CN112010978A - TrkA的抗体及其应用 - Google Patents

TrkA的抗体及其应用 Download PDF

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CN112010978A
CN112010978A CN202010465851.9A CN202010465851A CN112010978A CN 112010978 A CN112010978 A CN 112010978A CN 202010465851 A CN202010465851 A CN 202010465851A CN 112010978 A CN112010978 A CN 112010978A
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CN112010978B (zh
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陈超
任志衡
何转娣
卢杰联
林树珊
于婷婷
危喜玲
王旭昉
徐乐
董军纪
李想
张阔
杨雪尧
郭林峰
李晓平
陈小锋
李文佳
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Guangdong HEC Pharmaceutical
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Sunshine Lake Pharma Co Ltd
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Abstract

本发明提出了一种能够特异性识别TrkA的抗体或其抗原结合片段及其用途。该抗体含有选自下列至少之一的CDR序列或与其具有至少95％同一性的氨基酸序列：重链可变区CDR序列：SEQ ID NO:1～27，轻链可变区CDR序列：SEQ IN NO:28～54。根据本发明实施例的上述抗体能够特异性的靶向结合TrkA受体，阻断NGF和TrkA结合。

Description

TrkA的抗体及其应用
技术领域
本发明涉及生物技术领域，具体地，本发明涉及TrkA的抗体及其应用，更具体地，本发明涉及能够特异性识别TrkA的抗体或其抗原结合片段、核酸分子、表达载体、重组细胞、药物组合物、制药用途、检测TrkA的试剂盒以及小鼠B细胞。
背景技术
目前临床上对于轻至中度疼痛，主要使用非阿片类镇痛药，如非甾体类抗炎药(NSAIDs)；对于中至重度疼痛，主要使用阿片类镇痛药。然而，NSAIDs存在“封顶效应”，阿片仅能使不到30％的非肿瘤性慢性疼痛得到有效缓解，20％的癌痛患者存在阿片类药物耐药。此外，NSAIDs存在胃肠道和心血管安全性隐患，长期用药过程中尤其明显。对于阿片类镇痛药，多年的药物改进实验未能有效的降低其成瘾性和其它诸多副作用，患者期许新的更安全有效的药物。
神经生长因子(NGF，nerve growth factor)参与疼痛的病理生理过程，主要通过结合高亲和力的酪氨酸激酶(tyrosine-nase，TrkA)受体激活NGF/TrKA信号通路，影响炎症介质的释放、离子通道的开放以及促进神经纤维的生长，从而参与疼痛的发生，传导以及增敏过程。研究表明，阻断NGF-TrkA信号通路可以有效的缓解疼痛和痛觉过敏，NGF-TrkA信号通路是开发新型镇痛药物的有效靶点。对于NGF-TrkA靶点镇痛药物，目前在研的药物主要分为两大类：大分子抗NGF单克隆抗体和小分子TrkA激酶抑制剂。由于TrKA受体并不是NGF的唯一受体，NGF还存在一种低亲和力的受体P75，而NGF与P75的结合对于神经系统的功能以及损伤修复是有重要作用的，大分子抗NGF单克隆抗体作为一种新型的镇痛药物存在很大的安全性隐患。从目前的临床数据来看，最常见毒副作用包括骨组织坏死(潜在)、感官异常等。而小分子TrkA激酶抑制剂存在着更多的问题。首先，小分子药物的半衰期一般较短，给药频繁，病人的依从性差。其次，小分子Trka激酶抑制剂是受体酪氨酸激酶抑制剂，机体内存在多种受体酪氨酸激酶，因此，药物对于特定靶点的特异性和选择性很难保证，药物的不良反应多，毒副作用大。
因此，针对NGF-TrkA靶点的镇痛药物，仍然需要科研工作者不断的开发和改进。
发明内容
本申请是基于发明人对下列问题和事实的发现而提出的：
NGF-TrkA信号通路作为开发新型镇痛药物的有效靶点，如果通过TrkA单克隆抗体选择性的靶向结合TrkA受体，不仅可以阻断NGF对TrkA信号通路的激活，有效的抑制疼痛信号的传递，又不会发生像抑制NGF受体所造成的诸多神经毒性，以及使用抗NGF抗体过度中和NGF所带来的骨关节坏死等不可预测的毒副作用。但由于TrkA分子是受体膜蛋白，筛选阻断型抗TrkA单克隆抗体的难度比较大，其次，设计阻断TrkA受体抗体存在由于抗体介导免疫反应导致的安全性风险，因此，设计开发针对TrkA的单克隆抗体难度比较大。
然而本申请的发明人却成功地筛选到了一种新型的具有长效镇痛作用的抗TrkA单克隆抗体，同时，发明人通过选取特定的Fc亚型并对其进行改造有效避免了免疫毒性的风险。具体地，本研究通过杂交瘤技术筛选出一系列阻断NGF和TrkA结合的高亲和力抗TrkA单克隆抗体，并将鼠源抗TrkA单克隆抗体的恒定区人源化，保留鼠源抗TrkA单克隆抗体的可变区，得到一系列的抗TrkA嵌合单克隆抗体。发明人发现，本申请所获得的这些嵌合抗体候选物，不仅能够特异性的靶向结合TrkA受体，阻断NGF和TrkA结合，有效的抑制疼痛，而且具有免疫毒性低，基本没有抗体依赖的细胞介导的细胞毒性作用(ADCC)的特点。
在本发明的的第一方面，本发明提出了一种能够特异性识别TrkA的抗体或其抗原结合片段。根据本发明的实施例，所述抗体含有选自下列至少之一的CDR序列或与其具有至少95％同一性的氨基酸序列：重链可变区CDR序列：SEQ ID NO:1～27，轻链可变区CDR序列：SEQ IN NO:28～54。
VYSFTAYT(SEQ ID NO:1)。
INPHNGGT(SEQ ID NO:2)。
AISRYGSSSFYFDV(SEQ ID NO:3)。
GYAFTNYW(SEQ ID NO:4)。
FYPRTGNT(SEQ ID NO:5)。
ARAGTGFDY(SEQ ID NO:6)。
GYRFSSYW(SEQ ID NO:7)。
ILPGRGII(SEQ ID NO:8)。
ARTDPPYFGV(SEQ ID NO:9)。
GYTFSTYW(SEQ ID NO:10)。
NLPGRHIT(SEQ ID NO:11)。
ARGRGTYYFDY(SEQ ID NO:12)。
GYSFTGYT(SEQ ID NO:13)。
INPYNGGT(SEQ ID NO:14)。
AFSYYGSRGFYFDY(SEQ ID NO:15)。
GYSFTGYT(SEQ ID NO:16)。
INPYNGGT(SEQ ID NO:17)。
ASSSYRNDGNWYFDV(SEQ ID NO:18)。
GYSITGYT(SEQ ID NO:19)。
VNPYNGGT(SEQ ID NO:20)。
AISRYGSESWYFDV(SEQ ID NO:21)。
GYTFNIYW(SEQ ID NO:22)。
ILPGSGNT(SEQ ID NO:23)。
ARTDGRGYFDY(SEQ ID NO:24)。
GYTFSSYW(SEQ ID NO:25)。
FLPRSGKT(SEQ ID NO:26)。
ARTDPPYFGV(SEQ ID NO:27)。
SSISY(SEQ ID NO:28)。
ATS(SEQ ID NO:29)。
QQWSSNPPT(SEQ ID NO:30)。
ENVGGY(SEQ ID NO:31)。
GAS(SEQ ID NO:32)。
GQNYIYPFT(SEQ ID NO:33)。
TGAVTISNY(SEQ ID NO:34)。
GTN(SEQ ID NO:35)。
VLWYSNHWV(SEQ ID NO:36)。
QSLLHSNGITY(SEQ ID NO:37)。
QMS(SEQ ID NO:38)。
AQNLEFPFT(SEQ ID NO:39)。
SSVSY(SEQ ID NO:40)。
LTS(SEQ ID NO:41)。
QQWSSNPPT(SEQ ID NO:42)。
SSVSY(SEQ ID NO:43)。
DTS(SEQ ID NO:44)。
QQWSSNPPT(SEQ ID NO:45)。
SSVSY(SEQ ID NO:46)。
ATS(SEQ ID NO:47)。
QQWSSNPPT(SEQ ID NO:48)。
KSVSSSAYSY(SEQ ID NO:49)。
LAS(SEQ ID NO:50)。
QHSRELPFT(SEQ ID NO:51)。
SNISY(SEQ ID NO:52)。
DTS(SEQ ID NO:53)。
QQWSSVPLT(SEQ ID NO:54)。
根据本发明实施例的上述抗体能够特异性的靶向结合TrkA受体，阻断NGF和TrkA结合。
根据本发明的实施例，上述抗体或抗原结合片段还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述抗体包括：
分别如SEQ ID NO:1、2和3或者与SEQ ID NO:1、2和3具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:4、5和6或者与SEQ ID NO:4、5和6具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:7、8和9或者与SEQ ID NO:7、8和9具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:10、11和12或者与SEQ ID NO:10、11和12具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:13、14和15或者与SEQ ID NO:13、14和15具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:16、17和18或者与SEQ ID NO:16、17和18具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO19、20和21或者与SEQ ID NO:19、20和21具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:22、23和24或者与SEQ ID NO:22、23和24具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:25、26和27或者与SEQ ID NO:25、26和27具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列。
根据本发明的实施例，所述抗体包括：
分别如SEQ ID NO:28、29和30或者与SEQ ID NO:28、39和30具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:31、32和33或者与SEQ ID NO:31、32和33具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:34、35和36或者与SEQ ID NO:34、35和36具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:37、38和39或者与SEQ ID NO:37、38和39具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:40、41和42或者与SEQ ID NO:40、41和42具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:43、44和45或者与SEQ ID NO:43、44和45具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:46、47和48或者与SEQ ID NO:46、47和48具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:49、50和51或者与SEQ ID NO:49、50和51具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:52、53和54或者与SEQ ID NO:52、53和54具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列。
