CN114685669A - 结合trop2的抗体及其用途 - Google Patents

结合trop2的抗体及其用途 Download PDF

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CN114685669A
CN114685669A CN202011611968.XA CN202011611968A CN114685669A CN 114685669 A CN114685669 A CN 114685669A CN 202011611968 A CN202011611968 A CN 202011611968A CN 114685669 A CN114685669 A CN 114685669A
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吕强
黄冰
戎一平
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Heping Pharmaceutical Suzhou Co ltd
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Heping Pharmaceutical Suzhou Co ltd
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Priority to CN202011611968.XA priority Critical patent/CN114685669A/zh
Priority to TW110148873A priority patent/TWI824364B/zh
Priority to CN202180087709.8A priority patent/CN117177991A/zh
Priority to PCT/CN2021/142095 priority patent/WO2022143670A1/zh
Priority to EP21914404.5A priority patent/EP4273171A1/en
Priority to US18/260,074 priority patent/US20240067748A1/en
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Abstract

本发明提供了一种与Trop2特异结合的分离的单克隆抗体。还提供编码该抗体的核酸分子，以及用于表达该抗体的表达载体、宿主细胞和方法。本发明还提供包含该抗体的免疫偶联物、双特异性分子、嵌合抗原受体、溶瘤病毒和药物组合物，以及使用本发明抗体的诊断或治疗方法。

Description

结合TROP2的抗体及其用途
技术领域
本发明涉及一种与Trop2特异结合的抗体、及其制备和用途，尤其是其在诊断、预防和治疗与Trop2相关的疾病中的用途，疾病包括肿瘤，例如乳癌、胃癌、胰腺癌、卵巢癌、肠癌等。
背景技术
滋养层细胞表面抗原2(Trop2)，又称为肿瘤关联钙信号转导因子2(TACSTD2)、膜组分1表面标记1(M1S1)、上皮糖蛋白1(EGP1)或肠胃抗原733-1(GA733-1)，是一种I型跨膜细胞表面糖蛋白。
经无内含子基因的编码，Trop2共有323个氨基酸残基。该蛋白可以转导胞内钙信号，且含有保守的4,5-二磷酸磷脂酰肌醇(PIP(2))结合基序以及与蛋白激酶C相互作用的丝氨酸磷酸化位点。Trop2还与IGF-1、Claudin-1、Claudin-7、以及细胞周期蛋白D1相互作用。例如，Trop2经由周期蛋白D1和ERK-MAPK通路激活CREB1、NF-κB、STAT1和STAT3，促成肿瘤细胞发展。
Trop2在很多正常组织中表达，在器官发生和干细胞维持方面起到重要作用。同时，Trop2也在很多人的癌细胞中大量表达，例如乳癌、胃癌、胰腺癌和卵巢癌细胞，且Trop2的过表达与增加的肿瘤生长、肿瘤侵袭性、肿瘤转移以及较差的预后相关。例如，肠肿瘤细胞和卵巢肿瘤细胞中的Trop2低表达抑制细胞增殖。Trop2过表达增加胰腺癌细胞的迁移。因此，Trop2被认为是潜在的治疗靶点。
研究表明，Trop2抗体抑制体内肿瘤生长，减少肠癌细胞和乳癌细胞的体内细胞迁移。已经证实Sacituzumab govitecan(IMMU-132)，一种Trop2抗体-SN-38耦联药物，在一些肿瘤细胞系移植模型中有效果，且正在患有顽固三阴乳癌、转移性尿路上皮癌和耐受去势的转移前列腺癌的病人中做临床试验，中性粒细胞缺乏和腹泻是最常见的副作用。
因此，本领域迫切需要开发一种具有较佳的抗肿瘤效果并且毒性低的Trop2单克隆抗体。
发明内容
本发明的目的就是提供一种具有较佳的抗肿瘤效果并且毒性低的Trop2单克隆抗体。
在本发明的第一方面，提供了一种Trop-2的抗体或抗原结合片段，包含如下的互补决定区(CDR)：
(a)SEQ ID NO:192-220中任一所示的重链可变区(VH)中含有的HCDR1或其序列的变体、HCDR2或其序列的变体、以及HCDR3或其序列的变体；和/或
(b)SEQ ID NO:221-240中任一所示的轻链可变区(VL)中含有的LCDR1或其序列的变体、LCDR2或其序列的变体、以及LCDR3或其序列的变体；
优选地，所述序列的变体为与其来源CDR相比为具有一个或几个氨基酸的置换、缺失或添加(例如1个，2个或3个氨基酸的置换、缺失或添加)的CDR；优选地，所述的置换为保守置换。
在另一优选例中，所述抗体或抗原结合蛋白的重链可变区VH具有选自下组的互补决定区CDR组合：
(1)如SEQ ID NO:19所示的CDR1、如SEQ ID NO:47所示的CDR2，和如SEQ ID NO:86所示的CDR3；
(2)如SEQ ID NO:20所示的CDR1、如SEQ ID NO:48所示的CDR2，和如SEQ ID NO:87所示的CDR3；
(3)如SEQ ID NO:20所示的CDR1、如SEQ ID NO:48所示的CDR2，和如SEQ ID NO:87所示的CDR3；
(4)如SEQ ID NO:21所示的CDR1、如SEQ ID NO:49所示的CDR2，和如SEQ ID NO:88所示的CDR3；
(5)如SEQ ID NO:21所示的CDR1、如SEQ ID NO:50所示的CDR2，和如SEQ ID NO:89所示的CDR3；
(6)如SEQ ID NO:22所示的CDR1、如SEQ ID NO:51所示的CDR2，和如SEQ ID NO:90所示的CDR3；
(7)如SEQ ID NO:23所示的CDR1、如SEQ ID NO:52所示的CDR2，和如SEQ ID NO:91所示的CDR3；
(8)如SEQ ID NO:24所示的CDR1、如SEQ ID NO:53所示的CDR2，和如SEQ ID NO:92所示的CDR3；
(9)如SEQ ID NO:25所示的CDR1、如SEQ ID NO:54所示的CDR2，和如SEQ ID NO:87所示的CDR3；
(10)如SEQ ID NO:26所示的CDR1、如SEQ ID NO:51所示的CDR2，和如SEQ ID NO:93所示的CDR3；
(11)如SEQ ID NO:24所示的CDR1、如SEQ ID NO:53所示的CDR2，和如SEQ ID NO:92所示的CDR3；
(12)如SEQ ID NO:21所示的CDR1、如SEQ ID NO:50所示的CDR2，和如SEQ ID NO:89所示的CDR3；
(13)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:94所示的CDR3；
(14)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:95所示的CDR3；
(15)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:94所示的CDR3；
(16)如SEQ ID NO:19所示的CDR1、如SEQ ID NO:52所示的CDR2，和如SEQ ID NO:96所示的CDR3；
(17)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:95所示的CDR3；
(18)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:95所示的CDR3；
(19)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:97所示的CDR3；
(20)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:94所示的CDR3；
(21)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:94所示的CDR3；
(22)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:56所示的CDR2，和如SEQ ID NO:95所示的CDR3；
(23)如SEQ ID NO:28所示的CDR1、如SEQ ID NO:57所示的CDR2，和如SEQ ID NO:98所示的CDR3；
(24)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:95所示的CDR3；
(25)如SEQ ID NO:29所示的CDR1、如SEQ ID NO:58所示的CDR2，和如SEQ ID NO:99所示的CDR3；
(26)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:94所示的CDR3；
(27)如SEQ ID NO:23所示的CDR1、如SEQ ID NO:59所示的CDR2，和如SEQ ID NO:100所示的CDR3；
(28)如SEQ ID NO:30所示的CDR1、如SEQ ID NO:51所示的CDR2，和如SEQ ID NO:101所示的CDR3；
(29)如SEQ ID NO:31所示的CDR1、如SEQ ID NO:59所示的CDR2，和如SEQ ID NO:100所示的CDR3；或
(30)如SEQ ID NO:27所示的CDR1、如SEQ ID NO:55所示的CDR2，和如SEQ ID NO:94所示的CDR3；
其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸残基的，并能够保留Trop2结合亲和力的衍生序列。
在另一优选例中，所述重链可变区具有SEQ ID NO:192-220中任一项所示的氨基酸序列。
在另一优选例中，所述抗体或抗原结合蛋白的重链还包括重链恒定区。
在另一优选例中，所述重链恒定区为人源的。
在另一优选例中，所述重链恒定区为人抗体的重链恒定区Fc结构域；所述人抗体的重链恒定区Fc结构域包括人IgG1、IgG2、IgG3或IgG4的重链恒定区Fc结构域。
在另一优选例中，所述重链恒定区包含SEQ ID NO:292所示的氨基酸序列。
在另一优选例中，所述抗体或抗原结合蛋白的重链具有如SEQ ID NO:241-269中任一项所示的氨基酸序列。
在另一优选例中，所述抗体或抗原结合蛋白还包括一轻链，所述轻链中包含一轻链可变区，所述的轻链可变区具有选自下组的互补决定区CDR组合：
(1)如SEQ ID NO:115所示的CDR1、如SEQ ID NO:144所示的CDR2，和如SEQ ID NO:169所示的CDR3；
(2)如SEQ ID NO:116所示的CDR1、如SEQ ID NO:145所示的CDR2，和如SEQ ID NO:170所示的CDR3；
(3)如SEQ ID NO:117所示的CDR1、如SEQ ID NO:146所示的CDR2，和如SEQ ID NO:171所示的CDR3；
(4)如SEQ ID NO:118所示的CDR1、如SEQ ID NO:147所示的CDR2，和如SEQ ID NO:172所示的CDR3；
(5)如SEQ ID NO:119所示的CDR1、如SEQ ID NO:148所示的CDR2，和如SEQ ID NO:173所示的CDR3；
(6)如SEQ ID NO:120所示的CDR1、如SEQ ID NO:146所示的CDR2，和如SEQ ID NO:174所示的CDR3；
(7)如SEQ ID NO:121所示的CDR1、如SEQ ID NO:145所示的CDR2，和如SEQ ID NO:175所示的CDR3；
(8)如SEQ ID NO:122所示的CDR1、如SEQ ID NO:149所示的CDR2，和如SEQ ID NO:176所示的CDR3；
(9)如SEQ ID NO:123所示的CDR1、如SEQ ID NO:145所示的CDR2，和如SEQ ID NO:177所示的CDR3；
(10)如SEQ ID NO:120所示的CDR1、如SEQ ID NO:146所示的CDR2，和如SEQ IDNO:174所示的CDR3；
(11)如SEQ ID NO:124所示的CDR1、如SEQ ID NO:149所示的CDR2，和如SEQ IDNO:178所示的CDR3；
(12)如SEQ ID NO:125所示的CDR1、如SEQ ID NO:148所示的CDR2，和如SEQ IDNO:179所示的CDR3；
(13)如SEQ ID NO:126所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(14)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:151所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(15)如SEQ ID NO:126所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(16)如SEQ ID NO:128所示的CDR1、如SEQ ID NO:152所示的CDR2，和如SEQ IDNO:181所示的CDR3；
(17)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:151所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(18)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:151所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(19)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(20)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(21)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(22)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:151所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(23)如SEQ ID NO:129所示的CDR1、如SEQ ID NO:144所示的CDR2，和如SEQ IDNO:182所示的CDR3；
(24)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:151所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(25)如SEQ ID NO:130所示的CDR1、如SEQ ID NO:146所示的CDR2，和如SEQ IDNO:183所示的CDR3；
(26)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
(27)如SEQ ID NO:131所示的CDR1、如SEQ ID NO:153所示的CDR2，和如SEQ IDNO:184所示的CDR3；
(28)如SEQ ID NO:132所示的CDR1、如SEQ ID NO:154所示的CDR2，和如SEQ IDNO:185所示的CDR3；
(29)如SEQ ID NO:133所示的CDR1、如SEQ ID NO:153所示的CDR2，和如SEQ IDNO:184所示的CDR3；
(30)如SEQ ID NO:127所示的CDR1、如SEQ ID NO:150所示的CDR2，和如SEQ IDNO:180所示的CDR3；
其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸残基的，并能够保留Trop2结合亲和力的衍生序列。
在另一优选例中，所述轻链可变区具有SEQ ID NO:221-240任一项所示的氨基酸序列。
在另一优选例中，所述抗体或抗原结合蛋白的轻链还包括轻链恒定区。
在另一优选例中，所述轻链恒定区为人抗体恒定区。
在另一优选例中，所述轻链恒定区为人抗体轻链κ恒定区。
在另一优选例中，所述轻链恒定区包含SEQ ID NO:293的氨基酸序列。
在另一优选例中，所述的轻链具有如SEQ ID NO:270-289任一项所示的氨基酸序列。
在另一优选例中，所述抗体或抗原结合蛋白还包括上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留Trop2结合亲和力的衍生序列。
在另一优选例中，具有所述的衍生序列的抗体与TROP2结合的亲和力F1与相应非衍生的抗体与TROP2结合的亲和力F0之比(F1/F0)为0.5-2，较佳地为0.7-1.5，和更佳地0.8-1.2。
在另一优选例中，所述添加、缺失、修饰和/或取代的氨基酸数量为1-5个(如1-3个，较佳地1-2个，更佳地1个)。
在另一优选例中，所述的经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留TROP2结合亲和力的衍生序列为同源性或序列相同性为至少96％的氨基酸序列。
在另一优选例中，所述抗体的重链可变区还包括人源的框架区，和/或所述抗体的轻链可变区还包括人源的框架区。
在另一优选例中，所述抗体选自下组：动物源抗体、嵌合抗体、人源化抗体、全人抗体、或其组合。
在另一优选例中，所述的嵌合抗体在人中的免疫原性Z1与非嵌合的抗体(如鼠源抗体)在人中的免疫原性Z0之比(Z1/Z0)为0-0.5，较佳地0-0.2，更佳地0-0.05(如0.001-0.05)。
在另一优选例中，所述的抗体是全人的单克隆抗体。
在另一优选例中，所述的抗体为双链抗体。
在另一优选例中，所述抗体为抗体全长蛋白、或抗原结合片段。
在另一优选例中，所述的抗体为药物偶联物形式。
在另一优选例中，所述抗体具有选自下组的一个或多个特性：
(a)在FACS测定中，能与过表达人Trop2的HEK293细胞结合；
(b)在FACS测定中，能与过表达猴Trop2的CHO-K1结合；
(c)在ELISA测定中,能与小鼠Trop2蛋白结合；
(d)能内化到过表达人Trop2的HEK293细胞或BxPC-3细胞中；
(e)对BxPc-3细胞具有ADCC活性；和/或
(f)对BxPc-3细胞具有ADCP活性。
在另一优选例中，所述的抗体是具有表A所示的各重链VH-CDR1、VH-CDR2和VH-CDR3，以及轻链VL-CDR1、VL-CDR2和VL-CDR3的组合的抗体之一：
表A
抗体编号 抗体名称 LCDR1 LCDR2 LCDR3 HCDR1 HCDR2 HCDR3
(1) PR001128 115 144 169 19 47 86
(2) PR001130 116 145 170 20 48 87
(3) PR001131 117 146 171 20 48 87
(4) PR001132 118 147 172 21 49 88
(5) PR001133 119 148 173 21 50 89
(6) PR001134 120 146 174 22 51 90
(7) PR001138 121 145 175 23 52 91
(8) PR001139 122 149 176 24 53 92
(9) PR001142 123 145 177 25 54 87
(10) PR001143 120 146 174 26 51 93
(11) PR001145 124 149 178 24 53 92
(12) PR001147 125 148 179 21 50 89
(13) PR001150 126 150 180 27 55 94
(14) PR001151 127 151 180 27 55 95
(15) PR001152 126 150 180 27 55 94
(16) PR001153 128 152 181 19 52 96
(17) PR001154 127 151 180 27 55 95
(18) PR001155 127 151 180 27 55 95
(19) PR001156 127 150 180 27 55 97
(20) PR001158 127 150 180 27 55 94
(21) PR001159 127 150 180 27 55 94
(22) PR001160 127 151 180 27 56 95
(23) PR001162 129 144 182 28 57 98
(24) PR001163 127 151 180 27 55 95
(25) PR001164 130 146 183 29 58 99
(26) PR001165 127 150 180 27 55 94
(27) PR001166 131 153 184 23 59 100
(28) PR001168 132 154 185 30 51 101
(29) PR001170 133 153 184 31 59 100
(30) PR001171 127 150 180 27 55 94
其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留TROP2结合亲和力的衍生序列。
在另一优选例中，所述的抗体是表A中选自下组的编号的抗体：(1)、(2)、(5)、(8)、(9)、(23)、(26)或(27)。(分别对应于实施例中的抗体编号PR001128、PR001130、PR001133、PR001139、PR001142、PR001162、PR001165、PR001166)
在另一优选例中，所述的抗体是表A中选自下组的编号的抗体：(9)、(23)、(26)或(27)。(分别对应于实施例中的抗体编号PR001142、PR001162、PR001165、PR001166)
在另一优选例中，所述的抗体是表A中编号为(27)的抗体。(对应于实施例中的抗体编号PR001166)
在另一优选例中，所述重链可变区具有SEQ ID NO:192～220中任一项所示的氨基酸序列，或与其至少有80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的序列同源性的氨基酸序列；和/或所述轻链可变区具有SEQ ID NO:221～240中任一项所示的氨基酸序列，或与其至少有80％、85％、90％、91％、92％、93％、94％、95％、96％、97％、98％或99％的序列同源性的氨基酸序列。
在另一优选例中，所述的抗体是具有表B所示的各重链可变区、轻链可变区的组合的抗体之一：
表B
Figure BDA0002873116140000081
Figure BDA0002873116140000091
在另一优选例中，所述的抗体是表B中选自下组的编号的抗体：(1)、(2)、(5)、(8)、(9)、(23)、(26)或(27)。(分别对应于实施例中的抗体编号PR001128、PR001130、PR001133、PR001139、PR001142、PR001162、PR001165、PR001166)
在另一优选例中，所述的抗体是表B中选自下组的编号的抗体：(9)、(23)、(26)或(27)。(分别对应于实施例中的抗体编号PR001142、PR001162、PR001165、PR001166)
在另一优选例中，所述的抗体是表B中编号为(27)的抗体。(对应于实施例中的抗体编号PR001166)
在另一优选例中，所述的抗体是具有表C所示的各重链、轻链的组合的抗体之一：
表C
抗体编号 抗体名称 LC HC
(1) PR001128 270 241
(2) PR001130 271 242
(3) PR001131 272 242
(4) PR001132 273 243
(5) PR001133 274 244
(6) PR001134 275 245
(7) PR001138 276 246
(8) PR001139 277 247
(9) PR001142 278 248
(10) PR001143 275 249
(11) PR001145 279 250
(12) PR001147 280 251
(13) PR001150 281 252
(14) PR001151 282 253
(15) PR001152 281 254
(16) PR001153 283 255
(17) PR001154 282 256
(18) PR001155 282 257
(19) PR001156 284 258
(20) PR001158 284 259
(21) PR001159 284 260
(22) PR001160 282 261
(23) PR001162 285 262
(24) PR001163 282 263
(25) PR001164 286 264
(26) PR001165 284 265
(27) PR001166 287 266
(28) PR001168 288 267
(29) PR001170 289 268
(30) PR001171 284 269
在另一优选例中，所述的抗体是表B中选自下组的编号的抗体：(1)、(2)、(5)、(8)、(9)、(23)、(26)或(27)。(分别对应于实施例中的抗体编号PR001128、PR001130、PR001133、PR001139、PR001142、PR001162、PR001165、PR001166)
在另一优选例中，所述的抗体是表B中选自下组的编号的抗体：(9)、(23)、(26)或(27)。(分别对应于实施例中的抗体编号PR001142、PR001162、PR001165、PR001166)
在另一优选例中，所述的抗体是表B中编号为(27)的抗体。(对应于实施例中的抗体编号PR001166)
在本发明的第二方面，提供了一种重组蛋白，所述的重组蛋白包括：
(i)如本发明第一方面所述的抗体或抗原结合蛋白；和
(ii)任选的协助表达和/或纯化的标签序列。
在另一优选例中，所述的标签序列包括6His标签。
在另一优选例中，所述的重组蛋白(或多肽)包括融合蛋白。
在另一优选例中，所述的重组蛋白为单体、二聚体、或多聚体。
在本发明的第三方面，提供了一种多核苷酸，所述多核苷酸编码选自下组的多肽：如本发明第一方面所述的抗体或抗原结合蛋白，和/或如本发明第二方面所述的重组蛋白。
在本发明的第四方面，提供了一种载体，所述载体含有本发明第三方面所述的多核苷酸。
在另一优选例中，所述的载体包括：细菌质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒、或其他载体。
在另一优选例中，所述的载体为溶瘤病毒载体。
在本发明的第五方面，提供了一种遗传工程化的宿主细胞，所述宿主细胞含有本发明第四方面所述的载体或基因组中整合有本发明第三方面所述的多核苷酸。
在本发明的第六方面，提供了制备第一方面的抗体或其抗原结合片段的方法，其包括，在允许所述抗体或其抗原结合片段表达的条件下，培养第五方面的宿主细胞，和从培养的宿主细胞培养物中回收所述抗体或其抗原结合片段。
在本发明的第七方面，提供了一种抗体偶联物，所述抗体偶联物含有：
(a)抗体部分，所述抗体部分包括如本发明第一方面所述的抗体或抗原结合蛋白；和
(b)与所述抗体部分偶联的偶联部分，所述偶联部分选自下组：可检测标记物、药物、毒素、细胞因子、放射性核素、酶、或其组合。
在另一优选例中，所述的抗体部分与所述的偶联部分通过化学键或接头进行偶联。
在另一优选例中，偶联部分包括：细胞毒素、烷化剂、DNA小沟结合分子、DNA嵌入剂、DNA交联剂、组蛋白去乙酰化酶抑制剂、核输出抑制剂、蛋白酶体抑制剂、拓扑异构酶I或II的抑制剂、热激蛋白抑制剂、酪氨酸激酶抑制剂、抗生素和抗有丝分裂剂，优选为SN-38。
在本发明的第八方面，提供了一种嵌合抗原受体(CAR)，所述CAR包括本发明第一方面所述的抗体或抗原结合蛋白。
在本发明的第九方面，提供了一种免疫细胞，所述免疫细胞表达或在细胞膜外暴露有如本发明第一方面所述的抗体或抗原结合蛋白。在另一优选例中，所述的免疫细胞包括NK细胞、T细胞。
在另一优选例中，所述的免疫细胞来自人或非人哺乳动物(如鼠)。
在另一优选例中，所述的免疫细胞包括本发明第八方面所述的嵌合抗原受体，所述免疫细胞优选地是嵌合抗原受体T细胞(CAR-T细胞)或嵌合抗原受体NK细胞(CAR-NK细胞)。
在本发明的第十方面，提供了一种多特异性抗体，其包含本发明第一方面的结合Trop-2的抗体或抗原结合片段的，和另外的抗体或其片段或抗体类似物。
在另一优选例中，所述多特异性抗体由第一抗体或其抗原结合片段与其他抗体或其抗原结合片段或抗体类似物通过偶联形成，并且其中各抗体或其抗原结合片段或抗体类似物保持其原结合特异性，所述第一抗体或其抗原结合片段为本发明所述的抗体或其抗原结合片段。
在另一优选例中，所述多特异性抗体为双特异性抗体或三特异性抗体或四特异性抗体。
在本发明的第十一方面，提供了一种药物组合物，所述药物组合物含有：
(i)活性成分，所述活性成分选自下组：如本发明第一方面所述的抗体或抗原结合蛋白、如本发明第二方面所述的重组蛋白、如本发明第七方面所述的抗体偶联物、本发明第九方面所述的免疫细胞、本发明第十方面所述的多特异性抗体或其组合；以及
(ii)药学上可接受的载体。
在另一优选例中，所述的药物组合物为液态制剂。
在另一优选例中，所述的药物组合物为注射剂。
在另一优选例中，所述的药物组合物包括0.01～99.99％的如本发明第一方面所述的抗体或抗原结合蛋白、如本发明第二方面所述的重组蛋白、如本发明第六方面所述的抗体偶联物、如本发明第八方面所述的免疫细胞、或其组合和0.01～99.99％的药用载体，所述百分比为占所述药物组合物的质量百分比。
在另一优选例中，所述的药物组合物还包括第二治疗剂。
在另一优选例中，第二治疗剂包括第二抗体、或化疗剂。
在另一优选例中，所述第二抗体包括：LAG-3抗体、PD-1抗体或CTLA-4抗体等。
在另一优选例中，所述化疗剂选自下组：多西他赛、卡铂、卡培他滨、长春瑞滨，或其组合。
在另一优选例中，所述化疗剂是细胞毒性剂，包括：SN-38、表阿霉素、奥沙利铂，或5-FU。
在本发明的第十二方面，提供了一种治疗、预防或减轻肿瘤的方法，所述方法包括向有需要的受试者施用本发明第一方面所述的抗体或抗原结合蛋白，第二方面所述的重组蛋白，第七方面所述的抗体偶联物，第九方面所述的免疫细胞，第十方面所述的多特异性抗体及第十一方面所述的药物组合物。
在另一优选例中，所述肿瘤可以是实体瘤或非实体瘤。
在另一优选例中，所述实体瘤选自下组：乳癌、胃癌、胰腺癌、卵巢癌、肠癌、肺癌、子宫颈癌，或其他Trop2表达阳性的肿瘤。
在另一优选例中，所述方法进一步包括第二治疗剂的施用。
在另一优选例中，第二治疗剂包括第二抗体、或化疗剂。
在另一优选例中，所述第二抗体包括：LAG-3抗体、PD-1抗体或CTLA-4抗体。
在另一优选例中，所述化疗剂选自下组：多西他赛、卡铂、卡培他滨、长春瑞滨，或其组合。
在另一优选例中，所述化疗剂是细胞毒性剂，包括：SN-38、表阿霉素、奥沙利铂，或5-FU。
在本发明的第十三方面，提供了一种活性成分的用途，所述活性成分选自下组：如本发明第一方面所述的抗体或抗原结合蛋白、如本发明第二方面所述的重组蛋白、如本发明第七方面所述的抗体偶联物、如本发明第九方面所述的免疫细胞、第十方面所述的多特异性抗体或其组合，其中所述活性成分被用于：(a)制备与TROP2表达异常相关的疾病的诊断试剂或试剂盒；和/或(b)制备预防和/或治疗与TROP2表达异常相关的疾病的药物。
在另一优选例中，所述的诊断试剂为检测片或检测板。
在另一优选例中，所述TROP2表达或功能异常相关的疾病主要包括肿瘤。
