but is not limited to, TNF- related polypeptides such as TRAIL.

In certain embodiments, the specific-binding agent that IGF-1R Apoptosis is expressed in the combination and promotion of agonist ligand IGF-1 and/or IGF-2 and insulin-like growth factor-1 receptor (" IGF-1R ") (includes but is not limited to, anti- IGF-R1 antibody) with confrontation and so as to which the specific-binding agent for the Apoptosis for promoting to express TRAIL-R2 (includes but is not limited to, TRAIL and anti-TR-2 antibody) formulated in combination or administration.Exemplary anti-IGF-1 R antibodies are known in the art, and are disclosed in the WO 2006/069202 that on December 20th, 1 submits, and the patent is incorporated herein by reference for any purpose.

In certain embodiments, cancer therapeutic agent is the anti-angiogenic agent for reducing angiogenesis.Some such reagents include but is not limited to, ERBITUX^TM, IMC-C225；KDR (kinase domain receptor) inhibitor (for example, antibody and antigen binding regions for being specifically bound with kinase domain receptor)；Anti-vegf reagent (for example, specific binding VEGF antibody or antigen binding regions, or soluble VEGF-receptor or its ligand binding region) such as AVASTIN^TMOr VEGF-TRAP^TM；Anti-vegf receptor agents (for example, antibody or antigen binding regions for being specifically bound with it)；EGFR inhibitor (for example, antibody or antigen binding regions for being specifically bound with it) such as ABX-EGF, Victibix, IRESSA^TM, Gefitinib, TARCEVA^TM, Erlotinib, anti-Ang-1 and anti-Ang-2 reagents (for example, antibody or antigen binding regions for being specifically bound with it or with its acceptor such as Tie2/Tek)；With anti-Tie-2 kinase inhibitors (for example, antibody or antigen binding regions for being specifically bound with it).In certain embodiments, pharmaceutical composition can also include specific binding and suppress one or more reagents of growth factor activity (for example, antibody, antigen binding regions or soluble recepter), such as HGF (HGF, also referred to as dispersion factor) antagonist, and specifically bind the antibody or antigen binding regions of its acceptor " c-met ".

Exemplary anti-angiogenic agent includes but is not limited to, Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., U.S. Patent Application Publication No. 2003/0162712；U.S. Patent number 6,413,932)；Anti- TWEAK reagents (for example, specific binding antibody or antigen binding regions, or soluble T WEAK receptor antagonists；See, e.g., Wiley, U.S. Patent number 6,727,225)；The ADAM disintegrins domain (Fanslow et al., U.S. Patent Application Publication No. 2002/0042368) of the combination of antagonism integrin and its part；Specifically bind anti-eph acceptors and/or anti-ephrin antibody or antigen binding regions (U.S. Patent number 5,981,245；5,728,813；5,969,110；6,596,852；6,232,447；6,057,124；And its patent family member)；Anti- PDGF-BB antagonists are (for example, specific binding antibody or antigen binding regions) and the antibody or antigen binding regions that are combined with PDGF-BB ligand specificities, with PDGFR kinase inhibitors (for example, antibody or antigen binding regions for being specifically bound with it).

Exemplary anti-angiogenesis/antitumor agent includes but is not limited to, SF-7784 (Pfizer, USA)；Cilengitide (Merck KgaA, Germany, EPO 770622)；The sodium of piperazine Jia Tani eight (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：Anti-angiogenesis (ConjuChem, Canada)；Angiocidin (InKine Pharmaceutical, USA)；KM-2550 (Kyowa Hakko, Japan)；SU-0879 (Pfizer, USA)；CGP-79787 (Novartis, Switzerland, EP 970070)；ARGENT technology (Ariad, USA)；YIGSR-Strealth (Johnson＆Johnson, USA)；Fibrinogen-E fragments (BioActa, UK)；Angiogenesis inhibitors (Trigen, UK)；TBC-1635 (Encysive Pharmaceuticals, USA)；SC-236 (Pfizer, USA)；ABT-567 (Abbott, USA)；Metastatin (EntreMed, USA)；Angiogenesis inhibitors (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 (Fujisawa, Japan, JP 02233610)；Platelet factor 4 (RepliGen, USA, EP 407122)；Vegf antagonist (Borean, Denmark)；Temsirolimus (CCI-779) (University of South Carolina, USA)；Bevacizumab (pINN) (Genentech, USA)；Angiogenesis inhibitors (SUGEN, USA)；XL 784 (Exelixis, USA)；XL 647 (Exelixis, USA)；The integrins of 5 β of Mab, α 3, Vitaxin and second 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)；BC1 (Genoa Institute of Cancer Research, Italy)；Angiogenesis inhibitors (Alchemia, Australia)；VEGF antagonist (Regeneron, USA)；Anti-angiogenesis (XOMA, USA) derived from rBPI 21 and BPI；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, USA Universityof California)；PX 478 (Pro1X, USA)；Metastatin (EntreMed, USA)；Troponin I (Harvard University, USA)；SU 6668 (SUGEN, USA)；OXI 4503 (OXiGENE, USA)；O- guanidines (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 vaccines (EntreMed, USA)；Urokinase plasminogen activator inhibitor (Dendreon, USA)；Oglufanide (pINN) (Melmotte, USA)；HIF-1 alpha inhibitors (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)；The β 1 (Protein Design, USA) of Mab, α 5；KDR kinase inhibitors (Celltech Group, UK and Johnson＆Johnson, USA)；GFB 116 (South Florida University, USA and Yale University, USA)；CS 706 (Sankyo, Japan)；Combretastatin A4 pro-drugs (Arizona State University, USA)；Chondroitinase AC (IBEX, Cahada)；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, VEGF (Xenova, UK)；Irsogladine (INN) (Nippon Shinyaku, Japan)；RG 13577 (Aventis, France)；WX 360 (Wilex, Germany)；Squalamine (pINN) (Genaera, USA)；RPI 4610 (Sirna, USA)；Newborn fucan sulfate (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)；The antagonist of VE- cadherins -2 (ImClone Systems, USA)；Vasostatin (National Institutes of Health, USA)；Vaccine, Flk-1 (ImClone Systems, USA)；TZ 93 (Tsumura, Japan)；TumStatin (BethIsrael Hospital, USA)；The soluble FLT 1 (Vascular endothelial growth factor receptor-1) (Merck＆Co, USA) of truncation；Tie-2 parts (Regeneron, USA)；With platelet factor4 inhibitor (Allegheny Health, Education and Research Foundation, USA).

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

Bortezomib；Rapamycin；(the 8- acetyl group -7 of TAN 1120; 8; 9,10- tetrahydrochysenes -6,8; 11- trihydroxy -1- methoxyl groups -10- [[octahydro -5- hydroxyls -2-2- (hydroxypropyl)] -4; 10- dimethyl pyrans simultaneously [3,4-d] -1,3; 6- dioxa azepine Xin Ying -8- bases] epoxide) -5,12- aphthacenes diketone)；Suradista (7,7 '-[carbonyl is double [imino group (1- methyl isophthalic acid H- pyrroles -4,2- diyl) carbonylimino (1- methyl isophthalic acid H- pyrroles -4,2- diyl) carbonylimino]] double -1,3- naphthalenedisulfonic acids tetrasodium salt)；SU 302；SU 301；SU 1498 ((E) -2- cyano group -3- [double (1- Methylethyls) phenyl of 4- hydroxyls -3,5-]-N- (3- phenyl propyls) -2- acrylamides)；SU 1433 (4- (6,7- dimethyl -2- quinoxalinyls) -1,2- Benzenediols)；ST 1514；SR 25989；Soluble T ie-2；SERM derivatives, Pharmos；Semaxanib (pINN) (3- [(3,5- dimethyl -1H- pyrroles -2- bases) methylene] -1,3- dihydro -2H- indol-2-ones)；S 836；RG 8803；RESTIN；R 440 (3- (1- Methyl-1H-indole -3- bases) -4- (1- methyl -6- nitro -1H- indol-3-yls) -1H- pyrroles -2,5- diketone)；R 123942 (1- [6- (1,2,4- thiadiazoles -5- bases) -3- pyridazinyls]-N- [3- (trifluoromethyl) phenyl] -4- piperidinamines)；Prolyl hydroxylase inhibitors；Progressive raising (progression elevated) gene；Prinomastat (INN) ((S) -2,2- dimethyl -4- [[p- (4- pyridines epoxide) phenyl] sulfonyl] -3- thiomorpholine first hydroximic acid)；NV 1030；NM 3 (8- hydroxyl -6- methoxy-alpha-methyl -1- oxo -1H-2- chromene -3- acetic acid)；NF 681；NF 050；MIG；METH 2；METH 1；ManassantinB (α-[1- [4- [5- [4- [2- (3,4- Dimethoxyphenyls) -2- hydroxyl -1- methyl ethoxies] -3- methoxyphenyls] tetrahydrochysene -3,4- dimethyl -2- furyls] -2- methoxyphenoxies] ethyl] -1,3- benzodioxole -5- methanol)；KDR monoclonal antibodies；The integrin monoclonal antibodies of 5 β of α 3；LY 290293 (2- amino -4- (3- pyridine radicals) -4H- naphtho-s [1,2-b] pyrans -3- formonitrile HCNs)；KP0201448；KM 2550；Integrin specificity peptide；INGN 401；GYKI 66475；GYKI 66462；Greenstatin (101-354- plasminogens (people))；Rheumatoid arthritis, prostate cancer, oophoroma, glioma, endostatin, the gene therapy of colorectal cancer, ATF BTPI, anti-angiogenesis gene, angiogenesis inhibitors or angiogenesis；Gelatinase inhibitor, FR 111142 (4,5- dihydroxy -2- hexenoic acid 5- methoxyl groups -4- [2- methyl -3- (3- methyl-2-butenes base) Oxyranyle] -1- oxaspiros [2.5] octyl- 6- base esters)；Forfenimex (pINN) (S)-alpha-amido -3- hydroxyls -4- (hydroxymethyl) phenylacetic acid)；Fibrinogen antagonist (1- acetyl group-L- prolyl-L- histidyl--L- seryls-L- cysteinyls-altheine)；Fibroblast growth factor acceptor inhibitor；Fibroblast growth factor antagonist；FCE 27164 (7,7 '-[carbonyl is double [imino group (1- methyl isophthalic acid H- pyrroles -4,2- diyl) carbonylimino (1- methyl isophthalic acid H- pyrroles -4,2- diyl) carbonylimino]] double -1, the sodium salt of 3,5- naphthalene trisulfonic acid six)；FCE 26752 (8,8 '-[carbonyl is double [imino group (1- methyl isophthalic acid H- pyrroles -4,2- diyl) carbonylimino (1- methyl isophthalic acid H- pyrroles -4,2- diyl) carbonylimino]] double -1,3,6- naphthalene trisulfonic acids)；Endothelial mononuclear cell activating polypeptide II；VEGFR ASONs；Anti-angiogenesis and ANFs；ANCHOR angiostatins；Endostatin；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 collagens (people's gene COL15A1 α 1- chains 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 iminos glutaric acid (2- (1,3- dihydro -1- oxo -2H- iso-indoles -2- bases) -2- methylglutaric acids)；The monoclonal antibody BC-1 of Y90 mark；Semaxanib (3- (4,5- dimethyl pyrrole -2- methylenes) Indolin-2-one) (C15H14N2O)；PI 88 (phosphomamlose pentose sulfate)；Alvocidib (4H-1- benzopyran-4-ones, 2- (2- chlorphenyls) -5,7- dihydroxy -8- (3- hydroxyl -1- methyl -4- piperidyls)-cis- (-) -) (C21 H20 Cl N O5)；E 7820；SU11248 (5- [fluoro- 2- oxos -1, the 2- indylidenes of 3--(3Z)-ylmethyl] -2,4- dimethyl -1H- pyrroles -3- formic acid (2- diethylaminos ethyl) acid amides) (C22 H27 F N4 O2)；Squalamine (cholestane -7,24- glycol, and 3- [[3- [(4- aminobutyls) aminopropyl] amino] -, 24- (hydrogen sulfate), (3. β, 5. α, 7. α) -] (C34 H65 N3 O5 S)；Eriochrome Black T；(the carbamic acids of AGM 1470; (chloracetyl) -; 5- methoxyl groups -4- [2- methyl -3- (3- methyl-2-butenes base) Oxyranyle] -1- oxaspiros [2; 5] octyl- 6- base esters; [3R- [3 α, 4 α (2R, 3R); 5 β, 6 β]]) (C19 H28 Cl N O6)；AZD 9935；BIBF 1000；AZD 2171；ABT 828；KS- proleulzins；Uteroglobin；A 6；NSC 639366 (1- [3- (diethylamino) -2- hydroxypropyls] -4- (oxirane -2- vlmethyls) anthraquinone fumarate) (C24 H29 N3 O4.C4 H4 O4)；ISV 616；Anti- ED-B fusion proteins；HUI 77；Troponin 1；BC-1 monoclonal antibodies；SPV 5.2；ER 68203；(the 3- (3 of CKD 731,4,5- trimethoxyphenyls) -2 (E)-acrylic acid (3R, 4S, 5S, 6R) -4- [2 (R)-methyl -3 (R) -3 (R)-(3- methyl-2-butenes base) oxirane -2- bases] -5- methoxyl group -1- oxaspiros [2.5] octyl- 6- base esters) (C28 H38 O8)；IMC-1C11；aaATIII；SC 7；CM 101；Angiocol；Kringle 5；CKD 732 (3- [4- [2- (dimethylamino) ethyoxyl] phenyl] -2 (E)-acrylic acid) (C29 H41 N O6)；U 995；Canstatin；SQ 885；(the 1- [11- (dodecylamino) -10- hydroxyundecyls -3,7-dimethylxanthine] (C30 H55 N5 O3) of CT 2584；Salmosin；EMAP II；TX 1920 (1- (4- methyl piperazines) -2- (2- nitro -1H-1- imidazole radicals) -1- ethane ketone) (C10 H15 N5 O3)；α-v β-x inhibitor；CHIR 11509 (double (4- methoxyphenyls) formamides of N- (1- propinyls) glycyl-[N- (2- naphthyls)] glycyl-[N- (carbamo, lmethyl)] glycine) (C36 H37 N5 O6)；BST2002；BST 2001；B 0829；FR 111142；4,5- dihydroxy -2 (E)-hexenoic acid (3R, 4S, 5S, 6R) -4- [1 (R), 2 (R)-epoxy -1,5- dimethyl -4- hexenyls] -5- methoxyl group -1- oxaspiros [2.5] octane -6- base esters (C22 H34 O7)；And kinase inhibitor, including but not limited to N- (4- chlorphenyls) -4- (4- pyridylmethyls) -1- phthalazines amine (phthalazinamine)；4- [4- [[[[4- chloro- 3- (trifluoromethyl) phenyl] amino] carbonyl] amino] phenoxy group]-N- methyl -2- pyridine carboxamides；N- [2- (diethylamino) ethyl] -5- [(fluoro- 1, the 2- dihydro-2-oxos -3H- indylidenes -3- bases of 5-) methyl] -2,4- dimethyl -1H- pyrrole-3-carboxamides；3- [(bromo- 2, the 6- difluorophenyls of 4-) methoxyl group] -5- [[[[4- (1- pyrrolidinyls) butyl] amino] carbonyl] amino] -4- isothiazole formamides；N- (the bromo- 2- fluorophenyls of 4-) -6- methoxyl groups -7- [(1- methyl -4- piperidines) methoxyl group] -4- quinazoline amine；3- [5,6,7,13- tetrahydrochysene -9- [(1- methyl ethoxies) methyl] -5- oxo -12H- indenos [2,1-a] pyrrolo- [3,4-c] carbazole -12- bases] propyl diester DMG；N- [5- [[[5- (1,1- dimethyl ethyl) -2- oxazolyls] methyl] sulfenyl] -2- thiazolyls] -4- piperidine formamides；N- [the chloro- 4- of 3- [(3- fluorophenyls) methoxyl group] phenyl] -6- [5- [[[2- (methyl sulphonyl) ethyl] amino] methyl] -2- furyls] -4- quinazoline amine；4- [(4- methyl isophthalic acids-piperazinyl) methyl]-N- [4- methyl -3- [[4- (3- pyridine radicals) -2- pyrimidine radicals] amino]-phenyl] benzamide；N- (the chloro- 4- fluorophenyls of 3-) -7- methoxyl groups -6- [3- (4- morpholinyls) propoxyl group] -4- quinazoline amine；Double (2- the methoxy ethoxies) -4- quinazoline amine of N- (3- ethynyl phenyls) -6,7-；N- (3- ((((2R) -1- methyl -2- pyrrolidinyls) methyl) epoxide) -5- (trifluoromethyl) phenyl) -2- ((3- (1,3- oxazole -5- bases) phenyl) amino)-Niacinamide；2- (((4- fluorophenyls) methyl) amino)-N- (3- ((((2R) -1- methyl -2- pyrrolidinyls) methyl) epoxide) -5- (trifluoromethyl) phenyl)-Niacinamide；N- [3- (azetidine -3- ylmethoxies) -5- trifluoromethyl-phenyls] -2- (the fluoro- benzylaminos of 4-)-niacinamide；The fluoro- N- of 6- (4- (1- Methylethyls) phenyl) -2- ((4- pyridylmethyls) amino)-Niacinamide；2- ((4- pyridylmethyls) amino)-N- (3- (((2S) -2- pyrrolidinylmethyls) epoxide) -5- (trifluoromethyl) phenyl)-Niacinamide；N- (3- (1,1- dimethyl ethyl) -1H- pyrazoles -5- bases) -2- ((4- pyridylmethyls) amino)-Niacinamide；N- (3,3- dimethyl -2,3- dihydro -1- benzofuran -6- bases) -2- ((4- pyridylmethyls) amino)-Niacinamide；N- (3- ((((2S) -1- methyl -2- pyrrolidinyls) methyl) epoxide) -5- (trifluoromethyl) phenyl) -2- ((4- pyridylmethyls) amino)-Niacinamide；2- ((4- pyridylmethyls) amino)-N- (3- ((2- (1- pyrrolidinyls) ethyl) epoxide) -4- (trifluoromethyl) phenyl)-Niacinamide；N- (3,3- dimethyl -2,3- dihydro -1H- indoles -6- bases) -2- ((4- pyridylmethyls) amino)-Niacinamide；N- (4- (pentafluoroethyl group) -3- (((2S) -2- pyrrolidinylmethyls) epoxide) phenyl) -2- ((4- pyridylmethyls) amino)-Niacinamide；N- (3- ((3- azetidines ylmethyl) epoxide) -5- (trifluoromethyl) phenyl) -2- ((4- pyridylmethyls) amino)-Niacinamide；N- (3- (4- piperidyls epoxide) -5- (trifluoromethyl) phenyl) -2- ((2- (3- pyridine radicals) ethyl) amino)-Niacinamide；N- (4,4- dimethyl -1,2,3,4- tetrahydro-isoquinoline -7- bases) -2- (1H- indazole -6- bases amino)-niacinamide；2- (1H- indazole -6- bases amino)-N- [3- (1- methylpyrroline -2- ylmethoxies) -5- trifluoromethyl-phenyls]-niacinamide；N- [1- (2- dimethylaminos-acetyl group) -3,3- dimethyl -2,3- dihydro -1H- indoles -6- bases] -2- (1H- indazole -6- bases amino)-niacinamide；2- (1H- indazole -6- bases amino)-N- [3- (pyrrolin -2- ylmethoxies) -5- trifluoromethyl-phenyls]-niacinamide；N- (1- acetyl group -3,3- dimethyl -2,3- dihydro -1H- indoles -6- bases) -2- (1H- indazole -6- bases amino)-niacinamide；N- (4,4- dimethyl -1- oxos -1,2,3,4- tetrahydro-isoquinoline -7- bases) -2- (1H- indazole -6- bases amino)-niacinamide；N- [4- (tert-butyl group) -3- (3- piperidinylpropyls) phenyl] [2- (1H- indazole -6- bases amino) (3- pyridine radicals)] formamide；N- [5- (tert-butyl group) isoxazole -3-bases] [2- (1H- indazole -6- bases amino) (3- pyridine radicals)] formamide；With N- [4- (tert-butyl group) phenyl] [2- (1H- indazole -6- bases amino) (3- pyridine radicals)] formamide, and the kinase inhibitor being disclosed in following patents：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, the publication is each in order to which any purpose is hereby incorporated by reference.