根据本发明的实施例，所述抗体或其抗原结合片段特异性识别TrkA的胞外区。
根据本发明的实施例，所述抗体含有重链框架区序列和轻链框架区序列的至少之一，所述重链框架区序列和轻链框架区序列的至少之一的至少一部分来自于鼠源抗体、人源抗体、灵长目源抗体或其突变体的至少之一。
根据本发明的实施例，所述抗体具有如SEQ ID NO：55～63任一项所示氨基酸序列的重链可变区。
EVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSS(SEQ ID NO:55)。
QVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSS(SEQ ID NO:56)。
QVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSS(SEQ ID NO:57)。
QVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSS(SEQ ID NO:58)。
EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSS(SEQ ID NO:59)。
EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSS(SEQ ID NO:60)。
EVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSS(SEQ ID NO:61)。
QVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSS(SEQ ID NO:62)。
QVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSS(SEQ ID NO:63)。
根据本发明的实施例，所述抗体具有如SEQ ID NO：64～72任一项所示氨基酸序列的轻链可变区。
QIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIK(SEQ ID NO:64)。
SIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIK(SEQ ID NO:65)。
QAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVL(SEQ ID NO:66)。
DIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIK(SEQ ID NO:67)。
QIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIK(SEQ ID NO:68)。
DIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELK(SEQ ID NO:69)。
QIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKK(SEQ ID NO:70)。
DIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIK(SEQ ID NO:71)。
QIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIK(SEQ ID NO:72)。
根据本发明的实施例，所述抗体含有重链恒定区和轻链恒定区的至少之一，所述重链恒定区和轻链恒定区的至少之一的至少一部分来自于鼠源抗体、人源抗体、灵长目源抗体或其突变体的至少之一。
根据本发明的实施例，所述抗体的轻链恒定区和重链恒定区均来自于人源IgG抗体或其突变体。进而所述抗体的免疫原性可以得到有效降低。
根据本发明的实施例，所述抗体的轻链恒定区和重链恒定区均来自于人源IgG4。
根据本发明的实施例，所述抗体的Fc区域与人源IgG4野生型的Fc相比具有S10P，F16A，L17A，R191K突变以及229K缺失突变。其中，上述氨基酸位置的定位是以人源IgG4野生型Fc序列SEQ ID NO:73所示氨基酸序列进行定位的，如S10P，是指SEQ ID NO:75所示氨基酸序列的第10位S突变为P，依次类推。发明人发现，所述抗体的Fc区域具有上述突变以及缺失后，抗体的安全性、稳定性可以得到显著提高，抗体在体内的半衰期也显著延长。
ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQ ID NO:73)。
根据本发明的实施例，所述抗体恒定区的全长序列如SEQ ID NO:74或75所示。
ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ ID NO:74)。
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:75)。
其中，上述SEQ ID NO:74所示的抗体恒定区的全长序列包括IgG4重链恒定区和Fc区，其中，IgG4重链恒定区序列为ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRV，Fc区序列为ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG。上述SEQ ID NO:75所示的抗体恒定区的全长序列为IgG4轻链恒定区。
根据本发明的实施例，所述抗体具有SEQ ID NO:76～84任一项所示氨基酸序列的重链和具有SEQ ID NO:85～93任一项所示氨基酸序列的轻链。
EVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ ID NO:76)。
QIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:85)。
QVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ IDNO:77)。
SIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:86)。
QVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQID NO:78)。
QAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQID NO:87)。
QVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQID NO:79)。
DIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:88)。
EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ ID NO:80)。
QIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:89)。
EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ ID NO:81)。
DIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:90)。
EVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ ID NO:82)。
QIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:91)。
QVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQID NO:83)。
DIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:92)。
QVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQID NO:84)。
QIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:93)。
其中，在本申请中，上述SEQ ID NO:76和85所组成的抗体被称为20C8嵌合抗体，上述SEQ ID NO:77和86所组成的抗体被称为23E12嵌合抗体，上述SEQ ID NO:78和87所组成的抗体被称为27H3嵌合抗体，上述SEQ ID NO:79和88所组成的抗体被称为21E5嵌合抗体，上述SEQ ID NO:80和89所组成的抗体被称为2A5嵌合抗体，上述SEQ ID NO:81和SEQ IDNO:90所组成的抗体被称为4H4嵌合抗体，上述SEQ ID NO:82和SEQ ID NO:91所组成的抗体被称为22D12嵌合抗体，上述SEQ ID NO:83和SEQ ID NO:92所组成的抗体被称为1B9嵌合抗体，上述SEQ ID NO:84和SEQ ID NO:93所组成的抗体被称为15D4嵌合抗体。
根据本发明的实施例，所述抗体为单链抗体、多聚体抗体、CDR移植抗体或小分子抗体。
根据本发明的实施例，所述抗体为单链抗体，所述抗体具有SEQ ID NO:94～111所示的氨基酸序列。
QIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIKGGGGSGGGGSGGGGSEVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSS(SEQ ID NO:94)。
EVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIK(SEQ ID NO:95)。
SIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIKGGGGSGGGGSGGGGSQVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSS(SEQ ID NO:96)。
QVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSSGGGGSGGGGSGGGGSSIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIK(SEQ ID NO:97)。
QAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVLGGGGSGGGGSGGGGSQVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSS(SEQ ID NO:98)。
QVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSSGGGGSGGGGSGGGGSQAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVL(SEQ ID NO:99)。
DIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIKGGGGSGGGGSGGGGSQVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSS(SEQ ID NO:100)。
QVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSSGGGGSGGGGSGGGGSDIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIK(SEQ ID NO:101)。
QIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIKGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSS(SEQ ID NO:102)。
EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSSGGGGSGGGGSGGGGSQIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIK(SEQ ID NO:103)。
DIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELKGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSS(SEQ ID NO:104)。
EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSSGGGGSGGGGSGGGGSDIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELK(SEQ ID NO:105)。
QIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKKGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSS(SEQ ID NO:106)。
EVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKK(SEQ ID NO:107)。
DIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIKGGGGSGGGGSGGGGSQVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSS(SEQ ID NO:108)。
QVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSSGGGGSGGGGSGGGGSDIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIK(SEQ ID NO:109)。
QIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIKGGGGSGGGGSGGGGSQVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSS(SEQ ID NO:110)。
QVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSSGGGGSGGGGSGGGGSQIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIK(SEQ ID NO:111)。
其中，在本申请中，上述SEQ ID NO:94或95所示氨基酸序列的抗体被称为20C8单链抗体，上述SEQ ID NO:96或97所示氨基酸序列的抗体被称为23E12单链抗体，上述SEQ IDNO:98或99所示氨基酸序列的抗体被称为27H3单链抗体，上述SEQ ID NO:100或101所示氨基酸序列的抗体被称为21E5单链抗体，上述SEQ ID NO:102和103所示氨基酸序列的抗体被称为2A5单链抗体，上述SEQ ID NO:104和SEQ ID NO:105所示氨基酸序列的抗体被称为4H4单链抗体，上述SEQ ID NO:106和SEQ ID NO:107所示氨基酸序列的抗体被称为22D12单链抗体，上述SEQ ID NO:108和SEQ ID NO:109所示氨基酸序列的抗体被称为1B9单链抗体，上述SEQ ID NO:110和SEQ ID NO:111所示氨基酸序列的抗体被称为15D4单链抗体。其中，SEQID NO:94、96、98、100、102、104、106、108、110所示氨基酸序列的抗体从N端到C端可表示为VL-Link-VH(VL表示轻链可变区，VH表示重链可变区，Link表示连接VL和VH的连接链)，SEQID NO:95、97、99、101、103、105、107、109、111所示氨基酸序列的抗体从N端到C端可表示为VH-Link-VL(VL表示轻链可变区，VH表示重链可变区，Link表示连接VL和VH的连接链)。
根据本发明的实施例，所述小分子抗体包括Fab抗体、Fv抗体、单域抗体以及最小识别单位的至少之一。
在本发明的第二方面，本发明提出了一种核酸分子。根据本发明的实施例，所述核酸分子编码前面所述的抗体或其抗原结合片段。根据本发明实施例的核酸分子所编码的抗体或抗原结合片段可特异性靶向结合TrkA受体，阻断NGF和TrkA结合，且具有免疫原性低，ADCC低的优势。
根据本发明的实施例，上述核酸分子还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述核酸分子为DNA。
根据本发明的实施例，所述核酸分子具有如SEQ ID NO:112～120任一项所示核苷酸序列或具有SEQ ID NO:121～129任一项所示核苷酸序列或具有SEQ ID NO:130～147所示的核苷酸序列。
GAAGTGCTGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCTCCATGAAGATCAGCTGTAAGGCCTCCGTGTACAGCTTCACAGCCTACACAATGAACTGGGTGAAGCAGAGCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCACACAATGGCGGCACTAGGTACAACCAGAAGTTCAAGGGCAAGGCCACACTGACTCTGGATAAGTCCAGCAGCACTGCCTACATGGATCTGCTGTCTCTGACAAGCGAGGACAGCGCCGTCTACTATTGCGCCATCTCTAGGTACGGCAGCAGCAGCTTCTACTTCGATGTGTGGGGCGCCGGCACAACAGTGGCTGTGAGCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ IDNO:112)。
CAAGTGCAGCTCCAGCAGAGCGGAGCTGAGCTGGTGAGACCCGGCACTAGCGTGAAGATCAGCTGTAAGGCCAGCGGCTACGCCTTCACTAATTACTGGCTGGGCTGGATGAAGCAGAGACCCGGCCATGGACTGGAGTGGATCGGCGACTTCTACCCAAGGACTGGCAACACTTTCTACAACGAGAACTTTAAGGGCAAGGTGACTCTGACTGCCGATAAGTCCAGCAACACTGCCTACATGCAGCTGTCCTCTCTGACTAGCGAAGATAGCGCCGTGTATCTGTGTGCTAGGGCTGGCACTGGCTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGAGCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ ID NO:113)。
CAAGTGCAGCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCTTCCGTCAAGATCAGCTGCAAGACTACTGGCTATAGGTTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGCGAAATCCTCCCCGGAAGGGGCATCATCAACTACAATGAGAACTTTAGGGGCAAGGCCACATTCACAGCCGACACAAGCAGCAACACTGCCTACGTGCAGCTGAGCTCTCTGACTTCCGAGGACAGCGCCGTGTACTTCTGCGCCAGAACAGACCCTCCTTATTTCGGCGTGTGGGGCGCCGGAACTACTGTGACTGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ ID NO:114)。
CAAGTGCAGCTGCAGCAGAGCGGAGCCGAGCTGATGAAGCCCGGCGCCTCCATGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCACATACTGGATCGAGTGGGTCAAGCAGAGACCCGGCCACGGACTGGAGTGGATCGGAGAGAATCTGCCCGGAAGGCACATCACTAACTACAACGAGAAGTTCAAGGGCAAGGCCACTTTCACAGCCGACACTAGCAGCAACACTGCCTACATGCAGCTCAGCTCTCTGACAAGCGAAGATAGCGCCGTGTACTACTGTGCTAGGGGAAGGGGCACTTACTACTTCGATTACTGGGGCCAAGGCACTCCACTGACTGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ ID NO:115)。
GAGGTGCAGCTCCAACAGAGCGGACCAGAGCTGGTGAAACCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCTCCGGCTACAGCTTCACTGGCTACACTATGAACTGGGTGAAGCAGTCCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCATACAACGGCGGCACAAACTACAACCAGAAGTTCAAGGGCAAGGCCACTCTGACAGTCGATAAGAGCTCCAGCACTGCCTACATGGAGCTGCTGAGCCTCACTAGCGAGGACAGCGCTGTGTACTACTGTGCCTTCTCCTACTACGGCTCTAGGGGCTTCTACTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ IDNO:116)。
GAAGTGCAGCTGCAACAGAGCGGCCCAGAGCTCGTGAAGCCCGGCGCCAGCATGAAGATCAGCTGCAAGGCCAGCGGCTACAGCTTCACTGGCTACACAATGAACTGGGTCAAGCAGAGCCACGGAAAAAATCTGGAGTGGATCGGACTGATCAACCCTTACAACGGCGGCACAAGGTACAATCAGAAGTTCAAGGGCAAGGCCACTCTCACTGTGGATAAGAGCAGCAGCACTGCCTACATGGAGCTGCTGTCTCTGACAAGCGAGGATAGCGCCGTGTACTACTGCGCCAGCTCCTCCTATAGGAACGACGGCAACTGGTACTTCGATGTGTGGGGCGCCGGCACTACTGTGACAGTGAGCTCCGCCAGCACCAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQID NO:117)。
GAAGTGCAGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCAGCGGCTACAGCATCACTGGCTACACTATGAACTGGGTGAAGCAGAGCCACGGCAAGAACCTCGAGTGGATTGGCCTCGTGAACCCATACAACGGCGGCACTTCCTACAACCAGAAGTTCAAAGGCAAGGCCACACTCACAGTCGATAAGTCCAGCTCCACAGCCTACATGGAGCTGCTGTCTCTGAAGAGCGAGGATAGCGCTGTCTACTACTGTGCCATCAGCAGATACGGCAGCGAGAGCTGGTACTTCGACGTGTGGGGCGCCGGCACAACAGTGACAGTGAGCAGCGCCAGCACAAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ IDNO:118)。
CAAGTGCATCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCCAGCGTGAAGATTAGCTGCAAGGCCACTGGCTACACATTCAACATCTACTGGATCGACTGGGTGAAGCAGAGGCCCGGCCACGGACTGGAATGGATCGGCGAAATTCTGCCCGGCAGCGGCAACACTCACTACAACGAGAACTTCAAGGGCAAGGCCACAATGACAGCCGACACAAGCTCCAACACTGCTTACATGCAGCTGACTTCTCTGACTAGCGAGGACAGCGCCGTGTACTATTGCGCTAGGACAGACGGAAGGGGCTACTTCGATTACTGGGGCCAAGGCACTACACTCACAGTGAGCAGCGCCAGCACTAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ ID NO:119)。
CAGGTGCAGCTGCAGCAGTCCGGCGCTGAGCTCATGAAGCCCGGCGCCAGCGTGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGAGAGTTTCTGCCTAGAAGCGGCAAGACAAACTACAACGAGGAGTTTAGGGGCAAGGCTACATTCACTGCCGACACATCCAGCAACACTGCCTACATGCAGCTGAGCAGCCTCACAAGCGAGGATTCCGCCGTCTACTACTGTGCTAGGACTGATCCTCCTTACTTCGGAGTGTGGGGCGCTGGCACAATGGTGGCTGTGAGCAGCGCCTCCACTAAGGGCCCCAGCGTGTTCCCTCTGGCTCCTTGTAGCCGGTCCACCTCCGAGTCCACAGCTGCTCTGGGCTGCCTCGTGAAGGACTACTTTCCCGAACCCGTTACCGTGAGCTGGAATAGCGGCGCTTTAACCTCCGGAGTGCACACCTTCCCCGCTGTGCTCCAGTCCTCCGGTTTATACTCTTTATCCTCCGTGGTGACCGTGCCTTCCTCCAGCCTCGGCACCAAGACCTACACTTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGTCCAAGTACGGACCTCCTTGTCCCCCTTGCCCCGCCCCCGAGGCCGCTGGCGGACCCTCCGTGTTCCTCTTCCCCCCCAAACCCAAGGACACTTTAATGATCTCCCGGACCCCCGAAGTGACTTGTGTGGTGGTGGACGTGTCCCAAGAAGACCCCGAGGTGCAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTAGGGAGGAACAGTTCAACTCCACCTACCGGGTGGTGTCCGTGCTCACCGTGCTGCATCAAGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAACAAGGGACTGCCCAGCTCCATCGAGAAGACCATCAGCAAGGCCAAAGGCCAGCCCCGGGAACCTCAAGTTTATACACTGCCCCCCAGCCAAGAAGAGATGACCAAGAACCAAGTTTCTTTAACTTGTTTAGTGAAGGGCTTCTACCCTAGCGACATCGCTGTGGAGTGGGAGTCCAATGGCCAGCCCGAAAACAATTATAAGACCACCCCCCCCGTGCTGGACTCCGATGGTTCTTTTTTTTTATACTCCAAGCTGACAGTGGACAAGTCTCGTTGGCAAGAAGGCAACGTGTTCTCTTGTAGCGTGATGCACGAGGCTTTACACAACCACTACACCCAGAAGTCTTTATCTCTGTCTTTAGGCTGATGAGAATTC(SEQ ID NO:120)。
CAGATCGTGCTGAGCCAGAGCCCAGCTATTCTGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCAGCTCCAGCATCAGCTACATGCACTGGTACCAGCAGAAGCCCGGCTCCTCCCCAAAGCCTTGGATCAGCGCCACTAGCAATCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGAAGCGGCAGCGGCACTAGCTACTCTCTGACTATCTCCGGCGTGGAAGCTGAGGATGCCGCCACTTACTACTGCCAGCAGTGGTCCAGCAACCCTCCTACTTTCGGCGGCGGCACAAATCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:121)。
AGCATCGTCATGACACAAAGCCCTAAGAGCATGAGCATGAGCGTGGGCGAGAGAGTGACTCTGAGCTGTAAGGCCAGCGAGAATGTGGGCGGCTACGTGAGCTGGTATCAGCAGAAGCCAGATCAGAGCCCAAAGCTGCTGATCTACGGCGCCAGCAGCAGACACACTGGCGTGCCAGATAGGTTCACTGGCAGCGGCTCCGAGACAGACTTCACTCTGACTATCAGCAGCGTCCAAGCCGAAGATCTGGCCGCCTATCACTGCGGCCAGAACTACATCTACCCATTCACATTCGGCGGCGGCACAAAGCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:122)。
CAAGCCGTCGTGACACAAGAGTCCGCTCTGACAACTTCCCCCGGCGAGACTGTGACACTGACTTGTAGGAGCAGCACTGGCGCCGTGACTATCAGCAACTACGCCAACTGGGTCCAAGAGAAGCCAGATCATCTGTTCACTGGACTGATCGGCGGCACAAATAATAGGCCTCCCGGCGTGCCAGCCAGATTTAGCGGCTCTCTGATTGGCGATAAGGCTGCTCTGACAATCACTGGCGCCCAGACTGAGGACGAGGCCATCTACTTCTGCGTCCTCTGGTACAGCAACCACTGGGTGTTCGGCGGCGGCACTAAGCTGACTGTGCTGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:123)。
GATATCGTCATGACTCAAGCCGCCTTCAGCACTCCAGTCACTCTCGGCACAAGCGCCAGCATCAGCTGTAGGTCCAGCCAGTCTCTGCTGCACAGCAACGGCATCACTTATCTGTACTGGTATCTGCAGAAGCCCGGCCAAAGCCCACAGCTGCTGATCTACCAGATGAGCAATCTGGCCAGCGGCGTGCCAGATAGATTCAGCAGCAGCGGAAGCGGAACAGACTTCACACTGAGGATCTCTAGGGTGGAAGCCGAAGATGTGGGCGTCTATTACTGCGCCCAGAACCTCGAGTTCCCTTTCACATTCGGCAGCGGCACTAAGCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ IDNO:124)。
CAGATCGTGCTGACACAGAGCCCAGCTCTGATGAGCGCCAGCCCCGGCGAGAAGGTCACAATGACTTGCAGCGCCACATCCAGCGTGAGCTACATCTACTGGTACCAGCAGAAGCCTAGGAGCAGCCCTAAGCCTTGGATCTACCTCACAAGCAATCTGGCCAGCGGAGTGCCAGCTAGGTTCAGCGGAAGCGGCAGCGGCACAAGCTACTCTCTGACAATCTCCAGCATGGAAGCCGAAGATGCCGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCACCTACATTCGGAGGCGGCACTAAGCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:125)。
GATATCGTGATGACTCAGTCCCCAGCCATCATGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGCTCCGCCAGCAGCAGCGTGAGCTACATGCACTGGTACCAGCAGAAGAGCGGCACATCCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGCTCCGGCAGCGGAACAAGCTACTCTCTGACTATCAGCAGCATGGAGACAGAGGACGCTGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCTCCAACTTTCGGCGGAGGCACTAAGCTGGAGCTGAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:126)。
CAGATTGTGCTGTCCCAGTCCCCAGCCATTCTGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCACAAGCAGCGTGAGCTACATGTACTGGTACCAGCAGAAGCCCGGCAGCAGCCCTAAGCCTTGGATCTACGCCACAAGCAATCTGGCCAGCGGCGTCCCAGCTAGATTTAGCGGCAGCGGATCCGGCACTAGCTATTCTCTGACTATCTCTAGGGTCGAGGCCGAAGATGCCGCCACATACTACTGCCAGCAGTGGTCCTCCAATCCTCCAACATTCGGCGGCGGAACTAAGCTGGAGAAGAAGAGGACAGTGGCTGCCCCTTCCGTGTTCATCTTCCCTCCAAGCGACGAGCAGCTGAAGTCCGGCACTGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:127)。
GATATCGTGCTGACTCAGAGCCCAGCCTCTCTGGTGGTGTCTCTGGGACAGAGGGCCACAATCAGCTGTAGGACTTCCAAGAGCGTGAGCAGCTCCGCCTACAGCTACATGCACTGGTACCAGCAGAAGCCCGGCCAGCCTCCTAAGGTGCTGATCTATCTGGCCAGCAATCTGGAGAGCGGCGTCCCAGCTAGATTCAGCGGCTCCGGAAGCGGCACTGACTTCACTCTGAACATCCACCCAGTGGAAGAGGAGGATGCCGCCACATACTACTGCCAGCACTCTAGGGAGCTGCCTTTCACATTTGGCAGCGGAACTAAGCTGGAGATCAAGAGGACTGTCGCCGCCCCTAGCGTGTTCATCTTCCCTCCAAGCGATGAGCAGCTGAAGAGCGGCACTGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:128)。