在另一优选例中，所述肿瘤可以是实体瘤或非实体瘤。
在另一优选例中，所述实体瘤选自下组：乳癌、胃癌、胰腺癌、卵巢癌、肠癌、肺癌、子宫颈癌，或其他Trop2表达阳性的肿瘤。
在另一优选例中，所述诊断试剂或试剂盒用于：
(1)检测样品中TROP2蛋白；
(2)检测细胞中内源性的TROP2蛋白；和/或
(3)检测表达TROP2蛋白的细胞。
在另一优选例中，所述的抗体为药物偶联物(ADC)形式。
在本发明的第十四方面，提供了一种体外检测(包括诊断性或非诊断性)样品中TROP2蛋白的方法，所述方法包括步骤：
(1)在允许本发明第一方面所述的抗体或抗原结合片段与Trop-2之间形成复合物的条件下，将样品与所述的抗体或抗原结合蛋白接触；
(2)检测是否形成抗原-抗体复合物，其中形成复合物就表示样品中存在TROP2蛋白。
在另一优选例中，所述检测用于非诊断目的。在另一优选例中，所述非诊断目的包括将第一方面所述的抗体或抗原结合蛋白用于途径研究、药物筛选、组化分析等。
在本发明的第十五方面，提供了一种检测板，所述的检测板包括：基片(支撑板)和测试条，所述的测试条含有如本发明第一方面所述的抗体或抗原结合蛋白、如本发明第二方面所述的重组蛋白、如本发明第七方面所述的抗体偶联物、如本发明第九方面所述的免疫细胞、或其组合。
在本发明的第十六方面，提供了一种试剂盒，所述试剂盒中包括：
(1)第一容器，所述第一容器中含有本发明的抗体或抗原结合片段；和任选地
(2)第二容器，所述第二容器中含有抗本发明抗体的二抗；
在另一优选例中，所述试剂盒含有如本发明第十五方面所述的检测板。
在另一优选例中，所述试剂盒进一步含有使用说明书。
在本发明的第十七方面，提供了一种给药装置，所述装置包括：
(i)输注模块，所述输注模块用于对受试者施用包括具有一活性成分的药物组合物；
(ii)用于输注的药物组合物，所述药物组合物中含有一活性成分，所述活性成分选自下组：如本发明第一方面所述的抗体或抗原结合蛋白、如本发明第二方面所述的重组蛋白、如本发明第七方面所述的抗体偶联物、本发明第九方面所述的免疫细胞、或其组合；以及
(iii)任选的药效监控模块。
在另一优选例中，所述的施用包括肠道外施用和非肠道外施用。
在另一优选例中，所述肠道外施用包括注射施用，所用的注射的路径包括：静脉内、肌内、动脉内、膜内、囊内、眶内、心脏内、皮内、腹膜内、经气管、皮下、表皮下、关节内、囊下、蛛网膜下、脊柱内、硬脑膜上和胸骨内注射和推注。
在另一优选例中，所述非肠道外施用包括外用、表皮施用或粘膜施用，例如鼻内、经口、阴道、直肠、舌下、或局部外用。
在另一优选例中，所述的输注模块为无针皮下注射设备、微量输液泵、经皮给药设备、推注设备，或渗透设备。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1显示了本发明抗体与293T-人Trop2细胞的结合能力。
图2显示了本发明抗体经MMAF偶联后对BxPC-3肿瘤细胞的内化杀伤活性。
图3显示了本发明抗体与肿瘤细胞BxPC-3的结合能力。
图4显示了本发明抗体与过表达猴Trop2的CHO-K1细胞的结合能力。
图5显示了本发明抗体与鼠Trop2蛋白的结合能力。
图6显示了本发明抗体介导的ADCC活性。
图7显示了本发明抗体的ADCP活性。
图8显示了本发明抗体的CDC活性。
图9显示了本发明抗体分别在4℃和37℃的HEK293/hTrop2细胞上的内吞情况。
图10显示了本发明抗体分别在4℃和37℃的肿瘤细胞BxPC-3上的内吞情况。
具体实施方式
本发明人经过广泛而深入的研究，经过大量的筛选，首次开发了一种靶向Trop2的结合蛋白及其应用。在此基础上完成了本发明。
本发明所要解决的技术问题是为克服现有技术中抗体药物缺乏抗肿瘤疗效，以及安全性欠佳等缺陷，提供一种靶向Trop2结合蛋白及其应用。
为更好理解本发明，首先定义一些术语。其他定义则贯穿具体实施方式部分而列出。
术语“Trop2”是滋养层细胞表面抗原2。该术语包括变体、同源物、类似物、直向同源物和/或平行同源物。例如，对人Trop2特异的抗体可以在某些情况下与另一物种例如猴的Trop2蛋白交叉反应。在其他实施方式中，对人Trop2蛋白特异的抗体可以完全地对人Trop2蛋白特异而不与其他物种或其他类型的蛋白交叉反应，或者可以与一些其他物种而非所有其他物种的Trop2蛋白交叉反应。
术语“人Trop2”是指具有人氨基酸序列的Trop2蛋白，例如Genbank登录号为NP_002344.2的氨基酸序列。
术语“猴Trop2”是指具有猴氨基酸序列的Trop2蛋白，例如Genbank登录号为XP_005543292.1的氨基酸序列。
术语“小鼠Trop2”是指具有小鼠氨基酸序列的Trop2蛋白，例如Genbank登录号为NP_064431.2的氨基酸序列。
本文中的术语“抗体”意在包括全长抗体及其任何抗原结合片段(即，抗原结合部分)或单链。全长抗体是包含至少两条重(H)链和两条轻(L)链的糖蛋白，重链和轻链由二硫键连接。各重链由重链可变区(简称VH)和重链恒定区构成。重链恒定区由三个结构域构成，即CH1、CH2和CH3。各轻链由轻链可变区(简称VL)和轻链恒定区构成。轻链恒定区由一个结构域CL构成。VH和VL区还可以划分为称作互补决定区(CDR)的高变区，其由较为保守的框架区(FR)区分隔开。各VH和VL由三个CDR以及四个FR构成，从氨基端到羧基端以FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4的顺序排布。重链和轻链的可变区包含与抗原相互作用的结合域。抗体的恒定区可以介导免疫球蛋白与宿主组织或因子的结合，包括多种免疫系统细胞(例如，效应细胞)和传统补体系统的第一组分(C1q)。
本文中的术语，抗体的“抗原结合部分”(或简称为抗体部分)，是指抗体的保持有特异结合抗原(例如，Trop2蛋白)能力的一个或多个片段。已证实，抗体的抗原结合功能可以通过全长抗体的片段来实施。包含在抗体的“抗原结合部分”中的结合片段的例子包括(i)Fab片段，由VL、VH、CL和CH1构成的单价片段；(ii)F(ab’)2片段，包含铰链区二硫桥连接的两个Fab片段的二价片段；(iii)由VH和CH1构成的Fd片段；(iv)由抗体单臂VL和VH构成的Fv片段；(v)由VH构成的dAb片段；(vi)分离的互补决定区(CDR)；以及(vii)纳米抗体，一种包含单可变结构域和两个恒定结构域的重链可变区。此外，尽管Fv片段的两个结构域VL和VH由不同的基因编码，它们可以通过重组法经由使两者成为单蛋白链的合成接头而连接，其中VL和VH区配对形成单价分子(称为单链Fc(scFv))。这些单链抗体也意在包括在术语涵义中。这些抗体片段可以通过本领域技术人员已知的常用技术而得到，且片段可以通过与完整抗体相同的方式进行功能筛选。
本文所用的术语“分离的抗体”是指基本不含具有不同抗原特异性的其他抗体的抗体。例如，与Trop2蛋白特异结合的分离抗体基本不含特异结合Trop2蛋白之外抗原的抗体。但是，特异结合人Trop2蛋白的分离抗体可能对其他抗原例如其他物种的Trop2蛋白具有交叉结合性。此外，分离的抗体基本不含其他细胞材料和/或化学物质。
术语“单克隆抗体”或“单抗”或“单克隆抗体组成”是指单一分子组成的抗体分子制品。单克隆抗体组成呈现出对于特定表位的单一结合特异性和亲和力。
术语“人源抗体”是指可变区框架和CDR区得自人种系免疫球蛋白序列的抗体。此外，如果抗体包含恒定区，其也得自人种系免疫球蛋白序列。本发明的人源抗体可以包含不由人种系免疫球蛋白序列编码的氨基酸残基，例如通过体外随机突变或点突变或通过体内体细胞突变而导入的突变。然而，术语“人源抗体”不包括在人框架序列中插入得自其他哺乳动物物种的CDR序列的抗体。
术语“人源抗体”是指可变区框架和CDR区得自人种系免疫球蛋白序列的抗体。此外，如果抗体包含恒定区，其也得自人种系免疫球蛋白序列。本发明的人源抗体可以包含不由人种系免疫球蛋白序列编码的氨基酸残基，例如通过体外随机突变或点突变或通过体内体细胞突变而导入的突变。然而，术语“人源抗体”不包括在人框架序列中插入得自其他哺乳动物物种的CDR序列的抗体。
术语“识别抗原的抗体”以及“对抗原特异的抗体”在本文中与术语“特异结合抗原的抗体”交替使用。
在本文中，“特异结合人Trop2”的抗体是指与人Trop2(还可能是其他非人物种的Trop2)结合但是基本不与非Trop2蛋白结合的抗体。优选地，抗体以“高亲和力”结合人Trop2蛋白，即KD值为1.0×10-8M以下，更优选为5.0×10-9M以下。
术语“基本不结合”蛋白或细胞是指，不与蛋白或细胞结合，或者不以高亲和力与其结合，即结合蛋白或细胞的KD为1.0×10-6M以上，更优选1.0×10-5M以上，更优选1.0×10-4M以上、1.0×10-3M以上，更优选1.0×10-2M以上。
术语“高亲和性”对于IgG抗体而言，是指对于抗原的KD为1.0×10-6M以下，优选5.0×10-8M以下，更优选1.0×10-8M以下、5.0×10-9M以下，更优选1.0×10-9M以下。对于其他抗体亚型，“高亲和性”结合可能会变化。例如，IgM亚型的“高亲和性”结合是指KD为10-6M以下，优选10-7M以下，更优选10-8M以下。
术语“Kassoc”或“Ka”是指特定抗体-抗原相互作用的结合速率，而术语“Kdis”或“Kd”是指特定抗体-抗原相互作用的离解速率。术语“KD”是指解离常数，由Kd与Ka比(Kd/Ka)得到，并以摩尔浓度(M)表示。抗体的KD值可以通过领域内已知的方法确定。优选的确定抗体KD的方式是使用表面等离子共振仪(SPR)测得的，优选使用生物传感系统例如BiacoreTM或OctetRed96e系统测得。
术语“EC50”，又叫半最大效应浓度，是指引起50％最大效应的抗体浓度。
术语“抗体依赖的细胞毒性”、“抗体依赖的细胞介导的细胞毒性”或“ADCC”是指细胞介导的免疫防御，其中免疫系统效应细胞主动地将细胞膜表面抗原与抗体，例如Trop2抗体，结合的靶细胞例如癌细胞裂解。
术语“抗体依赖的细胞介导的吞噬作用”或“ADCP”是指抗体与靶细胞上相应的抗原结合后，抗体的Fc段与效应细胞如吞噬细胞上的Fc受体结合，从而诱导效应细胞吞噬靶细胞的作用。
术语“受试者”包括任何人或非人动物。术语“非人动物”包括所有脊椎动物，例如哺乳类和非哺乳类，例如非人灵长类、羊、狗、猫、牛、马、鸡、两栖类、和爬行类，优选哺乳动物，例如非人灵长类、羊、狗、猫、牛和马。
术语“治疗有效量”是指足以防止或减缓与疾病或病症(例如癌症)相关的症状的本发明抗体量。治疗有效量与被治疗的疾病相关，其中本领域技术人员可以方便地判别出实际的有效量。
本发明的多个方面在以下更加具体地加以描述。
Trop2抗体对Trop2的结合特异性以及其他有益的功能特征
本发明的抗体特异性地结合人或猴Trop2，也与小鼠Trop2结合。具体而言，本发明的抗体以与hRS7相当或更低的EC50值与人、猴Trop2结合。此外，本发明的抗体还具有与hRS7相当或更好的内化活性、ADCC或ADCP。
优选的本发明抗体是单克隆抗体。此外，抗体可以是例如鼠源的、嵌合的或人源的单克隆抗体，优选为人源抗体。
Trop2单克隆抗体
优选的本发明抗体是结构和化学特性在以下描述的单克隆抗体。Trop2抗体的VH包含SEQ ID NO:192-220中的任一氨基酸序列。Trop2抗体的VL包含SEQ ID NO:221-240中的任一氨基酸序列。抗体的重链/轻链可变区序列列于以下表1a和表1b中。所有抗体的重链恒定区和轻链恒定区分别包含SEQ ID NO:292和293的氨基酸序列。抗体还可以包含其他合适的重链恒定区和轻链恒定区序列。
表1a.重链/轻链可变区及CDR的氨基酸序列总结(SEQ ID NO)
Figure BDA0002873116140000171
Figure BDA0002873116140000181
表1b.重链/轻链可变区框架区的氨基酸序列总结(SEQ ID NO)
Figure BDA0002873116140000182
Figure BDA0002873116140000191
与人Trop2结合的其他Trop2抗体的VH和/或VL序列(或CDR序列)可以与本发明抗体的VH和/或VL序列(或CDR序列)“混合并配对”。优选地，当VH和VL(或其中的CDR)混合并配对时，特定VH/VL配对中的VH序列可以由结构近似的VH序列取代。相似地，优选特定VH/VL配对中的VL序列由结构近似的VL序列取代。
因此，在一个实施方式中，本发明的抗体或其抗原结合部分包括：
(a)包含列于表1a中氨基酸序列的重链可变区；以及
(b)包含列于表1a中氨基酸序列的轻链可变区，或者另一Trop2抗体的VL，其中该抗体特异结合人Trop2。
在另一实施方式中，本发明的抗体或其抗原结合部分包括：
(a)列于表1a中的重链可变区的CDR1、CDR2和CDR3；以及
(b)列于表1a中的轻链可变区的CDR1、CDR2和CDR3，或者另一Trop2抗体的CDR，其中该抗体特异结合人Trop2。
在另一实施方式中，本发明的抗体或其抗原结合部分包括Trop2抗体的重链可变区CDR以及其他结合人Trop2的抗体的CDR，例如重链可变区CDR1、CDR2和/或CDR3，和/或另一Trop2抗体的轻链可变区CDR1、CDR2和/或CDR3。
此外，领域内公知的是，CDR3结构域，独立于CDR1和/或CDR2，可单独确定抗体对同种抗原的结合特异性，且可以预测到基于该CDR3序列可生成具有相同结合特异性的多种抗体。
在另一实施方式中，本发明的抗体包含Trop2抗体的重链可变区的CDR2以及至少Trop2抗体的重链和/或轻链可变区的CDR3，或另一Trop2抗体的重链和/或轻链可变区的CDR3，其中该抗体能够特异结合人Trop2。优选这些抗体(a)竞争结合Trop2；(b)保留功能特性；(c)结合相同表位；和/或(d)具有与本发明Trop2抗体相似的结合亲和力。在另一实施方式中，抗体还可以包含Trop2抗体的轻链可变区CDR2，或者另一Trop2抗体的轻链可变区CDR2，其中该抗体特异结合人Trop2。在另一实施方式中，本发明的抗体可以包括Trop2抗体的重链/轻链可变区CDR1，或另一Trop2抗体的重链和/或轻链可变区CDR1，其中该抗体特异结合人Trop2。
保守修饰
在另一实施方式中，本发明的抗体包含与本发明Trop2抗体存在一个或多个保守修饰的重链和/或轻链可变区序列或CDR1、CDR2和CDR3序列。本领域知道，一些保守序列修改不会使抗原结合性消失。参见，例如，Brummell et al.,(1993)Biochem 32:1180-8；deWildt et al.,(1997)Prot.Eng.10:835-41；Komissarov et al.,(1997)J.Biol.Chem.272:26864-26870；Hall et al.,(1992)J.Immunol.149:1605-12；Kelleyand O’Connell(1993)Biochem.32:6862-35；Adib-Conquy et al.,(1998)Int.Immunol.10:341-6and Beers et al.,(2000)Clin.Can.Res.6:2835-43。
因此，在一个实施方式中，抗体包含重链可变区和/或轻链可变区，重链可变区和轻链可变区分别包含CDR1、CDR2和CDR3，其中：
(a)重链可变区CDR1包含表1a列出的序列，和/或其保守修改；和/或
(b)重链可变区CDR2包含表1a列出的序列，和/或其保守修改；和/或
(c)重链可变区CDR3包含表1a列出的序列，和/或其保守修改；和/或
(d)轻链可变区CDR1、和/或CDR2、和/或CDR3包含表1a列出的序列，和/或其保守修改；且
(e)该抗体特异结合人Trop2。
本发明的抗体具有一个或多个以下功能特征，例如对人Trop2的高亲和力，以及引发对Trop2表达细胞的ADCC或CDC的能力。
在多个实施方式中，抗体可以是例如鼠源、人源、嵌合或人源化抗体。
本文所用的术语“保守序列修饰”是指不会显著影响或改变抗体结合特性的氨基酸修饰。这样的保守修饰包括氨基酸替换、添加和删除。可以通过领域内已知的标准技术，例如点突变和PCR介导的突变，将修饰引入本发明抗体中。保守氨基酸替换是氨基酸残基用具有相似侧链的氨基酸残基进行替换。具有相似侧链的氨基酸残基组在领域内已知。这些氨基酸残基组包括具有碱性侧链(例如，赖氨酸、精氨酸、组氨酸)、酸性侧链(例如，天冬氨酸、谷氨酸)、不带电极性侧链(例如，甘氨酸、天冬酰胺、谷氨酰胺、丝氨酸、苏氨酸、酪氨酸、半胱氨酸、色氨酸)、非极性侧链(例如，丙氨酸、缬氨酸、亮氨酸、异亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸)、β-支链侧链(例如，苏氨酸、缬氨酸、异亮氨酸)和芳香族侧链(例如，酪氨酸、苯丙氨酸、色氨酸、组氨酸)的氨基酸。因此，本发明抗体的CDR区中的一个或多个氨基酸残基可以用同侧链组的其他氨基酸残基替换，且得到的抗体可以使用本文所述的功能检测对其进行保留功能(即，上述的功能)的测试。
基因修饰的抗体
本发明的抗体可以以具备本发明Trop2抗体的一个或多个VH/VL序列的抗体作为起始材料，制备成基因修饰的抗体。抗体可以通过修饰一个或两个可变区(即，VH和/或VL)内(例如，在一个或多个CDR区和/或一个或多个框架区)的一个或多个残基来进行基因修饰，以改善结合亲和力和/或增加与某些物种天然产生的抗体的相似性。例如，框架区经修饰成提供人源化的抗体。此外，或者抗体可以通过修饰恒定区中的残基进行基因修饰，例如改变抗体的效应功能。
在某些实施方式中，CDR区植入可以用来基因修饰抗体的可变区。抗体主要通过位于六个重链和轻链互补决定区(CDR)中的氨基酸残基与靶标抗原进行相互作用。出于这个原因，CDR内的氨基酸残基比起CDR外的序列在个体抗体之间更加地多样。因为CDR序列负责主要的抗体-抗原相互作用，可以通过构建含有特定天然抗体的CDR序列植入到不同特性的不同抗体的框架序列的表达载体，来表达模拟特定天然抗体的特性的重组抗体(Riechmannet al.,(1998)Nature 332:323-327；Jones et al.,(1986)Nature 321:522-525；Queenet al.,(1989)Proc.Natl.Acad；U.S.A.86:10029-10033；U.S.Pat.Nos.5,225,539；5,530,101；5,585,089；5,693,762和6,180,370)。
因此，本发明的另一实施方式涉及分离的单克隆抗体或其抗原结合部分，其包含重链可变区和/或轻链可变区，重链可变区包含具有本发明上述序列的CDR1、CDR2和CDR3，轻链可变区包含具有本发明上述序列的CDR1、CDR2和CDR3。尽管这些抗体包含本发明单克隆抗体的VH和VL CDR序列，它们可以含有不同的框架序列。
这样的框架序列可以从包括种系抗体基因序列的公开DNA数据库或公开参考文献中获得。例如，用于人重链和轻链可变区基因的种系DNA序列可以在Vbase人种系序列数据库(www.mrc-cpe.cam.ac.uk/vbase)以及Kabat et al.,(1991),同上；Tomlinson et al.,(1992)J.Mol.Biol.227:776-798；和Cox et al.,(1994)Eur.J.Immunol.24:827-836中获得。作为另一实施方式，用于人重链和轻链可变区基因的种系DNA序列可以在Genbank数据库中得到。例如，下列HCo7 HuMAb小鼠中的重链种系序列的Genbank登录号为1-69(NG--0010109,NT--024637&BC070333)、3-33(NG--0010109&NT--024637)和3-7(NG--0010109&NT--024637)。作为另一例子，以下来自Hco12 HuMAb小鼠的重链种系序列的Genbank登录号为1-69(NG--0010109,NT--024637&BC070333)、5-51(NG--0010109&NT--024637)、4-34(NG--0010109&NT--024637)、3-30.3(CAJ556644)和3-23(AJ406678)。
通过使用本领域公知的称为空格(gap)BLAST的序列相似性搜索方法之一(Altschul et al.,(1997),同上)，将抗体蛋白序列与蛋白序列数据库进行比较。
用于本发明抗体的优选框架序列是结构上与本发明抗体所用的框架序列相似的那些。VH CDR1、CDR2、和CDR3序列可以植入到与得到该框架序列的种系免疫球蛋白基因具有相同序列的框架区中，或者CDR序列可以植入到包含有与种系序列相比具有一个或多个突变的框架区中。例如，在一些情况下，将框架区中的残基进行突变是有益的，以保持或增强抗体的抗原结合性(参见例如U.S.Pat.Nos.5,530,101；5,585,089；5,693,762和6,180,370)。
另一类的可变区修饰是将VH和/或VL CDR1、CDR2和/或CDR3区内的氨基酸残基进行突变，从而改进目标抗体的一种或多种结合特性(例如，亲和力)。可以进行点突变或PCR介导的突变来引入突变，且其对于抗体结合或其他功能特性的影响可以在本领域所知的体外或体内检测中进行评价。优选地，引入本领域所知的保守修饰。突变可以是氨基酸替换、添加或缺失，但优选为替换。此外，通常改变CDR区内的不多于一个、两个、三个、四个或五个的残基。
此外，在另一实施方式中，本发明提供分离的Trop2单克隆抗体或其抗原结合部分，包含重链可变区和轻链可变区，其包含：(a)VH CDR1区，包含本发明的序列，或一个、两个、三个、四个或五个氨基酸替换、缺失或添加的氨基酸序列；(b)VH CDR2区，包含本发明的序列，或一个、两个、三个、四个或五个氨基酸替换、缺失或添加的氨基酸序列；(c)VH CDR3区，包含本发明的序列，或一个、两个、三个、四个或五个氨基酸替换、缺失或添加的氨基酸序列；(d)VL CDR1区，包含本发明的序列，或一个、两个、三个、四个或五个氨基酸替换、缺失或添加的氨基酸序列；(e)VL CDR2区，包含本发明的序列，或一个、两个、三个、四个或五个氨基酸替换、缺失或添加的氨基酸序列；和(f)VL CDR3区，包含本发明的序列，或一个、两个、三个、四个或五个氨基酸替换、缺失或添加的氨基酸序列。
本发明的基因改造抗体包括在VH和/或VL的框架残基中做出基因修饰以例如改变抗体特性的那些。通常而言，这些框架修饰用来降低抗体的免疫原性。例如，一种方法是将一个或多个框架残基“回复突变”成相应的种系序列。更加具体而言，经历体细胞突变的抗体可能包含不同于得到抗体的种系序列的框架残基。这些残基可以通过将抗体框架序列与得到抗体的种系序列相比较而识别出来。
另一类的框架修饰包括对框架区的、或者甚至一个或多个CDR区的一个或多个残基进行突变，以去除T细胞表位，从而减少抗体的可能导致的免疫原性。该方法也称为“去免疫化”，在美国专利公开20030153043中有更加详细的描述。
本发明还包括抗Trop2抗体的活性片段、衍生物和类似物。如本文所用，术语“片段”、“衍生物”和“类似物”是指基本上保持Trop2结合亲和力功能或活性的多肽。本发明的多肽片段、衍生物或类似物可以是(i)有一个或几个保守或非保守性氨基酸残基(优选保守性氨基酸残基)被取代的多肽，或(ii)在一个或多个氨基酸残基中具有取代基团的多肽，或(iii)CDR3-P1多肽与另一个化合物(比如延长多肽半衰期的化合物，例如聚乙二醇)融合所形成的多肽，或(iv)附加的氨基酸序列融合于此多肽序列而形成的多肽(与前导序列、分泌序列或6His等标签序列融合而形成的然后蛋白)。根据本文的教导，这些片段、衍生物和类似物属于本领域熟练技术人员公知的范围。
一类优选的活性衍生物指与表1的氨基酸序列相比，有至多3个，较佳地至多2个，更佳地至多1个氨基酸被性质相似或相近的氨基酸所替换而形成多肽。这些保守性变异多肽最好根据表2进行氨基酸替换而产生。
表2
Figure BDA0002873116140000221
Figure BDA0002873116140000231
此外，作为框架或CDR区内修饰之外的另一种选择，本发明的抗体可以基因改造成在Fc区包括基因修饰，通常来改变抗体的一个或多个功能特性，例如血清半衰期、补体结合、Fc受体结合、和/或抗体依赖的细胞毒性。此外，本发明的抗体可以进行化学修饰(例如，可以向抗体附加一个或多个化学功能基团)，或者修饰成改变其糖基化，来改变抗体的一个或多个功能特性。
在一个实施方式中，CH1的铰链区进行修饰，改变，例如增加或减少铰链区的半胱氨酸残基的数量。该方法在美国专利5,677,425中进一步描述。改变CH1铰链区的半胱氨酸残基，来例如促进重链轻链的组装或增加/降低抗体的稳定性。
在另一个实施方式中，对抗体的Fc铰链区进行突变，以降低抗体的生物半衰期。更加具体地，将一个或多个氨基酸突变引入Fc铰链片段的CH2-CH3连接区，从而抗体相对于天然Fc-铰链结构域SpA结合而言，具有减弱的SpA结合力。该方法在美国专利6,165,745中有更详细的描述。
在另一实施方式中，修饰抗体的糖基化。例如，可以制备去糖基化的抗体(即，抗体缺少糖基化)。可以改变糖基化，来例如增加抗体对抗原的亲和性。这样的糖化修饰可以通过例如改变抗体序列中的一个或多个糖基化位点来达成。例如，可以做出一个或多个氨基酸替换，以消除一个或多个可变区框架糖基化位点，从而消除该位置的糖基化。这样的去糖基化可以增加抗体对抗原的亲和性。参见，例如美国专利5,714,350和6,350,861。
此外，可以制备具有改变的糖基化类型的抗体，例如岩藻糖残基量减少的低岩藻糖基抗体，或者具有增加的平分型GlcNac结构的抗体。改变的糖基化形式被证明能增加抗体的ADCC活性。这样的糖化修饰可以通过例如在糖基化系统改变的宿主细胞中表达抗体而进行。具有改变的糖基化系统的细胞在领域中已知，且可以用作表达本发明重组抗体的宿主细胞，以制备具有改变的糖基化的抗体。例如，细胞系Ms704、Ms705和Ms709缺少岩藻糖基转移酶基因FUT8(α(1,6)-岩藻糖基转移酶)，从而在Ms704、Ms705和Ms709细胞系中表达的抗体在其糖中缺失岩藻糖。Ms704、Ms705和Ms709 FUT8-/-细胞系通过在CHO/DG44细胞中使用两种替换载体定向破坏FUT8基因而制备(参见美国专利公开20040110704和Yamane-Ohnuki et al.,(2004)Biotechnol Bioeng 87:614-22)。作为另一个例子，EP 1,176,195记载了FUT8基因功能破坏的细胞系，其编码岩藻糖基转移酶，从而在该细胞系中表达的抗体通过降低或消除α-1,6键相关酶而表现出低岩藻糖基化。EP 1,176,195也描述了一种细胞系，其具有较低的用于向结合抗体Fc区的N-乙酰葡糖胺添加岩藻糖的酶活性，或者不具有这种酶的活性，例如大鼠骨髓瘤细胞系YB2/0(ATCC CRL 1662)。WO 03/035835描述了CHO变体细胞系，Lec13细胞，其具有降低的向Asn(297)-相关糖添加岩藻糖的能力，从而使得宿主细胞中表达的抗体的低岩藻糖基化(参见Shields et al.,(2002)J.Biol.Chem.277:26733-26740)。具有改变的糖基化图谱的抗体也可以在鸡蛋中制备，如WO 06/089231中所述的。或者，具有改变的糖基化图谱的抗体可以在植物细胞如浮萍中制备。WO 99/54342公开了一种细胞系，其基因改造成表达修饰糖蛋白的糖基转移酶(例如，β(1,4)-N-乙酰葡糖胺转移酶III(GnTIII))，从而在基因改造细胞系中表达的抗体表现出增加的平分型GlcNac结构，其引起抗体增强的ADCC活性(Umana et al.,(1999)Nat.Biotech.17:176-180)。或者，抗体的岩藻糖残基可以使用岩藻糖苷酶来切除抗体的岩藻糖残基，例如α-L-岩藻糖苷酶从抗体移除岩藻糖残基(Tarentino et al.,(1975)Biochem.14:5516-23)。
本文抗体的另一修饰是聚乙二醇化(PEG化)。抗体可以PEG化，例如来增加抗体的生物(例如，血清)半衰期。为使抗体PEG化，抗体或其片段通常与聚乙二醇(PEG)，例如PEG的反应性酯或醛类衍生物，在使一个或多个PEG基团附于抗体或抗体片段的条件下反应。优选地，PEG化通过与反应性PEG分子(或类似的有反应性的水溶性聚合物)的酰化反应或烷化反应进行。本文中所用的术语“聚乙二醇”包括任何形式的用于衍生其他蛋白的PEG，例如单(C1-C10)烷氧基-或芳氧基聚乙二醇或聚乙二醇马来酰亚胺。在某些实施方式中，需要PEG化的抗体是去糖基化的抗体。PEG化蛋白的方法在领域内已知，且可以应用到本发明的抗体。参见，例如EPO 154 316和EP 0 401 384。
抗体的物理特性
本发明的抗体可以用它们的多种物理特性进行表征，以检测和/或区别其分类。
例如，抗体可以在轻链或重链可变区包含一个或多个糖基化位点。这些糖基化位点可能引起增加的抗体免疫原性，或由于改变的抗原结合而引起改变的抗体pK值(Marshall et al(1972)Annu Rev Biochem 41:673-702；Gala and Morrison(2004)JImmunol 172:5489-94；Wallick et al(1988)J Exp Med 168:1099-109；Spiro(2002)Glycobiology 12:43R-56R；Parekh et al(1985)Nature 316:452-7；Mimura et al.,(2000)Mol Immunol 37:697-706)。糖基化已知发生在含有N-X-S/T序列的基序中。在一些情况下，优选Trop2抗体不包含可变区糖基化。这可以通过选择不在可变区包含糖基化基序的抗体或通过突变糖基化区域的残基来实现。
在优选实施方式中，抗体不包含天冬酰胺异构位点。天冬酰胺的脱酰胺可能出现在N-G或D-G序列，创建出异天冬氨酸残基，其向多肽链中引入扭结并降低其稳定性(异天冬氨酸效果)。
各抗体将具有独特的等电点(pI)，基本落在6-9.5的pH范围内。IgG1抗体的pI通常落在7-9.5的pH范围内，而IgG4抗体的pI基本落在6-8的pH范围内。推测pI在正常范围外的抗体可能在体内条件下具有一些展开结构且不稳定。因此，优选Trop2抗体的pI值落在正常范围内。这可以通过选择pI在正常范围内的抗体或通过突变不带电的表面残基来实现。
编码本发明抗体的核酸分子
在另一方面，本发明提供编码本发明抗体的重链和/或轻链可变区或CDR的核酸分子。核酸可以存在整细胞中，在细胞裂解液中，或处于部分纯化或基本纯的形式。当通过标准技术从其他细胞组分或其他污染物例如其他细胞核酸或蛋白中纯化出来后，核酸是“分离的”或“基本纯的”。本发明的核酸可以为例如DNA或RNA，且可能包含或可能不包含内含子序列。在优选实施方式中，核酸是cDNA分子。
本发明的核酸可以使用标准的分子生物学技术获得。对于由杂交瘤(例如，由携带人免疫球蛋白基因的转基因小鼠制备的杂交瘤)表达的抗体，编码杂交瘤制备的抗体的轻链和重链的cDNA可以通过标准PCR扩增或cDNA克隆技术获得。对于(例如使用噬菌体展示技术)从免疫球蛋白基因库获得的抗体，编码这类抗体的核酸可以从基因库中收集。
优选的本发明核酸分子包括编码Trop2单克隆抗体的VH和VL序列或CDR的那些。一旦获得了编码VH和VL的DNA片段，这些DNA片段可以进一步通过标准的重组DNA技术进行操作，例如将可变区基因转变为全长抗体链基因、Fab片段基因或scFv基因。在这些操作中，编码VH或VL的DNA片段与编码另一蛋白的另一DNA片段，例如抗体恒定区或柔性接头，可操作地连接。术语“可操作地连接”是指两个DNA片段连接在一起，从而两个DNA片段编码的氨基酸序列都在阅读框内。
编码VH区的分离DNA可以通过可操作地连接VH编码DNA与编码重链恒定区(CH1、CH2和CH3)的另一DNA分子而转变成全长重链基因。人重链恒定区基因的序列在领域内已知，且包括这些区域的DNA片段可以通过标准PCR扩增而获得。重链恒定区可以是IgG1、IgG2、IgG3、IgG4、IgA、IgE、IgM或IgD恒定区，但是优选为IgG1或IgG4恒定区。对于Fab片段重链基因，编码VH区的DNA可以可操作地与仅编码重链CH1恒定区的另一DNA分子连接。
编码VL区的分离DNA可以通过可操作地连接VL编码DNA与编码轻链恒定区CL的另一DNA分子而转变成全长轻链基因。人轻链恒定区基因的序列在领域内已知，且包括这些区域的DNA片段可以通过标准PCR扩增而获得。在优选实施方式中，轻链恒定区可以是κ和λ恒定区。