TR-2 is expressed in various cells, including liver, brain, kidney, colon, mammary gland, lung, spleen, thymus gland, PBLC, pancreas, prostate, testis, ovary, uterus and the various tissues along intestines and stomach.Example T R-2 associated cancers include but is not limited to, liver cancer, the cancer of the brain, kidney, breast cancer, cancer of pancreas, colorectal cancer, lung cancer (ED-SCLC and non-small cell lung cancer), spleen cancer, thymus gland or haemocyte cancer (that is, leukaemia), prostate cancer, carcinoma of testis, oophoroma, uterine cancer, stomach cancer, H＆N squamous cell carcinoma, melanoma and lymthoma.

In certain embodiments, anti-TR-2 antibody can be used together individually or with least one other therapeutic agent for treating cancer.In certain embodiments, anti-TR-2 antibody is used in combination with the other therapeutic agent of therapeutically effective amount.The exemplary treatment agent that can be applied together with anti-TR-2 antibody includes but is not limited to, the geldanamycin family member of anisamycin (suspected of anisomycin, anisomycin) antibiotic；Pro-HGF；NK2；C-Met inhibitor peptides；The antagonist of Grb2Src homologues 2；Gab1 conditioning agents；Dominant Src；Von-Hippel-Landau inhibitor, includes but is not limited to, wortmannin；P13 kinase inhibitors, other anti-receptor therapies, anti-EGFr, cox 2 inhibitor, Celebrex^TM, celecoxib, Vioxx^TM, rofecoxib；VEGF (VEGF), VEGF conditioning agents, fibroblast growth factor (FGF), FGF conditioning agents, EGF (EGF)；EGF conditioning agents；Keratinocyte growth factor (KGF), KGF correlation molecules, KGF conditioning agents；With matrix metalloproteinase (MMP) conditioning agent.

In certain embodiments, anti-TR-2 antibody is used together to treat various cancers with particular therapeutic agent.In certain embodiments, it is contemplated that symptom and required treatment level, 2 kinds, 3 kinds or more kind reagents can be applied.When compound is used together with one or more other components, compound and other one or more combinations together, separately or sequentially (for example, with medicament forms) can be applied.In certain embodiments, such reagent can together be provided by being included in same preparation.In certain embodiments, such reagent and anti-TR-2 antibody can together be provided by being included in same preparation.In certain embodiments, such reagent can be with separately formulated and provided together by being included in therapeutic reagent box.In certain embodiments, such reagent and anti-TR-2 antibody can be with separately formulated and provided together by being included in therapeutic reagent box.In certain embodiments, such reagent can be provided separately.

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

In certain embodiments, anti-TR-2 antibody can be used for treating non-human animal, such as pet (dog, pig, bird, primate), and raise and train farm-animals (horse, ox, sheep, pig, bird etc.).It is some it is such in the case of, suitable dosage can be determined according to the body weight of animal.For example, in certain embodiments, 0.2-1mg/kg dosage can be used.In certain embodiments, dosage can determine that exemplary dose is 0.1-20mg/in according to the surface area of animal², or 5-12mg/m².For toy, such as dog or cat, in certain embodiments, suitable dosage is 0.4mg/kg.In certain embodiments, anti-TR-2 antibody can apply one or many weekly by injection or other suitable pathways, until the symptom of animal is improved, or it can indefinitely be applied.

It should be understood that response of the single patient to foregoing pharmaceutical or combination treatment can be different, and it can be determined for the suitable medicine efficient combination of each patient by his or her doctor.

Machin provides the useful model on some diseases.Exemplary diseases include but is not limited to, graft rejection syndrome and inflammatory bowel disease sample disease.When tester MAb effect in machin human disease's model, whether in certain embodiments, it is useful with comparable horizontal integration TR-2 in people and machin to determine anti-TR-2MAb.

In certain embodiments, anti-TR-2 antibody can be the part of the conjugate molecules comprising all or part of anti-TR-2 antibody and cytotoxic agent.Term " cytotoxic agent ", which refers to, to be suppressed or prevents cell function and/or cause the material of cell death or destruction.The term includes but is not limited to, and radio isotope is (for example, I¹³¹、I¹²⁵、Y⁹⁰And Re¹⁸⁶), chemotherapeutant, and toxin such as bacterium, fungi, plant or animal origin enzymatic activity toxin, or its fragment.Exemplary cytotoxins agent includes but is not limited to, Doxorubicin, adriamycin, 5 FU 5 fluorouracil, cytarabin (" AraC "), endoxan, thiotepa, taxotere (docetaxel), busulfan, Cytoxin, taxol, amethopterin, cis-platinum, melphalan, vincaleukoblastinum, bleomycin, Etoposide, ifosfamide, mitomycin C, mitoxantrone, vincristine, vinorelbine, carboplatin, Teniposide, daunomycin, carminomycin, aminopterin, dactinomycin D, mitomycin, ai sibo mycin, melphalan and other related mustargen.

In certain embodiments, anti-TR-2 antibody can be the part of the conjugate molecules comprising all or part of anti-TR-2 antibody and pro-drug.In certain embodiments, term " pro-drug " refers to the precursor or derivative form of pharmaceutically active substances.In certain embodiments, compared with parent drug, pro-drug is less to the cytotoxicity of cell, and can activate or be transformed into more active cytotoxicity parent fo by enzymatic.Exemplary precursors medicine includes but is not limited to, phosphate-containing pro-drug, containing thio phosphate precursor medicine, containing sulfate pro-drug, medicine containing peptide precursor, pro-drug amino acid modified D-, glycosylated prodrugs, pro-drug containing beta-lactam, containing optionally substituted phenoxy acetamide pro-drug and containing optionally substituted phenyl acetamide pro-drug, can be transformed into the 5-flurocytosine and other 5-FUD pro-drugs of more active cytotoxic free drug.The example that the cytotoxic drug of pro-drug can be derivatized to includes but is not limited to, those described above cytotoxic agent.See, e.g., U.S. Patent number 6,702,705.

In certain embodiments, antibody conjugates are worked by making the antibody moiety of molecule target the specific cell precursor in the prodrug moieties targeting patient of cytotoxic moieties or molecule.In the case of anti-TR-2 antibody, such conjugate molecules can be used for, for example, in certain embodiments, destroying the cell of abnormality proliferation, such as cancer cell.

In certain embodiments there is provided the method for the treatment of patient, methods described includes applying the anti-TR-2 antibody of therapeutically effective amount.In certain embodiments there is provided the method for the treatment of patient, methods described includes applying the antibody conjugates of therapeutically effective amount.In certain embodiments, as described above, antibody is used in combination with least one other therapeutic agent of therapeutically effective amount.

As described above, in certain embodiments, anti-TR-2 antibody can be administered simultaneously with being applied to one or more other drugs of same patient, every kind of medicine is applied according to the scheme for being suitable for that medicine.Such treatment is comprising pretreatment, while treatment, sequentially treatment and alternate scheme.The other example of such medicine includes but is not limited to, antivirotic, antibiotic, anodyne, corticosteroid, inflammatory cytokine antagonist, DMARDs, non-steroidal anti-inflammatory drugs, chemotherapy, angiogenesis inhibitors and angiogenesis stimulant.

In certain embodiments, various medical conditions are treated with the anti-TR-2 antibody combined with another apoptotic stimulus thing.For example, in certain embodiments, anti-TR-2 antibody can be administered in the composition, the composition also includes the compound of the one or more Apoptosis of stimulation.In certain embodiments, anti-TR-2 antibody and apoptotic stimulus thing can be administered as separated composition, and these can be administered by identical or different approach.

In certain embodiments there is provided pharmaceutical composition, antibody of the described pharmaceutical composition comprising therapeutically effective amount is together with pharmaceutically acceptable diluent, carrier, solubilizer, emulsifying agent, preservative and/or adjuvant.

In certain embodiments, there is provided pharmaceutical composition, at least one other therapeutic agent of antibody of the described pharmaceutical composition comprising therapeutically effective amount and therapeutically effective amount, together with pharmaceutically acceptable diluent, carrier, solubilizer, emulsifying agent, preservative and/or adjuvant.

In certain embodiments, acceptable formulation materials are nontoxic to acceptor preferably on used dosage and concentration.In certain embodiments, antibody of the invention is provided in the preparation without buffer being such as disclosed in PCT/US06/22599, and the patent was submitted on June 8th, 2006, in order to which any purpose is incorporated herein by reference.

In certain embodiments, pharmaceutical composition, which can be included, is used for pH, permeability, viscosity, transparency, color, isotonicity, smell, aseptic, stability, dissolving or the rate of release of modifying, maintain or preserve such as composition, the formulation materials for absorbing or permeating.In certain embodiments, suitable formulation materials include but is not limited to, amino acid (such as glycine, glutamine, asparagine, arginine or lysine)；Antimicrobial；Antioxidant (such as ascorbic acid, sodium sulfite or sodium hydrogensulfite)；Buffer (for example, borate, bicarbonate, Tris-HCl, citrate, phosphate or other organic acids)；Swelling agent (such as mannitol or glycine)；Chelating agent (such as ethylenediamine tetra-acetic acid (EDTA))；Complexing agent (such as caffeine, polyvinylpyrrolidone, beta-schardinger dextrin or hydroxypropyl-β-cyclodextrin)；Filler；Monose；Disaccharides；With other carbohydrate (such as glucose, mannose or dextrin)；Protein (such as seralbumin, gelatin or immunoglobulin)；Colouring agent, flavor enhancement and diluent；Emulsifying agent；Hydrophilic polymer (such as polyvinylpyrrolidone)；Low molecular weight polypeptide；Salt-forming counterion (such as sodium)；Preservative (such as Benasept, benzoic acid, salicylic acid, thimerosal, benzyl carbinol, methyl p-hydroxybenzoate, propylparaben, Chlorhexidine, sorbic acid or hydrogen peroxide)；Solvent (such as glycerine, propane diols or polyethylene glycol)；Sugar alcohol (such as mannitol or D-sorbite)；Suspending agent；Surfactant or wetting agent (such as polyethers, EG, sorbitan ester, polysorbate such as polysorbate20, polysorbate80, trinitrotoluene, trometamol, lecithin, cholesterol, tyloxapal (tyloxapol))；Stability enhancer (such as sucrose or D-sorbite)；Tension-elevating agent (such as alkali metal halide, preferably sodium chloride or potassium, mannitol, D-sorbite)；Delivery vehicle；Diluent；Excipient and/or pharmaceutical adjuvants.(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 medium known in the art.Such medium includes but is not limited to, Fc domains, polyethylene glycol and glucan.Such medium is described in such as U.S. Patent number 6,660,843 and disclosed PCT Application No. WO 99/25044.

In certain embodiments, optimal drug composition will be depended on for example being expected route of administration, delivering mode and required dosage determining by those skilled in the art.See, e.g., Remington ' s Pharmaceutical Sciences, ibid.In certain embodiments, such composition may influence condition, the stability of antibody, internal rate of release and internal clearance rate.

In certain embodiments, the main media thing or carrier in pharmaceutical composition can be water or non-aqueous in nature.For example, in certain embodiments, suitable medium or carrier can be water for injection, normal saline solution or artificial cerebrospinal fluid, may be supplemented with other materials common in the composition for parenteral administration.In certain embodiments, neutral buffered saline or the salt solution mixed with seralbumin are further exemplary media things.In certain embodiments, pharmaceutical composition includes about pH 7.0-8.5 Tris buffer solutions, or about pH4.0-5.5 acetate buffer, it may further include D-sorbite or its suitable substituent.In certain embodiments, pharmaceutical composition is the water or liquid preparation of the acetate buffer comprising about pH 4.0-5.5, polyol (polyalcohol) and optional ground surfactant, wherein said composition does not include salt, for example, sodium chloride, and wherein said composition is isotonic for patient.Exemplary polyol includes but is not limited to, sucrose, glucose, D-sorbite and mannitol.Exemplary surfactants include but is not limited to, polysorbate.In certain embodiments, pharmaceutical composition is the water or liquid preparation of the acetate buffer comprising about pH 5.0, D-sorbite and polysorbate, and wherein said composition does not include salt, for example, sodium chloride, and wherein said composition is isotonic for patient.Some exemplary compositions are found in such as U.S. Patent number 6,171,586.Other pharmaceutical carrier includes but is not limited to, oil, including oil, animal oil, vegetable oil, peanut oil, soybean oil, mineral oil, sesame oil etc..In certain embodiments, glucose and glycerine water solution are also used as liquid-carrier, especially for Injectable solution.In certain embodiments, by the selected composition with required purity and lyophilized agglomerate or the optional preparaton (Remington ' sPharmaceutical Sciences of aqueous solution form, ibid) mix, can prepare comprising antibody, together with or together with the composition of at least one other therapeutic agent be not used to store.In addition, in certain embodiments, using suitable excipient solution (for example, sucrose) as diluent can by comprising antibody, together with or together with the composition of at least one other therapeutic agent be not formulated as lyophilized products.