CAAATTGTTCTCACACAGAGCCCTCCTATCATGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTTCCGCCAGCAGCAACATCAGCTACATGCACTGGTACCAGCAGAAGTCCGGCACAAGCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTTAGCGGCTCCGGAAGCGGCACTAGCTACTCTCTGACAATCAGCAGCATGGAAGCCGAGGACGCCGCTACATACTACTGCCAGCAGTGGAGCTCCGTCCCACTGACTTTCGGCGCTGGCACTAAGCTGGAGATCAAGAGGACTGTGGCCGCCCCTTCCGTGTTCATCTTCCCTCCTAGCGACGAACAGCTCAAGAGCGGCACTGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO:129)。
CAGATCGTGCTGAGCCAGAGCCCAGCTATTCTGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCAGCTCCAGCATCAGCTACATGCACTGGTACCAGCAGAAGCCCGGCTCCTCCCCAAAGCCTTGGATCAGCGCCACTAGCAATCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGAAGCGGCAGCGGCACTAGCTACTCTCTGACTATCTCCGGCGTGGAAGCTGAGGATGCCGCCACTTACTACTGCCAGCAGTGGTCCAGCAACCCTCCTACTTTCGGCGGCGGCACAAATCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAAGTGCTGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCTCCATGAAGATCAGCTGTAAGGCCTCCGTGTACAGCTTCACAGCCTACACAATGAACTGGGTGAAGCAGAGCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCACACAATGGCGGCACTAGGTACAACCAGAAGTTCAAGGGCAAGGCCACACTGACTCTGGATAAGTCCAGCAGCACTGCCTACATGGATCTGCTGTCTCTGACAAGCGAGGACAGCGCCGTCTACTATTGCGCCATCTCTAGGTACGGCAGCAGCAGCTTCTACTTCGATGTGTGGGGCGCCGGCACAACAGTGGCTGTGAGCAGC(SEQ ID NO:130)。
GAAGTGCTGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCTCCATGAAGATCAGCTGTAAGGCCTCCGTGTACAGCTTCACAGCCTACACAATGAACTGGGTGAAGCAGAGCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCACACAATGGCGGCACTAGGTACAACCAGAAGTTCAAGGGCAAGGCCACACTGACTCTGGATAAGTCCAGCAGCACTGCCTACATGGATCTGCTGTCTCTGACAAGCGAGGACAGCGCCGTCTACTATTGCGCCATCTCTAGGTACGGCAGCAGCAGCTTCTACTTCGATGTGTGGGGCGCCGGCACAACAGTGGCTGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGATCGTGCTGAGCCAGAGCCCAGCTATTCTGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCAGCTCCAGCATCAGCTACATGCACTGGTACCAGCAGAAGCCCGGCTCCTCCCCAAAGCCTTGGATCAGCGCCACTAGCAATCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGAAGCGGCAGCGGCACTAGCTACTCTCTGACTATCTCCGGCGTGGAAGCTGAGGATGCCGCCACTTACTACTGCCAGCAGTGGTCCAGCAACCCTCCTACTTTCGGCGGCGGCACAAATCTGGAGATCAAG(SEQ ID NO:131)。
AGCATCGTCATGACACAAAGCCCTAAGAGCATGAGCATGAGCGTGGGCGAGAGAGTGACTCTGAGCTGTAAGGCCAGCGAGAATGTGGGCGGCTACGTGAGCTGGTATCAGCAGAAGCCAGATCAGAGCCCAAAGCTGCTGATCTACGGCGCCAGCAGCAGACACACTGGCGTGCCAGATAGGTTCACTGGCAGCGGCTCCGAGACAGACTTCACTCTGACTATCAGCAGCGTCCAAGCCGAAGATCTGGCCGCCTATCACTGCGGCCAGAACTACATCTACCCATTCACATTCGGCGGCGGCACAAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGTGCAGCTCCAGCAGAGCGGAGCTGAGCTGGTGAGACCCGGCACTAGCGTGAAGATCAGCTGTAAGGCCAGCGGCTACGCCTTCACTAATTACTGGCTGGGCTGGATGAAGCAGAGACCCGGCCATGGACTGGAGTGGATCGGCGACTTCTACCCAAGGACTGGCAACACTTTCTACAACGAGAACTTTAAGGGCAAGGTGACTCTGACTGCCGATAAGTCCAGCAACACTGCCTACATGCAGCTGTCCTCTCTGACTAGCGAAGATAGCGCCGTGTATCTGTGTGCTAGGGCTGGCACTGGCTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGAGCAGC(SEQ ID NO:132)。
CAAGTGCAGCTCCAGCAGAGCGGAGCTGAGCTGGTGAGACCCGGCACTAGCGTGAAGATCAGCTGTAAGGCCAGCGGCTACGCCTTCACTAATTACTGGCTGGGCTGGATGAAGCAGAGACCCGGCCATGGACTGGAGTGGATCGGCGACTTCTACCCAAGGACTGGCAACACTTTCTACAACGAGAACTTTAAGGGCAAGGTGACTCTGACTGCCGATAAGTCCAGCAACACTGCCTACATGCAGCTGTCCTCTCTGACTAGCGAAGATAGCGCCGTGTATCTGTGTGCTAGGGCTGGCACTGGCTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTAGCATCGTCATGACACAAAGCCCTAAGAGCATGAGCATGAGCGTGGGCGAGAGAGTGACTCTGAGCTGTAAGGCCAGCGAGAATGTGGGCGGCTACGTGAGCTGGTATCAGCAGAAGCCAGATCAGAGCCCAAAGCTGCTGATCTACGGCGCCAGCAGCAGACACACTGGCGTGCCAGATAGGTTCACTGGCAGCGGCTCCGAGACAGACTTCACTCTGACTATCAGCAGCGTCCAAGCCGAAGATCTGGCCGCCTATCACTGCGGCCAGAACTACATCTACCCATTCACATTCGGCGGCGGCACAAAGCTGGAGATCAAG(SEQ ID NO:133)。
CAAGCCGTCGTGACACAAGAGTCCGCTCTGACAACTTCCCCCGGCGAGACTGTGACACTGACTTGTAGGAGCAGCACTGGCGCCGTGACTATCAGCAACTACGCCAACTGGGTCCAAGAGAAGCCAGATCATCTGTTCACTGGACTGATCGGCGGCACAAATAATAGGCCTCCCGGCGTGCCAGCCAGATTTAGCGGCTCTCTGATTGGCGATAAGGCTGCTCTGACAATCACTGGCGCCCAGACTGAGGACGAGGCCATCTACTTCTGCGTCCTCTGGTACAGCAACCACTGGGTGTTCGGCGGCGGCACTAAGCTGACTGTGCTGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGTGCAGCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCTTCCGTCAAGATCAGCTGCAAGACTACTGGCTATAGGTTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGCGAAATCCTCCCCGGAAGGGGCATCATCAACTACAATGAGAACTTTAGGGGCAAGGCCACATTCACAGCCGACACAAGCAGCAACACTGCCTACGTGCAGCTGAGCTCTCTGACTTCCGAGGACAGCGCCGTGTACTTCTGCGCCAGAACAGACCCTCCTTATTTCGGCGTGTGGGGCGCCGGAACTACTGTGACTGTGTCCAGC(SEQ ID NO:134)。
CAAGTGCAGCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCTTCCGTCAAGATCAGCTGCAAGACTACTGGCTATAGGTTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGCGAAATCCTCCCCGGAAGGGGCATCATCAACTACAATGAGAACTTTAGGGGCAAGGCCACATTCACAGCCGACACAAGCAGCAACACTGCCTACGTGCAGCTGAGCTCTCTGACTTCCGAGGACAGCGCCGTGTACTTCTGCGCCAGAACAGACCCTCCTTATTTCGGCGTGTGGGGCGCCGGAACTACTGTGACTGTGTCCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGCCGTCGTGACACAAGAGTCCGCTCTGACAACTTCCCCCGGCGAGACTGTGACACTGACTTGTAGGAGCAGCACTGGCGCCGTGACTATCAGCAACTACGCCAACTGGGTCCAAGAGAAGCCAGATCATCTGTTCACTGGACTGATCGGCGGCACAAATAATAGGCCTCCCGGCGTGCCAGCCAGATTTAGCGGCTCTCTGATTGGCGATAAGGCTGCTCTGACAATCACTGGCGCCCAGACTGAGGACGAGGCCATCTACTTCTGCGTCCTCTGGTACAGCAACCACTGGGTGTTCGGCGGCGGCACTAAGCTGACTGTGCTG(SEQ ID NO:135)。