为创建scFv基因，编码VH和VL的DNA片段可以可操作地与编码柔性接头例如编码氨基酸序列(Gly4-Ser)3的另一片段连接，从而VH和VL序列可以作为连续的单链蛋白进行表达，其中VH和VL区域通过该柔性接头连接(参见，例如Bird et al.,(1988)Science 242:423-426；Huston et al.,(1988)Proc.Natl.Acad.Sci.USA 85:5879-5883；McCafferty etal.,(1990)Nature 348:552-554)。
本发明单克隆抗体的制备
本发明使用和铂生物H2L2转基因小鼠来制备具有全人源可变区的两重链和两轻链免疫球蛋白抗体。利用基因工程改造，在小鼠体内引入人源重链和轻链转基因位点以产生人源抗体。根据引入的抗体重链基因座的设计不同，特定抗原的免疫刺激可以产生一类具有特定功能效应的传统抗体(如IgG或IgA)或多类抗体(如IgM和IgG)。
抗原特异性的H2L2单克隆抗体可以使用Kohler and Milstein(1975)Nature256:495的体细胞杂交(杂交瘤)技术进行制备。制备单克隆抗体的其他实施方式包括B淋巴细胞的病毒或致癌性转化以及噬菌体展示技术。嵌合或人源化抗体也在领域内熟知。参见，例如美国专利4,816,567；5,225,539；5,530,101；5,585,089；5,693,762和6,180,370。
制备本发明单克隆抗体的转染瘤的生成
本发明的抗体还可以使用例如重组DNA技术结合基因转染方法，在宿主细胞转染瘤中生成(例如Morrison,S.(1985)Science 229:1202)。在一个实施方式中，将由标准分子生物技术得到的编码部分或全长轻链和重链的DNA插入一个或多个表达载体中，从而基因与转录和翻译调控序列可操作地连接。在该情况下，术语“可操作地连接”是指抗体基因连接到载体中，从而载体内的转录和翻译控制序列行使它们既定的调控抗体基因转录和翻译的功能。
术语“调控序列”包括控制抗体基因转录或翻译的启动子、增强子和其他表达控制元件(例如，多腺苷酸化信号)。这样的调控序列在例如Goeddel(Gene ExpressionTechnology.Methods in Enzymology 185,Academic Press,San Diego,Calif.(1990))中有过描述。优选的用于哺乳动物宿主细胞表达的调控序列包括引导在哺乳动物细胞中的高水平蛋白表达的病毒元件，例如得自巨细胞病毒(CMV)、猿猴病毒40(SV40)、腺病毒的启动子和/或增强子，如腺病毒主要晚期启动子(AdMLP)和多瘤病毒。或者，可以使用非病毒调控序列，例如泛素启动子或β-珠蛋白启动子。另外，调控元件由不同来源的序列构成，例如SRα启动子系统，其包含来自SV40早期启动子的序列和人T细胞白血病I型病毒的长末端重复(Takebe et al.,(1988)Mol.Cell.Biol.8:466-472)。表达载体和表达控制序列选为与所使用的表达宿主细胞相容。
抗体轻链基因和抗体重链基因可以插入到同一或不同的表达载体中。在优选实施方式中，可变区通过插入到已经编码所需亚型的重链恒定区和轻链恒定区的表达载体中而构建全长抗体基因，从而VH与载体中的CH可操作地连接，VL与载体中的CL可操作地连接。或者，重组表达载体可以编码促进抗体链从宿主细胞分泌的信号肽。抗体链基因可以克隆到载体中，从而信号肽在阅读框内连接到抗体链基因的氨基端。信号肽可以是免疫球蛋白信号肽或异源信号肽(即，来自非免疫球蛋白的信号肽)。
除抗体链基因和调控序列外，本发明的重组表达载体可以携带其他序列，例如调控载体在宿主细胞中复制的序列(例如，复制起始点)和可选择标记物基因。可选择标记物基因可用于选择已导入载体的宿主细胞(参见，例如，美国专利4,399,216；4,634,665和5,179,017)。例如，通常可选择标记物基因赋予已导入载体的宿主细胞以药物抗性，例如G418、潮霉素、或氨甲喋呤抗性。优选的可选择标记物基因包括二氢叶酸还原酶(DHFR)基因(用于dhfr宿主细胞的氨甲喋呤选择/扩增)和neo基因(用于G418选择)。
对于轻链和重链的表达，编码重链和轻链的表达载体通过标准技术转染到宿主细胞中。多个形式的术语“转染”包括多种常用于将外源DNA导入原核或真核宿主细胞的技术，例如，电穿孔、磷酸钙沉淀、DEAE-右旋糖转染等。尽管在原核或真核宿主细胞中表达本发明抗体在理论上是可行的，优选抗体在真核细胞中表达，最优选在哺乳动物宿主细胞中表达，因为真核细胞，特别是哺乳动物细胞，比原核细胞更可能组装并分泌适当折叠且有免疫活性的抗体。
优选的用于表达本发明重组抗体的哺乳动物宿主细胞包括中国仓鼠卵巢(CHO细胞)(包括与DHFR可选择标记物一起施用的dhfr-CHO细胞，在Urlaub and Chasin,(1980)Proc.Natl.Acad.Sci.USA 77:4216-4220中有过描述，DHFR可选择标记物在例如R.J.Kaufman and P.A.Sharp(1982)J.Mol.Biol.159:601-621中描述)、NSO骨髓瘤细胞、COS细胞和SP2细胞。特别在使用NSO骨髓瘤细胞时，另一优选的表达系统是GS基因表达系统，记载于WO 87/04462、WO 89/01036和EP 338,841。当编码抗体基因的重组表达载体导入哺乳动物宿主细胞时，通过将宿主细胞培养足以使得宿主细胞中抗体表达、或优选地足以使得使抗体分泌到宿主细胞生长的培养基中的一段时间，从而制备抗体。抗体可以使用蛋白纯化方法从培养基中回收。
重组蛋白及其制备方法
本发明还涉及包含本发明抗体或抗原结合蛋白，或包含与本发明抗体或抗原结合蛋白具有至少50％，较佳地至少70％，更佳地至少80％序列相同性的氨基酸序列的重组蛋白。此外，本发明的重组蛋白还可包括本发明纳米抗体与表达标签(如6His)融合在一起所形成的融合蛋白。
一旦获得了有关的序列，就可以用重组法来大批量地获得有关序列。这通常是将其克隆入载体，再转入细胞，然后通过常规方法从增殖后的宿主细胞中分离得到有关序列。本发明所涉及的生物分子(核酸、蛋白等)包括以分离的形式存在的生物分子。
目前，已经可以完全通过化学合成来得到编码本发明重组蛋白(或其片段，或其衍生物)的DNA序列。然后可将该DNA序列引入本领域中已知的各种现有的DNA分子(或如载体)和细胞中。此外，还可通过化学合成将突变引入本发明重组蛋白序列中。
本发明还涉及包含上述的适当DNA序列以及适当启动子或者控制序列的载体。这些载体可以用于转化适当的宿主细胞，以使其能够表达蛋白质。
宿主细胞可以是原核细胞，如细菌细胞；或是低等真核细胞，如酵母细胞；或是高等真核细胞，如哺乳动物细胞。代表性例子有：大肠杆菌，链霉菌属；鼠伤寒沙门氏菌的细菌细胞；真菌细胞如酵母；果蝇S2或Sf9的昆虫细胞；CHO、COS7、293细胞的动物细胞等。
用重组DNA转化宿主细胞可用本领域技术人员熟知的常规技术进行。当宿主为原核生物如大肠杆菌时，能吸收DNA的感受态细胞可在指数生长期后收获，用CaCl2法处理，所用的步骤在本领域众所周知。另一种方法是使用MgCl2。如果需要，转化也可用电穿孔的方法进行。当宿主是真核生物，可选用如下的DNA转染方法：磷酸钙共沉淀法，常规机械方法如显微注射、电穿孔、脂质体包装等。
获得的转化子可以用常规方法培养，表达本发明的基因所编码的多肽。根据所用的宿主细胞，培养中所用的培养基可选自各种常规培养基。在适于宿主细胞生长的条件下进行培养。当宿主细胞生长到适当的细胞密度后，用合适的方法(如温度转换或化学诱导)诱导选择的启动子，将细胞再培养一段时间。
在上面的方法中的重组多肽可在细胞内、或在细胞膜上表达、或分泌到细胞外。如果需要，可利用其物理的、化学的和其它特性通过各种分离方法分离和纯化重组的蛋白。这些方法是本领域技术人员所熟知的。这些方法的例子包括但并不限于：常规的复性处理、用蛋白沉淀剂处理(盐析方法)、离心、渗透破菌、超处理、超离心、分子筛层析(凝胶过滤)、吸附层析、离子交换层析、高效液相层析(HPLC)和其它各种液相层析技术及这些方法的结合。
免疫偶联物
本发明的抗体可以与治疗剂交联，形成免疫偶联物，例如抗体-药物偶联物(ADC)。合适的治疗剂包括细胞毒素、烷化剂、DNA小沟结合分子、DNA嵌入剂、DNA交联剂、组蛋白去乙酰化酶抑制剂、核输出抑制剂、蛋白酶体抑制剂、拓扑异构酶I或II的抑制剂、热激蛋白抑制剂、酪氨酸激酶抑制剂、抗生素和抗有丝分裂剂，优选为SN-38。在ADC中，抗体和治疗剂优选地通过接头交联，该接头可切割，例如肽类接头、二硫类接头或腙类接头。更优选地，接头是肽类接头，例如Val-Cit、Ala-Val、Val-Ala-Val、Lys-Lys、Pro-Val-Gly-Val-Val、Ala-Asn-Val、Val-Leu-Lys、Ala-Ala-Asn、Cit-Cit、Val-Lys、Lys、Cit、Ser或Glu。ADC可以如美国专利7,087,600；6,989,452；和7,129,261；PCT公开WO 02/096910；WO 07/038,658；WO07/051,081；WO 07/059,404；WO 08/083,312；和WO 08/103,693；美国专利公开20060024317；20060004081；和20060247295中描述般进行制备。
多特异性抗体
另一方面，本发明涉及包含与至少一个其他功能分子如另一种肽或蛋白(例如，另一抗体或受体配体)相连接的一种或多种本发明抗体的多特异性抗体，以生成与至少两个不同结合位点或靶向分子结合的多特异性抗体。术语“多特异性抗体”包括具有三种或更多种特异性的抗体。
在实施方式中，除Fc结合特异性和Trop2结合特异性外，双特异性分子还具有第三特异性。第三特异性可以是针对增强因子(EF)，例如与参与细胞毒性活性的表面蛋白结合并从而增加针对靶细胞的免疫应答的分子。例如，增强因子抗体可以与细胞毒性T细胞(例如，通过CD2、CD3、CD8、CD28、CD4、CD40或ICAM-1)或其他免疫细胞结合，引起增强的针对靶细胞的免疫应答。
多特异性抗体可以以多种形式和尺寸出现。在尺寸谱的一端，多特异性抗体保持传统抗体形式，除其具有两个结合臂且各臂具有不同特异性外，替代具有两个特异性相同的结合臂的情况。在另一极端的是双特异性分子，由两个经肽链连接的单链抗体片段(scFv)构成，称为Bs(scFv)2构建体。中间尺寸的双特异性分子包括由肽类接头连接的两个不同的F(ab)片段。这些和其他形式的双特异性分子可以通过基因改造、体细胞杂交或化学法进行制备。参见，例如Kufer et al,cited supra；Cao and Suresh,BioconjugateChemistry,9(6),635-644(1998)；和van Spriel et al.,Immunology Today,21(8),391-397(2000)。
编码抗体或携带抗体的溶瘤病毒
溶瘤病毒偏好地侵染并杀灭癌细胞。本发明的抗体与溶瘤病毒一起使用。此外，编码本发明抗体的溶瘤病毒可以引入人体中。
药物组合物
在另一方面，本发明提供一种药物组合物，其包含本发明的一种或多种抗体，与药学上可接受的载体配制在一起。组合物可以任选地包含一种或多种其他药学上的有效成分，例如另一抗体、化疗剂等。本发明的药学组合物可以在与例如另一抗癌剂、另一抗炎剂或疫苗的联合疗法中进行施用。
药学组合物可以包含任何数量的赋形剂。可以使用的赋形剂包括载体、表面活性剂、增稠或乳化剂、固体粘合剂、分散或混悬剂、增溶剂、染色剂、矫味剂、涂层、崩解剂、润滑剂、甜味剂、防腐剂、等渗剂及其组合。合适赋形剂的选择和使用在Gennaro,ed.,Remington:The Science and Practice of Pharmacy,20th Ed.(Lippincott Williams&Wilkins 2003)中有教导。
优选地，药物组合物适合于静脉内、肌内、皮下、肠道外、脊柱或表皮施用(例如通过注射或推注)。基于施用途径的不同，有效成分可以包在材料中，以保护其不受酸和可能使其失活的其他自然条件的影响。“肠道外施用”是指不同于肠道和局部外用的方式，通常通过注射进行，包括但不限于静脉内、肌内、动脉内、膜内、囊内、眶内、心脏内、皮内、腹膜内、经气管、皮下、表皮下、关节内、囊下、蛛网膜下、脊柱内、硬脑膜上和胸骨内注射和推注。或者，本发明的抗体可以通过非肠道外路径施用，例如外用、表皮施用或粘膜施用，例如鼻内、经口、阴道、直肠、舌下、或局部外用。
药物组合物可以为无菌水溶液或分散液的形式。它们也可以配制在微乳剂、脂质体或其他适于高浓度药物的有序结构中。
与载体材料一起制备成单剂型的有效成分的量将随着治疗主体和特定施用模式而变，且基本上而言是产生疗效的组合物的量。以百分比计，该量为约0.01-约99％的与药学上可接受载体结合的有效成分，优选为约0.1％-约70％，最优选为约1％-约30％的有效成分。
给药方案经调整提供最佳的所需反应(例如，治疗反应)。例如，可以施用快速灌注剂，可以随时间推移施用多个分剂量，或者剂量可以随治疗情况的危急程度成比例降低或提高。特别有利的是，以方便施用和剂量均匀的剂量单位型配置肠道外组合物。剂量单位型是指物理上分开的单位，适于治疗主体的单次给药；各单位包含计算出来与药学载体一起产生所需疗效的预定量的有效成分。或者，抗体可以以缓释剂施用，这种情况下所需的施用频率降低。
对于抗体的施用，剂量可以为约0.001-100mg/kg宿主体重，更常见的为0.01-5mg/kg。例如，剂量可以为0.3mg/kg体重、1mg/kg体重、3mg/kg体重、5mg/kg体重或10mg/kg体重，或在1-10mg/kg范围内。示例性的治疗方案涉及每周施用一次、两周一次、三周一次、四周一次、一个月一次、3个月一次、或3-6个月一次。本发明Trop2的优选给药方案包括静脉内施用，1mg/kg体重或3mg/kg体重，抗体以以下给药时间表中的一个进行给药：(i)每四周给药六次，然后每三个月一次；(ii)每三周一次；(iii)3mg/kg体重一次，之后1mg/kg体重每三周一次。在一些方法中，剂量调整成实现约1-1000μg/ml的血药浓度，在一些方法中为约25-300μg/ml。
“治疗有效量”的本发明Trop2抗体引起疾病症状严重程度的降低、无症状期频率和持续时间的增加、或对感病性引起的损伤或无能的预防能力。例如，对于带瘤受试者的治疗，“治疗有效量”优选地，与未治疗受试者相比，将肿瘤生长抑制至少约20％、更优选抑制至少约40％，甚至更优选地抑制至少约60％，且更优选地抑制至少约80％。治疗有效量的治疗抗体可以减小肿瘤尺寸，或者减轻受试者的症状，受试者可以是人或另一哺乳动物。
药物组合物可以是缓释试剂，包括植入体、透皮贴剂、和微胶囊递送系统。可以使用生物可降解、生物相容的聚合物，例如乙烯-醋酸乙烯、聚酸酐、聚乙醇酸、胶原蛋白、聚原酸酯、和聚乳酸。参见，例如，Sustained and Controlled Release Drug DeliverySystems,J.R.Robinson,ed.,Marcel Dekker,Inc.,New York,1978。
药学组合物可以经医学设备来给药，例如(1)无针皮下注射设备(例如，美国专利5,399,163；5,383,851；5,312,335；5,064,413；4,941,880；4,790,824；和4,596,556)；(2)微量输液泵(美国专利4,487,603)；(3)经皮给药设备(美国专利4,486,194)；(4)推注设备(美国专利4,447,233和4,447,224)；和(5)渗透设备(美国专利4,439,196和4,475,196)。
在某些实施方式中，本发明的单抗可以经配制，以确保合适的体内分布。例如，为确保本发明的治疗抗体穿越血脑屏障，抗体可以配制在脂质体中，其还可以额外地包含靶向功能基团，以增强对特定细胞或器官的选择性输送。参见，例如美国专利4,522,811；5,374,548；5,416,016；和5,399,331；V.V.Ranade(1989)J.Clin.Pharmacol.29:685；Umezawaet al.,(1988)Biochem.Biophys.Res.Commun.153:1038；Bloeman et al.,(1995)FEBSLett.357:140；M.Owais et al.,(1995)Antimicrob.Agents Chemother.39:180；Briscoeet al.,(1995)Am.J.Physiol.1233:134；Schreier et al.,(1994)J.Biol.Chem.269:9090；Keinanen and Laukkanen(1994)FEBS Lett.346:123；和Killion and Fidler(1994)Immunomethods 4:273。
本发明的用途和方法
本发明的抗体(组合物、双特异性分子和免疫偶联物)具有多种体外和外内应用，涉及例如癌症的诊断、和/或治疗。抗体可以施用至人受试者，以例如体内抑制肿瘤生长。在癌症的诊断中，可以收集目标组织样本，并与本发明抗体接触，其中，如果在某些区域或细胞类中检测到高水平的Trop2，则诊断该受试者可能患有癌症，且Trop2表达的增加/减少表示癌症发展/减轻。
考虑到本发明Trop2抗体的抑制肿瘤细胞增殖和存活的能力，本发明提供抑制受试者中肿瘤细胞生长的方法，包括向该受试者施用本发明的抗体，从而在该受试者中肿瘤生长被抑制。可以由本发明抗体治疗的肿瘤的非限制性例子包括但不限于乳癌、胃癌、胰腺癌、卵巢癌、和肠癌，原发或转移的。此外，难治的或复发的恶性肿瘤可能可以用本发明的抗体抑制。
本发明的这些和其他方法在以下进一步讨论。
联合疗法
本发明提供本发明Trop2抗体或其抗原结合部分与一种或多种其他抗体一起施用的联合疗法，其能有效抑制受试者中的肿瘤生长。在一个实施方式中，本发明提供在受试者中抑制肿瘤生长的方法，包括向受试者施用Trop2抗体以及一种或多种其他抗体，例如LAG-3抗体、PD-1抗体和/或CTLA-4抗体。在某些实施方式中，受试者是人。在另一个方面，本发明提供癌症治疗方法，其中本发明的Trop2抗体或其抗原结合部分与化疗剂一起施用，化疗剂可以是细胞毒性剂。例如，SN-38、表阿霉素、奥沙利铂、和/或5-FU可以施用给接受Trop2抗体疗法的患者。
其他可以与Trop2抗体联合的疗法包括但不限于免疫原性剂施用、白介素2(IL-2)施用、放疗、手术或激素去除。
本文讨论的治疗剂的组合可以作为在药学可接受载体中的单一组合物同时施用，或者作为分开的组合物同时施用，其中各药剂处于药学可接受载体中。在另一个实施方式中，治疗剂的组合可以按序施用。
此外，如果进行多次联合疗法施用，且药剂按序施用，则在各时间点的按序施用的次序可以反转或保持相同，按序施用可以与同时施用或其任何组合相结合。
试剂盒
本发明还提供了一种含有本发明的抗体(或其片段)或检测板的试剂盒，在本发明的一个优选例中，所述的试剂盒还包括容器、使用说明书、缓冲剂等。
本发明还提供了用于检测Trop2水平的检测试剂盒，该试剂盒包括识别Trop2蛋白的抗体，用于溶解样本的裂解介质，检测所需的通用试剂和缓冲液，如各种缓冲液、检测标记、检测底物等。该检测试剂盒可以是体外诊断装置。
本发明的主要优点包括：
本申请提供了一种抗Trop2结合蛋白的全人源抗体，其具有至少一种下述性质：
1)FACS测定中，能够特异性结合过表达人Trop2的HEK293细胞、过表达猴Trop2的CHO-K1细胞或BxPC-3细胞表面的Trop2蛋白。
2)经MMAF偶联后能够内化杀伤BxPC-3细胞。
3)对BxPC-3细胞具有ADCP活性。
4)对BxPC-3细胞具有ADCC活性。
5)结合表位与参照抗体hRS7不同。
6)能够在较短的时间被肿瘤细胞BxPC-3或者过表达人Trop2的HEK293细胞内吞。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，例如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring HarborLaboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。
实施例1：H2L2鼠免疫，杂交瘤细胞融合和抗体筛选
H2L2转基因小鼠(WO2010/070263Al)能够产生与野生型小鼠(如BALB/C)相当的免疫应答和抗体滴度。抗人Trop2的抗体分别由两种免疫原(人Trop2 ECD-hFc蛋白及HEK293T/hTrop2细胞株)免疫H2L2小鼠产生。
1.1 Trop2 ECD-hFc蛋白免疫
用免疫原为重组的糖基化的人Trop2 ECD-hFc蛋白(Chempartner,Shanghai)免疫6-8周龄的Harbour H2L2转基因小鼠,并将其养殖于无特定的病原条件(SPF)下。在第一次免疫中，每只小鼠分别在腹腔和腋下淋巴结及腹股沟淋巴结处注射50μg免疫原蛋白和0.2ml完全弗氏佐剂(CFA,Sigma,#F5881)。为了增强免疫应答，在第一次免疫接种两周后，将25μg免疫原蛋白连同200μl Ribi(Sigma佐剂系统,Sigma,#S6322)，注射到每只小鼠的腹腔内和皮下淋巴结处，随后每只老鼠每隔2周注射一次25μg免疫原和200μl Ribi佐剂，加上第一次免疫共计6次。分别在此免疫期间的第四针及第六针的一周后，采集小鼠血液，对血液进行10倍稀释取6个浓度(1:100、1:1000、1:10000、1:100000、1:1000000)，在包被有人Trop2 Fc蛋白(Chempartner,Shanghai)的ELISA板进行ELISA检测来确定小鼠血液中抗人Trop2的滴度，并经流式细胞术检测2个浓度的小鼠血液(1:100、1:1000)对Trop2高表达的CHO-K1/hTrop2细胞(Chempartner,Shanghai)和CHO-K1母细胞的特异反应性。空白对照组(PB)为免疫前老鼠的血清。
1.2 Trop2的稳转细胞株HEK293/hTrop2免疫
用免疫原为高表达人Trop2的稳转细胞株HEK293/hTrop2(Chempartner,Shanghai)免疫6-8周龄的Harbour H2L2转基因小鼠,并将其养殖于无特定的病原条件(SPF)下。在第一次免疫中，每只小鼠分别在腹腔和腋下及腹股沟共注射2×106免疫原HEK293/Trop2细胞和0.2ml完全弗氏佐剂(CFA,Sigma,#F5881)。为了增强免疫应答，在第一次免疫接种两周后，将2×10^6免疫原HEK293/Trop2细胞连同200μl Ribi(Sigma佐剂系统,Sigma,#S6322)，注射到每只小鼠的腹腔内和皮下淋巴结处，随后每只老鼠每隔2周注射一次2×106免疫原和200μl Ribi佐剂，加上第一次免疫共计6次。分别在此免疫期间的第四针及第六针的一周后，采集小鼠血液，对血液进行10倍稀释取6个浓度(1:100、1:1000、1:10000、1:100000、1:1000000)，在包被有人Trop2 Fc蛋白(Chempartner,Shanghai)的ELISA板进行ELISA检测来确定小鼠血液中抗人Trop2的滴度，并经流式细胞术检测2个浓度的小鼠血液(1:100、1:1000)对Trop2高表达的CHO-K1/hTrop2细胞(Chempartner,Shanghai)和CHO-K1母细胞的特异反应性。空白对照组(PB)为免疫前老鼠的血清。
在完成上述步骤之后，选择具有针对人Trop2的特异性免疫应答的小鼠进行融合之前，在会阴内注射100μg纯化的人Trop2 Fc(Chempartner,Shanghai，#151215003)蛋白加强免疫。三天后，处死小鼠，并收集其脾细胞和淋巴结细胞。将NH4OH添加到脾细胞和淋巴结样品中，最终浓度为1％(w/w)，以溶解样品中的红细胞。样品以1000转/分的速度离心，用DMEM培养基洗涤三次，测定细胞的存活率和数量。小鼠骨髓瘤细胞sp2/0(ATCC,#CRL-1581)用无血清的DMEM洗涤两次，测定细胞的存活率和数量。然后用高效电融合法以4:1的比例将活脾细胞与小鼠骨髓瘤细胞sp2/0(ATCC,#CRL-1581)融合。
将融合细胞重新悬浮于并在含有20％超低IgG FBS(ultra-low IgG,FetalBovine Serum,#16250086,Life Technologies)并添加有1×次黄嘌呤、氨蝶呤和胸苷(50×HAT supplement,#21060017,Life Technologies)的培养基(Hybridoma-SFM,#12045084,Life Technologies)将浓度调整为105个细胞/200μl。在96孔板的每个孔中加入200μl的融合细胞，在37摄氏度、5％CO2条件下培养。细胞融合后14天，免疫原为糖基化Trop2 Fc蛋白得到的杂交瘤上清液用酶联免疫吸附试验(ELISA)测定其与人Trop2 Fc蛋白结合的能力来筛选；免疫原为HEK293/hTrop2得到的杂交瘤上清液用Acumen荧光微孔板进行检测其与CHO-K1/hTrop2及CHO-K1母细胞的结合能力。
筛选出的阳性克隆(OD450>2)或者在Acumen上阳性百分比>80％的克隆再通过流式细胞仪筛选与CHO-K1/人Trop2细胞,CHO-K1/猴Trop2细胞及CHO-K1/小鼠Trop2细胞(Chempartner,Shanghai)特异结合的克隆。在荧光强度评分后，选择荧光强度最强的70个杂交瘤母克隆，通过有限稀释法进行亚克隆，经筛选生长起来的亚单克隆通过ELISA和Acumen及流式细胞术筛选，找出与人，猴及小鼠Trop2细胞均结合的最强的亚单克隆。经小鼠Ig分型Ready-SET-GO！ELISA(Life technologies,#88-50640-88)确定为IgG亚型的亚单克隆将进行测序分析。
实施例2：单克隆Trop2抗体的测序、表达和纯化
对单克隆Trop2抗体进行测序，序列总结在表1a和表1b中。
将Trop2抗体的重链可变区序列亚克隆到含有信号肽和人重链IgG1恒定区的pTT5表达载体中。将Trop2抗体的轻链可变区序列亚克隆到含有信号肽和人抗体轻链kappa恒定区的表达载体中。重组质粒经测序证实后用大抽试剂盒(Macherey-Nagel，
Figure BDA0002873116140000351
Xtra Midi)抽提质粒，以提高重组质粒的纯度和质量，质粒通过0.22μm过滤器(millpore)过滤。纯化后的质粒用于转染。
293-6E细胞(Genescript,Nanjing,China)在FreeStyle 293培养基(invitrogen,#12338026)中37℃，130rpm，5％CO2培养箱培养。调整HEK293-6E细胞到1-1.5×106/ml细胞密度后进行转染。用重链质粒和轻链质粒通过PEI(sigma)共转染HEK293-6E细胞，持续一周。大约在第5-7天，测定抗体的滴度。约在第6-7天，对HEK293E培养物进行离心(30分钟，3500rpm)，收集上清液，并通过0.22μm过滤器过滤纯化。
蛋白A柱(GE)用0.1M NaOH洗涤30分钟或用5倍柱体积的0.5M NaOH洗涤以去除内毒素。长期未使用的蛋白A柱先在1mNaOH中浸泡至少1小时，再用无内毒素水清洗至pH为中性值，最后用10倍柱体积的1％Triton X100进行清洗。然后用5倍柱体积的磷酸盐缓冲液PBS(pH7.4)平衡蛋白柱。将上述收集的上清液装载到柱上，必要时收集流过的液体。用5倍柱体积的PBS洗涤柱，然后用5倍柱体积的0.1M甘氨酸-HCl(pH3.0)洗脱。含有Trop2抗体的洗脱液用0.5倍柱体积的1M Tris-HCl(Nacl 1.5M)pH8.5中和。人抗Trop2抗体在1×PBS中透析4小时，以避免内毒素污染。透析后用分光光度法或试剂盒测定抗Trop2的抗体浓度，用高效液相色谱-质谱法测定抗体纯度，用便携式内毒素测试仪器(
Figure BDA0002873116140000352
nexgen-PTSTM，Charles River)测定内毒素含量。
经上述实验，共得到29种具有独特序列的Trop2单克隆抗体，抗体的可变区序列列于表1a。得到的29种抗体均为IgG1亚型，两条链的种系不同于参照抗体Tab(hRS7)。
此外，准备了两种对比例抗体，用于后续实施例，一种是对比例1抗体PR001131(自制)，另一种是对比例2抗体Tab(hRS7，ChemPartner，#T07-001)。
具体而言，对比例1抗体PR001131的恒定区序列，可变区序列，具体地，其VH的氨基酸序列如NO.193所示，VL的氨基酸序列如NO.223所示，重链的氨基酸序列如NO.242所示，轻链的氨基酸序列如NO.272所示。
对比例2抗体Tab(hRS7)是在Immunomedics,Inc.和Seattle Genetics公司的sacituzumab govitecan基础上制备的对比例抗体，其重链和轻链的氨基酸序列分别如SEQNO ID:290和SEQ NO ID:291所示。
实施例3：重组Trop2抗体与293T-人Trop2结合能力
用3ml TrypLE酶(Life technologies,#12604-013)消化293T-人Trop2细胞(KyInno，#KC-0995)，用7ml相应培养基终止消化。确定细胞密度，用PBS重悬细胞至1×106/ml,每孔取100ul细胞悬液至96孔V型板内。96孔V型板1000rpm离心5分钟，用PBS+0.5％BSA(VWR,#0332-100G)清洗一次。将细胞重悬于100μl梯度稀释的抗体中(5倍稀释，起始浓度66.67nM)，并于4℃孵育1小时，hRS7(ChemPartner，#T07-001)和hlgG1分别作为参考对照和阴性对照。1小时后用PBS+0.5％BSA清洗两次。将细胞与100μl Alexa Fluor 488山羊抗人IgG(Life technologies，1:1000，#A-11013)避光4℃孵育1小时，用PBS+0.5％BSA清洗两次。将细胞重新悬浮在200μl PBS+0.5％BSA中。用FACS verse流式细胞仪分析细胞的荧光强度。所有操作均在冰上进行。
如图1和表3所示，本发明的29株Trop2抗体均能特异性地结合人Trop2，且检测到的抗体结合能力与抗体浓度成正相关关系递增。与之相反，对比例1抗体PR001131与人Trop2的结合则很弱。与对比例1抗体Tab(hRS7)相比，在相同浓度下，本发明的26个Trop2抗体表现出比参照抗体hRS7更高或者相当的Emax，表明了抗体能在HEK293/hTrop2细胞上结合更多的hTrop2蛋白。其中14个抗体的EC50比参照抗体更低，说明这些抗体能以较低的浓度更灵敏得结合Trop2，其中以PR001142,PR001162和PR001166为最佳，其EC50均小于0.3nM,比参照抗体的EC50小约三倍。
表3.Trop2抗体与293T-人Trop2的结合力
Figure BDA0002873116140000361
Figure BDA0002873116140000371
对Trop2抗体与人Trop2稳转株的结合亲和力的EC50和中位荧光强度(MFI)最高值(MFI max)进行排序，选择EC50<4ug/ml且MFI max>4000的29个Trop2抗体进行后续检测。
实施例4：Trop2抗体对重组人或猴Trop2蛋白的结合亲和力和解离常数的测定
按照制造商提供的详细操作和方法，使用Octet RED96仪器(Fortiebio)和抗人IgG Fc亲和素传感器(AHC传感器，Pall ForteBio，#18-5060)测定亲和力。具体地，用含有0.1％(w/w)BSA和0.02％(v/v)吐温20的PBS缓冲液(pH7.4)将人TROP2蛋白(SinoBiological，#10428-H08H)或者猴Trop2蛋白(Sino Biological，#90893-C08H)稀释至200nM，分别与AHC传感器孵育。将40nM的Trop2抗体分别与负载人Trop2蛋白或猴蛋白的AHC传感器在30℃孵育3分钟。该反应混合物在含有0.1％(v/w)BSA和0.02％(v/v)吐温20的PBS缓冲液(pH7.4)中于30℃继续孵育5分钟。Octet Red 96实时记录Trop2抗体与人Trop2蛋白或者猴Trop2蛋白的结合和分离信号。亲和力、关联和解离常数由Octet使用软件确定，结果如表4、表5和表6所示，测试的29个Trop2抗体中4个抗体(PR001128，PR001130，PR001166，PR001170)与人Trop2蛋白和猴Trop2蛋白的KD值均低于参照抗体Tab(hRS7)，表明其更强的人和猴Trop2结合亲和力。在与人Trop2蛋白结合亲和力方面，除上述四种抗体外，PR001132、PR001138、PR001150、PR001159、PR001153、PR001165的KD值约低于参照抗体10倍；而这些抗体与猴蛋白的亲和力与参照抗体相当。