In certain embodiments, anti-TR-2 antibody is applied in the form of the acceptable composition of physiology, and the acceptable composition of physiology includes the recombinant protein of the purifying combined with the acceptable carrier of physiology, excipient or diluent.In certain embodiments, examples of such carriers is nontoxic to acceptor on used dosage and concentration.In certain embodiments, the preparation of such composition, which can be related to, makes anti-TR-2 antibody be combined with following substances：Buffer, antioxidant such as ascorbic acid, low molecular weight polypeptide (for example, those with less than 10 amino acid), protein, amino acid, carbohydrate such as glucose, sucrose or dextrin, chelating agent such as EDTA, glutathione and/or other stabilizers and excipient.In certain embodiments, suitable dosage is determined in standard medicine-feeding test, and can be become according to selected route of administration.In certain embodiments, according to suitable industrial standard, preservative can also be added, this includes but is not limited to, phenmethylol.In certain embodiments, the amount and frequency of administration can based on such factor as disease to be treated property and severity, needed for response, the age of patient and condition etc. determine.

In certain embodiments, pharmaceutical composition can select to be used for potential delivery.The preparation of some such pharmaceutically acceptable compositions is in art technology.

In certain embodiments, formulation components exist to apply the acceptable concentration in site.In certain embodiments, using buffer solution so that composition is maintained at physiology pH or slightly lower pH, typically about 5- about 8 pH.

In certain embodiments, when expected parenteral administration, pharmaceutical composition can in pharmaceutically acceptable medium, comprising required antibody, together with or not together with the pyrogen-free of other therapeutic agent, the acceptable aqueous solution form of parenteral.In certain embodiments, the medium for parenteral injection is sterile distilled water, wherein antibody, together with or be not configured to together with other therapeutic agent that appropriate corrosion-resistant is sterile, isotonic solution.In certain embodiments, preparation can be related to molecule needed for the microsphere, bio-erodible particles, polymerizable compound (such as PLA or polyglycolic acid) of reagent such as injectable, pearl or liposome formulation, the reagent may provide control or the sustained release of product, and the product can then be delivered via depot injection.In certain embodiments, hyaluronic acid can also be used, and may have the effect for promoting the duration in the circulating cycle.In certain embodiments, implantable drug delivery device can be used to introduce required molecule.

In certain embodiments, pharmaceutical composition can be formulated for suction.In certain embodiments, antibody, together with or not together with least one other therapeutic agent, can be configured to dry powder be used for suck.In certain embodiments, comprising antibody, together with or together with the inhalation solution of at least one other therapeutic agent can not be formulated for aerosol delivery with propellant.In certain embodiments, solution can be vaporific.Pulmonary administration is further described in PCT Publication WO94/20069, and the patent describes the pulmonary delivery of the protein of chemical modification.

In certain embodiments, it is contemplated that preparation can be applied with oral area.In certain embodiments, the antibody applied by this way, together with or not together with least one other therapeutic agent, can together with or those carriers for commonly using in not coordinating together with solid dosage forms such as tablet and capsule prepare.In certain embodiments, capsule can be designed as in the gastrointestinal tract when bioavailability maximizes the active part with delivery formulations on the point degraded before system when minimizing.In certain embodiments, at least one other reagent can be included to promote the absorption of antibody and/or any other therapeutic agent.In certain embodiments, diluent, flavor enhancement, low melt wax, vegetable oil, lubricant, suspending agent, tablet disintegrant and/or adhesive can also be used.

In certain embodiments, pharmaceutical composition can be related to the antibody of effective dose, together with or not together with least one other therapeutic agent, mixed with the non-toxic excipients for being suitable for tablet preparation.In certain embodiments, by making tablet dissolved in sterilized water or another suitable medium, the solution of unit dosage forms can be prepared.Suitable excipient includes but is not limited to, inert diluent, such as calcium carbonate, sodium carbonate or sodium acid carbonate, lactose or calcium phosphate；With adhesive such as starch, gelatin and Arabic gum；With lubricant such as magnesium stearate, stearic acid and talcum.

Other pharmaceutical composition will be apparent for those skilled in the art, be included in continue or control delivering preparation in be related to antibody, together with or the not preparation together with least one other therapeutic agent.Include but is not limited in some exemplary persistents or control release preparation, liposome vectors, bio-erodible microparticles, porous bead and depot injection.Some example techniques for preparing some preparations are well known by persons skilled in the art.See, for example, PCT Publication WO93/15722, the patent describes the control release of the porous polymerizing particulate for delivering pharmaceutical composition.In certain embodiments, extended release preparation can include the semi-permeable polymer matrices of formed article form such as film or microencapsulation form.Sustained-release matrix includes but is not limited to, polyester, hydrogel, polyactide (U.S. Patent number 3,773,919 and EP 058,481), Pidolidone and γ ethyl-L-glutamate salt copolymers (Sidman et al., Biopolymers, 22：547-556 (1983)), poly- (2- ethoxys-methacrylate) (Langer et al., J.Biomed.Mater.Res., 15：167-277 (1981) and Langer, Chem.Tech., 12：98-105 (1982)), ethylene vinyl acetate (Langer et al., ibid) and poly- D (-) -3- hydroxybutyric acids (EP 133,988).In certain embodiments, sustained-release composition can also include liposome, and in certain embodiments, the liposome can be prepared by any one of several method known in the art.See, for example, Eppstein et al., Proc.Natl.Acad.Sci.USA, 82：3688-3692(1985)；EP 036,676；EP 088,046 and EP 143,949.

In certain embodiments, the pharmaceutical composition for applying in vivo is sterile.In certain embodiments, the pharmaceutical composition for applying in vivo is sterile to be made by being filtered via sterilised membrane filter.In certain embodiments, when composition is lyophilized, it can be carried out using the sterilizing of sterilised membrane filter before or after lyophilized and reconstruct.In certain embodiments, the composition for parenteral administration can be stored with lyophilized form or in the solution.In certain embodiments, parenteral composition is typically disposed in the container with sterile access port, the container for example with can by the plug of hypodermic injection needle-penetration intravenous solution bag or phial.

In certain embodiments, after pharmaceutical composition has been prepared, it can be stored in sterile phial as solution, suspension, gel, emulsion, solid, or as dehydration or freeze-dried powder.In certain embodiments, such preparation can the storage (for example, lyophilized form) in the form of being reconstructed with form or before administration.

There is provided the kit that unit is applied for producing single dose in certain embodiments.In certain embodiments, the kit can each self-contained the first container with drying protein and second of container with water formulation.In certain embodiments, the kit of the pre-filled syringe (such as fluid injector and lyosyringes) comprising list and/or multicompartmented is included.

In certain embodiments, the effective dose of the pharmaceutical composition used in the treatment will depend on such as treatment background and purpose, and described pharmaceutical composition includes antibody, together with or not together with least one other therapeutic agent.It should be understood by those skilled in the art that according to some embodiments, Suitable dosage levels for treatment are therefore by molecule of the part according to delivering, antibody, together with or not together with least one other therapeutic agent by for its indication, route of administration, and patient's size (body weight, body surface or organ size) and/or condition (age and general health) and become.In certain embodiments, clinician can be stepped up dosage and change route of administration to obtain optimum curative effect.In certain embodiments, depending on above-mentioned factor, general dosage can be about 0.1 μ g/kg- and be up to about 100mg/kg or more.In certain embodiments, dosage can be up to about 100mg/kg for 0.1 μ g/kg-；Or 1 μ g/kg- be up to about 100mg/kg；Or 5 μ g/kg- be up to about 100mg/kg；Or 0.1mg/kg- is up to about 100mg/kg.

In certain embodiments, the pharmacokinetic parameter of antibody and/or any other therapeutic agent in the preparation that administration frequency uses consideration.In certain embodiments, clinician will apply composition until reaching the dosage for obtaining required effect.In certain embodiments, therefore said composition can be used as single dose, or as time go by as 2 times or more time dosage (this can be included or not comprising same amount of required molecule), or applied as continuous infusion via implanted device or conduit.Some methods of suitable dosage are further improved in art technology.In certain embodiments, suitable dosage can be determined by using suitable dose response data.

In certain embodiments, the route of administration of pharmaceutical composition is according to known method, such as oral area, by intravenous, intraperitoneal, big intracerebral (essence in), interior, the intramuscular ventricles of the brain, intraocular, intra-arterial, portal vein is interior or intralesional routes are via injection；By sustained release system or pass through implanted device.In certain embodiments, said composition can be by injecting or continuous infusion or being applied by implanted device.

As described above, in certain embodiments, any effective route of administration can be used for applying anti-TR-2 antibody.If injection, then in certain embodiments, anti-TR-2 antibody can for example via it is intra-articular, intravenous, intramuscular, damage in, intraperitoneal, encephalic, it is intranasal, suction or by injecting or being applied by the subcutaneous route of continuous infusion.Exemplary application process includes but is not limited to, the sustained release from implant, aerosol suction, eye drops, oral preparations, including pill, syrup, lozenge and chewing gum, and local preparation example such as lotion, gel, spray, ointment and other suitable technologies.

In certain embodiments, it is favourable by sucking administration when treating the disease relevant with lung conditions.In certain embodiments, anti-TR-2 antibody can be applied by being implanted into the culture cell of expression antibody.In certain embodiments, by using the own cells of the in vivo or in vitro transfection induction production patient of the one or more carriers for encoding anti-TR-2 antibody.In certain embodiments, the naked DNA or liposome encapsulated dna of anti-TR-2 antibody are encoded by injecting, or by other transfection methods, this carrier can be introduced the intracellular of patient.When anti-TR-2 antibody is administered in combination with one or more other biologically active cpds, in certain embodiments, these can be by identical or applied by different approaches, and can together, separately or sequentially apply.

In certain embodiments, film, sponge or another suitable material of molecule carry out local application needed for said composition can thereon absorb or encapsulate via implantation.In certain embodiments, when using implanted device, the device can be implanted in any suitable tissue or organ, and the delivering of required molecule can be via diffusion, time controlled released bolus or continuous administration.

In some embodiments it may be desirable to used with ex vivo comprising antibody, together with or the not pharmaceutical composition together with least one other therapeutic agent.In such embodiment, the cell, tissue and/or organ that have been taken out from patient is set to be exposed to pharmaceutical composition, described pharmaceutical composition includes antibody, together with or cell, tissue and/or organ then planted not together with least one other therapeutic agent, after this returned in patient.

In certain embodiments, antibody and any other therapeutic agent can be delivered by being implanted into some cells, the cell application method for example it is described herein those carry out genetic modifications, with express and secrete polypeptide.In certain embodiments, such cell can be animal or people's cell, and can be Autologous, heterologous or xenogenesis.In certain embodiments, the cell can be immortalized.In certain embodiments, in order to reduce the probability of immune response, the cell can be packaged to avoid the infiltration of peripheral organization.In certain embodiments, encapsulating material is usually bio-compatible, semipermeable polymeric shell or film, and it allows one or more proteins to discharge but prevents cell by the immune system of patient or other injurious factors from peripheral organization are destroyed.

Embodiment

Embodiment 1

The production of some human monoclonal antibodies

The anti-TR-2 antibody of people is produced in one of 2 kinds of methods.Make the transgenic mice of expression human immunoglobulin geneExposed to people TR-2.Using hybridoma technology some anti-TR-2 monoclonal antibodies of people are produced by these mouse.Some other anti-TR-2 monoclonal antibodies of people are produced by those mouse using XenoMax technologies, the XenoMax technologies incorporate selection lymphocyte antibody method (" SLAM ") technology (see, e.g., U.S. Patent number 5,627,052；With Babcook et al., Proc.Natl.Acad.Sci.USA 93：7843-7848(1996)).

Method for the production anti-TR-2 monoclonal antibodies of people in the transgenic mice of expression human immunoglobulin gene is as follows.As shown in fig. 1,5 groups of recombined human TR-2 (ripe amino acid sequence ALITQQDLAPQQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWND LLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMV KVGDCTPWSDIECVHKESGTKHSGEAPAVEETVTSSPGTPASRSGSSHHHHHH (the SEQ ID NO of mouse with the histidine mark of C-terminal six (TR-2-His)：140)) (Genbank reference number NM-003842) is immunized.Mouse in group 1, group 3, group 4 and group 5 is transformed to produce the antibody (Fig. 2) of IgG2 isotypes.Mouse in group 2 is transformed to produce the antibody (Fig. 2) of IgG4 isotypes.Group 1 includes 7 mouse, and group 2 includes 8 mouse, and group 3 includes 8 mouse, and group 4 includes 10 mouse, and group 5 includes 5 mouse.Mouse in group 1, group 2 and group 3 carries out immune (10 μ g/ injections) by the way that TR-2-His is expelled in palmula, and the mouse organized in 4 and group 5 carries out Intraperitoneal immunization (10 μ g/ injections) with TR-2-His.At the 0th day, 10 μ g antigens are applied by the approach.In appointed interval, booster shots are applied to mouse.1 mouse of group has 9 booster shots, the 5th, 11,14,18,24,28,34,42 and 46 days when.Group 2 and 3 mouse of group have 7 booster shots；On group 2 those the 3rd, 7,10,14,17,24 and 27 days when, and on group 3 those the 5th, 8,15,21,26,30 and 33 days when.Group 4 and 5 mouse of group have 5 booster shots, the 14th, 28,42,56 and 72 days when.10 μ g TR-2-His and adjuvant are each included with each booster shots first, the adjuvant is Titermax Gold (group 1,2 and 3), alum gel (group 1,2 and 3), complete Freund's adjuvant (CFA) (group 4 and 5), incomplete Freund's adjuvant (IFA) (group 4 and 5) or DulbeccoShi phosphate buffered saline (PBS)s (D-PBS) (group 1,2,3,4 and 5) (referring to Fig. 1).(group 4 and 5) after being injected at 3 times, injected at 4 times after (group 1,2 and 3), injected at 6 times after (group 1 and 2) and (group 1) takes blood to mouse after being injected at 10 times.As shown in Figure 2, blood is taken to be estimated TR-2-His reactivity by ELISA every time.

ELISA determination methods are carried out as follows.Porous flat plate is coated with by staying overnight Passive intake soluble T R-2-His (0.5 μ g/mL) in 4.Hole is coated with breast washing and is closed 30 minutes.The 10 every kind of mice serums of μ L is combined with 40 μ L breasts, and incubated 1 hour, 1.25 hours or 2 hours in the hole of different flat boards.Flat board is washed with water 5 times.Flat board is then set to be incubated 1 hour at room temperature together with the specific horseradish peroxidase conjugation of antibodies (Pierce) of the μ g/mL of final concentration 1 Goat anti human IgG Fc.Flat board is washed with water 5 times.Flat board is set to be incubated 30 minutes together with the blue substrates (Neogen) of K.Negative control includes lacking TR-2-His blank well, and the hole incubated including TR-2-His but together with the expected natural G2 serum for lacking anti-TR-2 antibody.

Method for producing the anti-TR-2 monoclonal antibodies of people is as follows.For XenoMax technologies, CD19+B cells are separated from high immunizing transgenic mice, 37th day (mouse M712-7 from group 3) of the high immunizing transgenic mice after immunity inoculation starting or the 76th day (mouse M564-1 from group 4, and mouse M564-3, M564-5 and M563-5 from group 5) when harvest.By B cell culture 1 week to allow it to expand and then break up plasmablast.The thick liquid cell that supernatant comprising secretory antibody is preserved in further analysis, and each hole is frozen in -80celcius degree (suspected of celsius, degree Celsius) in the culture medium comprising 10%DMSO and 90%FCS.For hybridoma technology, as shown in fig. 1, the cell from remaining high immunizing transgenic mice is the 31st, 37 or 46 days when harvest for further analyzing.

For XenoMax technologies, the supernatant from B cell culture is screened in the presence by ELISA just for TR-2 antibody.Anti-human IgG antibodies' detection reagent is used as described below anti-TR-2 antibody is detected by assessment and immobilization TR-2-His combination.By staying overnight Passive intake soluble T R-2-His (0.5 μ g/mL) coating flat boards in 4 DEG C.After the hole in flat board is closed 30 minutes with water by Plate wash 5 times and with breast, the 10 μ L cell supernatants from each single hybridoma are made to be combined with 40 μ L breasts, and incubated 1 hour, 1.25 hours or 2 hours in the hole of different flat boards.Flat board is washed with water 5 times, and is incubated 1 hour at room temperature together with specific horseradish peroxidase (Pierce) conjugation of antibodies of the μ g/mL of final concentration 1 Goat anti human IgG Fc.Flat board is washed with water 5 times.Flat board is set to be incubated 30 minutes together with the blue substrates (Neogen) of K.Negative control includes lacking TR-2-His blank well, and uses the hole of the expected natural G2 serum for lacking anti-TR-2 antibody.Positive is screened for TR-2-His by ELISA for the second time, to confirm the cell identity for producing the antibody special to TR-2.