GATATCGTCATGACTCAAGCCGCCTTCAGCACTCCAGTCACTCTCGGCACAAGCGCCAGCATCAGCTGTAGGTCCAGCCAGTCTCTGCTGCACAGCAACGGCATCACTTATCTGTACTGGTATCTGCAGAAGCCCGGCCAAAGCCCACAGCTGCTGATCTACCAGATGAGCAATCTGGCCAGCGGCGTGCCAGATAGATTCAGCAGCAGCGGAAGCGGAACAGACTTCACACTGAGGATCTCTAGGGTGGAAGCCGAAGATGTGGGCGTCTATTACTGCGCCCAGAACCTCGAGTTCCCTTTCACATTCGGCAGCGGCACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGTGCAGCTGCAGCAGAGCGGAGCCGAGCTGATGAAGCCCGGCGCCTCCATGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCACATACTGGATCGAGTGGGTCAAGCAGAGACCCGGCCACGGACTGGAGTGGATCGGAGAGAATCTGCCCGGAAGGCACATCACTAACTACAACGAGAAGTTCAAGGGCAAGGCCACTTTCACAGCCGACACTAGCAGCAACACTGCCTACATGCAGCTCAGCTCTCTGACAAGCGAAGATAGCGCCGTGTACTACTGTGCTAGGGGAAGGGGCACTTACTACTTCGATTACTGGGGCCAAGGCACTCCACTGACTGTGTCCAGC(SEQ IDNO:136)。
CAAGTGCAGCTGCAGCAGAGCGGAGCCGAGCTGATGAAGCCCGGCGCCTCCATGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCACATACTGGATCGAGTGGGTCAAGCAGAGACCCGGCCACGGACTGGAGTGGATCGGAGAGAATCTGCCCGGAAGGCACATCACTAACTACAACGAGAAGTTCAAGGGCAAGGCCACTTTCACAGCCGACACTAGCAGCAACACTGCCTACATGCAGCTCAGCTCTCTGACAAGCGAAGATAGCGCCGTGTACTACTGTGCTAGGGGAAGGGGCACTTACTACTTCGATTACTGGGGCCAAGGCACTCCACTGACTGTGTCCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGATATCGTCATGACTCAAGCCGCCTTCAGCACTCCAGTCACTCTCGGCACAAGCGCCAGCATCAGCTGTAGGTCCAGCCAGTCTCTGCTGCACAGCAACGGCATCACTTATCTGTACTGGTATCTGCAGAAGCCCGGCCAAAGCCCACAGCTGCTGATCTACCAGATGAGCAATCTGGCCAGCGGCGTGCCAGATAGATTCAGCAGCAGCGGAAGCGGAACAGACTTCACACTGAGGATCTCTAGGGTGGAAGCCGAAGATGTGGGCGTCTATTACTGCGCCCAGAACCTCGAGTTCCCTTTCACATTCGGCAGCGGCACTAAGCTGGAGATCAAG(SEQ IDNO:137)。
CAGATCGTGCTGACACAGAGCCCAGCTCTGATGAGCGCCAGCCCCGGCGAGAAGGTCACAATGACTTGCAGCGCCACATCCAGCGTGAGCTACATCTACTGGTACCAGCAGAAGCCTAGGAGCAGCCCTAAGCCTTGGATCTACCTCACAAGCAATCTGGCCAGCGGAGTGCCAGCTAGGTTCAGCGGAAGCGGCAGCGGCACAAGCTACTCTCTGACAATCTCCAGCATGGAAGCCGAAGATGCCGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCACCTACATTCGGAGGCGGCACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAGGTGCAGCTCCAACAGAGCGGACCAGAGCTGGTGAAACCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCTCCGGCTACAGCTTCACTGGCTACACTATGAACTGGGTGAAGCAGTCCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCATACAACGGCGGCACAAACTACAACCAGAAGTTCAAGGGCAAGGCCACTCTGACAGTCGATAAGAGCTCCAGCACTGCCTACATGGAGCTGCTGAGCCTCACTAGCGAGGACAGCGCTGTGTACTACTGTGCCTTCTCCTACTACGGCTCTAGGGGCTTCTACTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGTCCAGC(SEQ ID NO:138)。
GAGGTGCAGCTCCAACAGAGCGGACCAGAGCTGGTGAAACCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCTCCGGCTACAGCTTCACTGGCTACACTATGAACTGGGTGAAGCAGTCCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCATACAACGGCGGCACAAACTACAACCAGAAGTTCAAGGGCAAGGCCACTCTGACAGTCGATAAGAGCTCCAGCACTGCCTACATGGAGCTGCTGAGCCTCACTAGCGAGGACAGCGCTGTGTACTACTGTGCCTTCTCCTACTACGGCTCTAGGGGCTTCTACTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGTCCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGATCGTGCTGACACAGAGCCCAGCTCTGATGAGCGCCAGCCCCGGCGAGAAGGTCACAATGACTTGCAGCGCCACATCCAGCGTGAGCTACATCTACTGGTACCAGCAGAAGCCTAGGAGCAGCCCTAAGCCTTGGATCTACCTCACAAGCAATCTGGCCAGCGGAGTGCCAGCTAGGTTCAGCGGAAGCGGCAGCGGCACAAGCTACTCTCTGACAATCTCCAGCATGGAAGCCGAAGATGCCGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCACCTACATTCGGAGGCGGCACTAAGCTGGAGATCAAG(SEQ ID NO:139)。
GATATCGTGATGACTCAGTCCCCAGCCATCATGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGCTCCGCCAGCAGCAGCGTGAGCTACATGCACTGGTACCAGCAGAAGAGCGGCACATCCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGCTCCGGCAGCGGAACAAGCTACTCTCTGACTATCAGCAGCATGGAGACAGAGGACGCTGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCTCCAACTTTCGGCGGAGGCACTAAGCTGGAGCTGAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAAGTGCAGCTGCAACAGAGCGGCCCAGAGCTCGTGAAGCCCGGCGCCAGCATGAAGATCAGCTGCAAGGCCAGCGGCTACAGCTTCACTGGCTACACAATGAACTGGGTCAAGCAGAGCCACGGAAAAAATCTGGAGTGGATCGGACTGATCAACCCTTACAACGGCGGCACAAGGTACAATCAGAAGTTCAAGGGCAAGGCCACTCTCACTGTGGATAAGAGCAGCAGCACTGCCTACATGGAGCTGCTGTCTCTGACAAGCGAGGATAGCGCCGTGTACTACTGCGCCAGCTCCTCCTATAGGAACGACGGCAACTGGTACTTCGATGTGTGGGGCGCCGGCACTACTGTGACAGTGAGCTCC(SEQ ID NO:140)。
GAAGTGCAGCTGCAACAGAGCGGCCCAGAGCTCGTGAAGCCCGGCGCCAGCATGAAGATCAGCTGCAAGGCCAGCGGCTACAGCTTCACTGGCTACACAATGAACTGGGTCAAGCAGAGCCACGGAAAAAATCTGGAGTGGATCGGACTGATCAACCCTTACAACGGCGGCACAAGGTACAATCAGAAGTTCAAGGGCAAGGCCACTCTCACTGTGGATAAGAGCAGCAGCACTGCCTACATGGAGCTGCTGTCTCTGACAAGCGAGGATAGCGCCGTGTACTACTGCGCCAGCTCCTCCTATAGGAACGACGGCAACTGGTACTTCGATGTGTGGGGCGCCGGCACTACTGTGACAGTGAGCTCCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGATATCGTGATGACTCAGTCCCCAGCCATCATGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGCTCCGCCAGCAGCAGCGTGAGCTACATGCACTGGTACCAGCAGAAGAGCGGCACATCCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGCTCCGGCAGCGGAACAAGCTACTCTCTGACTATCAGCAGCATGGAGACAGAGGACGCTGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCTCCAACTTTCGGCGGAGGCACTAAGCTGGAGCTGAAG(SEQ ID NO:141)。
CAGATTGTGCTGTCCCAGTCCCCAGCCATTCTGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCACAAGCAGCGTGAGCTACATGTACTGGTACCAGCAGAAGCCCGGCAGCAGCCCTAAGCCTTGGATCTACGCCACAAGCAATCTGGCCAGCGGCGTCCCAGCTAGATTTAGCGGCAGCGGATCCGGCACTAGCTATTCTCTGACTATCTCTAGGGTCGAGGCCGAAGATGCCGCCACATACTACTGCCAGCAGTGGTCCTCCAATCCTCCAACATTCGGCGGCGGAACTAAGCTGGAGAAGAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAAGTGCAGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCAGCGGCTACAGCATCACTGGCTACACTATGAACTGGGTGAAGCAGAGCCACGGCAAGAACCTCGAGTGGATTGGCCTCGTGAACCCATACAACGGCGGCACTTCCTACAACCAGAAGTTCAAAGGCAAGGCCACACTCACAGTCGATAAGTCCAGCTCCACAGCCTACATGGAGCTGCTGTCTCTGAAGAGCGAGGATAGCGCTGTCTACTACTGTGCCATCAGCAGATACGGCAGCGAGAGCTGGTACTTCGACGTGTGGGGCGCCGGCACAACAGTGACAGTGAGCAGC(SEQ ID NO:142)。
GAAGTGCAGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCAGCGGCTACAGCATCACTGGCTACACTATGAACTGGGTGAAGCAGAGCCACGGCAAGAACCTCGAGTGGATTGGCCTCGTGAACCCATACAACGGCGGCACTTCCTACAACCAGAAGTTCAAAGGCAAGGCCACACTCACAGTCGATAAGTCCAGCTCCACAGCCTACATGGAGCTGCTGTCTCTGAAGAGCGAGGATAGCGCTGTCTACTACTGTGCCATCAGCAGATACGGCAGCGAGAGCTGGTACTTCGACGTGTGGGGCGCCGGCACAACAGTGACAGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGATTGTGCTGTCCCAGTCCCCAGCCATTCTGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCACAAGCAGCGTGAGCTACATGTACTGGTACCAGCAGAAGCCCGGCAGCAGCCCTAAGCCTTGGATCTACGCCACAAGCAATCTGGCCAGCGGCGTCCCAGCTAGATTTAGCGGCAGCGGATCCGGCACTAGCTATTCTCTGACTATCTCTAGGGTCGAGGCCGAAGATGCCGCCACATACTACTGCCAGCAGTGGTCCTCCAATCCTCCAACATTCGGCGGCGGAACTAAGCTGGAGAAGAAG(SEQ ID NO:143)。
GATATCGTGCTGACTCAGAGCCCAGCCTCTCTGGTGGTGTCTCTGGGACAGAGGGCCACAATCAGCTGTAGGACTTCCAAGAGCGTGAGCAGCTCCGCCTACAGCTACATGCACTGGTACCAGCAGAAGCCCGGCCAGCCTCCTAAGGTGCTGATCTATCTGGCCAGCAATCTGGAGAGCGGCGTCCCAGCTAGATTCAGCGGCTCCGGAAGCGGCACTGACTTCACTCTGAACATCCACCCAGTGGAAGAGGAGGATGCCGCCACATACTACTGCCAGCACTCTAGGGAGCTGCCTTTCACATTTGGCAGCGGAACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGTGCATCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCCAGCGTGAAGATTAGCTGCAAGGCCACTGGCTACACATTCAACATCTACTGGATCGACTGGGTGAAGCAGAGGCCCGGCCACGGACTGGAATGGATCGGCGAAATTCTGCCCGGCAGCGGCAACACTCACTACAACGAGAACTTCAAGGGCAAGGCCACAATGACAGCCGACACAAGCTCCAACACTGCTTACATGCAGCTGACTTCTCTGACTAGCGAGGACAGCGCCGTGTACTATTGCGCTAGGACAGACGGAAGGGGCTACTTCGATTACTGGGGCCAAGGCACTACACTCACAGTGAGCAGC(SEQ ID NO:144)。
CAAGTGCATCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCCAGCGTGAAGATTAGCTGCAAGGCCACTGGCTACACATTCAACATCTACTGGATCGACTGGGTGAAGCAGAGGCCCGGCCACGGACTGGAATGGATCGGCGAAATTCTGCCCGGCAGCGGCAACACTCACTACAACGAGAACTTCAAGGGCAAGGCCACAATGACAGCCGACACAAGCTCCAACACTGCTTACATGCAGCTGACTTCTCTGACTAGCGAGGACAGCGCCGTGTACTATTGCGCTAGGACAGACGGAAGGGGCTACTTCGATTACTGGGGCCAAGGCACTACACTCACAGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGATATCGTGCTGACTCAGAGCCCAGCCTCTCTGGTGGTGTCTCTGGGACAGAGGGCCACAATCAGCTGTAGGACTTCCAAGAGCGTGAGCAGCTCCGCCTACAGCTACATGCACTGGTACCAGCAGAAGCCCGGCCAGCCTCCTAAGGTGCTGATCTATCTGGCCAGCAATCTGGAGAGCGGCGTCCCAGCTAGATTCAGCGGCTCCGGAAGCGGCACTGACTTCACTCTGAACATCCACCCAGTGGAAGAGGAGGATGCCGCCACATACTACTGCCAGCACTCTAGGGAGCTGCCTTTCACATTTGGCAGCGGAACTAAGCTGGAGATCAAG(SEQ ID NO:145)。
CAAATTGTTCTCACACAGAGCCCTCCTATCATGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTTCCGCCAGCAGCAACATCAGCTACATGCACTGGTACCAGCAGAAGTCCGGCACAAGCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTTAGCGGCTCCGGAAGCGGCACTAGCTACTCTCTGACAATCAGCAGCATGGAAGCCGAGGACGCCGCTACATACTACTGCCAGCAGTGGAGCTCCGTCCCACTGACTTTCGGCGCTGGCACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGGTTCAGCTGCAGCAGTCCGGCGCTGAGCTCATGAAGCCCGGCGCCAGCGTGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGAGAGTTTCTGCCTAGAAGCGGCAAGACAAACTACAACGAGGAGTTTAGGGGCAAGGCTACATTCACTGCCGACACATCCAGCAACACTGCCTACATGCAGCTGAGCAGCCTCACAAGCGAGGATTCCGCCGTCTACTACTGTGCTAGGACTGATCCTCCTTACTTCGGAGTGTGGGGCGCTGGCACAATGGTGGCTGTGAGCAGC(SEQ ID NO:146)。
CAGGTTCAGCTGCAGCAGTCCGGCGCTGAGCTCATGAAGCCCGGCGCCAGCGTGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGAGAGTTTCTGCCTAGAAGCGGCAAGACAAACTACAACGAGGAGTTTAGGGGCAAGGCTACATTCACTGCCGACACATCCAGCAACACTGCCTACATGCAGCTGAGCAGCCTCACAAGCGAGGATTCCGCCGTCTACTACTGTGCTAGGACTGATCCTCCTTACTTCGGAGTGTGGGGCGCTGGCACAATGGTGGCTGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAATTGTTCTCACACAGAGCCCTCCTATCATGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTTCCGCCAGCAGCAACATCAGCTACATGCACTGGTACCAGCAGAAGTCCGGCACAAGCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTTAGCGGCTCCGGAAGCGGCACTAGCTACTCTCTGACAATCAGCAGCATGGAAGCCGAGGACGCCGCTACATACTACTGCCAGCAGTGGAGCTCCGTCCCACTGACTTTCGGCGCTGGCACTAAGCTGGAGATCAAG(SEQ ID NO:147)。
其中，在本申请中，上述SEQ ID NO:112和121所示核苷酸序列分别编码20C8嵌合抗体的重链和轻链，上述SEQ ID NO:113和122所示核苷酸序列分别编码23E12嵌合抗体的重链和轻链，上述SEQ ID NO:114和123所示核苷酸序列分别编码27H3嵌合抗体的重链和轻链，上述SEQ ID NO:115和124所示核苷酸序列分别编码21E5嵌合抗体的重链和轻链，上述SEQ ID NO:116和125所示核苷酸序列分别编码2A5嵌合抗体的重链和轻链，上述SEQ IDNO:117和SEQ ID NO:126所示核苷酸序列分别编码4H4嵌合抗体的重链和轻链，上述SEQ IDNO:118和SEQ ID NO:127所示核苷酸序列分别编码22D12嵌合抗体的重链和轻链，上述SEQID NO:119和SEQ ID NO:128所示核苷酸序列分别编码1B9嵌合抗体的重链和轻链，上述SEQID NO:120和SEQ ID NO:129所示核苷酸序列分别编码15D4嵌合抗体的重链和轻链。SEQ IDNO:130、132、134、136、138、140、142、144、146所示核苷酸序列分别编码SEQ ID NO:94、96、98、100、102、104、106、108、110所示氨基酸序列的单链抗体，SEQ ID NO:131、133、135、137、139、141、143、145、147所示核苷酸序列分别编码SEQ ID NO:95、97、99、101、103、105、107、109、111所示氨基酸序列的单链抗体。
在本发明的第三方面，本发明提出了一种表达载体。根据本发明的实施例，所述表达载体携带前面所述的核酸分子。根据本发明实施例的表达载体导入合适的受体细胞后，可在调控系统的介导下，有效实现前面所述的特异性识别TrkA的抗体或其抗原结合片段表达，进而实现所述抗体或抗原结合片段的体外大量获得。
根据本发明的实施例，上述表达载体还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述表达载体为真核表达载体。进而实现前面所述的特异性识别TrkA的抗体或其抗原结合片段在真核细胞中的表达，如CHO细胞。
在本发明的第四方面，本发明提出了一种重组细胞。根据本发明的实施例，所述重组细胞携带前面所述的核酸分子，或者表达前面所述的抗体或其抗原结合片段。根据本发明实施例的重组细胞可用于前面所述的特异性识别TrkA的抗体或其抗原结合片段体外表达和大量获得。
根据本发明的实施例，上述重组细胞还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述重组细胞是通过将前面所述的表达载体引入至宿主细胞中而获得的。
根据本发明的实施例，通过电转导的方法将所述表达载体引入所述宿主细胞中。
根据本发明的实施例，所述重组细胞为真核细胞。
根据本发明的实施例，所述重组细胞为哺乳动物细胞。