表4 Trop2抗体与人Trop2蛋白的结合与解离常数
Figure BDA0002873116140000381
Figure BDA0002873116140000391
表5 Trop2抗体与猴Trop2蛋白的结合与解离常数
抗原 抗体 应答 KD(M) kon(1/Ms) kdis(1/s)
猴Trop2 PR001128 0.3835 <1.0E-12 8.35E+04 <1.0E-07
猴Trop2 PR001130 0.3145 <1.0E-12 3.77E+04 <1.0E-07
猴Trop2 PR001132 0.4181 1.64E-10 8.69E+04 1.43E-05
猴Trop2 PR001133 0.3691 2.49E-09 5.98E+04 1.49E-04
猴Trop2 PR001134 0.335 9.33E-09 5.49E+04 5.12E-04
猴Trop2 PR001138 0.3289 4.09E-10 9.74E+04 3.98E-05
猴Trop2 PR001139 0.3278 7.75E-09 6.81E+04 5.28E-04
猴Trop2 PR001142 0.3824 2.60E-10 1.65E+05 4.29E-05
猴Trop2 PR001143 0.3078 1.24E-08 5.21E+04 6.45E-04
猴Trop2 PR001145 0.2767 1.91E-08 7.01E+04 1.34E-03
猴Trop2 PR001147 0.3405 1.92E-09 6.09E+04 1.17E-04
猴Trop2 PR001150 0.3441 3.19E-10 1.07E+05 3.41E-05
猴Trop2 PR001151 0.3512 2.38E-09 7.58E+04 1.80E-04
猴Trop2 PR001152 0.3432 1.02E-09 1.03E+05 1.05E-04
猴Trop2 PR001153 0.3366 1.81E-10 5.93E+04 1.07E-05
猴Trop2 PR001154 0.3792 1.91E-09 7.27E+04 1.39E-04
猴Trop2 PR001155 0.3561 4.26E-09 5.15E+04 2.19E-04
猴Trop2 PR001156 0.3494 5.36E-10 1.15E+05 6.15E-05
猴Trop2 PR001158 0.4055 7.08E-10 8.79E+04 6.23E-05
猴Trop2 PR001159 0.3716 3.07E-10 1.11E+05 3.39E-05
猴Trop2 PR001160 0.3603 1.82E-09 7.43E+04 1.35E-04
猴Trop2 PR001162 0.3701 3.66E-09 2.00E+05 7.32E-04
猴Trop2 PR001163 0.3721 1.96E-09 6.84E+04 1.34E-04
猴Trop2 PR001164 0.173 2.04E-09 7.94E+04 1.62E-04
猴Trop2 PR001165 0.3925 2.60E-10 1.01E+05 2.62E-05
猴Trop2 PR001166 0.3224 <1.0E-12 3.16E+05 <1.0E-07
猴Trop2 PR001168 0.3111 2.45E-09 5.93E+04 1.45E-04
猴Trop2 PR001170 0.3865 <1.0E-12 3.07E+05 <1.0E-07
猴Trop2 PR001171 0.3194 1.51E-10 8.83E+04 1.34E-05
猴Trop2 Tab(hRS7) 0.3224 1.37E-10 1.08E+05 1.48E-05
表6抗体与猴Trop2蛋白和人Trop2蛋白的解离常数的比较
Figure BDA0002873116140000401
实施例5：MMAF偶联抗体内化对靶细胞的杀伤
Trop2抗体可以通过结合Trop2的胞外部介导细胞表面表达的Trop2蛋白的内化。在本实施例中，测试Trop2抗体内化后Trop2+细胞对于Trop2抗体杀伤的易感性。
BxPC-3细胞在T-75培养瓶中用含10％血清(BI,胎牛血清，#04-002-1A)的DMEM培养基(Life technologies，#11995-065)培养和扩增，达到90％的融合后吸取培养基，用PBS洗涤细胞两次。细胞用胰酶(Invitrogen，#15050065)处理1分钟左右，再用培养基中和胰酶。将细胞转移到15ml无菌离心管中，1000rpm室温离心5分钟，使细胞成团。吸去培养基，将细胞重新悬浮于各自的培养基。轻柔地吹打细胞，得到单细胞混悬液。用细胞计数板进行计数，之后将2×103BxPC-3细胞加至黑色ViewPlate-96 TC(Perkin Elmer,#6005225)板中。37℃、5％CO2孵育箱中孵育过夜。
第二天，用不含FBS的培养基制备10×起始浓度(100nM)的抗体溶液，5倍稀释，制备6个抗体浓度。将10μl各梯度的抗体样品移至上述细胞板内，各孔的终体积为100μl。每孔加入2μl的50μg/mlαHFc-CL-MMAF培养基(Moradec,αHFc-CL-MMAF试剂盒，#AH-102AF)，使其终浓度为1μg/ml。37℃、5％CO2孵育4天。
在第六天，每孔加入100μl
Figure BDA0002873116140000412
发光细胞活性试剂(Promega,USA,#G7570)，在摇床上混合2分钟，诱导细胞裂解。96孔板在室温孵育10分钟来稳定光信号。使用PE Enspire酶标仪(Perkin Elmer,EnSpire)记录发光情况，存活率＝(1-(IgG1发光值平均值-抗体样品发光值)/IgG1发光值平均值)*100％。根据存活率确定EC50值。
图2和表7示出抗体处理下BxPC-3细胞的存活率。本发明的27个抗体的内化活性的EC50与hRS7相当或更好，除了PR001163和PR001164。具体而言，绝大部分的Trop2抗体的EC50值低于参照抗体，说明其能够以较低浓度实现最大抗体内化杀伤效应，而PR001166抗体处理下的细胞存活率与参照抗体Tab(hRS7)相比，最高存活率明显低于Tab处理下的细胞存活率，说明其在较低浓度下具有最好的抗体内化杀伤效应。
表7 Trop2抗体的内化杀伤靶细胞活性
Figure BDA0002873116140000411
Figure BDA0002873116140000421
根据抗体与HEK293/hTrop2的结合EC50的高低、抗体序列的种系多样性和翻译后修饰的位点多少(表8)、与人猴蛋白亲和力的高低以及内吞效应的优劣，选择出表8中的8个抗体做后续分析。
表8 Trop2抗体序列的种系和翻译后修饰位点
抗体编号 重链种系 轻链种系 重链翻译后修饰 轻链翻译后修饰
PR001128 VH4_4*02 Vk3_11*01
PR001130 VH4_4*02 Vk1_5*01
PR001133 VH3_30*18 Vk1_39*01 DG(HCDR2),DG(HFR3) NS(LCDR1)
PR001139 VH3_30*01 Vk3_15*01 DG(HCDR2)
PR001142 VH4_4*02 Vk1_5*01
PR001145 VH3_30*01 Vk3_15*01 DG(HCDR2)
PR001162 VH3_30*01 Vk3_11*01 DG(HCDR1),DG(HCDR2)
PR001165 VH3_30*01 Vk1_9*01 DG(HCDR2),DG(HFR3) NS(LCDR3)
PR001166 VH4_38-2*01 Vk3_15*01
实施例6：重组抗体与BxPC-3肿瘤细胞系结合
用3mL TrypLE消化BxPC-3肿瘤细胞(中国科学院细胞库，#TCHu 12)，用7mL相应培养基终止消化。确定细胞密度，为每孔的测试样品收集1×105细胞。细胞液离心，用PBS+0.5％BSA缓冲液清洗一次。将细胞悬浮在100μl梯度稀释的抗体(5倍稀释，起始浓度100nM)，并于4℃孵育1小时，hRS7和hIgG1用作对照。用PBS+0.5％BSA清洗一次。将细胞与100μl 1:1000稀释的Alexa Fluor 488山羊抗人IgG(Life technologies，#A-11013)避光4℃孵育1小时，用PBS+0.5％BSA清洗两次。将细胞重新悬浮在200μl PBS+0.5％BSA中。2000rpm、4℃离心5分钟，用FACS verse流式细胞仪或NovoCyte流式细胞仪(ACEABiosciences)测试。所有操作均在冰上进行。
抗体与BxPC肿瘤细胞的结合如图3和表9所示。与对照hRS7相比，本发明的这8个抗体表现出与参照抗体相当的或者更高的BxPC肿瘤细胞的结合力。本发明中的3个抗体(PR001142,PR001162和PR001166)EC50比参照抗体更低，其EC50比参照抗体的EC50小约二倍，说明这些抗体能以较低的浓度更灵敏地结合Trop2。
表9 Trop2抗体与BxPC-3肿瘤细胞的结合力
Figure BDA0002873116140000431
实施例7：重组抗体与CHO-K1/猴Trop2结合
分别用3mL TrypLE消化CHO-K1/猴Trop2细胞(Chempartner,Shanghai,China)和CHO-K1母细胞，分别用7mL相应培养基终止消化。测定细胞密度，为每孔测试样品收集2×105细胞。细胞液离心，用PBS+0.5％BSA清洗一次。将细胞悬浮在100μl梯度稀释的抗体中(5倍稀释，起始浓度100nM)，并于4℃孵育1小时，hRS7和hlgG1用作对照。用PBS+0.5％BSA清洗一次。将细胞与1:1000稀释的100μl Alexa Fluor 488山羊抗人IgG(Life technologies，#A-11013)避光4℃孵育1小时，用PBS+0.5％BSA清洗两次。将细胞重新悬浮在200μl PBS+0.5％BSA中。2000rpm、4℃离心5分钟，用FACS verse流式细胞仪或NovoCyte流式细胞仪(ACEA Biosciences)测试。所有操作均在冰上进行。
如图4和表10所示，本发明的8个抗体与猴Trop2的结合力与参照抗体hRS7相当，其中PR001166的EC50比参照抗体的EC50小，说明该抗体能以较低的浓度更灵敏地结合Trop2。但PR001130与猴Trop2的结合力稍弱一些。
表10.Trop2抗体与CHO-K1/猴Trop2及CHO-K1的结合力
Figure BDA0002873116140000432
Figure BDA0002873116140000441
Figure BDA0002873116140000442
实施例7：重组抗体与小鼠Trop2结合
用100μl溶于PBS的1μg/ml小鼠Trop2-his蛋白(Sino Biological,#50922-M08H)对ELISA板进行包被，4℃过夜。用200μl溶于PBST(PBS+0.05％吐温20)+2％BSA进行封闭，37℃，1小时。用PBST清洗4遍。向ELISA板加入100μl梯度稀释的Trop2抗体(5倍稀释，起始浓度33.33nM)，37℃孵育1小时。用PBST清洗4遍。加入100μl 1:4000稀释的抗人IgG(H+L)-HRP(Sigma，#A8667)，37℃孵育1小时。用PBST清洗4遍。加入100μl TMB底物(Invitrogen，#88-7025-88)，室温孵育5分钟，随后加入50μl终止缓冲液(BBI life sciences，#E661006-0200)来终止反应。用酶标仪(Perkin Elmer,EnSpire)记录OD 450nm读值。
结果图5显示，抗体PR001139、PR001166与小鼠Trop2蛋白结合，参照抗体hRS7及其他6种Trop2抗体均没有小鼠Trop2结合活性。
根据上述各种实验结果选择3种抗体，即PR001142、PR001165和PR001166，进一步测试其抗体依赖的细胞介导的细胞毒性作用、补体依赖的细胞毒性作用和抗体依赖的细胞介导的细胞吞噬作用。
实施例8：Trop2抗体介导的ADCC活性
细胞裂解后细胞内的乳酸脱氢酶(lactate dehydrogenase,LDH)会随之增加，因此可以通过检测LDH-释放量来判定肿瘤细胞被杀伤的效果。先用2％FBS的RPMI1640稀释抗体至40nM，再从40nM做5倍稀释，稀释为不同的7个浓度。取25μl加至100μl的终体积，得到终浓度为10.0、2.0、0.4、0.08、0.016、0.0032、0.00064nM的7个浓度。收集BxPC-3和外周血单核细胞(PBMC，Miaotong,#PB050F)，1200rpm离心5min，弃上清，并分别再悬浮于含2％FBS的无酚红的RPMI-1640培养基中，PBMC密度调整为4×106/ml，取50μl/孔加入96孔U形板(Corning,#3799)，BxPC-3细胞密度调整为4×105/ml，取25μl/孔加入同一96孔板内，效靶比为20:1，设三个重复。在同一板内加入25μl相应的稀释抗体(此为ER:BxPC-3+PBMC+mAb)，37度二氧化碳培养箱孵育4小时。同时在板内设置不同的对照组：ER(0)：BxPC-3+PBMC+2％FBS+1640；ESR：PBMC+2％FBS+1640；TSR:BxPC-3+2％FBS+1640；CMB:2％FBS+1640；TMR:BxPC-3+2％FBS+1640+裂解缓冲液(裂解缓冲液于孵育3小时后加入)；VCC：2％FBS 1640+裂解缓冲液(裂解缓冲液于孵育3小时后加入)。孵育完成后，取50μl上清液，加入96孔板(Corning,#3599)，并加入50μl细胞毒性检测试剂(CytoTox
Figure BDA0002873116140000452
非放射性细胞毒性检测试剂盒，Promega,USA，#G1780)，室温孵育30分钟，加入终止溶液停止反应并用Enspire(Perkin Elmer)读取OD490值。该读值按照以下公式来计算ADCC效果，ADCC特异性杀伤％＝{(ER-CMB)-[(ESR-CMB)+(TSR-CMB)]}/(TMR-VCC)*100％，重复2个不同的PBMC供体(Lot#1152和Lot#2153)。最高特异性杀伤％和EC50值总结在表9中。
图6显示3种Trop2抗体在不同的供体的PBMC中介导对BxPC-3的细胞杀伤作用。从表11可以看出，3种抗体均显示出对BxPC-3细胞的特异杀伤性。其中，在使用两种不同的供体1152#和2153#的PBMC的情况下，从特异性杀伤率的均值和标准误差看，相比，抗体PR001166和参照抗体Tab(hRS7)的最高杀伤率相当，但EC50值更低，说明PR001166抗体能够在更低浓度下达到与参照抗体相当的杀伤力。而其他两种抗体PR001142和PR001165与参照抗体的杀伤程度相当。
表11. 3种Trop2抗体对BxPC-3的ADCC效果
Figure BDA0002873116140000451
实施例9：Trop2抗体的ADCP活性
抗体依赖的细胞介导的细胞吞噬作用是治疗性抗体对抗病毒感染或者肿瘤疾病的一种重要作用机制。
从PBMC(Miaotong,#PB050F)中用人CD14分选磁珠(Meltenyi Biotech,#130-050-201)分离CD14+单核细胞，以密度为1×106/mL重悬于含10％FBS的RPMI1640培养基，加入100ng/ml GM-CSF(PeproTech,#300-03-A)。取2×106个单核细胞每孔于6孔板内，37度二氧化碳培养箱培养9天，使其分化为巨噬细胞。每隔3-4天换一次液(含100ng/ml GM-CSF)。9天后用胰酶消化巨噬细胞，用含10％FBS的RPMI1640终止胰酶反应。收集细胞，用PBS洗涤一次，用PBS重悬为密度为1×106/ml。同样收集BxPC-3细胞，用PBS重悬为密度为1×106/ml。巨噬细胞用0.1μM Far-red(溶于PBS)，BxPC-3细胞用0.5μM CFSE(溶于PBS)在4度染色10分钟。离心染色后的细胞，用>20ml的RPMI1640+10％FBS培养基洗涤一次。重悬洗涤后的细胞于1％BSA+RPMI1640培养基中，并调整细胞密度均为1.6×106/ml。在96孔V型板内(Corning,#3894)每孔加入25μl BxPC-3细胞(每孔细胞数为4×104)和25μl巨噬细胞(每孔细胞数为4×104)。用1％BSA+RPMI1640稀释抗体至中间浓度20nM，再从20nM 5倍稀释成7个梯度。含有BxPC-3和巨噬细胞的同一96孔V型板内每孔加入50μl的稀释抗体，混匀完全。37℃孵育1小时。使用NovoCyte流式细胞仪(ACEA Biosciences)经流式细胞术识别FITC+BxPC-3细胞和Alexa647+巨噬细胞双阳性的比例。数据用FlowJo软件(Tree Star,Ashland,OR)分析，双染细胞的百分比用于确定ADCP介导的细胞吞噬。
图7显示3种Trop2抗体在不同的供体的巨噬细胞中介导的对BxPC-3的吞噬作用。如表12所示，3种抗体均显示出对BxPC-3细胞的ADCP效应。其中，在使用供体A1925144#的PBMC的情况下，从特异性杀伤率的均值和标准误看，3种抗体的最高吞噬％均接近于参照抗体Tab，而这些抗体的EC50值均小于Tab，表明其能够在较低浓度下达到与参照抗体相当或更高的最大杀伤力。而在使用供体A19143143#的PBMC的情况下，从特异性杀伤率的均值和标准误看，3种抗体最高吞噬％与Tab没有显著差异，但EC50值更低，表明其能够在较低浓度下达到与参照抗体相当或更高的最大杀伤力。这两种供体均表明这3种抗体能够在更低浓度下达到与参照抗体相当的杀伤力。
表12 3种Trop2抗体对BxPC-3的ADCP效果
Figure BDA0002873116140000461
实施例10：Trop2抗体的CDC活性
通过补体介导的细胞杀伤检测来评估Trop2抗体的CDC活性。用胰酶消化处理BxPC-3细胞或HEK293/hTrop2细胞，终止胰酶反应后用无血清的RPMI-1640重悬BxPC-3细胞或HEK293/hTrop2细胞，在黑色底透96孔板(ViewPlate-96 TC,black，Perkin Elmer，#6005225)铺上每孔25μl的1×104BxPC-3细胞。用无血清的RPMI-1640稀释抗体，每孔加入25μl不同梯度的抗体。每孔再加入50ul的人血清，混匀完全。在37度二氧化碳培养箱培养24小时。根据制造商提供的方法，使用Celltiter Glo发光细胞活力试剂盒(Promega，#G7573)测试细胞活力。将培养皿在微孔板摇床上以200的转速摇动2分钟，然后在室温下培养10分钟。用读板机(Perkin Elmer,EnSpire)读取发光信号。为了进行数据分析，使用graphpadprism 7.0计算细胞毒性百分比。
图8结果表明3种抗体以及参照抗体Tab(hRS7)在BxPC-3和HEK293/hTrop2细胞上均未显示出CDC活性(CDC≤20％)。
实施例11：BIACORE测试的抗体与人及猴Trop2蛋白的结合亲和力
整个测试过程中使用HBS-EP+(10mM HEPES，150mM NaCl，3mM EDTA和0.05％P20，pH7.4，GE Healthcare，BR-1001-88)作为运行缓冲液。先使用手动运行模式通过以下程序将抗原人Trop2蛋白或猴Trop2蛋白固定在芯片表面上，该程序为：1)用新混合的50mM N-羟基琥珀酰亚胺(NHS)和200mM 1-乙基-3-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDC)以10μl/分钟活化系列S CM5传感器芯片(GE Healthcare，#29-1275-56)的流动池3,该过程持续420秒。2)然后将稀释于10mM NaOAc(pH 5.0)中的0.2μg/ml人Trop2或者猴Trop2以10μl/分钟注入活化流动池3中，该过程持续120秒。3)用1M乙醇胺(pH8.5)以10μL/min封闭剩余的活性酯基团120秒。再将不同浓度的稀释后的抗体(浓度分别为0、3.125、6.25、12.5、25.0、50.0、100nM)以30μl/分钟注入将流体池1,2,3和4内，结合时间设定为180秒。PR001142、PR001165和hRS7的解离时间为900s，PR001166解离时间设为1800s。为了从表面除去测试的抗体，将10mM甘氨酸-HCl pH1.5(GE Life Sciences,#BR-1003-54)以30μL/min注入流动池1,2,3和4中30秒。使用Biacore T200(GE Healthcare,T200 model,#BIAC-B20-03)评估3.1软件来分析数据。
如表13所示BIACORE测试结果，结果显示不管是与人还是猴的Trop2蛋白的结合亲和力上，抗体PR001142和PR001165的结合亲和力与hRS7相当，而PR001166的结合亲和力高于hRS7。
表13.BIACORE测定的Trop2抗体的与人和猴Trop2蛋白的结合亲和力
Figure BDA0002873116140000471
Figure BDA0002873116140000481
实施例12：抗原结合蛋白的表位鉴定(Epitope binning)
使用ForteBio Octet平台对得到的抗原结合蛋白(PR001142,PR001165,PR001166)和hRS7进行表位鉴定。生物素分子(Sulfo-NHS-LC-Biotin,Thermo Fisher,#A39257)与人Trop2蛋白(Sino biological，#10428-H08H)按摩尔数3:1的比例在室温孵育2小时。孵育后的生物素化混合物用PBS及Zeba离心脱盐柱(Thermo Fisher,#89882)洗涤三次去除多余的生物素。生物素化Trop2蛋白由SA传感器(Pall ForteBio，#18-5019)捕获，上样基线设置为0.3nm。传感器浸入含有50nM的第一抗体的测定缓冲液的孔，持续300秒。再将传感器转移至含有50nM的第一抗体的测定缓冲液的孔，持续300秒。如果第二抗体表现出明显结合，那么就将它视为非竞争剂(即，在与第一抗体不同的表位区间中)。如果第二抗体不表现明显结合，那么就将它视为竞争剂(即，在与第一抗体相同的表位区间中)。通过比较第二抗体在第一抗体存在下与Trop2的结合与第一抗体阻断本身来进行结合测定。
结果如下表14所示，PR001142和PR001165与hRS7在Trop2的结合位点重叠性高，但是PR001166与hRS7及PR001165的结合位点重叠性低。
表14本申请抗原结合蛋白与临床抗体的表位鉴定
Figure BDA0002873116140000482
实施例13：Trop2抗体在靶细胞上的短时间内吞效应
Trop2抗体可以通过结合Trop2的胞外部介导细胞表面表达的Trop2蛋白的内化。在本实施例中，测试Trop2抗体在短时间(3小时以内)不同时间点内化的情况。BxPC-3或HEK293/hTrop2细胞在T-75培养瓶中用含10％血清(BI,胎牛血清，#04-002-1A)的DMEM培养基(Life technologies，#11995-065)培养和扩增，达到90％的融合后吸取培养基，用PBS洗涤细胞两次。细胞用胰酶(Invitrogen，#15050065)处理1分钟左右，再用培养基中和胰酶。将细胞转移到15ml无菌离心管中，1000rpm室温离心5分钟，使细胞成团。吸去培养基，将细胞重新悬浮于各自的培养基。轻柔地吹打细胞，得到单细胞混悬液。用细胞计数板进行计数，之后用冰冷的FACS缓冲液(PBS+2％FBS)将BxPC-3细胞重悬成2×106个细胞/ml,取100ul每孔细胞悬液至96孔V型板(Corning,#3894)内,400g离心5分钟。弃上清后，对于无内吞作用的测定按如下流程操作：分别加入100ul含终浓度为100nM的PR001142或PR001165或PR001166或hIgG1的FACS缓冲液，在4℃孵育一小时。用冰冷的FACS缓冲液洗涤两次去除未结合的抗体。而对于内吞作用的测定，则将加入抗体后的细胞分别在37℃放置0/15min/30min/45min/1hr/1.5hr/2hr/3hr，然后再将细胞转移至4℃，加入预冷的PBS来阻止抗体的内吞。用冰冷的FACS缓冲液洗涤两次去除未结合的抗体。两种情况下，细胞用2ug/ml的Alexa Fluor 488goat anti-human lgG(H+L)(Jackson ImmunoResearch，#109-545-003)在冰冷的FACS缓冲液中于4℃放置半小时。用冰冷的FACS缓冲液洗涤两次去除未结合的二抗。将细胞重新悬浮在200μl FACS缓冲液中。用FACS canto II流式细胞仪分析细胞的荧光强度。内吞率％(＝(1-MFI 37℃/MFI4℃)*100％。
结果如图9和图10所示，PR001142/PR001165/PR001166在饱和的抗体结合浓度下，在重组细胞系HEK293/hTrop2或者Trop2阳性的肿瘤细胞BxPC-3中的内吞速率彼此相差不大。然而在1小时以内，PR001166与其他抗体包括参照抗体相比，具有更好的内吞效应。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
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<110> 和铂医药（苏州）有限公司
<120> 结合TROP2的抗体及其用途
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<210> 53
<211> 6
<212> PRT
<213> 人工序列(artificial sequence)
<400> 53
Trp Phe Asp Gly Ser Asn
1 5
<210> 54
<211> 5
<212> PRT
<213> 人工序列(artificial sequence)
<400> 54
Tyr Pro Gly Gly Asn
1 5
<210> 55
<211> 6
<212> PRT
<213> 人工序列(artificial sequence)
<400> 55
Trp Tyr Asp Gly Ile Lys
1 5
<210> 56
<211> 6
<212> PRT
<213> 人工序列(artificial sequence)
<400> 56
Trp Tyr Asp Gly Ile Met
1 5
<210> 57
<211> 6
<212> PRT
<213> 人工序列(artificial sequence)
<400> 57
Trp Phe Asp Gly Gly Ser
1 5
<210> 58
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 58
Cys Tyr Arg Ser Lys Trp Tyr
1 5
<210> 59
<211> 5
<212> PRT
<213> 人工序列(artificial sequence)
<400> 59
Tyr His Ser Gly Arg
1 5
<210> 60
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 60
Thr Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Leu Val Asp
1 5 10 15
Lys Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 61
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 61
Thr Ser Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp
1 5 10 15
Lys Ser Lys Asn Gln Phe Ser Leu Lys Leu Asn Ser Val Thr Ala Ala
20 25 30
Asp Thr Ala Met Tyr Tyr Cys Ala Arg
35 40
<210> 62
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 62
Lys Tyr Tyr Ala Asn Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu
20 25 30
Asp Thr Ala Leu Tyr Tyr Cys Ala Lys
35 40
<210> 63
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 63
Arg Tyr Tyr Thr Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Ser Leu Gln Met Lys Ser Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Lys
35 40
<210> 64
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 64
Glu Asp Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Gly Glu
20 25 30
Asp Thr Ala Met Tyr Tyr Cys Ala Arg
35 40
<210> 65
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 65
Thr Tyr Cys Asn Pro Ser Leu Lys Ser Arg Val Thr Met Ser Val Asp
1 5 10 15
Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Arg Ser Val Thr Ala Ala
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 66
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 66
Lys Tyr Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 67
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 67
Pro Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp
1 5 10 15
Lys Ser Lys Asn Gln Phe Ser Leu Lys Leu Asn Ser Val Thr Ala Ala
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 68
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 68
Glu Asp Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Gly Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 69
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 69
Arg Tyr Tyr Pro Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asp Ser Arg Asn Thr Val Ser Leu Gln Met Asn Ser Leu Arg Pro Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 70
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 70
Arg Tyr Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 71
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 71