It is induced to the antibody reacted with TR-2 that the ability screening of WM-266 melanoma cells (ATCC catalog number (Cat.No.) CRL-1676) apoptosis is identified above using apoptosis determination method.The WM-266 cells such as advised by ATCC in microtiter plate with the density of 4500 cells/wells in normal cell culture medium overnight incubation.For B cell culture, 20 μ L antigen-specific b cells culture supernatants or control B cell culture supernatant are added into 180 μ L apoptosis culture medium mixtures (cell culture medium comprising 1 μ g/mL cycloheximides (CHX) and 0.5% hyclone (" FCS ")).Remove the culture medium from WM-266 cells and antibody-apoptosis culture medium mixture is added into cell, every time 1 row.Cell is set to incubate 20 hours to allow apoptosis together with antibody-apoptosis culture medium.Add DNA binding fluorescent dyes propidium iodide (Sigma) and Hoechst 33342 (Molecular Probes) into each hole with 0.5 μ g/mL and 2.5 μ g/mL final concentration respectively.Hoechst 33342 is to pass through film, and therefore marks live and dead cells；Propidium iodide can not pass through film, and therefore can only mark dead cell.In 37 DEG C after 1 hour, capture the image in each hole and analyze TCS mesh (by the amount for assessing Hoechst marks) and dead cell total number (by the amount for assessing propidium iodide mark).Apoptosis percentage test is (propidium iodide-positive cell/Hoechst- positive cells) × 100.

For XenoMax technologies, the antibody from the several holes for showing optimal apoptosis induction is selected to be used to rescue using haemolysis plaque assay.Make TR-2-His biotinylations and be coated on the coated sheep red blood cell (SRBC) of Streptavidin.The thick liquid cell of correspondence antigen-specific well is thawed, and incubated in the presence of complement and guinea pig anti-human IgG enhancing serum together with the red blood cell of antigen coat.Production causes the sheep red blood cell (SRBC) around them to crack for the thick liquid cell of TR-2-His antibody, and therefore allows to identify the antigen-specific plasma cell in mixture.Those thick liquid cells are separated by single celled micromanipulation from mixture.

After single thick liquid cell needed for separation, mRNA is extracted from those cells.The mRNAs of coding weight and light chain variable sequence is transformed into cDNA, and expanded by reverse transcriptase PCR, use the merger antisense primer of the homing sequence and constant region to human IgG2 and people κ mRNA specifically.Primer sequence is provided in table 2 below：

Primer introduces above-mentioned restriction site in heavy chain cDNA 5 ' ends (BglII) and 3 ' ends (Xba1) place.Similarly, primer introduces above-mentioned restriction site in κ chains cDNA 5 ' ends (BglII) and 3 ' ends (NheI) place.

Variable heavy chain cDNA amplicons are digested with to the suitable restriction enzyme of restriction site, and the restriction site is added during PCR reacts.By the product cloning of that digestion in each in IgG1, IgG2 and IgG4 expression vector for the compatible jag of clone.IgG2 and IgG4 expression vectors are digested with BamHI and XbaI, to produce the compatible jag for being subcloned.IgG1 expression constructs are digested with BamHI and NheI, to produce the compatible jag for being subcloned.By human IgG1, IgG2 and IgG4 constant domain being cloned into carrier pcDNA3.1+/Hygro (Invitrogen) multiple cloning sites produce these carriers.

Variable light cDNA amplicons are equally digested with to the suitable restriction enzyme of restriction site, and the restriction site is added during PCR reacts.By in the product cloning of that digestion to IgK expression vectors, the IgK expression vectors are digested with BamHI and NheI, to produce the compatible jag for being subcloned.By the constant domain of people's IgK genes being cloned into carrier pcDNA4.1+/Neo (Invitrogen) multiple cloning sites produce that carrier.

Then by heavy chain and light chain expression vector altogether lipofection in the 60mm wares of the 70% people's tire kidney 293 cell (ATCC catalog number (Cat.No.) CRL-1573) converged.For 24 hours, it is allowed to which transfectional cell secretion has and the specific recombinant antibodies of Plasmablast identical.The supernatant (3mL) from the cells of HEK 293 is harvested, and confirms the secretion of complete antibody with specific detection human IgG with sandwich ELISA.As for combining flat board, control flat board is coated with 2mg/mL Goat anti human IgG H+L O/N.Flat board is washed with water 5 times.For 7 holes from undiluted lipofection supernatant, recombinant antibodies carry out 1: 2 and titrated.Flat board is washed 5 times with dH2O.For secretion and 2 kinds of binding assays, the μ g/mL of final concentration 1 Goat anti human IgG Fc specificity HRP conjugation of antibodies is added in room temperature 1 hour.Flat board is washed 5 times with dH2O.Flat board is set to develop the color within 30 minutes by adding tetramethyl benzidine (TMB), and stop ELISA by adding 1M phosphoric acid.

In addition to above-described XenoMax methods, some antibody are obtained using hybridoma technology.Immune mouse is put to death by cervical dislocation, and harvests from per share draining lymph node and merges.Grinding separates lymphoid cell to discharge cell from tissue in by improveing Eagle culture mediums (" DMEM ") in Dulbecco.Cell resuspension will be reclaimed in DMEM.Cell is counted, and adds into cell mass the lymphocytes of 0.9mL DMEM/100 million, so that cell gently but completely resuspension.Make the cell and 100 μ L CD90 of resuspension⁺The cell one of magnetic bead/100,000,000 arises from 4 DEG C and incubated 15 minutes.The cell suspending liquid of magnetic marker (is included up to 10⁸Positive cell (or up to 2 × 10⁹Total cell)) it is loaded into LS⁺On post.The post is washed with DMEM.Total efflux is collected as CD90^-Female one, this is expected mainly to include B cell.

By with 1: 1 ratio by the enriched B cell from washing above and non-secreting myeloma P3X63Ag*.653 cells (ATCC (CRL 1580, see, for example, Kearney et al., J.Immunol.123,1979,1548-1550)) mixing is merged.Cell mixture lightly forms agglomerate by being centrifuged under 800x g.Removed completely after supernatant from cell, by cell 2-4ml pronase solutions (CalBiochem；It is dissolved in phosphate buffered saline (PBS) (" PBS ") 0.5mg/ml) handle no more than 2 minutes.3-5mL hyclones (" FBS ") are added to stop enzymatic activity, and promote cell fusion solution (" ECFS ") (0.3M sucrose, 0.1mM magnesium acetates, 0.1mM calcium acetates) using electricity to adjust suspension to 40mL cumulative volumes.Supernatant is removed after centrifugation and by Cell resuspension in 40mL ECFS.Repeat this washing step and again by Cell resuspension in 40mL ECFS to 2 × 10⁶Cell/mL concentration.

Electricity promotees cell fusion using fusion generator (model ECM2001, Genetronic, Inc.) to carry out.The fusion room size used is 2.0mL, is set using following instruments：Comparison condition：50V, 50 seconds；Film is broken under 3000V, 30 microseconds；Retention time after fusion：3 seconds.

Electricity promotees after cell fusion generation, aseptically carefully take out cell suspending liquid from fusion room and be transferred in the sterile tube of the Hybridoma medium comprising same volume, the Hybridoma medium includes DMEM, (JRH Biosciences), 15%FBS (Hyclone), is supplemented with Glu, penicillin/streptomycin, OPI (oxaloacetate, acetonate, bovine insulin) and IL-6 (Boehringer Mannheim).Cell is incubated 15-30 minutes in 37 DEG C, and then centrifuged 5 minutes under 400x g (1000rpm).Cell is lightly resuspended in the hybridoma Selective agar medium of small size (being supplemented with 0.5x hyaluronic acids (Sigma) Hybridoma medium).Based on altogether 5 × 10⁶The hole plate of B cell/96 and the final Each FLIPR in 200 μ L/ holes, cumulative volume is suitably adjusted with more hybridoma Selective agar mediums.Cell is gently mixed, is drawn in 96 hole plates, and allows growth.At the 7th or 10 day, half culture medium is removed, and cell is resupplied with fresh hybridoma Selective agar medium.

After culture 14 days, doma supernatant is screened with regard to TR-2 monoclonal antibody specifics by ELISA.In first screening, ELISA flat boards (Fisher catalog number (Cat.No.) 12-565-136) are with 50 μ L/ holes TR-2 protein (2 μ g/mL) in coating buffer solution (0.1M carbonate buffer solutions, pH 9.6, NaHCO₃It is coated with, is incubated overnight with after 4 DEG C in 8.4g/L).After incubation, flat board is washed 1 time with lavation buffer solution (0.05%Tween 20 for being dissolved in PBS).200 μ L/ holes Block buffers (being dissolved in 1x PBS 0.5%BSA, 0.1%Tween 20,0.01% thimerosal) are added, and flat board is incubated at room temperature 1 hour.After incubation, flat board is washed 1 time with lavation buffer solution.The aliquot (50uL/ holes) of doma supernatant and positive and negative control is added, and flat board is incubated at room temperature 2 hours.The positive control used from beginning to end is the serum from high immune XenoMouse animals, and negative control is the serum from the KLH- XenoMouse animals being immunized.After incubation, flat board is washed 3 times with lavation buffer solution.The 100 μ L/ holes detection anti-huIgGFc-HRP of antibodies Goat (Caltag Inc. catalog number (Cat.No.) H10507, concentration is 1: 2000 dilution factor) is added, and flat board is incubated at room temperature 1 hour.After incubation, flat board is washed 3 times with lavation buffer solution.100 μ L/ hole TMB (BioFX Lab. catalog number (Cat.No.) TMSK-0100-01) are added, and allow flat board to develop the color about 10 minutes (until negative control hole just starts display color).It is subsequently added 50 μ L/ holes and stops solution (TMB Stop Solution (BioFX Lab. catalog number (Cat.No.) STPR-0100-01), and the plate read at 450nm wavelength on ELISA flat bed readers.

The antibody produced by hybridoma is analyzed using above-described identical apoptosis determination method.WM-266 cells overnight incubation in microtiter plate in normal incubation medium with the density of 4500 cells/wells.Using without FCS 2x apoptosis culture medium mixtures are prepared with the cell culture medium for comprising additionally in 1.8 μ g/mL cycloheximides and 0.9%FCS.The parallel titration doma supernatant 1: 2 of the anti-KLH antibody of negative control matched using separated microtiter plate with isotype (in 2x apoptosis culture medium mixtures).Culture medium is removed from WM-266 cells, and 100 μ L antibody-apoptosis culture medium mixture is added to wrapping in celliferous each hole, every time 1 row.Microtiter plate is set to incubate 20 hours to allow apoptosis.Add DNA binding fluorescent dyes propidium iodide (Sigma) and Hoechst33342 (Molecular Probes) into each hole with 0.5 μ g/mL and 2.5 μ g/mL final concentration respectively.In 37 DEG C after 1 hour, capture the fluoroscopic image in each hole and analyze dead cell total number (PI) and TCS mesh (Hoechst).Apoptosis percentage test is (PI- positive cells/Hoechst- positive cells) × 100.

17 kinds of different anti-TR-2 antibody (antibody A-Q) are obtained using XenoMax or hybridoma method.All antibody are sequenced, and identify the sequence of weight and light chain variable district (referring to Fig. 3-19).The heavy chain and light chain of 17 kinds of antibody, which are compared, to be shown in Figure 20 and 21.

Using to a kind of above-described similar apoptosis determination method, with regard to its some antibody of apoptosis-induced efficiency test in cell.WM-266 melanoma cells in microtiter plate with the density of 4500 cells/wells in normal incubation medium overnight incubation.In separated microtiter plate, recombinant antibodies to be tested are titrated, (M413, the anti-TR-2 antibody of mouse IgG 1 has variable heavy chain sequence to suitable positive control：MEVQLVESGGGLVQPGGSLKLSCAASGFTFSTYGMSWVRQTPDKRLELVALINSQGGSTYNSDSVKGRFTISRDNARNTLYLQMSSLKSEDTAMYYCARRDYESLDSWGQGTSVTVSSG(SEQ ID NO：And light chain variable sequence 141)：DIVLTQSPASLPVSLGQRATISCRASESVEYSGTSLIQWYRQKPGQPPKLLIYAASNVDSEVPARFSGSGSGTDFSLYIHPVEEDDIAMYFCQQSRKVPWTFGGGTKLEIKRTDAAPGLEAA(SEQ ID NO：142)) and isotype matching the negative control antibody potential anti-TR-2 antibody of activity (can not show) so that the ultimate density of antibody will include 0.0001 μ g/mL-5 μ g/mL scope.The mixed antibody in the apoptosis culture medium comprising final concentration 0.9 μ g/mL CHX and 0.45%FCS.Culture medium, and addition antibody-apoptosis culture medium mixture into cell are removed from WM-266 cells.After culture 20 hours, with propidium iodide (Sigma) and Hoechst 33342 (Molecular Probes) staining cell.In 37 DEG C after 1 hour, capture the image in each hole and analyze dead cell total number (PI) and TCS mesh (Hoechst).Apoptosis percentage test is (PI- positive cells/Hoechst- positive cells) × 100.Significant cell death is observed in the cell handled with M413 or with above-described some anti-TR-2 antibody.

Embodiment 2

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

Use

The dynamics that the anti-TR-2 antibody As of 2000 Instrumental Analysis are combined to Q with TR-2.Using conventional amine coupling in CM-5

Highdensity goat anti-human antibody surface is prepared on chip.The anti-TR-2 antibody of every kind of purifying is diluted to about 1 μ g/ml in the HBS-P running buffers comprising 100 μ g/ml BSA.Every kind of anti-TR-2 antibody is captured on separated surface using 2 minute contact times and washing in 5 minutes, to stablize the anti-TR-2 antibody surfaces on chip.

In order to analyze TR-2 and every kind of single anti-TR-2 antibody bindings dynamics, 226nM recombined humans TR-2-His (is described into) Dynamic Injection on every kind of anti-TR-2 surfaces 1 minute (using kinject) in embodiment 1 in 25 DEG C, is then 5 minutes dissociation phases.Baseline shift is subtracted from the combination each observed in other surfaces, the baseline shift is due to the buffer injection for lacking TR-2 on anti-TR-2 antibody surfaces.In addition, the amount of monoclonal antibody of the data on TR-2 and anti-TR-2 antibody bindings for being captured on every kind of surface is standardized.Each data set is totally fitted to determine binding kineticses to 1: 1 interaction model.The k obtained for every kind of antibody_a、k_dAnd K_dValue is shown in table 3.

Table 3：In 25 DEG C of TR-2 and the dynamics of anti-TR-2 antibody bindings

Antibody	k_a(M^-1s^-1)	k_d(s^-1)	K_d(nM)
Antibody	k_a(M^-1s^-1)	k_d(s^-1)	K_d(nM)	A	5.3×10⁵	3.7×10^-3	6.9
B	5.7×10⁵	1.1×10^-2	19	A	5.3×10⁵	3.7×10^-3	6.9
B	5.7×10⁵	1.1×10^-2	19	C	6.8×10⁵	2.6×10^-3	3.9
D	6.2×10⁵	2.7×10^-3	4.5	C	6.8×10⁵	2.6×10^-3	3.9
D	6.2×10⁵	2.7×10^-3	4.5	E	8.7×10⁵	1.8×10^-3	2.1
F	3.8×10⁵	5.0×10^-3	13	E	8.7×10⁵	1.8×10^-3	2.1
F	3.8×10⁵	5.0×10^-3	13	G	6.0×10⁵	1.9×10^-2	31
H	8.6×10⁵	8.4×10^-3	9.8	G	6.0×10⁵	1.9×10^-2	31
H	8.6×10⁵	8.4×10^-3	9.8	I	2.9×10⁵	1.3×10^-3	4.4
J	5.7×10⁵	7.1×10^-3	12	I	2.9×10⁵	1.3×10^-3	4.4
J	5.7×10⁵	7.1×10^-3	12	K	6.8×10⁵	1.2×10^-2	18
L	6.0×10⁵	1.1×10^-2	18	K	6.8×10⁵	1.2×10^-2	18
L	6.0×10⁵	1.1×10^-2	18	M	3.4×10⁵	1.2×10^-2	37
N	8.1×10⁵	5.5×10^-2	68^*	M	3.4×10⁵	1.2×10^-2	37
N	8.1×10⁵	5.5×10^-2	68^*	O	4.4×10⁵	8.4×10^-3	19
P	8.1×10⁵	2.7×10^-2	33^*	O	4.4×10⁵	8.4×10^-3	19
P	8.1×10⁵	2.7×10^-2	33^*	Q	1.2×10⁶	1.6×10^-2	13^*

^*Data display on that sample is heterogeneous and very poor to 1: 1 models fitting.