Lys Asp Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 72
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 72
Ser Tyr Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asp Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 73
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 73
Thr Asp Tyr Asn Pro Ser Leu Lys Ser Arg Val Ile Ile Ser Val Asp
1 5 10 15
Lys Ser Lys Asn Gln Phe Ser Leu Lys Val Arg Ser Val Thr Ala Ala
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 74
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 74
Lys Tyr Tyr Ile Glu Ser Val Lys Gly Arg Phe Ile Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 75
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 75
Lys Tyr Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 76
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 76
Lys Asn Tyr Val Asp Ser Leu Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 77
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 77
Lys Tyr Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 78
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 78
Thr Asn Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 79
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 79
Lys Asn Tyr Ile Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 80
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 80
Lys Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Asn Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 81
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 81
Asn Asp Tyr Ala Val Ser Val Lys Ser Arg Ile Thr Ile Thr Pro Asp
1 5 10 15
Thr Ser Lys Asn Gln Phe Ser Leu Arg Leu Asn Pro Val Thr Pro Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 82
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 82
Thr Tyr Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 83
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 83
Thr His Tyr Asn Pro Ser Leu Lys Ser Arg Val Ile Ile Ser Leu Asp
1 5 10 15
Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Arg Ser Val Thr Ala Ala
20 25 30
Asp Ser Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 84
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 84
Lys Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Val Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 85
<211> 41
<212> PRT
<213> 人工序列(artificial sequence)
<400> 85
Lys Asn Tyr Val Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
1 5 10 15
Asn Ser Lys Asn Thr Leu Asn Leu Gln Met Asn Arg Leu Arg Ala Glu
20 25 30
Asp Thr Ala Val Tyr Tyr Cys Ala Arg
35 40
<210> 86
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 86
Asp Asn Thr Lys Ser Trp Asp His Phe Asp His
1 5 10
<210> 87
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 87
Gly Ser Gly Val Ile Ser Tyr Phe Asp Tyr
1 5 10
<210> 88
<211> 12
<212> PRT
<213> 人工序列(artificial sequence)
<400> 88
Asp Gly Trp Gly Ser Tyr Gly Glu Tyr Phe Gln His
1 5 10
<210> 89
<211> 12
<212> PRT
<213> 人工序列(artificial sequence)
<400> 89
Asp Gly Trp Gly Thr Tyr Gly Glu Tyr Phe Gln His
1 5 10
<210> 90
<211> 12
<212> PRT
<213> 人工序列(artificial sequence)
<400> 90
Gly Gly Tyr Asn Ser Gly Trp Trp Ala Phe Asp Ile
1 5 10
<210> 91
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 91
Asp Gly Ala Ala Ala Pro Phe Asp Tyr
1 5
<210> 92
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 92
Leu Asp Phe Ser Gly Trp Pro Leu Ile Asp Tyr
1 5 10
<210> 93
<211> 12
<212> PRT
<213> 人工序列(artificial sequence)
<400> 93
Gly Gly Tyr Asn Ser Gly Trp Trp Ala Phe Asp Val
1 5 10
<210> 94
<211> 14
<212> PRT
<213> 人工序列(artificial sequence)
<400> 94
Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
1 5 10
<210> 95
<211> 14
<212> PRT
<213> 人工序列(artificial sequence)
<400> 95
Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
1 5 10
<210> 96
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 96
Asp Asn Trp Gly Phe Asp Tyr
1 5
<210> 97
<211> 14
<212> PRT
<213> 人工序列(artificial sequence)
<400> 97
Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Met Asp Val
1 5 10
<210> 98
<211> 13
<212> PRT
<213> 人工序列(artificial sequence)
<400> 98
Asp His Pro His Tyr Leu Gly Ser Gly Ser Phe Asp Tyr
1 5 10
<210> 99
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 99
Gly Asp Phe Arg Phe Asp Ser
1 5
<210> 100
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 100
Glu Gly Tyr Ala Ser Gly Ser Tyr Tyr
1 5
<210> 101
<211> 12
<212> PRT
<213> 人工序列(artificial sequence)
<400> 101
Glu Thr Ser Ser Trp Tyr Val Gly Tyr Leu Gln His
1 5 10
<210> 102
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 102
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
1 5 10
<210> 103
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 103
Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser
1 5 10
<210> 104
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 104
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
1 5 10
<210> 105
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 105
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys
20
<210> 106
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 106
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys
20
<210> 107
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 107
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys
20
<210> 108
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 108
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Glu Arg Val Thr Ile Thr Cys
20
<210> 109
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 109
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
20
<210> 110
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 110
Gln Val Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys
20
<210> 111
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 111
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys
20
<210> 112
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 112
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys
20
<210> 113
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 113
Glu Ile Val Leu Thr Gln Phe Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys
20
<210> 114
<211> 23
<212> PRT
<213> 人工序列(artificial sequence)
<400> 114
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Leu Ser Cys
20
<210> 115
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 115
Arg Ala Ser Gln Ser Val Gly Ser Tyr Leu Ala
1 5 10
<210> 116
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 116
Arg Ala Ser Gln Ser Ile Gly Asn Trp Leu Ala
1 5 10
<210> 117
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 117
Arg Ala Thr Gln Gly Ile Ser Asn Tyr Leu Ala
1 5 10
<210> 118
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 118
Arg Ala Ser Gln Asn Ile Asn Ser Tyr Leu Ser
1 5 10
<210> 119
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 119
Arg Ala Ser Gln Asn Ile Asn Ser Tyr Leu His
1 5 10
<210> 120
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 120
Arg Ala Ser Gln Gly Ile Ser His Phe Leu Ala
1 5 10
<210> 121
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 121
Arg Ala Ser Gln Ser Ile Ser Asn Trp Leu Ala
1 5 10
<210> 122
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 122
Arg Ala Ser Gln Ser Val Ser Ser Lys Leu Ala
1 5 10
<210> 123
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 123
Arg Ala Ser Gln Ser Ile Gly His Trp Leu Ala
1 5 10
<210> 124
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 124
Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala
1 5 10
<210> 125
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 125
Arg Ala Ser Gln Ser Ile Asn Ser Tyr Leu His
1 5 10
<210> 126
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 126
Arg Ala Ser Gln Gly Ile Asn Ser Tyr Leu Gly
1 5 10
<210> 127
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 127
Arg Ala Ser Gln Gly Ile Ser Ser Tyr Leu Gly
1 5 10
<210> 128
<211> 12
<212> PRT
<213> 人工序列(artificial sequence)
<400> 128
Arg Ser Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala
1 5 10
<210> 129
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 129
Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala
1 5 10
<210> 130
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 130
Arg Ala Ser Gln Gly Ile Ser Ser Tyr Leu Ala
1 5 10
<210> 131
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 131
Arg Ser Ser Gln Ser Val Asn Asn Tyr Leu Ala
1 5 10
<210> 132
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 132
Arg Ala Ser Gln Ser Val Asn Gly Asn Val Ala
1 5 10
<210> 133
<211> 11
<212> PRT
<213> 人工序列(artificial sequence)
<400> 133
Arg Ser Ser Gln Ser Val Asn Ser Tyr Leu Ala
1 5 10
<210> 134
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 134
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ser Leu Leu Ile Tyr
1 5 10 15
<210> 135
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 135
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr
1 5 10 15
<210> 136
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 136
Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Asn Leu Leu Ile Tyr
1 5 10 15
<210> 137
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 137
Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile Tyr
1 5 10 15
<210> 138
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 138
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile Tyr
1 5 10 15
<210> 139
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 139
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Ile Ile Tyr
1 5 10 15
<210> 140
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 140
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr
1 5 10 15
<210> 141
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 141
Trp Tyr Gln Arg Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr
1 5 10 15
<210> 142
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 142
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Ser
1 5 10 15
<210> 143
<211> 15
<212> PRT
<213> 人工序列(artificial sequence)
<400> 143
Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr
1 5 10 15
<210> 144
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 144
Asp Ala Ser Asn Arg Ala Thr
1 5
<210> 145
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 145
Lys Ala Ser Ser Leu Glu Ser
1 5
<210> 146
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 146
Ala Ala Ser Thr Leu Gln Ser
1 5
<210> 147
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 147
Ala Ala Ser Ser Leu Gln Asn
1 5
<210> 148
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 148
Ala Ala Ser Ser Leu Gln Ser
1 5
<210> 149
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 149
Gly Ala Ser Thr Arg Ala Thr
1 5
<210> 150
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 150
Ile Ala Ser Thr Leu Gln Ser
1 5
<210> 151
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 151
Val Ala Ser Thr Leu Gln Ser
1 5
<210> 152
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 152
Gly Ala Ser Ser Arg Ala Thr
1 5
<210> 153
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 153
Asp Ser Ser Thr Arg Ala Thr
1 5
<210> 154
<211> 7
<212> PRT
<213> 人工序列(artificial sequence)
<400> 154
Ala Ala Ser Thr Arg Ala Thr
1 5
<210> 155
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 155
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
20 25 30
<210> 156
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 156
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys
20 25 30
<210> 157
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 157
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Leu Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys
20 25 30
<210> 158
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 158
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys
20 25 30
<210> 159
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 159
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Ile Tyr Tyr Cys
20 25 30
<210> 160
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 160
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys
20 25 30
<210> 161
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 161
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Asn Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys
20 25 30
<210> 162
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 162
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys
20 25 30
<210> 163
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 163
Gly Val Pro Ser Arg Phe Arg Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys
20 25 30
<210> 164
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 164
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys
20 25 30
<210> 165
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 165
Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
20 25 30
<210> 166
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 166
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys
20 25 30
<210> 167
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 167
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Phe Ala Phe Tyr Phe Cys
20 25 30
<210> 168
<211> 32
<212> PRT
<213> 人工序列(artificial sequence)
<400> 168
Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Ser Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ser Glu Asp Cys Ala Val Tyr Tyr Cys
20 25 30
<210> 169
<211> 8
<212> PRT
<213> 人工序列(artificial sequence)
<400> 169
Gln Gln Arg Thr Asn Trp Tyr Thr
1 5
<210> 170
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 170
Gln Gln Tyr Asp Ser Tyr Ser Pro Ile Thr
1 5 10
<210> 171
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 171
Gln Lys Cys Asn Ser Ala Pro Arg Thr
1 5
<210> 172
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 172
Gln Gln Ser Phe Ser Thr Pro Pro Ile Thr
1 5 10
<210> 173
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 173
Gln Gln Ser Tyr Ser Ile Pro Pro Ile Thr
1 5 10
<210> 174
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 174
Gln Lys Phe Asn Ser Ala Pro Leu Thr
1 5
<210> 175
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 175
Gln Gln Tyr Asp Ser Tyr Ser Ile Thr
1 5
<210> 176
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 176
Gln Gln Tyr Asp Asn Trp Pro Pro Ile Thr
1 5 10
<210> 177
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 177
Gln Gln Tyr Ser Ser Tyr Ser Pro Ile Thr