Embodiment 3

Cell kills determination method

Cell is carried out with the anti-TR-2 antibody of some people described in embodiment 2 and kills determination method, to determine the degree of every kind of antibody triggering apoptosis and cell death.The some anti-TR-2 antibody of people and the anti-TR-2 antibody M412 and M413 of mouse are fixed in the difference hole of 96 PFP matter G coatings flat board (reactin conjugated proteins G is coated with flat board, Pierce catalog number (Cat.No.)s 15131).M412 is that the anti-TR-2 antibody of mouse IgG 1 has variable heavy chain sequence：KVQLQQSGTELVKPGASVKLSCKASGYTFTEYIIHWVKQRSGQGLEWIGWFYPGSGYIKYNEKFKDKATMTADKSSSTVYMELSRLTSEDSAVYFCTRHEEDGYYAAYWGQGTLVTVSA(SEQ ID NO：And light chain variable sequence 143)：DIVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDYTLTISSVQAEDLALYYCQQHYSTPYTFGGGTKLEIKR(SEQ ID NO：144).

M412 is that the anti-TR-2 antibody of mouse IgG 1 has variable heavy chain sequence：KVQLQQSGTELVKPGASVKLSCKASGYTFTEYIIHWVKQRSGQGLEWIGWFYPGSGYIKYNEKFKDKATMTADKSSSTVYMELSRLTSEDSAVYFCTRHEEDGYYAAYWGQGTLVTVSA(SEQ ID NO：And light chain variable sequence 143)：DIVMTQSHKFMSTSVGDRVSITCKASQDVSSAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDYTLTISSVQAEDLALYYCQQHYSTPYTFGGGTKLEIKR(SEQ ID NO：144).M413 is the anti-TR-2 antibody of mouse IgG 1 as described in Example 1.Every kind of antibody is added in first hole with 50 μ g/ml concentration, and is carried out 1: 3x in each of 7 other holes and be serially diluted.Every kind of antibody dilution is carried out in triplicate.Flat board is incubated 24 hours using preceding in 4 DEG C.After each hole is washed with culture medium (RPMI adds 10%FBS), by one of 4 kinds of different cell lines bed board to every kind of sessile antibody, density is 50,000 cells/well in 200 μ L cumulative volume.Test cell system is the cells of COLO 205 (human colon adenocarcinoma), MDA-231 cells (human breast carcinoma), WM35 cells (human melanoma) and WM793 cells (human melanoma).Cell is in 37 DEG C/6%CO₂It is lower incubate 24 hours, then with³H- thymidines are incubated 6 hours together.What survivaling cell percentage was mixed by determining in processing cell³H- thymidines are horizontally relative to mixing in untreated cell³H- thymidine levels are estimated.The survival of processing cell is reduced by 50% antibody concentration relative to untreated cell by determining, the ED of every kind of antibody is obtained by cell survival titration curve₅₀.ED of the human antibody for the cells of COLO 205₅₀Scope is 0 μ g/ml-3.25 μ g/ml.Mouse antibodies M412 and M413 has 1.85 μ g/ml and 0.07 μ g/ml ED for those cells respectively₅₀s.ED of the human antibody for MDA-231 cells₅₀Scope is 0.05 μ g/ml-0.5 μ g/ml.Mouse antibodies M412 and M413 has 0.6 μ g/ml and 0.07 μ g/ml ED for those cells respectively₅₀s.ED of the human antibody for WM35 cells₅₀Scope is 0.1 μ g/ml-0.6 μ g/ml.Mouse antibodies M412 and M413 has 1.85 μ g/ml and 0.07 μ g/ml ED for those cells respectively₅₀s.ED of the human antibody for WM793 cells₅₀Scope is 0.2 μ g/ml-0.2 μ g/ml.Mouse antibodies M412 and M413 has 1.85 μ g/ml and 0.05 μ g/ml ED for those cells respectively₅₀s。

Embodiment 4

People TR-2 expression in tumor cell line

With regard to TR-2 expression screening human tumor cell line.The cell line used includes those from mammary gland, central nervous system, colon, liver, lung, cervix, uterus, ovary, pancreas, prostate and kidney, and leukaemia and melanoma.

The TR-2 expression on human tumor cells is determined using the array based on cell.In short, will be in 4 × 10 in 100M1CBA buffer solutions (PBS, 3%FBS, 0.02% azide)⁵Cell is assigned in each hole of 20 hole plates of V-Bottom 96.CBA buffer solutions (150 μ L) are added into each hole and make flat board centrifugation so that cell is rotated down.Discard culture medium and the μ g/ml of 100 μ L 10 antibody-solutions (one of antibody A to Q) are added into cell mass, the cell mass is resuspended in the PBS comprising 2%PBS (" measure buffer solution ").After incubated on ice 25 minutes, cell is washed 1 time in buffer solution is determined.Xiang Kongzhong adds the specific horseradish peroxidases (HRP, Pierce) of 100 μ L secondary goat anti-human igg Fc, and makes flat board in incubated on ice 20 minutes.Flat board is washed 2 times with measure buffer solution, and adds 100 μ L tmb substrates (ZYMED) at room temperature 10 minutes.Flat board is set to centrifuge and be transferred to 50 μ L every kind of supernatant in the clean flat board for stopping solution (BioFX Laboratories) comprising 50 μ L.Optical density reading is carried out at 450nm using SpectraMax/plus readers (Molecular Devices).The OD value of Isotype control antibodies is derived from by subtracting makes data normalization.

Several cell lines have the OD more than 0.1 in the determination method₄₅₀, including 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), liver cancer cell lines 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), RPMI 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).Maximum TR-2 is expressed in colon carcinoma cell line KM-12 and HT-29 in the tumor cell line of research, and is found in Leukemia Cell Lines HL-60.None in central nervous system, cellule liver, cervix, uterus or the ovarian cancer cell line of research has the OD450 more than background.

In order to determine TR-2 expression characteristics that human tumor cells are fastened, above-mentioned human tumor cell line is determined with the anti-TR-2 antibody M412 of mouse.The TR-2 expression on human tumor cells is determined using the determination method based on cell.In short, will be in 4 × 10 in 100Ml CBA buffer solutions (PBS, 3%FBS, 0.02% azide)⁵Cell is assigned in each hole of 20 hole plates of V-Bottom 96.CBA buffer solutions (150 μ L) are added into each hole and make flat board centrifugation so that cell is rotated down.Discard culture medium and the μ g/ml of the 100 μ L 10 anti-TR-2 monoclonal antibodies M412 of mouse is added into cell mass, the cell mass is resuspended in the PBS comprising 2%PBS (" measure buffer solution ").After incubated on ice 25 minutes, cell is washed 1 time in buffer solution is determined.Xiang Kongzhong adds the specific horseradish peroxidases (HRP, Pierce) of 100 μ L secondary goat anti-mouse IgG Fc, and makes flat board in incubated on ice 20 minutes.Flat board is washed 2 times with measure buffer solution, and adds 100 μ L tmb substrates (ZYMED) at room temperature 10 minutes.Flat board is set to centrifuge and be transferred to 50 μ L every kind of supernatant in the clean flat board for stopping solution (BioFX Laboratories) comprising 50 μ L.Optical density reading is carried out at 450nm using SpectraMax/plus readers (Molecular Devices).The OD value of Isotype control antibodies is derived from by subtracting makes data normalization.

Many cell lines have TR-2 expression.Highest expresser (those with the OD450nm more than 0.3) includes 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 lines 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), liver cancer cell lines 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), ED-SCLCCell line 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).Maximum TR-2 is expressed in renal carcinoma cell line Caki-1 in the tumor cell line handled with the anti-TR-2 antibody of mouse, and is found in Colon cancer cell line HT-29 and KM-12.

Embodiment 5

Antibody cross reaction

Such as Jia et al., J.Immunol.Methods 288：Described in 91-98 (2004), the ability that some anti-other antibody of TR-2 antibody blockings of people are combined with TR-2 is assessed.Pearl is set to be conjugated with anti-human IgG antibodies using the COUPLING PROCEDURE for being directly derived from User ' the s Manual, Version 1.7 of Luminex 100.After integument activation, according to the specification of manufacturer, them are made to be coupled with the anti-hIgG mAb of Pharmingen mouse.Carry out 2 experiments.In first time tests, coating pearl is set to incubate at room temperature 2 hours.In second is tested, coating pearl is set to be incubated overnight in 4 DEG C.At the end of incubation, coating pearl is set to close and then be counted using Coulter cell counters.Conjugated pearl immediately using or be stored in 4 DEG C of dark be used for use in the future.

Anti- TR-2 antibody classifications based on epitope cross reactivity are carried out by following step.First, make to arise from 4 DEG C with reference antibody (" reference antibody ") one respectively from the anti-hIgG compounds of every group of pearl-mouse above and be incubated overnight on circulator.Reference antibody is selected from above-described anti-TR-2 antibody As-Q.After antibody capture, 2000 every kind of anti-hIgG of pearl-mouse are merged into 1 pipe together with reference to Ab compounds, and then in the tight each hole for adding 96 hole plates and aspirated.TR-2 (50ng) is added into each hole and is incubated 1 hour at room temperature.After washing hole, the anti-TR-2 antibody of another people of 100-500ng/mL (" detection antibody ") is added into each hole and is incubated 2 hours at room temperature.After the washing of hole, reference antibody is captured using 1 μ g/ml biotinylation forms and detects the detection antibody combined using the anti-hIgG of identical Monoclonal mouse.Incubate with after washing hole, add 0.5 μ g/ml SA-PEs.Mixture is incubated at room temperature 30 minutes, and then detect phycoerythrin signal using Luminex 100.Using the hole of another group of shortage antigen as negative control to help data analysis.

Data are analyzed during 2 steps.First, data normalization is made using negative control value.Secondly, anti-TR-2 antibody is clustered according to the ability of other one or more anti-TR-2 antibody bindings of its obstruction.For clustering, dissimilarity matrix is produced by the intensity matrix standardized.Antibody is clustered based on the value in average dissimilarity matrix, is assembled nested hierarchical cluster subprogram using SPLUS 2000 and is measured with Manhattan, uses the input dissimilarity matrix of actual average dissimilarity matrix.

Based on the discovery, antibody is put into 4 different epitopes groups.In any one group, the combination of another member and TR-2 of one of group membership and TR-2 identical group of combination blocking.However, for example, group one of 1 member and TR-2 combination not one of

blocking group

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

Embodiment 6

Epitope mapping

In order to identify a pair TR-2 specific regions important with some anti-TR-2 antibody bindings, epitope mapping research is carried out.N- avidin-TR-2 constructs are prepared by following methods：PCR amplification come self-template resource on mature T R-2 (MacFarlane, 1997) coded sequence, and be cloned into it pCEP4 carriers (Invitrogen) comprising chicken avidin sequence Nei with such direction, so that after being inserted at HindIII sites, TR-2 sequences are connected at the C-terminal of avidin sequence.Forward primer on mature T R-2 coded sequence is GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQID NO：, and reverse primer is GATTAGGGATCCAGAGGCAGGAGTCCCTGG (SEQ ID NO 145)：146).The amino acid sequence of resulting avidin-TR-2 fusion proteins is LSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGTYTTAVTATSNEIK ESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRNGKEVLKTMWLLRS SVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAPQQRAAPQQKRSSP SEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTT RNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHS GEAPAVEETVTSSPGTPAS (SEQ ID NO：69).

Synthesis is proceeded as described below comprising 12 kinds of truncate molecules of N- avidins and people TR-2.Only the C-terminal with people TR-2 is truncate (TR-2-1 to TR-2-3) for 3 kinds of molecules, and 9 kinds of molecules have truncate (TR-2-4 to TR-2-13) (the schematically showing in fig 23) at people TR-2 N and C-terminal.Polynucleotides truncate encoding human TR-2 are prepared by PCR amplifications using primer described below.In order to form each in 12 kinds of molecules, by the people TR-2 insertions pCEP4 carriers (Invitrogen) as described above comprising chicken avidin sequence of the truncation produced by amplification.Use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO：145) with reverse primer TAGTTGGGATCCTCAGGAGATGCAATCTCT ACCGT (SEQ ID NO：147) polynucleotides of amplification coding mature T R-2 amino acid/11-43.TR-2-1 amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCPPGHHISEDGRDCIS (SEQ ID NO：70).

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

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

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

Use forward primer GTAATGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO：150) with reverse primer GGTAGTGGATCCTCACTGACACACTGTGTTTCTGG (SEQ ID NO：148) amplification coding mature T R-2 amino acid/11 6-85 polynucleotides.TR-2-5 amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRN TVCQ (SEQ ID NO：74).

Use forward primer GTAATGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO：150) with reverse primer GTAATGGGATCCTCAGACACATTCGATGTCACTCC (SEQ ID NO：149) amplification coding mature T R-2 amino acid/11 6-126 polynucleotides.TR-2-6 amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTAVSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRN TVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECV (SEQ ID NO：75).

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

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

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

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

Use forward primer TGATTGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO：150) with reverse primer GATGGAGGATCCTCAACACCTGGTGCAGCGCAAG (SEQ ID NO：153) amplification coding mature T R-2 amino acid/11 6-66 polynucleotides.TR-2-12 amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTASNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRNG KEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSEG LCPPGHHISEDGRDCISYKYGQDYSTHWNDLLFCLRCTRC (SEQ ID NO：80).

Use forward primer TGATTGAAGCTTGCCACAACAAAAGAGGTCCAG (SEQ ID NO：150) with reverse primer GTAAGTGGATCCTCAGCAGGGACTTAGCTCCACT (SEQ ID NO：154) amplification coding mature T R-2 amino acid/11 6-74 polynucleotides.TR-2-13 amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPSE GLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELS (SEQ ID NO：81).4 kinds of truncate TR-2 comprising N- avidins and from machin molecules proceed as described below synthesis.Use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQID NO：155) with reverse primer GTTGATGGATCCTTCTTTGTGGACACTCGAT (SEQ ID NO：156) polynucleotides of amplification coding maturation cyno TR-2 amino acid/11-132.Cyno TR-2, the amino acid sequence of (short) is PLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGTYTTAVT ATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRNGKEVLK TMWLLRS SVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDPQRRAAPQQKRSSP TEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKCDSGEVEVSSCTTT RNTVCQCEEGTFREEDSPEICRKCRTGCPRGMVKVKDCTPWSDIECPQRRIQT, (SEQ ID NO：82).

Use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQID NO：155) with reverse primer GTAGTTGGATCCTCAAGAAGCAGGAGTCCCAGGG (SEQ ID NO：157) polynucleotides of amplification coding maturation cynoTR-2 amino acid/11-154.Cyno TR-2 (length) amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVSSCTTTRNTVCQCEEGTFREEDSPEICRKCRTGCPRGMVKVKDCTPWSD IECVHKESGTKHTGEVPAVEKTVTTSPGTPAS (SEQ ID NO：83).

Use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQID NO：155) with reverse primer GTATGAGGGATCCTCACTGACACACCGTGTTTCTGG (SEQ ID NO：158) polynucleotides of amplification coding maturation cynoTR-2 amino acid/11-85.Cyno TR-2 (length) amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVSSCTTTRNTVCQ (SEQ ID NO：84).

Use forward primer GTATGGAAGCTTGCCACAACAAAAGAGATCCAGC (SEQ ID NO：159) with reverse primer GTATGAGGGATCCTCACTGACACACCGTGTTTCTGG (SEQ ID NO：158) amplification maturation cyno TR-2 amino acid/11 6-85 polynucleotides.Cyno 16-85 amino acid sequence is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLPQQKRSSPIE GLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKCDSGEVEVSSCTTTRN TVCQ (SEQ ID NO：85).

As shown in Figure 25, user TR-2 and cyno TR-2 different piece equally prepare 4 kinds of N- Avidin-Fusion chimeras.Every kind of construct is built by preparing 2 kinds of PCR primers with overlapping end, the PCR primer is then expanded together using the primer of identical 5 ' and 3 '.In order to form every kind of chimera, then the polynucleotides of amplification are subcloned into the pCEP4 carriers (Invitrogen) as described above comprising chicken avidin sequence.People, cyno (short) and mouse TR-2 sequences comparison are shown in Figure 26.

Cyno/ people's chimera #1 is prepared by following：Use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQ ID NO：155) with reverse primer GGACCTCTTTTGTTGTGGAGCCGCTCTTCGCTGG (SEQ IDNO：159), amplification orresponding amino acid 1-16 ripe cyno TR-2 regions, and use forward primer CAGCGAAGAGCGGCTCCACAACAAAAG AGGTCCAG (SEQ ID NO：160) with reverse primer GGTAGTGGATCCTCACTGACACACTGTGTTTCTGG (SEQ ID NO：148), amplification orresponding amino acid 17-85 into acquaintance TR-2 regions.Using the forward primer on the fragment of cyno TR-2 amino acid/11s -16, (SEQ ID NO above：155), and the reverse primer on people's TR-2 amino acid/11 7-85 fragments, (SEQ ID NO above：148) cyno and the over-lap PCR of people's TR-2 fragments, are carried out.Amino acid sequence on cyno/ people's chimera #1 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQ (SEQ ID NO：86).

Cyno/ people's chimera #2 is prepared by following：Use forward primer GTTAGTAAGCTTGGCTCCAATCACCCGAC (SEQ ID NO：155) with reverse primer GGACCTCTTTTGTTGTGGAGCCGCTCTTCGCTGG (SEQ IDNO：159), amplification orresponding amino acid 1-16 ripe cyno TR-2 regions, and use forward primer CAGCGAAGAGCGGCTCCACAACAAAA GAGGTCCAG (SEQ ID NO：160) with reverse primer GATTAGGGATCCTCAAGAGGCAGGAGTCCCTGG (SEQ ID NO：146), amplification orresponding amino acid 17-154 into acquaintance TR-2 regions.Using the forward primer on the fragment of cyno TR-2 amino acid/11s -16, (SEQ ID NO above：155), and the reverse primer on people's TR-2 amino acid/11 7-154 fragments, (SEQ ID NO above：146) cyno and the over-lap PCR of people's TR-2 fragments, are carried out.Amino acid sequence on cyno/ people's chimera #2 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLAPITRQSLDP QRRAAPQQKRSSPSEGLCPPGHHISEDGRDYISCKYGQDYSTHWNDLLFCLRCTRC DSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSD IECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQID NO：87).

Cyno/ people's chimera #3 is prepared by following：Use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO：145) with reverse primer GGATCTCTTTTGTTGTGGGGCCGCTCTCTGCTGG G (SEQ ID NO：161), amplification orresponding amino acid 1-16 into acquaintance TR-2 regions, and use forward primer CAGCAGAGAGCGGCCCCACAACAAAAGAGATCCAGC (SEQ ID NO：162) with reverse primer GTATGAGGGATCCTCACTGACACACCGTGTTTCTGG (SEQ ID NO：158), amplification orresponding amino acid 17-85 ripe cyno TR-2 regions.Using the forward primer on the fragment of people TR-2 amino acid/11s -16, (SEQ ID NO above：145), and the reverse primer on cyno TR-2 amino acid/11 7-85 fragments, (SEQ ID NO above：158) cyno and the over-lap PCR of people's TR-2 fragments, are carried out.Amino acid sequence on cyno/ people's chimera #3 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVSSCTTTRNTVCQ (SEQ ID NO：88).

Cyno/ people's chimera #4 is prepared by following：Use forward primer GTAAGCAAGCTTGGCTCTGATCACCCAACAAGA (SEQ ID NO：145) with reverse primer GGATCTCTTTTGTTGTGGGGCCGCTCTCTGCTGG G (SEQ ID NO：161), amplification orresponding amino acid 1-16 into acquaintance TR-2 regions, and use forward primer CAGCAGAGAGCGGCCCCACAACAAAAGAGATCCAGC (SEQ ID NO：162) with reverse primer GTAGTTGGATCCTCAAGAAGCAGGAGTCCCAGGG (SEQ ID NO：157), amplification orresponding amino acid 17-154 ripe cyno TR-2 regions.Using the forward primer on the fragment of people TR-2 amino acid/11s -16, (SEQ ID NO above：145), and the reverse primer on cyno TR-2 amino acid/11 7-154 fragments, (SEQ ID NO above：157) cyno and the over-lap PCR of people's TR-2 fragments, are carried out.Amino acid sequence on cyno/ people's chimera #4 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPTEGLCPPGHHISEDSRDCISCKYGQDYSTHWNDFLFCLRCTKC DSGEVEVSSCTTTRNTVCQCEEGTFREEDSPEICRKCRTGCPRGMVKVKDCTPWSD IECVHKESGTKHTGEVPAVEKTVTTSPGTPAS (SEQ ID NO：89).Under the background of N- Avidin fusions, the short region for replacing people TR-2 by using corresponding mouse TR-2 sequences builds the TR-2 protein of 4 kinds of other modifications.People/mouse TR-2#1 includes the mouse TR-2 sequences from amino acid/11-22 and people's TR-2 sequences from amino acid 23-150.People/mouse TR-2#2 includes the people TR-2 sequences from amino acid/11-28 and the mouse TR-2 sequences from amino acid 29-34.People/mouse TR-2#3 includes the people TR-2 sequences from amino acid/11-53 and the mouse TR-2 sequences from amino acid 54-59.People/mouse TR-2#4 includes the people TR-2 sequences from amino acid/11-66 and the mouse TR-2 sequences from amino acid 67-75.In order to form the protein of every kind of modification, then the polynucleotides of amplification are subcloned into the pCEP4 carriers (Invitrogen) as described above comprising chicken avidin sequence.

People/mouse TR-2#1 is prepared by following：Use forward primer CAGCGGCCGGAGGAGAGCCCCTCAGAGGGATTGT (SEQ ID NO：163) with reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO：164), amplification orresponding amino acid 23-150 into acquaintance TR-2 regions, and use forward primer TGAATGAAGCTTGGTTCCAGTAACAGCTAACCCA (SEQ ID NO：165) with reverse primer TCCCTCTGAGGGGCTCTCCTCCGGCCGCTGTAG (SEQ ID NO：166), amplification orresponding amino acid 1-22 ripe mouse TR-2 regions.Using the forward primer on the fragment of mouse TR-2 amino acid/11s -22, (SEQ ID NO above：165), and the reverse primer on people's TR-2 amino acid 23-150 fragments, (SEQ ID NO above：164) over-lap PCR of pedestrian and mouse TR-2 fragments, is entered.Amino acid sequence on people/mouse TR-2#1 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT 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) with reverse primer CAGGTACTGGCCTGCTAGACACAATCCCTCTGAGGGG (SEQ ID NO：167), amplification orresponding amino acid 1-28 into acquaintance TR-2 regions, use forward primer CTAGCAGGCCAGTACCTGTCAGAAGACGGTAGAGATTGC (SEQ ID NO：168) with reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO：164), amplification orresponding amino acid 35-150 into acquaintance TR-2 regions, and use forward primer CAGGTACTGGCCTGCTAGACACAATCCCTCTGAGGGG (SEQ IDNO：169) with reverse primer CTAGCAGGCCAGTACCTGTCAGAAGACGGTAGAGATTGC (SEQ ID NO：170), amplification orresponding amino acid 29-34 ripe mouse TR-2 regions.Using the forward primer on the fragment of people TR-2 amino acid/11s -28, (SEQ ID NO above：145), and the reverse primer on people's TR-2 amino acid 35-150 fragments, (SEQ ID NO above：170) over-lap PCR of pedestrian and mouse TR-2 fragments, is entered.Amino acid sequence on 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) with reverse primer TGAATCCAGAGAATGGTTGGAGTGAGTGCTATAGTCCTG TC (SEQID NO：171), amplification orresponding amino acid 1-53 into acquaintance TR-2 regions, and use forward primer TCCAACCATTCTCTGGATTCA TGCTTGCGCTGCACCAGG (SEQ ID NO：172) with reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO：173), amplification orresponding amino acid 60-154 into acquaintance TR-2 regions.Above-mentioned primer includes coding orresponding amino acid 54-59 mouse TR-2 nucleotides.Using the forward primer on the fragment of people TR-2 amino acid/11s -53, (SEQ ID NO above：145), and the reverse primer on people's TR-2 amino acid 60-154 fragments, (SEQ ID NO above：173) over-lap PCR of pedestrian and mouse TR-2 fragments, is entered.Amino acid sequence on 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) with reverse primer TCGGGTTTCTACGACTTTATCTTCCTTACACCTGGTGCAGCGCAAG (SEQ ID NO：174), amplification orresponding amino acid 1-66 into acquaintance TR-2 regions, and use forward primer AAGGAAGATAAAGTCGTAGAAACCCGATGCACCACGACCAGAAAC AC (SEQID NO：175) with reverse primer GATTGAGGATCCCTAAGAGGCAGGAGTCCCTGG (SEQ ID NO：176), amplification orresponding amino acid 76-154 into acquaintance TR-2 regions.Above-mentioned primer includes coding orresponding amino acid 67-75 mouse TR-2 nucleotides.Using the forward primer on the fragment of people TR-2 amino acid/11s -66, (SEQID NO above：145), and the reverse primer on people's TR-2 amino acid 76-154 fragments, (SEQ ID NO above：176) over-lap PCR of pedestrian and mouse TR-2 fragments, is entered.Amino acid sequence on people/mouse TR-2#4 is MVHATSPLLLLLLLSLALVAPGLSARKCSLTGKWTNDLGSNMTIGAVNSKGEFTGT YTTAVTATSNEIKESPLHGTQNTINKRTQPTFGFTVNWKFSESTTVFTGQCFIDRN GKEVLKTMWLLRSSVNDIGDDWKATRVGINIFTRLRTQKEQLLASLALITQQDLAP QQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHSNHSLDSCLRCTR CDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWS DIECVHKESGTKHSGEAPAVEETVTSSPGTPAS (SEQ ID NO：93).

By transiently transfecting the expression that people 293T adherent cells carry out Avidin fusion proteins in ventilation T75 tissue culture flasks.Cell is in the DMEM of the FBS and 1x pen-step- glutamine containing 10% dialysis in 37 DEG C and 5%CO₂Lower growth is simultaneously maintained., will about 3 × 10 in order to have preparation to transfection⁶293T cells be inoculated into a series of clean T75 flasks comprising 15ml growth mediums each in, and all flasks growth overnight about 20 hours.PCEP4-Avidin (N)-TR-2 constructs are each transfected into as follows different intracellular.In the presence of Opti-MEM culture mediums (Invitrogen), 15 μ g DNA are made to be mixed with 75 μ L Lipofectamine2000 (Invitrogen), to form DNA-Lipofectamine compounds.Compound is set to incubate 20 minutes.During incubating, growth medium is suctioned out from T75 flasks and is replaced with 15mLOpti-MEM.After incubation, every kind of transfection composite is inoculated into different flasks, and incubated 4-5 hours in 37 DEG C.At the end of incubation period, the Opti-MEM culture mediums in each flask are replaced with into fresh growth medium.About 48 hours after transfection, harvest conditioned medium and be transferred to 50ml pipes (Falcon).Pipe is centrifuged 10 minutes in 4 DEG C under 2000x g to remove cell and fragment, and be subsequently transferred to clean 50mL pipes.The control flask for lacking transfection DNA is equally prepared according to same approach, the negative control condition culture medium for Binding experiment is produced.

The concentration of every kind of N- avidins-TR-2 fusion proteins is determined using the determination method based on quantitative FACS.Avidin fusion proteins are captured on 6.7 μm of biotin polystyrene beads (Spherotech, Inc.).2 kinds of samples are prepared for every kind of fusion protein：5 μ L (about 3.5 × 10⁵) pearl suspension adds 20 μ L 1x conditioned mediums, and 5 μ L pearl suspension to add 200 μ L1x conditioned mediums.Accompanying rotation is incubated 1 hour all samples at room temperature.By centrifuging and being washed with the PBS (BPBS) comprising 0.5%BSA conditioned medium is removed from every kind of sample.Avidin pearl is dyed with the μ g/mL goats FITC of 200 μ L 0.5 for being dissolved in BPBS anti-anti-avidin antibody (Vector Labs, Burlingame, CA) solution marked.Reaction is allowed to carry out 45 minutes at room temperature, wherein reaction tube is covered by paper tinsel.After incubation, collection pearl is washed again by centrifugation and with BPBS, and be resuspended in 0.5ml BPBS for analyzing.FITC fluorescence is detected using FACScan (Becton Dickinson Bioscience).Protein quality is transposed the signals into using the standard curve as derived from restructuring avidin.

Assess the anti-TR-2 antibody of 2 kinds of people truncate, people TR-2 and respective combinations of TR-2 from machin with people TR-2.Binding assay is carried out as follows.Biotin pearl as described above loads one of about 100ng N- avidin TR-2 fusion proteins/3.5 × 10⁵Pearl and reach volume with growth medium.Pearl is set to be mixed with the 1 μ g FITC anti-TR-2 monoclonal antibodies of people being conjugated in 0.2mLBPBS.After incubating 1 hour at room temperature, add 3mL BPBS and collect antibody-pearl compound by centrifuging 5 minutes under 750x g.Agglomerate is washed in 3mL BPBS.The antibody combined with avidin-pearl compound is detected by facs analysis.For every kind of sample record average fluorescent strength.Using with lacking TR-2 those antibody for being combined of conditioned medium as negative control (" Neg CM ").As a result it is shown in Figure 24.

The binding pattern that 2 kinds of antibody are observed is similar.It was observed that most strong combine be that wherein average fluorescent strength is 7349 on positive control people TR-2.It was observed that antibody (as with fluorescence intensity measurement) and truncate TR-2-2 combination be 6561-6693, and truncate TR-2-3 and TR-2-5 combination is 3158-3866, and truncate TR-2-6 combination is 1959-2202, and and truncate TR-2-1 combination be 662-759.The antibody and total length TR-2 from machin combination (as with fluorescence intensity measurement) are 666-764.As by with reference to the background classes on experiment it is true determine, the antibody not with mouse or rat TR-2, or do not combined with truncate TR-2-4, TR-2-7, TR-2-9, TR-2-10, TR-2-11, TR-2-12 or TR-2-13.

TR-2-1 is that C-terminals of the TR-2 after amino acid 43 is truncate, and TR-2-2, -3, -5 and -6 all include at least amino acid/11 6-85.(referring to the result on TR-2-2) in the presence of the whole region from amino acid/11-85, with reference to generation.Amino acid 86-126 addition, which makes to combine, reduces by about 2 times (comparing the result on TR-2-2 and TR-2-3).Being not present for amino acid/11-15 from TR-2N ends in TR-2-2 makes to combine about 2 times (comparing the result on TR-2-2 and TR-2-5) of reduction.Simultaneous amino acid/11-15 be not present and amino acid 86-126 addition make combine reduction about 3 times (comparing the result on TR-2-2 and TR-2-6).Residue 44-85 elimination (TR-2-1), which makes to combine, to be reduced to for about the 11% of those observed of TR-2-2.Those results point out (the SEQ ID NO of amino acid/11-15：94；) and 44-85 (SEQ ID NO ALITQQDLAPQQRAA：95；CKYGQDYSTHWNDLL FCLRCTRCDSGEVE LSPCTTTRNTVCQ) one or more of region residue is important for that anti-TR-2 antibody of 2 kinds of people and people TR-2 combination.

Also the anti-TR-2 antibody of evaluator and cyno TR-2 be truncate, people/cyno chimeras and the respective combinations of people TR-2 comprising some mouse TR-2 domains.Anti- TR-2 antibody is combined strongly with total length people TR-2 (fluorescence intensity (" FI ") 5681).Anti- TR-2 antibody and the total length cynoTR-2 of long form combination for that of total length people TR-2 than reducing by about 5 times (FI 1573).Total length cyno TR-2 (FI 209) for short-form and only observe the background levels combined for the truncate 17-154 of cyno TR-2 (FI51), cyno 1-85 (FI 11) and cyno 17-85 (FI 8).

It also have evaluated the combination of the anti-TR-2 antibody of some people and cyno/ people's TR-2 chimeras (referring to Figure 27).It was observed that antibody and 4 kinds of chimeras combination (FI) it is as follows：Cyno/ people's chimera #1：FI 5977；Cyno/ people's chimera #2：FI 47；Cyno/ people's chimera #3：FI 12；Cyno/ people's chimera #4：FI 1507.As above, it was observed that antibody and total length people TR-2 combination be 5681, and antibody and total length cyno TR-2 combination are 1573 (long forms) and 209 (short-forms).