1 5 10
<210> 178
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 178
Gln Gln Tyr Asn Asn Trp Pro Pro Ile Thr
1 5 10
<210> 179
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 179
Gln Gln Ser Tyr Ser Thr Pro Pro Ile Thr
1 5 10
<210> 180
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 180
Gln Gln Leu Asn Ser Tyr Pro Tyr Thr
1 5
<210> 181
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 181
Gln Gln Tyr Gly Ser Ser Pro Arg Thr
1 5
<210> 182
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 182
Gln Gln Tyr Tyr Ser Thr Pro Tyr Thr
1 5
<210> 183
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 183
Gln Gln Val Lys Ile Tyr Pro Leu Thr
1 5
<210> 184
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 184
Gln Gln Tyr Glu Asn Trp Pro Leu Thr
1 5
<210> 185
<211> 9
<212> PRT
<213> 人工序列(artificial sequence)
<400> 185
Gln Gln Tyr Asn Asn Trp Pro Ser Thr
1 5
<210> 186
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 186
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
1 5 10
<210> 187
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 187
Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
1 5 10
<210> 188
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 188
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
1 5 10
<210> 189
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 189
Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
1 5 10
<210> 190
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 190
Phe Gly Gln Gly Ser Arg Leu Glu Ile Lys
1 5 10
<210> 191
<211> 10
<212> PRT
<213> 人工序列(artificial sequence)
<400> 191
Phe Gly Pro Gly Thr Lys Leu Glu Val Thr
1 5 10
<210> 192
<211> 120
<212> PRT
<213> 人工序列(artificial sequence)
<400> 192
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Phe His Ser Gly Thr Thr Tyr Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Leu Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Asn Thr Lys Ser Trp Asp His Phe Asp His Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 193
<211> 119
<212> PRT
<213> 人工序列(artificial sequence)
<400> 193
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Ile Ser Ser His
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr His Ser Gly Ile Thr Ser Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Gly Ser Gly Val Ile Ser Tyr Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 194
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 194
Gln Val Glu Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Gly Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asn Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Trp Asp Asp Gly Asn Asn Lys Tyr Tyr Ala Asn 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 Thr Glu Asp Thr Ala Leu Tyr Tyr Cys
85 90 95
Ala Lys Asp Gly Trp Gly Ser Tyr Gly Glu Tyr Phe Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 195
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 195
Gln Val Asn Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Lys 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 Leu Ile Trp Ser Asp Gly Thr Asn Arg Tyr Tyr Thr Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Ser
65 70 75 80
Leu Gln Met Lys Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Asp Gly Trp Gly Thr Tyr Gly Glu Tyr Phe Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 196
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 196
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Val Ser Cys Ile Ala Ser Gly Phe Phe 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 Glu Asp Tyr Val 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 Gly Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Tyr Asn Ser Gly Trp Trp Ala Phe Asp Ile Trp Gly
100 105 110
Gln Gly Thr Met Val Thr Val Ser Ser
115 120
<210> 197
<211> 118
<212> PRT
<213> 人工序列(artificial sequence)
<400> 197
Gln Val Gln Leu Gln Glu Ser Gly Pro Arg Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly
20 25 30
Tyr Tyr Trp Gly Trp Ile Arg Gln Ser Pro Gly Met Gly Leu Glu Trp
35 40 45
Ile Ala Tyr Ile Tyr His Asp Gly Ser Thr Tyr Cys Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Ala Ala Ala Pro Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 198
<211> 120
<212> PRT
<213> 人工序列(artificial sequence)
<400> 198
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 Phe
20 25 30
Gly Met Gln Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Met Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Val 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 Leu Asp Phe Ser Gly Trp Pro Leu Ile Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 199
<211> 119
<212> PRT
<213> 人工序列(artificial sequence)
<400> 199
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Ile Ser Ser Asn
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr Pro Gly Gly Asn Pro Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Ser Gly Val Ile Ser Tyr Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 200
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 200
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Val Ser Cys Ala Ala Ser Gly Phe Ile 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 Glu Asp Tyr Val 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 Gly Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Tyr Asn Ser Gly Trp Trp Ala Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 201
<211> 120
<212> PRT
<213> 人工序列(artificial sequence)
<400> 201
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 Phe
20 25 30
Gly Met Gln Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Ile Met Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Val 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 Leu Asp Phe Ser Gly Trp Pro Leu Ile Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 202
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 202
Gln Val Asn Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Lys 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 Phe Ile Trp Ser Asp Gly Thr Asn Arg Tyr Tyr Pro Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Arg Asn Thr Val Ser
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Trp Gly Thr Tyr Gly Glu Tyr Phe Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 203
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 203
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Glu Lys
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Tyr
20 25 30
Gly Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Trp Tyr Asp Gly Ile Lys Arg Tyr Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 204
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 204
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 His 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 Ile Lys Lys Asp Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 205
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 205
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Ser Tyr Tyr Val Glu 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 Asp Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 206
<211> 116
<212> PRT
<213> 人工序列(artificial sequence)
<400> 206
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr His Asp Gly Ser Thr Asp Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ile Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Val Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Asn Trp Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 207
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 207
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Lys Tyr Tyr Ile Glu Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 208
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 208
Gln Val Gln Leu Val Gln 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 His 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 Ile Lys Lys Tyr Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 209
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 209
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 Glu Gly Ser Gly Phe Thr Phe Ser His 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 Ile Lys Lys Asn Tyr Val Asp Ser Leu
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 Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Met Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 210
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 210
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 Val Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Lys Tyr Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 211
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 211
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Thr Asn Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 212
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 212
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Met Lys Asn Tyr Ile Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 213
<211> 122
<212> PRT
<213> 人工序列(artificial sequence)
<400> 213
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Thr
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asn Asp
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Met Gly Leu Glu Trp Val
35 40 45
Ala Ile Met Trp Phe Asp Gly Gly Ser 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 Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp His Pro His Tyr Leu Gly Ser Gly Ser Phe Asp Tyr Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 214
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 214
Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser His 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 Val Trp Tyr Asp Gly Ile Lys Lys Tyr Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 215
<211> 119
<212> PRT
<213> 人工序列(artificial sequence)
<400> 215
Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Thr
20 25 30
Ser Ala Val Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu
35 40 45
Trp Leu Gly Arg Thr Cys Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala
50 55 60
Val Ser Val Lys Ser Arg Ile Thr Ile Thr Pro Asp Thr Ser Lys Asn
65 70 75 80
Gln Phe Ser Leu Arg Leu Asn Pro Val Thr Pro Glu Asp Thr Ala Val
85 90 95
Tyr Tyr Cys Ala Arg Gly Asp Phe Arg Phe Asp Ser Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 216
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 216
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Thr Tyr Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 217
<211> 118
<212> PRT
<213> 人工序列(artificial sequence)
<400> 217
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 Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly
20 25 30
Tyr Phe Trp Ala Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Ser Ile Tyr His Ser Gly Arg Thr His Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ile Ile Ser Leu Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Arg Ser Val Thr Ala Ala Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Gly Tyr Ala Ser Gly Ser Tyr Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 218
<211> 121
<212> PRT
<213> 人工序列(artificial sequence)
<400> 218
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 Thr Ser Gly Phe Ile Phe Ser Ser Tyr
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Asp 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 Val Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Thr Ser Ser Trp Tyr Val Gly Tyr Leu Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 219
<211> 118
<212> PRT
<213> 人工序列(artificial sequence)
<400> 219
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 Ala Val Ser Gly Tyr Ser Ile Asn Ser Gly
20 25 30
Tyr Phe Trp Ala Trp Ile Arg Gln Pro Pro Gly Met Gly Leu Glu Trp
35 40 45
Ile Gly Ser Ile Tyr His Ser Gly Arg Thr His Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ile Ile Ser Leu Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Arg Ser Val Thr Ala Ala Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Gly Tyr Ala Ser Gly Ser Tyr Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 220
<211> 123
<212> PRT
<213> 人工序列(artificial sequence)
<400> 220
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 His 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 Ile Lys Lys Asn Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 221
<211> 106
<212> PRT
<213> 人工序列(artificial sequence)
<400> 221
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ser Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Thr Asn Trp Tyr Thr
85 90 95
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 222
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 222
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly Asn Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Ser Tyr Ser Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 223
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 223
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Thr Gln Gly Ile Ser Asn Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Asn 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 Leu Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Lys Cys Asn Ser Ala Pro Arg
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 224
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 224
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Glu Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Ser Tyr
20 25 30
Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Ser Leu Gln Asn 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 Phe Ser Thr Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 225
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 225
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Ser Tyr
20 25 30
Leu His 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 Ile Tyr Tyr Cys Gln Gln Ser Tyr Ser Ile Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 226
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 226
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 His Phe
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 Phe Asn Ser Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 227
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 227
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Asn Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile
35 40 45
Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro
65 70 75 80
Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Ser Tyr Ser Ile
85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 228
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 228
Gln Val Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Lys
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Ile Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Asn Trp Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 229
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 229
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly His Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Arg Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Ser Pro
85 90 95
Ile Thr Phe Gly Gln Gly Ser Arg Leu Glu Ile Lys
100 105
<210> 230
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 230
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 231
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 231
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 Asn Ser Tyr
20 25 30
Leu His 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 Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105
<210> 232
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 232
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asn Ser Tyr
20 25 30
Leu Gly Trp Tyr Gln Arg Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ile Ala Ser Thr 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 Gln Gln Leu Asn Ser Tyr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 233
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 233
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Val Ala Ser Thr 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 Gln Gln Leu Asn Ser Tyr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 234
<211> 108
<212> PRT
<213> 人工序列(artificial sequence)
<400> 234
Glu Ile Val Leu Thr Gln Phe Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ser 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 Ser 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
Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 235
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 235
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ile Ala Ser Thr 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 Gln Gln Leu Asn Ser Tyr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 236
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 236
Glu Ile Val Leu Thr Gln Ser Pro Ala 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 Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 237
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 237
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr
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 Thr 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 Gln Gln Val Lys Ile Tyr Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 238
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 238
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ser Ser Gln Ser Val Asn Asn Tyr
20 25 30
Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ser Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Phe Tyr Phe Cys Gln Gln Tyr Glu Asn Trp Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 239
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 239
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Leu Ser Cys Arg Ala Ser Gln Ser Val Asn Gly Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Ser Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Cys Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Ser
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Val Thr
100 105
<210> 240
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 240
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ser Ser Gln Ser Val Asn Ser Tyr
20 25 30
Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ser Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Phe Tyr Phe Cys Gln Gln Tyr Glu Asn Trp Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 241
<211> 450
<212> PRT
<213> 人工序列(artificial sequence)
<400> 241
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Phe His Ser Gly Thr Thr Tyr Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Leu Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Asn Thr Lys Ser Trp Asp His Phe Asp His Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 242
<211> 449
<212> PRT
<213> 人工序列(artificial sequence)
<400> 242
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Ile Ser Ser His
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr His Ser Gly Ile Thr Ser Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Gly Ser Gly Val Ile Ser Tyr Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 243
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 243
Gln Val Glu Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Gly Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asn Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Gly Ile Trp Asp Asp Gly Asn Asn Lys Tyr Tyr Ala Asn 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 Thr Glu Asp Thr Ala Leu Tyr Tyr Cys
85 90 95
Ala Lys Asp Gly Trp Gly Ser Tyr Gly Glu Tyr Phe Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 244
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 244
Gln Val Asn Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Lys 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 Leu Ile Trp Ser Asp Gly Thr Asn Arg Tyr Tyr Thr Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Ser
65 70 75 80
Leu Gln Met Lys Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Asp Gly Trp Gly Thr Tyr Gly Glu Tyr Phe Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 245
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 245
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Val Ser Cys Ile Ala Ser Gly Phe Phe 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 Glu Asp Tyr Val 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 Gly Glu Asp Thr Ala Met Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Tyr Asn Ser Gly Trp Trp Ala Phe Asp Ile Trp Gly
100 105 110
Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 246
<211> 448
<212> PRT
<213> 人工序列(artificial sequence)
<400> 246
Gln Val Gln Leu Gln Glu Ser Gly Pro Arg Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly
20 25 30
Tyr Tyr Trp Gly Trp Ile Arg Gln Ser Pro Gly Met Gly Leu Glu Trp
35 40 45
Ile Ala Tyr Ile Tyr His Asp Gly Ser Thr Tyr Cys Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Ala Ala Ala Pro Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 247
<211> 450
<212> PRT
<213> 人工序列(artificial sequence)
<400> 247
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 Phe
20 25 30
Gly Met Gln Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Met Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Val 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 Leu Asp Phe Ser Gly Trp Pro Leu Ile Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 248
<211> 449
<212> PRT
<213> 人工序列(artificial sequence)
<400> 248
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Asp Ser Ile Ser Ser Asn
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr Pro Gly Gly Asn Pro Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Ser Gly Val Ile Ser Tyr Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 249
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 249
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Val Ser Cys Ala Ala Ser Gly Phe Ile 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 Glu Asp Tyr Val 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 Gly Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Tyr Asn Ser Gly Trp Trp Ala Phe Asp Val Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 250
<211> 450
<212> PRT
<213> 人工序列(artificial sequence)
<400> 250
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 Phe
20 25 30
Gly Met Gln Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Ile Met Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Val 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 Leu Asp Phe Ser Gly Trp Pro Leu Ile Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 251
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 251
Gln Val Asn Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Lys 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 Phe Ile Trp Ser Asp Gly Thr Asn Arg Tyr Tyr Pro Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Arg Asn Thr Val Ser
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Trp Gly Thr Tyr Gly Glu Tyr Phe Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 252
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 252
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Glu Lys
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Tyr
20 25 30
Gly Met His Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Trp Tyr Asp Gly Ile Lys Arg Tyr Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 253
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 253
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 His 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 Ile Lys Lys Asp Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 254
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 254
Gln Val Gln Leu Val Glu Ser Gly Gly Ala Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Ser Tyr Tyr Val Glu 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 Asp Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 255
<211> 446
<212> PRT
<213> 人工序列(artificial sequence)
<400> 255
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr His Asp Gly Ser Thr Asp Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ile Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Val Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Asn Trp Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 256
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 256
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Lys Tyr Tyr Ile Glu Ser Val
50 55 60
Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 257
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 257
Gln Val Gln Leu Val Gln 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 His 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 Ile Lys Lys Tyr Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 258
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 258
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 Glu Gly Ser Gly Phe Thr Phe Ser His 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 Ile Lys Lys Asn Tyr Val Asp Ser Leu
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 Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Met Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 259
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 259
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 Val Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Lys Tyr Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 260
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 260
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Thr Asn Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 261
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 261
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Met Lys Asn Tyr Ile Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 262
<211> 452
<212> PRT
<213> 人工序列(artificial sequence)
<400> 262
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Thr
1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asn Asp
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Met Gly Leu Glu Trp Val
35 40 45
Ala Ile Met Trp Phe Asp Gly Gly Ser 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 Asn Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp His Pro His Tyr Leu Gly Ser Gly Ser Phe Asp Tyr Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
225 230 235 240
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
245 250 255
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
260 265 270
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
275 280 285
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
290 295 300
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
305 310 315 320
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
325 330 335
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
340 345 350
Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val
355 360 365
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
370 375 380
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
385 390 395 400
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
405 410 415
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
420 425 430
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
435 440 445
Ser Pro Gly Lys
450
<210> 263
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 263
Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser His 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 Val Trp Tyr Asp Gly Ile Lys Lys Tyr Tyr Val Glu 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 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Leu Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 264
<211> 449
<212> PRT
<213> 人工序列(artificial sequence)
<400> 264
Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Thr
20 25 30
Ser Ala Val Trp Asn Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu
35 40 45
Trp Leu Gly Arg Thr Cys Tyr Arg Ser Lys Trp Tyr Asn Asp Tyr Ala
50 55 60
Val Ser Val Lys Ser Arg Ile Thr Ile Thr Pro Asp Thr Ser Lys Asn
65 70 75 80
Gln Phe Ser Leu Arg Leu Asn Pro Val Thr Pro Glu Asp Thr Ala Val
85 90 95
Tyr Tyr Cys Ala Arg Gly Asp Phe Arg Phe Asp Ser Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 265
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 265
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 Glu Ala Ser Gly Phe Thr Phe Ser His 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 Ile Lys Thr Tyr Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 266
<211> 448
<212> PRT
<213> 人工序列(artificial sequence)
<400> 266
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 Ala Val Ser Gly Tyr Ser Ile Ser Ser Gly
20 25 30