Because antibody and cyno/ people's chimera #1 combination are similar to that with truncate TR-2-5, under human amino acid 17-85 background, replace amino acid/11-16 with corresponding cyno sequences and obviously do not interfere with antibody binding.However, under total length people TR-2 (cyno/ people #2) background, replacing amino acid/11-16 with corresponding cyno sequences and substantially cancelling combination, it was demonstrated that in the part from least one of 1-16 regions Amino acid profile epitope.And that of cyno/ people's chimera #3 and #4 combination ratio and total length people TR-2 are obviously reduced, the amino acid/11 7-85 for implying human sequence is important for combining.In a word, human sequence 1-85 regions (SEQ ID NO：96；ALITQQDLAPQQRAAPQQKRSSPSEGLCPPGHHISEDGRDCISCKYGQDYSTHWND LLFCLRCTRCDSGEVELSPCTTTRNTVCQ) in one or more amino acid be related to epitope combination.Similarly, the various human sequences in the region of amino acid/11-85 are replaced with corresponding mouse sequence and is obviously reduced antibody binding (referring to Figure 27), further confirm that one or more amino acid in that region are related to epitope 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 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>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 Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

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

85 90 95

Ala Arg Gly Arg 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 Ser

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 tcgagttacc 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>The description of artificial sequence：Synthesis

<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>The description of artificial sequence：Synthesis

<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>The 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>The 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>The 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>The 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 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

225

<210>75

<211>269

<212>PRT

<213>Artificial sequence

<220>

<223>The 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 Ash 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>The 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

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

195 200

<210>77

<211>243

<212>PRT

<213>Artificial sequence

<220>

<223>The 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>The 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>The 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>The 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>The 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>The 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>The 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>The 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>The 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>The 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

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 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>The 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>The 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>The 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>The 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>The 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

260 265 270

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>92

<211>312

<212>PRT

<213>Artificial sequence

<220>

<223>The 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 Ash 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>The 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>The description of artificial sequence：Synthetic primer

<400>97

gtaggtgctg tcct 14

<210>98

<211>14

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>98

tgagttccac gaca 14

<210>99

<211>13

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>99

cttccaagcc act 13

<210>100

<211>12

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>100

cargcactgt ca 12

<210>101

<211>18

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>101

gtaggtgctg tccttgct 18

<210>102

<211>21

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>102

ctctgtgaca ctctcctggg a 21

<210>103

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>103

gctctagatt ggagggcgtt atccaccttc cact 34

<210>104

<211>39

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>104

aactagctag cagttccaga tttcaactgc tcatcagat 39

<210>105

<211>18

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>105

gctcccgggt agaagtca 18

<210>106

<211>22

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>106

acyagtgtgg ccttgttggc tt 22

<210>107

<211>26

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>107

gctctagagg gygggaacag agtgac 26

<210>108

<211>18

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>108

acgacaccgt caccggtt 18

<210>109

<211>25

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>109

aagtagtcct tgaccaggca gccca 25

<210>110

<211>30

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>110

gctctagagg gtgccagggg gaagaccgat 30

<210>111

<211>28

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>111

gctctagagc agggcgccag ggggaaga 28

<210>112

<211>20

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>112

atgaggstcc cygctcagct 20

<210>113

<211>21

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>113

atggaarccc cagckcagct t 21

<210>114

<211>22

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>114

atggtgttgc agacccaggt ct 22

<210>115

<211>35

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>115

gaagatctca ccatgaggst cccygctcag ctyct 35

<210>116

<211>38

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>116

gaagatctca ccatggaarc cccagckcag cttctctt 38

<210>117

<211>38

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>117

gaagatctca ccatggtgtt gcagacccag gtcttcat 38

<210>118

<211>18

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>118

crtcwccacc atgrcmwg 18

<210>119

<211>19

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>119

caccatgrcc wgstyccct 19

<210>120

<211>20

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>120

accatggcct ggrctcykct 20

<210>121

<211>19

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>121

caccatggcm tggrycvyt 19

<210>122

<211>21

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>122

caccatggcy tggryccmay t 21

<210>123

<211>29

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>123

gaagatctca ccatgrccwg styccctct 29

<210>124

<211>32

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>124

gaagatctca ccatggcctg grctcykcts yt 32

<210>125

<211>30

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>125

gaagatctca ccatggcmtg grycvytctc 30

<210>126

<211>30

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>126

gaagatctca ccatggcytg gryccmaytc 30

<210>127

<211>25

<212>DNA

<213>Artificial sequence

<220>

<223>The 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>The description of artificial sequence：Synthetic primer

<400>128

caccatggac atactttgyt ccacgctc 28

<210>129

<211>23

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>129

caccatggar ttkggrctbh gct 23

<210>130

<211>30

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>130

caccatgaar cayctgtggt tcttcctyct 30

<210>131

<211>24

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>131

caccatgggg tcaaccgyca tcct 24

<210>132

<211>28

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>132

caccatgtct gtctccttcc tcatcttc 28

<210>133

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The 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>The description of artificial sequence：Synthetic primer

<400>134

gaagatctca ccatggacat actttgytcc acgctcc 37

<210>135

<211>41

<212>DNA

<213>Artificial sequence

<220>

<223>The 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>The description of artificial sequence：Synthetic primer

<400>136

gaagatctca ccatgaarca yctgtggttc ttcctyctc 39

<210>137

<211>32

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>137

gaagatctca ccatggggtc aaccgycatc ct 32

<210>138

<211>37

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>138

gaagatctca ccatgtctgt ctccttcctc atcttct 37

<210>139

<211>44

<212>DNA

<213>Artificial sequence

<220>

<223>The 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>Mice species

<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>Mice species

<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>Mice species

<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>Mice species

<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>The description of artificial sequence：Synthetic primer

<400>145

gtaagcaagc ttggctctga tcacccaaca aga 33

<210>146

<211>30

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>146

gattagggat ccagaggcag gagtccctgg 30

<210>147

<211>35

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>147

tagttgggat cctcaggaga tgcaatctct accgt 35

<210>148

<211>35

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>148

ggtagtggat cctcactgac acactgtgtt tctgg 35

<210>149

<211>35

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>149

gtaatgggat cctcagacac attcgatgtc actcc 35

<210>150

<211>33

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>150

gtaatgaagc ttgccacaac aaaagaggtc cag 33

<210>151

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>151

gattgaaagc ttgatctcct gcaaatatgg acag 34

<210>152

<211>32

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>152

gtaatgaagc ttgcagtgcg aagaaggcac ct 32

<210>153

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>153

gatggaggat cctcaacacc tggtgcagcg caag 34

<210>154

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>154

gtaagtggat cctcagcagg gacttagctc cact 34

<210>155

<211>29

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>155

gttagtaagc ttggctccaa tcacccgac 29

<210>156

<211>31

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>156

gttgatggat ccttctttgt ggacactcga t 31

<210>157

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>157

gtagttggat cctcaagaag caggagtccc aggg 34

<210>158

<211>36

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>158

gtatgaggga tcctcactga cacaccgtgt ttctgg 36

<210>159

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>159

gtatggaagc ttgccacaac aaaagagatc cagc 34

<210>160

<211>35

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>160

cagcgaagag cggctccaca acaaaagagg tccag 35

<210>161

<211>35

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>161

ggatctcttt tgttgtgggg ccgctctctg ctggg 35

<210>162

<211>36

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>162

cagcagagag cggccccaca acaaaagaga tccagc 36

<210>163

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>163

cagcggccgg aggagagccc ctcagaggga ttgt 34

<210>164

<211>33

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>164

gattgaggat ccctaagagg caggagtccc tgg 33

<210>165

<211>34

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>165

tgaatgaagc ttggttccag taacagctaa ccca 34

<210>166

<211>33

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>166

tccctctgag gggctctcct ccggccgctg tag 33

<210>167

<211>37

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>167

caggtactgg cctgctagac acaatccctc tgagggg 37

<210>168

<211>39

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>168

ctagcaggcc agtacctgtc agaagacggt agagattgc 39

<210>169

<211>37

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>169

caggtactgg cctgctagac acaatccctc tgagggg 37

<210>170

<211>39

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>170

ctagcaggcc agtacctgtc agaagacggt agagattgc 39

<210>171

<211>41

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>171

tgaatccaga gaatggttgg agtgagtgct atagtcctgt c 41

<210>172

<211>39

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>172

tccaaccatt ctctggattc atgcttgcgc tgcaccagg 39

<210>173

<211>33

<212>DNA

<213>Artificial sequence

<220>

<223>The description of artificial sequence：Synthetic primer

<400>173

gattgaggat ccctaagagg caggagtccc tgg 33

<210>174

<211>46

<212>DNA

<213>Artificial sequence

<220>

<223>The 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>The 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>The 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 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 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>Machin

<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>Mice species

<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 kind of isolated polypeptide, it includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a

Wherein CDR1a includes amino acid sequence abcdefghijkl, 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

Wherein CDR2a includes amino acid sequence mnopqrstuvwxyza ' b ' c ', and wherein amino acid m is selected from tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；Wherein CDR3a includes amino acid 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 tryptophan, aspartic acid, glycine, serine or glutamic acid；Amino acid e ' is selected from asparagine, aspartic acid, glycine, arginine, serine, valine or leucine；Amino acid f ' is selected from histidine, serine, alanine, tyrosine, proline, asparagine, glycine or threonine；Amino acid g ' is selected from tyrosine, serine, alanine, arginine, tryptophan, glycine or valine；Amino acid h ' is selected from glycine, alanine, serine, asparagine, methionine, tyrosine, tryptophan, cysteine or aspartic acid；Amino acid i ' is selected from serine, tryptophan, glycine, phenylalanine, aspartic acid, tyrosine or threonine；Amino acid j ' is selected from glycine, threonine, serine, leucine, valine, asparagine, tryptophan or tyrosine；Amino acid k ' is selected from serine, phenylalanine, aspartic acid, tryptophan, glycine or tyrosine or is not present；Amino acid l ' is selected from histidine, aspartic acid, alanine, tryptophan, tyrosine, serine, phenylalanine, valine or glycine or is not present；Amino acid m ' is selected from phenylalanine, tyrosine, glutamic acid, proline, aspartic acid, cysteine, isoleucine or methionine or is not present；Amino acid n ' is selected from aspartic acid, phenylalanine, alanine, leucine or serine or is not present；Amino acid o ' is selected from tyrosine, leucine, aspartic acid, phenylalanine, proline or valine or is not present；Amino acid p ' is selected from leucine, aspartic acid or tyrosine or is not present；Amino acid q ' is selected from serine or tyrosine or is not present；Amino acid r ' is tyrosine or is not present；Amino acid s ' is selected from glycine or tyrosine or is not present；Amino acid t ' is selected from glycine or methionine or is not present；Amino acid u ' is selected from methionine or aspartic acid or is not present；Amino acid v ' is selected from aspartic acid or valine or is not present；It is valine with amino acid w ' or is not present；With

2. the isolated polypeptide of claim 1, wherein the polypeptide includes heavy chain of antibody variable region.

3. the isolated polypeptide of claim 2, wherein the heavy chain of antibody variable region is human antibody heavy chain variable region.

4. the isolated polypeptide of claim 2, it further includes heavy chain constant region.

5. the isolated polypeptide of claim 4, wherein the heavy chain constant region is human antibody heavy chain's constant region.

6. the isolated polypeptide of claim 3, it further includes heavy chain constant region.

7. the isolated polypeptide of claim 6, wherein the heavy chain constant region is human antibody heavy chain's constant region.

8. the isolated polypeptide of claim 7, wherein the heavy chain of antibody variable region and the heavy chain constant region include such as 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：Amino acid sequence shown in 34.

9. the isolated polypeptide of claim 1, it includes at least two complementary determining region (CDRs) of CDR1a, CDR2a and CDR3a selected from claim 1.

10. the isolated polypeptide of claim 1, it includes CDR1a, CDR2a and CDR3a of claim 1.

11. antibody fragment of the one kind selected from Fab, Fab ', F (ab ') 2, Fv and single-chain antibody, wherein the antibody fragment includes the isolated polypeptide of claim 1.

12. a kind of isolated polypeptide, it, which is included, is selected from least one following complementary determining regions (CDR)：

SEQ ID NO：2 amino acid 26-35；

SEQ ID NO：2 amino acid 50-66；

SEQ ID NO：2 amino acid 99-110；

SEQ ID NO：4 amino acid 26-37；

SEQ ID NO：4 amino acid 52-67；

SEQ ID NO：4 amino acid/11 00-109；

SEQ ID NO：6 amino acid 26-37；

SEQ ID NO：6 amino acid 52-67；

SEQ ID NO：6 amino acid/11 00-109；

SEQ ID NO：8 amino acid 26-37；

SEQ ID NO：8 amino acid 52-67；

SEQ ID NO：8 amino acid/11 00-109；

SEQ ID NO：10 amino acid 26-35；

SEQ ID NO：10 amino acid 50-66；

SEQ ID NO：10 amino acid 99-110；

SEQ ID NO：12 amino acid 26-35；

SEQ ID NO：12 amino acid 50-66；

SEQ ID NO：12 amino acid 99-111；

SEQ ID NO：14 amino acid 26-35；

SEQ ID NO：14 amino acid 50-65；

SEQ ID NO：14 amino acid 98-111；

SEQ ID NO：16 amino acid 26-37；

SEQ ID NO：16 amino acid 52-67；

SEQ ID NO：16 amino acid/11 00-109；

SEQ ID NO：18 amino acid 26-35；

SEQ ID NO：18 amino acid 50-66；

SEQ ID NO：18 amino acid 99-105；

SEQ ID NO：20 amino acid 26-35；

SEQ ID NO：20 amino acid 50-66；

SEQ ID NO：20 amino acid 99-118；

SEQ ID NO：22 amino acid 26-35；

SEQ ID NO：22 amino acid 50-66；

SEQ ID NO：22 amino acid 99-118；

SEQ ID NO：24 amino acid 26-35；

SEQ ID NO：24 amino acid 50-65；

SEQ ID NO：24 amino acid 98-108；

SEQ ID NO：26 amino acid 26-35；

SEQ ID NO：26 amino acid 50-66；

SEQ ID NO：26 amino acid 99-110；

SEQ ID NO：28 amino acid 26-35；

SEQ ID NO：28 amino acid 50-66；

SEQ ID NO：28 amino acid 99-117；

SEQ ID NO：30 amino acid 26-37；

SEQ ID NO：30 amino acid 52-67；

SEQ ID NO：30 amino acid/11 00-111；

SEQ ID NO：32 amino acid 26-37；

SEQ ID NO：32 amino acid 52-67；

SEQ ID NO：32 amino acid/11 00-111；

SEQ ID NO：34 amino acid 26-37；

SEQ ID NO：34 amino acid 52-67；With

SEQ ID NO：34 amino acid/11 00-111；

The polypeptide combination TR-2 wherein combined with antibody light chain.

13. the isolated polypeptide of claim 12, complementary determining region (CDRs) described in its at least two comprising claim 12.

14. the isolated polypeptide of claim 12, complementary determining region (CDRs) described in its at least three comprising claim 12.

15. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：2 amino acid 26-35, SEQ ID NO：2 amino acid 50-66 and SEQ ID NO：2 amino acid 99-110.

16. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：4 amino acid 26-37, SEQ ID NO：4 amino acid 52-67 and SEQ ID NO：4 amino acid/11 00-109.

17. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：6 amino acid 26-37, SEQ ID NO：6 amino acid 52-67 and SEQ ID NO：6 amino acid/11 00-109.

18. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：8 amino acid 26-37, SEQ ID NO：8 amino acid 52-67 and SEQ ID NO：8 amino acid/11 00-109.

19. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：10 amino acid 26-35, SEQ ID NO：10 amino acid 50-66 and SEQ IDNO：10 amino acid 99-110.

20. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：12 amino acid 26-35, SEQ ID NO：12 amino acid 50-66 and SEQ IDNO：12 amino acid 99-111.

21. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：14 amino acid 26-35, SEQ ID NO：14 amino acid 50-65 and SEQ IDNO：14 amino acid 98-111.

22. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：16 amino acid 26-37, SEQ ID NO：16 amino acid 52-67 and SEQ IDNO：16 amino acid/11 00-109.

23. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：18 amino acid 26-35, SEQ ID NO：18 amino acid 50-66 and SEQ IDNO：18 amino acid 99-105.

24. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：20 amino acid 26-35, SEQ ID NO：20 amino acid 50-66 and SEQ IDNO：20 amino acid 99-118.

25. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：22 amino acid 26-35, SEQ ID NO：22 amino acid 50-66 and SEQ IDNO：22 amino acid 99-118.

26. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：24 amino acid 26-35, SEQ ID NO：24 amino acid 50-65 and SEQ IDNO：24 amino acid 98-108.

27. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：26 amino acid 26-35, SEQ ID NO：26 amino acid 50-66 and SEQ IDNO：26 amino acid 99-110.

28. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：28 amino acid 26-35, SEQ ID NO：28 amino acid 50-66 and SEQ IDNO：28 amino acid 99-117.

29. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：30 amino acid 26-37, SEQ ID NO：30 amino acid 52-67 and SEQ IDNO：30 amino acid/11 00-111.

30. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：32 amino acid 26-37, SEQ ID NO：32 amino acid 52-67 and SEQ IDNO：32 amino acid/11 00-111.

31. the isolated polypeptide of claim 12, wherein the isolated polypeptide includes SEQ IDNO：34 amino acid 26-37, SEQ ID NO：34 amino acid 52-67 and SEQ IDNO：34 amino acid/11 00-111.

32. a kind of isolated polypeptide, it includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b

The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

33. the isolated polypeptide of claim 32, wherein the polypeptide includes antibody light chain variable region.