Tyr Phe Trp Ala Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Ser Ile Tyr His Ser Gly Arg Thr His Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ile Ile Ser Leu Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Arg Ser Val Thr Ala Ala Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Gly Tyr Ala Ser Gly Ser Tyr Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 267
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 267
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 Thr Ser Gly Phe Ile Phe Ser Ser Tyr
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Asp 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 Val Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Thr Ser Ser Trp Tyr Val Gly Tyr Leu Gln His Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 268
<211> 448
<212> PRT
<213> 人工序列(artificial sequence)
<400> 268
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 Ala Val Ser Gly Tyr Ser Ile Asn Ser Gly
20 25 30
Tyr Phe Trp Ala Trp Ile Arg Gln Pro Pro Gly Met Gly Leu Glu Trp
35 40 45
Ile Gly Ser Ile Tyr His Ser Gly Arg Thr His Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ile Ile Ser Leu Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Arg Ser Val Thr Ala Ala Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Gly Tyr Ala Ser Gly Ser Tyr Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 269
<211> 453
<212> PRT
<213> 人工序列(artificial sequence)
<400> 269
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 His 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 Ile Lys Lys Asn Tyr Val Glu Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Asn
65 70 75 80
Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gly Gly Phe Gly Glu Ser Asn Tyr Tyr Gly Leu Asp Val
100 105 110
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
145 150 155 160
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val
195 200 205
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys
210 215 220
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
225 230 235 240
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
260 265 270
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
275 280 285
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
305 310 315 320
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
340 345 350
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
355 360 365
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
385 390 395 400
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
420 425 430
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
435 440 445
Leu Ser Pro Gly Lys
450
<210> 270
<211> 213
<212> PRT
<213> 人工序列(artificial sequence)
<400> 270
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ser Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Thr Asn Trp Tyr Thr
85 90 95
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro
100 105 110
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
115 120 125
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
130 135 140
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
145 150 155 160
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
165 170 175
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
180 185 190
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
195 200 205
Asn Arg Gly Glu Cys
210
<210> 271
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 271
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly Asn Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Ser Tyr Ser Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 272
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 272
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Thr Gln Gly Ile Ser Asn Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Asn 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 Leu Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Lys Cys Asn Ser Ala Pro Arg
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 273
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 273
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Glu Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Ser Tyr
20 25 30
Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Ser Leu Gln Asn 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 Phe Ser Thr Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 274
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 274
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Ser Tyr
20 25 30
Leu His 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 Ile Tyr Tyr Cys Gln Gln Ser Tyr Ser Ile Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 275
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 275
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 His Phe
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 Phe Asn Ser Ala Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 276
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 276
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Asn Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile
35 40 45
Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro
65 70 75 80
Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Ser Tyr Ser Ile
85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 277
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 277
Gln Val Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Lys
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Ile Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp Asn Trp Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 278
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 278
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly His Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Arg Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Ser Pro
85 90 95
Ile Thr Phe Gly Gln Gly Ser Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 279
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 279
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 280
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 280
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 Asn Ser Tyr
20 25 30
Leu His 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 Pro
85 90 95
Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 281
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 281
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asn Ser Tyr
20 25 30
Leu Gly Trp Tyr Gln Arg Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ile Ala Ser Thr 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 Gln Gln Leu Asn Ser Tyr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 282
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 282
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Val Ala Ser Thr 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 Gln Gln Leu Asn Ser Tyr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 283
<211> 215
<212> PRT
<213> 人工序列(artificial sequence)
<400> 283
Glu Ile Val Leu Thr Gln Phe Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ser 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 Ser 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
Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 284
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 284
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ile Ala Ser Thr 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 Gln Gln Leu Asn Ser Tyr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 285
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 285
Glu Ile Val Leu Thr Gln Ser Pro Ala 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 Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 286
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 286
Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr
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 Thr 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 Gln Gln Val Lys Ile Tyr Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 287
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 287
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ser Ser Gln Ser Val Asn Asn Tyr
20 25 30
Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ser Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Phe Tyr Phe Cys Gln Gln Tyr Glu Asn Trp Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 288
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 288
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Leu Ser Cys Arg Ala Ser Gln Ser Val Asn Gly Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Ser Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Cys Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Ser
85 90 95
Thr Phe Gly Pro Gly Thr Lys Leu Glu Val Thr Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 289
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 289
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ser Ser Gln Ser Val Asn Ser Tyr
20 25 30
Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ser Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Phe Tyr Phe Cys Gln Gln Tyr Glu Asn Trp Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 290
<211> 451
<212> PRT
<213> 人工序列(artificial sequence)
<400> 290
Gln Val Gln Leu Gln Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
20 25 30
Gly Met Asn Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Lys Trp Met
35 40 45
Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Thr Asp Asp Phe
50 55 60
Lys Gly Arg Phe Ala Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr
65 70 75 80
Leu Gln Ile Ser Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Gly Phe Gly Ser Ser Tyr Trp Tyr Phe Asp Val Trp Gly
100 105 110
Gln Gly Ser Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 291
<211> 214
<212> PRT
<213> 人工序列(artificial sequence)
<400> 291
Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Ile Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp 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 Val Tyr Tyr Cys Gln Gln His Tyr Ile Thr Pro Leu
85 90 95
Thr Phe Gly Ala Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 292
<211> 330
<212> PRT
<213> 人工序列(artificial sequence)
<400> 292
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
225 230 235 240
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 293
<211> 107
<212> PRT
<213> 人工序列(artificial sequence)
<400> 293
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
1 5 10 15
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
20 25 30
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
35 40 45
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
50 55 60
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
65 70 75 80
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
85 90 95
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105

Claims (20)

1.一种Trop-2的抗体或抗原结合片段，所述抗体或其抗原结合片段包含如下的互补决定区(CDR)：
(a)SEQ ID NO:192-220中任一所示的重链可变区(VH)中含有的HCDR1或其序列的变体、HCDR2或其序列的变体、以及HCDR3或其序列的变体；
和/或
(b)SEQ ID NO:221-240中任一所示的轻链可变区(VL)中含有的LCDR1或其序列的变体、LCDR2或其序列的变体、以及LCDR3或其序列的变体；
优选地，所述序列的变体为与其来源CDR相比为具有一个或几个氨基酸的置换、缺失或添加的CDR；优选地，所述的置换为保守置换。
2.如权利要求1所述的抗体或抗原结合蛋白，其特征在于，所述的抗体或抗原结合蛋白是具有表A所示的各重链VH-CDR1、VH-CDR2和VH-CDR3，以及轻链VL-CDR1、VL-CDR2和VL-CDR3的组合的抗体之一：
表A
抗体编号 抗体名称 LCDR1 LCDR2 LCDR3 HCDR1 HCDR2 HCDR3 (1) PR001128 115 144 169 19 47 86 (2) PR001130 116 145 170 20 48 87 (3) PR001131 117 146 171 20 48 87 (4) PR001132 118 147 172 21 49 88 (5) PR001133 119 148 173 21 50 89 (6) PR001134 120 146 174 22 51 90 (7) PR001138 121 145 175 23 52 91 (8) PR001139 122 149 176 24 53 92 (9) PR001142 123 145 177 25 54 87 (10) PR001143 120 146 174 26 51 93 (11) PR001145 124 149 178 24 53 92 (12) PR001147 125 148 179 21 50 89 (13) PR001150 126 150 180 27 55 94 (14) PR001151 127 151 180 27 55 95 (15) PR001152 126 150 180 27 55 94 (16) PR001153 128 152 181 19 52 96 (17) PR001154 127 151 180 27 55 95 (18) PR001155 127 151 180 27 55 95 (19) PR001156 127 150 180 27 55 97 (20) PR001158 127 150 180 27 55 94 (21) PR001159 127 150 180 27 55 94 (22) PR001160 127 151 180 27 56 95 (23) PR001162 129 144 182 28 57 98 (24) PR001163 127 151 180 27 55 95 (25) PR001164 130 146 183 29 58 99 (26) PR001165 127 150 180 27 55 94 (27) PR001166 131 153 184 23 59 100 (28) PR001168 132 154 185 30 51 101 (29) PR001170 133 153 184 31 59 100 (30) PR001171 127 150 180 27 55 94
其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留TROP2结合亲和力的衍生序列。
3.根据权利要求1或2所述的抗体或抗原结合片段，其特征在于，所述抗体或其抗原结合片段选自scFv、Fab、Fab’、(Fab’)2、Fv片段、二硫键连接的Fv(dsFv)和双抗体(diabody)。
4.一种重组蛋白，其特征在于，所述的重组蛋白包括：
(i)如权利要求1所述的抗体或抗原结合蛋白；和
(ii)任选的协助表达和/或纯化的标签序列。
5.一种多核苷酸，其特征在于，所述多核苷酸编码选自下组的多肽：如权利要求1-3任一项所述的抗体或抗原结合蛋白，和/或如权利要求4所述的重组蛋白。
6.一种载体，其特征在于，所述载体含有权利要求5所述的多核苷酸。
7.一种遗传工程化的宿主细胞，其特征在于，所述宿主细胞含有权利要求6所述的载体或基因组中整合有权利要求5所述的多核苷酸。
8.制备权利要求1-3中任一项所述的抗体或其抗原结合片段的方法，其包括，在允许所述抗体或其抗原结合片段表达的条件下，培养权利要求7所述的宿主细胞，和从培养的宿主细胞培养物中回收所述抗体或其抗原结合片段。
9.一种抗体偶联物，其特征在于，所述抗体偶联物含有：
(a)抗体部分，所述抗体部分包括如权利要求1-3所述的抗体或抗原结合蛋白；和
(b)与所述抗体部分偶联的偶联部分，所述偶联部分选自下组：可检测标记物、药物、毒素、细胞因子、放射性核素、酶、或其组合；
所述抗体部分和偶联部分通过化学键或接头进行偶联。
10.一种嵌合抗原受体(CAR)，其特征在于，所述CAR包括权利要求1-3所述的抗体或抗原结合蛋白。
11.一种免疫细胞，其特征在于，所述免疫细胞表达权利要求10所述的嵌合抗原受体(CAR)。
12.一种多特异性抗体，其特征在于，包含权利要求1-3任一项所述的Trop-2的抗体或抗原结合片段，和另外的抗体或其片段或抗体类似物；优选地，所述多特异性为双特异性、三特异性或四特异性。
13.一种药物组合物，其特征在于，所述药物组合物含有：
(i)活性成分，所述活性成分选自下组：如权利要求1-3所述的抗体或抗原结合蛋白、如权利要求4所述的重组蛋白、如权利要求9所述的抗体偶联物、权利要求11所述的免疫细胞、权利要求12所述的多特异性抗体或其组合；以及
(ii)药学上可接受的载体。
14.一种治疗、预防或减轻肿瘤的方法，其特征在于，所述方法包括向有需要的受试者施用权利要求1-3所述的抗体或抗原结合蛋白，权利要求4所述的重组蛋白，权利要求9所述的抗体偶联物，权利要求11所述的免疫细胞，权利要求12所述的多特异性抗体及权利要求13所述的药物组合物，优选地所述肿瘤为乳癌、胃癌、胰腺癌、卵巢癌、肠癌、肺癌、子宫颈癌，或其他Trop2表达阳性的肿瘤。
15.如权利要求14所述的方法，其特征在于，进一步包括第二治疗剂的施用。
16.如权利要求1-3所述的抗体或抗原结合蛋白、如权利要求4所述的重组蛋白、如权利要求9所述的抗体偶联物、如权利要求11所述的免疫细胞、如权利要求12所述的多特异性抗体和/或如权利要求13所述的药物组合物在制备预防和/或治疗肿瘤的药物中的用途，其特征在于，所述肿瘤为乳癌、胃癌、胰腺癌、卵巢癌、肠癌、肺癌、子宫颈癌，或其他Trop2表达阳性的肿瘤。
17.一种检测Trop-2在样品中存在或其水平的方法，其特征在于，包括在允许权利要求1-3所述的抗体或抗原结合片段与Trop-2之间形成复合物的条件下，使所述样品与所述抗体或其抗原结合片段接触，并且检测所述复合物的形成；优选地，所述检测用于非诊断目的。
18.如权利要求1-3所述的抗体或抗原结合片段在制备Trop-2检测试剂或试剂盒中的用途。
19.一种试剂盒，其特征在于，包含权利要求1-3所述的抗体或抗原结合蛋白、权利要求4所述的重组蛋白、权利要求9所述的抗体偶联物、权利要求11所述的免疫细胞、权利要求12所述的多特异性抗体，和/或权利要求13所述的药物组合物，以及任选的使用说明书。
20.一种给药装置，其特征在于，包含：
(i)输注模块，所述输注模块用于对受试者施用包括具有一活性成分的药物组合物；
(ii)用于输注的药物组合物，所述药物组合物中含有一活性成分，所述活性成分选自下组：如本发明第一方面所述的抗体或抗原结合蛋白、如本发明第二方面所述的重组蛋白、如本发明第七方面所述的抗体偶联物、本发明第九方面所述的免疫细胞、或其组合；以及
(iii)任选的药效监控模块。
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