34. the isolated polypeptide of claim 33, wherein the antibody light chain variable region is human antibody light chain variable region.

35. the isolated polypeptide of claim 33, it further includes antibody light chain constant region.

36. the isolated polypeptide of claim 35, wherein the antibody light chain constant region is human antibody light chain constant region.

37. the isolated polypeptide of claim 34, it further includes antibody light chain constant region.

38. the isolated polypeptide of claim 37, wherein the antibody light chain constant region is human antibody light chain constant region.

39. the isolated polypeptide of claim 38, wherein the antibody light chain variable region and the antibody light chain constant region include such as 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：Amino acid sequence shown in 68.

40. the isolated polypeptide of claim 32, it includes at least two complementary determining region (CDRs) of CDR1b, CDR2b and CDR3b selected from claim 32.

41. the isolated polypeptide of claim 32, it includes CDR1b, CDR2b and CDR3b of claim 32.

42. antibody fragment of the one kind selected from Fab, Fab ', F (ab ') 2, Fv and single-chain antibody, wherein the antibody fragment includes the isolated polypeptide of claim 32.

43. a kind of isolated polypeptide, it, which is included, is selected from least one following complementary determining regions (CDR)：

SEQ ID NO：36 amino acid 24-34；

SEQ ID NO：36 amino acid 50-56；

SEQ ID NO：36 amino acid 89-97；

SEQ ID NO：38 amino acid 24-34；

SEQ ID NO：38 amino acid 50-56；

SEQ ID NO：38 amino acid 89-97；

SEQ ID NO：40 amino acid 24-34；

SEQ ID NO：40 amino acid 50-56；

SEQ ID NO：40 amino acid 89-97；

SEQ ID NO：42 amino acid 24-34；

SEQ ID NO：42 amino acid 50-56；

SEQ ID NO：42 amino acid 89-97；

SEQ ID NO：44 amino acid 24-34；

SEQ ID NO：44 amino acid 50-56；

SEQ ID NO：44 amino acid 89-97；

SEQ ID NO：46 amino acid 24-34；

SEQ ID NO：46 amino acid 50-56；

SEQ ID NO：46 amino acid 89-97；

SEQ ID NO：48 amino acid 24-40；

SEQ ID NO：48 amino acid 56-62；

SEQ ID NO：48 amino acid 95-103；

SEQ ID NO：50 amino acid 24-39；

SEQ ID NO：50 amino acid 55-61；

SEQ ID NO：50 amino acid 94-102；

SEQ ID NO：52 amino acid 24-40；

SEQ ID NO：52 amino acid 56-62；

SEQ ID NO：52 amino acid 95-103；

SEQ ID NO：54 amino acid 24-34；

SEQ ID NO：54 amino acid 50-56；

SEQ ID NO：54 amino acid 89-97；

SEQ ID NO：56 amino acid 24-34,

SEQ ID NO：56 amino acid 50-56；

SEQ ID NO：56 amino acid 89-97；

SEQ ID NO：58 amino acid 24-40；

SEQ ID NO：58 amino acid 56-62；

SEQ ID NO：58 amino acid 95-103；

SEQ ID NO：60 amino acid 24-34；

SEQ ID NO：60 amino acid 50-56；

SEQ ID NO：60 amino acid 89-97；

SEQ ID NO：62 amino acid 24-34；

SEQ ID NO：62 amino acid 50-56；

SEQ ID NO：62 amino acid 89-97；

SEQ ID NO：64 amino acid 24-35；

SEQ ID NO：64 amino acid 51-57；

SEQ ID NO：64 amino acid 90-88；

SEQ ID NO：66 amino acid 24-34；

SEQ ID NO：66 amino acid 50-57；

SEQ ID NO：66 amino acid 89-97；

SEQ ID NO：68 amino acid 24-34；

SEQ ID NO：68 amino acid 50-56；With

SEQ ID NO：68 amino acid 89-97；

The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

44. the isolated polypeptide of claim 43, complementary determining region (CDRs) described in its at least two comprising claim 43.

45. the isolated polypeptide of claim 43, complementary determining region (CDRs) described in its at least three comprising claim 43.

46. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：36 amino acid 24-34, SEQ ID NO：36 amino acid 50-56 and SEQ IDNO：36 amino acid 89-97.

47. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：38 amino acid 24-34, SEQ ID NO：38 amino acid 50-56 and SEQ IDNO：38 amino acid 89-97.

48. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：40 amino acid 24-34, SEQ ID NO：40 amino acid 50-56 and SEQ IDNO：40 amino acid 89-97.

49. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：42 amino acid 24-34, SEQ ID NO：42 amino acid 50-56 and SEQ IDNO：42 amino acid 89-97.

50. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：44 amino acid 24-34, SEQ ID NO：44 amino acid 50-56 and SEQ IDNO：44 amino acid 89-97.

51. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：46 amino acid 24-34, SEQ ID NO：46 amino acid 50-56 and SEQ IDNO：46 amino acid 89-97.

52. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：48 amino acid 24-40, SEQ ID NO：48 amino acid 56-62 and SEQ IDNO：48 amino acid 95-103.

53. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：50 amino acid 24-39, SEQ ID NO：50 amino acid 55-61 and SEQ IDNO：50 amino acid 94-102.

54. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：52 amino acid 24-40, SEQ ID NO：52 amino acid 56-62 and SEQ IDNO：52 amino acid 95-103.

55. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：54 amino acid 24-34, SEQ ID NO：54 amino acid 50-56 and SEQ IDNO：54 amino acid 89-97.

56. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：56 amino acid 24-34, SEQ ID NO：56 amino acid 50-56 and SEQ IDNO：56 amino acid 89-97.

57. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：58 amino acid 24-40, SEQ ID NO：58 amino acid 56-62 and SEQ IDNO：58 amino acid 95-103.

58. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：60 amino acid 24-34, SEQ ID NO 60 amino acid 50-56 and SEQ IDNO：60 amino acid 89-97.

59. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：62 amino acid 24-34, SEQ ID NO：62 amino acid 50-56 and SEQ IDNO：62 amino acid 89-97.

60. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：64 amino acid 24-35, SEQ ID NO：64 amino acid 51-57 and SEQ IDNO：64 amino acid 90-88.

61. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：66 amino acid 24-34, SEQ ID NO：66 amino acid 50-57 and SEQ IDNO：66 amino acid 89-97.

62. the isolated polypeptide of claim 43, wherein the isolated polypeptide includes SEQ IDNO：68 amino acid 24-34, SEQ ID NO：68 amino acid 50-56 and SEQ IDNO：68 amino acid 89-97.

63. one kind separation polynucleotides, it includes the sequence of coded polypeptide, and the polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a

Wherein CDR1a includes amino acid sequence abcdefghijkl, and wherein amino acid a is glycine, amino acid b 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

Wherein CDR2a includes amino acid sequence mnopqrstuvwxyza ' b ' c ', and wherein amino acid m is selected from tryptophan, tyrosine, histidine, valine, glutamic acid or serine；Amino acid n is selected from methionine or isoleucine；Amino acid o is selected from asparagine, tyrosine, serine, tryptophan or histidine；Amino acid p is selected from proline, tyrosine, serine, arginine, histidine or asparagine；Amino acid q is selected from 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 asparagine, threonine, alanine, isoleucine or tyrosine；Amino acid u is selected from threonine, tyrosine, leucine, lysine, asparagine or isoleucine；Amino acid v is selected from glycine, tyrosine, aspartic acid or cysteine；Amino acid w is selected from tyrosine or asparagine；Amino acid x is selected from alanine or proline；Amino acid y is selected from glutamine, serine or aspartic acid；Amino acid z is selected from lysine, leucine or serine；Amino acid a ' is selected from phenylalanine, lysine or valine；Amino acid b ' is selected from glutamine, serine or lysine；It is glycine with amino acid c ' or is not present；

The polypeptide combination TR-2 wherein combined with antibody light chain.

64. the sequence of the separation polynucleotides of claim 63, wherein coded polypeptide is the sequence of encoding antibody heavy variable region.

65. the sequence of the separation polynucleotides of claim 64, wherein encoding antibody heavy variable region is the sequence of encoding human antibody's weight chain variable district.

66. the separation polynucleotides of claim 63, wherein the separation polynucleotide encoding single-chain antibody.

67. the separation polynucleotides of claim 64, it further includes the polynucleotide sequence of encoding antibody heavy constant region.

68. the polynucleotide sequence of the separation polynucleotides of claim 67, wherein encoding antibody heavy constant region includes human antibody heavy chain's constant region.

69. the separation polynucleotides of claim 65, it further includes the polynucleotide sequence of encoding antibody heavy constant region.

70. the polynucleotide sequence coding human antibody heavy chain constant region of the separation polynucleotides of claim 69, wherein encoding antibody heavy constant region.

71. the separation polynucleotides of claim 70, wherein the separation polynucleotides include the sequence of coded polypeptide, the polypeptide includes such as SEQ ID NO：2、SEQ ID NO：4、SEQID NO：6、SEQ ID NO：8、SEQ ID NO：10、SEQ ID NO：12、SEQ IDNO：14、SEQ ID NO：16、SEQ ID NO：18、SEQ ID NO：20、SEQ IDNO：22、SEQ ID NO：24、SEQ ID NO：26、SEQ ID NO：28、SEQ IDNO：30、SEQ ID NO：32 or SEQ ID NO：Amino acid sequence shown in 34.

72. the separation polynucleotides of claim 70, wherein the separation polynucleotides include such as SEQ ID NO：1；SEQ ID NO：3、SEQ ID NO：5；SEQ ID NO：7、SEQID NO：9；SEQ ID NO：11、SEQ ID NO：13；SEQ ID NO：15、SEQID NO：17；SEQ ID NO：19、SEQ ID NO：21、SEQ ID NO：23、SEQID NO：25、SEQ ID NO：27、SEQ ID NO：29、SEQ ID NO：31 or SEQID NO：Nucleotide sequence shown in 33.

73. one kind separation polynucleotides, it includes the sequence of coded polypeptide, and the polypeptide includes at least one complementary determining region (CDR) selected from CDR1b, CDR2b and CDR3b

The polypeptide combination TR-2 wherein combined with heavy chain of antibody.

74. the sequence of the separation polynucleotides of claim 73, wherein coded polypeptide is the sequence of encoding antibody light variable region.

75. the sequence of the separation polynucleotides of claim 74, wherein encoding said antibody light chain variable district is the sequence of encoding human antibody's light chain variable district.

76. the separation polynucleotides of claim 73, wherein the separation polynucleotide encoding single-chain antibody.

77. the separation polynucleotides of claim 74, it further includes the polynucleotide sequence of encoding antibody light constant region.

78. the polynucleotide sequence coding human antibody light chain constant region of the separation polynucleotides of claim 77, wherein encoding antibody light constant region.

79. the separation polynucleotides of claim 75, it further includes the polynucleotide sequence of encoding antibody light constant region.

80. the polynucleotide sequence coding human antibody light chain constant region of the separation polynucleotides of claim 79, wherein encoding antibody light constant region.

81. the separation polynucleotides of claim 80, wherein the separation polynucleotides include the sequence of coded polypeptide, the polypeptide includes such as 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：Amino acid sequence shown in 68.

82. the separation polynucleotides of claim 80, wherein the separation polynucleotides include such as SEQ ID NO：35、SEQ ID NO：37、SEQ ID NO：39、SEQ ID NO：41、SEQ ID NO：43、SEQ ID NO：45、SEQ ID NO：47、SEQ ID NO：49、SEQ ID NO：51、SEQ ID NO：53、SEQ ID NO：55、SEQ ID NO：57、SEQ ID NO：59、SEQ ID NO：61、SEQ ID NO：63、SEQ ID NO：65 or SEQ ID NO：Nucleotide sequence shown in 67.

83. the anti-TR-2 antibody of one kind separation, it includes variable region and constant region, wherein the antibody is included：

Wherein CDR1a includes amino acid sequence abcdefghijkl, 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

84. the separation antibody of claim 83, wherein the antibody is human antibody.

85. the separation antibody of claim 83, wherein the antibody is chimeric antibody.

86. the separation antibody of claim 85, it further includes detectable label.

87. the separation antibody of claim 86, wherein the detectable label be enzymatic, fluorescence, it is chemiluminescent or radioactive.

88. the anti-TR-2 antibody of one kind separation, it includes variable region and constant region, wherein the antibody is included：

89. a kind of be used to detect the present or absent methods of TR-2 in sample, it includes：

A) antibody and the sample combination of claim 83 are made；

B) separate antibody and uncombined antibody with antigen binding；With

C) detection and the existence or non-existence of the antibody of the antigen binding.

90. the method for claim 89, wherein methods described use enzyme-linked immunosorbent assay (ELISA).

91. a kind of be used to detect the present or absent kits of TR-2 in sample, it is included：

A) antibody of claim 83；With

B) it is used for the reagent for detecting the antibody.

92. a kind of method for separating TR-2, it includes：

A) antibody and matrix for making claim 83 adhere to；

B) sample comprising TR-2 is made to be exposed to the part a) antibody；With

C) TR-2 is separated.

93. a kind of kit for being used to separate TR-2, it is included：

A) antibody for the claim 83 adhered to matrix；With

B) it is used for the reagent for separating TR-2.

94. a kind of method for being used to treat the cancer in patient, it includes the antibody of the claim 83 to patient therapeuticallv's effective dose.

95. the method for claim 94, wherein the cancer is selected from least one of liver cancer, the cancer of the brain, kidney, colorectal cancer, lung cancer, spleen cancer, thymus gland or haemocyte cancer (i.e. leukaemia), prostate cancer, carcinoma of testis, oophoroma, uterine cancer, breast cancer, cancer of pancreas, stomach cancer, H＆N squamous cell carcinoma and lymthoma.

96. a kind of expression vector, it includes the polynucleotides of claim 63.

97. a kind of expression vector, it includes the polynucleotides of claim 73.

98. a kind of cell, its at least one expression vector comprising claim 96 or claim 97.

99. a kind of cell, it is included：

(a) the first polynucleotides for the sequence for encoding the first polypeptide are included, the first described polypeptide includes at least one complementary determining region (CDR) selected from CDR1a, CDR2a and CDR3a, wherein CDR1a includes amino acid sequence abcdefghijkl, and wherein amino acid a is glycine, amino acid b 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 asparagine；Amino acid f is selected from serine, aspartic acid, tyrosine, 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, asparagine or serine；Amino acid i is selected from tyrosine, isoleucine, histidine, methionine or tryptophan；Amino acid j is selected from asparagine, tyrosine, histidine, serine or phenylalanine；Amino acid k is tryptophan or is not present；It is serine with amino acid l or is not present；

100. the cell of claim 99, wherein the first described polynucleotides and second of polynucleotides are in separated expression vector.

101. the cell of claim 99, wherein the first described polynucleotides and second of polynucleotides are in single expression vector.

102. a kind of method for preparing polypeptide, it is included in the polynucleotides for being suitable for wherein including and expresses that under conditions of producing the polypeptide, the polypeptide is produced in the cell of the expression vector comprising claim 96.

103. a kind of method for preparing polypeptide, it is included in the polynucleotides for being suitable for wherein including and expresses that under conditions of producing the polypeptide, the polypeptide is produced in the cell of the expression vector comprising claim 97.

104. a kind of method for preparing anti-TR-2 antibody, it, which is included in, is suitable for the polynucleotides that wherein include and expresses that under conditions of producing the antibody, the antibody is produced in the cell of the expression vector comprising claim 96 or the expression vector of claim 97.

105. a kind of method for preparing anti-TR-2 antibody, it, which is included in the cell of claim 99, claim 100 or claim 101, produces the antibody.

106. a kind of pharmaceutical composition, its antibody comprising claim 83 and pharmaceutically acceptable carrier.

107. a kind of separation antibody, it is combined with epitope specificity, and the epitope is by selected from following at least one antibody specificity combinations：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.

108. a kind of polypeptide, it, which is included, is selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：96 at least one amino acid sequence.

109. a kind of polypeptide, it is substantially by selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：96 at least one amino acid sequence composition.

110. a kind of antibody or antigen-binding domains, it, which is combined, is selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：96 at least one amino acid sequence.

111. a kind of method for obtaining the antibody for combining TR-2, it, which includes applying to animal, is selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：96 at least one polypeptide, and the antibody for combining TR-2 is obtained from the animal.

112. a kind of method for reducing or preventing antibody to be combined with TR-2, it is by applying comprising selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：The polypeptide of 96 at least one amino acid sequence.

113. a kind of method for reducing or preventing antibody to be combined with TR-2, it is by applying by selected from SEQ ID NO：94、SEQ ID NO：95 and SEQ ID NO：The polypeptide of 96 at least one amino acid sequence composition.