在本发明的第五方面，本发明提出了一种药物组合物。根据本发明的实施例，所述药物组合物含有前面所述的抗体，前面所述的核酸分子，前面所述的表达载体或前面所述的重组细胞。根据本发明实施例的药物组合物中所包含的抗体或表达的抗体不仅能够特异性的靶向结合TrkA受体，阻断NGF和TrkA结合，有效的抑制疼痛，而且具有免疫原性低，基本没有抗体依赖的细胞介导的细胞毒性作用(ADCC)的特点。
在本发明的第六方面，本发明提出了前面所述的抗体、前面所述的核酸分子、前面所述的表达载体或前面所述的重组细胞、前面所述的药物组合物在制备药物中的用途，所述药物用于治疗或者预防疼痛、癌症、炎症或炎性疾病、神经变性疾病、舍格伦氏综合征、子宫内膜异位、糖尿病性周围神经病变、前列腺炎、盆腔疼痛综合征、与骨重塑调节失衡相关的疾病以及由结缔组织生长因子异常信号传导引起的疾病。
根据本发明的实施例，上述用途还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述药物用于治疗或预防神经性疼痛、炎性疼痛、与癌症有关的疼痛、与骨折有关的疼痛、与手术有关的疼痛、炎性肺病、间质性膀胱炎、痛性膀胱综合征、炎性肠疾病、炎性皮肤病、雷诺氏综合征、特发性肺纤维化、瘢痕(肥大型、瘢痕瘤型和其他形式)、硬化、心内膜心肌纤维化、心房纤维化、骨髓纤维化、进行性块状纤维化(肺)、肾源性系统性纤维化、硬皮病、系统性硬化、关节纤维化、眼部纤维化、非小细胞肺癌、乳头状甲状腺癌、多形性成胶质细胞瘤、结肠直肠癌、黑色素瘤、胆管癌或肉瘤、急性骨髓性白血病、大细胞神经内分泌癌、成神经细胞瘤、前列腺癌、成神经细胞瘤、胰腺癌、黑色素瘤、头颈鳞状细胞癌或胃癌。
在本发明的第七方面，本发明提出了一种检测TrkA的试剂盒。根据本发明的实施例，所述试剂盒包括前面任一所述的抗体。前面所述的TrkA抗体能够特异性靶向结合TrkA，根据本发明实施例的试剂盒可以实现TrkA的特异性检测，如当抗体结合有荧光基团时，可以采用荧光检测装置实现对TrkA的定位或实时检测。
在本发明的第八方面，本发明提出了前面所述的抗体、前面所述的核酸分子、前面所述的表达载体或前面所述的重组细胞在制备试剂盒中的用途，所述试剂盒用于检测TrkA或者诊断TrkA相关的疾病。
在本发明的第九方面，本发明提出了一种小鼠B细胞。根据本发明的实施例，所述B细胞的基因组中携带编码恒定区的序列，所述恒定区序列与人IgG4的恒定区相比，存在S108P，F114A，L115A，R289K突变以及327K缺失突变。其中，人IgG4的恒定区具有SEQ ID NO:148所示的氨基酸序列。
ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQ ID NO:148)。
根据本发明实施例的小鼠B细胞可用于分泌鼠源抗体，所分泌的鼠源抗体在人体内的免疫原性进一步降低，稳定性得到显著提高，在体内的半衰期显著延长。
在本发明的第十方面，本发明提出了前面所述的小鼠B细胞在制备单克隆抗体中的用途。
附图说明
图1是根据本发明实施例的通过ELISA方法筛选得到的9株能产生分子水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的单克隆杂交瘤细胞株的实验结果图；
图2是根据本发明实施例的流式筛选得到的9株能产生细胞水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的单克隆杂交瘤细胞株的实验结果图，％parent(百分比)；
图3是根据本发明实施例的ELISA方法检测各阳性克隆上清产生的单克隆抗体与Mouse-TrKA受体的结合情况的实验结果图；
图4是根据本发明实施例的应用HEK-293T-TrkA细胞模型评价受试抗体的阻断活性的结果图，％parent(百分比)；
图5是根据本发明实施例的Tanezumab验证NIH-3T3-TrkA细胞模型是否可以用于评价抑制NGF-TrkA通路药物(受试药物)的体外活性的结果图；
图6是根据本发明实施例的通过AlphaLISA的方法检测受试抗体及阳性抗体MNAC13作用下，TrkA蛋白的酪氨酸磷酸化水平的实验结果图，p-TrkA level(TrkA蛋白的酪氨酸磷酸化水平)；
图7是根据本发明实施例的通过流式细胞术检测受试抗体亲和力EC50的结果图；
图8A～8G是根据本发明实施例的ELISA方法评价受试抗体与靶标TrKA结合的特异性的结果图；
图9是根据本发明实施例的应用ELISA方法检测受试抗体对Mouse-TrKA蛋白的结合能力的结果图；
图10是根据本发明实施例的应用流式细胞术检测受试抗体与Mouse-TrKA蛋白的结合能力的结果图；
图11是根据本发明实施例的应用ELISA方法检测受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用的结果图；
图12是根据本发明实施例的应用流式细胞术检测受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用的结果图；
图13是根据本发明实施例的应用ELISA方法检测受试抗体对Human-NGF和Human-TrKA结合的抑制作用的结果图；
图14是根据本发明实施例的应用福尔马林致痛模型评价受试抗体的体内镇痛活性的结果图；以及
图15是根据本发明实施例的应用完全弗氏佐剂诱导炎疼痛模型评价受试抗体的体内镇痛活性的结果图。
具体实施方式
下面详细描述本发明的实施例，所述实施例的示例在附图中示出，其中自始至终相同或类似的标号表示相同或类似的元件或具有相同或类似功能的元件。下面通过参考附图描述的实施例是示例性的，旨在用于解释本发明，而不能理解为对本发明的限制。
在对本发明描述的过程中，对于本文中有关的术语进行了解释和说明，这些解释和说明仅仅是为了方便对于方案的理解，并不能看做是对本发明保护方案的限制。
抗体
本文中，术语“抗体”是能够与特异性抗原结合的免疫球蛋白分子。包括两条分子量较轻的轻链和两条分子量较重的重链，重链(H链)和轻链(L链)由二硫键连接形成一个四肽链分子。其中，肽链的氨基端(N端)氨基酸序列变化很大，称为可变区(V区)，羧基端(C端)相对稳定，变化很小，称为恒定区(C区)。L链和H链的V区分别称为VL和VH。
在可变区中某些区域氨基酸组成和排列顺序具有更高的变化程度，称为高变区(Hypervariable region，HVR)，高变区为抗原和抗体结合的位置，因此也称为决定簇互补区(complementarity-determining region，CDR)。重链可变区和轻链可变区上均有三个CDR区。
本发明利用TrkA胞外段，通过免疫获得了高特异性的高亲和力的抗TrkA的Fab(antigen-binding fragment)抗体片段。利用该抗体片段能够与TrkA抗原特异性结合，从而可以靶向性治疗疼痛或肿瘤等疾病。
在一些实施方案中，本发明提供了一种能够特异性识别TrkA的抗体或者抗原结合片段，所述抗体含有选自下列至少之一的CDR序列或与其具有至少95％同一性的氨基酸序列：重链可变区CDR序列：SEQ ID NO:1～27，轻链可变区CDR序列：SEQ IN NO:28～54。在另一些实施方案中，本发明所提供的抗体或者抗原结合片段与上述重链和轻链相比，具有保守氨基酸取代。“抗原结合片段”是指保持特异性结合抗原(ROR2)能力的抗体片段。“保守氨基酸取代”指的是氨基酸被另一氨基酸发生生物学上、化学上或者结构上相似的残基所取代。生物学上相似的指的是该取代不破坏TrkA抗体或者与TrkA抗原的生物学活性。结构上相似指的是氨基酸具有相似长度的侧链，如丙氨酸、甘氨酸或丝氨酸，或具有相似大小的侧链。化学相似性指的是氨基酸具有相同的荷电或者都是亲水或者疏水的。例如疏水残基异亮氨酸、缬氨酸、亮氨酸或者甲硫氨酸相互取代。或者极性氨基酸相互取代，例如用精氨酸取代赖氨酸、谷氨酸取代天冬氨酸、谷氨酰胺取代天冬酰胺，丝氨酸取代苏氨酸等等。
在一些实施方案中，本发明提供了一种抗体或抗原结合片段，所述抗体或抗原结合片段具有SEQ ID NO：55～63任一项所示氨基酸序列的重链可变区和具有如SEQ ID NO：64～72任一项所示氨基酸序列的轻链可变区。发明人通过抗体序列比对数据库(NCBI、IMGT)可得到上述抗重链可变区序列的CDR区(如SEQ ID NO:1～27所示)和轻链可变区序列的CDR区(如SEQ ID NO:28～54所示)。在另一些实施方案中，所述抗体或抗原结合片段的重链可变区序列与SEQ ID NO:55～63所示氨基酸序列相比，具有保守氨基酸取代。在一些实施方案中，所述抗体或抗原结合片段的轻链可变区序列与SEQ ID NO:64～72任一项所示氨基酸序列相比，具有保守氨基酸取代。当然，这些保守氨基酸取代不会对抗体或者抗原结合片段的生物学功能带来改变。在一些具体方式中，这些保守氨基酸取代可以发生在重链可变区和轻链可变区中除了CDR区之外的氨基酸上。
在一些优选方案中，本发明提供了一种抗TrkA抗体，该抗体具有SEQ ID NO:76～84任一项所示氨基酸序列的重链和具有SEQ ID NO:85～93任一项所示氨基酸序列的轻链。
在一些优选方案中，本发明提供了一种抗TrkA单链抗体，该抗体具有SEQ ID NO:94～111所示的氨基酸序列。
核酸分子、表达载体、重组细胞
在制备或者获取这些抗体的过程中，可以利用表达这些抗体的核酸分子，与不同的载体连接，然后在不同细胞中表达，来获得相应抗体。
为此，本发明还提供了一种分离的核酸分子，所述核酸分子编码上述所述的抗体或抗原结合片段。
在一些实施方案中，所述分离核酸分子具有如SEQ ID NO:112～120任一项所示核苷酸序列或具有SEQ ID NO:121～129任一项所示核苷酸序列或具有SEQ ID NO:130～147所示的核苷酸序列.
在一些实施方案中，所述分离的核酸分子与上述SEQ ID NO:112～120所示的核苷酸序列至少具有90％以上的同源性，优选具有95％以上的同源性，更优选具有98％、99％以上的同源性。在至少一些实施方案中，所述分离的多核苷酸与所述SEQ ID NO:121～129所示的核苷酸序列至少具有90％以上的同源性，优选具有95％以上的同源性，更优选具有98％、99％以上的同源性。在至少一些实施方案中，所述分离的多核苷酸与所述SEQ ID NO:130～147所示的核苷酸序列至少具有90％以上的同源性，优选具有95％以上的同源性，更优选具有98％、99％以上的同源性。这些与SEQ ID NO:112～120或者SEQ ID NO:121～129或SEQ ID NO:130～147所示核苷酸序列具有同源性的序列，能够表达与SEQ ID NO:76～84和SEQ ID NO:85～93相似的氨基酸或与SEQ ID NO:94～111相似的氨基酸序列，从而能够与TrkA抗原特异性结合，实现抗体的靶向性功能。
在一些优选实施方式中，所述分离的核酸分子具有SEQ ID NO:112～120所示的重链核苷酸序列和SEQ ID NO:121～129所示的轻链核苷酸序列。这些核苷酸序列经过种属优化，更易在哺乳动物细胞中表达。
本发明还提供了一种表达载体，所述表达载体包含上述分离的核酸分子。在将上述分离的多核苷酸连接到载体上时，可以将多核苷酸与载体上的控制元件直接或者间接相连，只要这些控制元件能够控制多核苷酸的翻译和表达等即可。当然这些控制元件可以直接来自于载体本身，也可以是外源性的，即并非来自于载体本身。当然，多核苷酸与控制元件进行可操作地连接即可。本文中“可操作地连接”是指将外源基因连接到载体上，使得载体内的控制元件，例如转录控制序列和翻译控制序列等等，能够发挥其预期的调节外源基因的转录和翻译的功能。当然用来编码抗体重链和轻链的多核苷酸，可以分别独立的插入到不同的载体上，常见的是插入到同一载体上。常用的载体例如可以为质粒、噬菌体等等。例如Plasmid-X质粒。
本发明还提供了一种重组细胞，该重组细胞中包含有该表达载体。可以将表达载体导入到哺乳动物细胞中，构建获得重组细胞，然后利用这些重组细胞表达本发明提供的抗体或者抗原结合片段。通过该重组细胞进行培养，即可以获得相应抗体。这些可用的哺乳动物细胞例如可以为CHO细胞等。
药物组合物、试剂盒及制药用途和在制备试剂盒中的用途。
本发明还提供了一种药物组合物，所述药物组合物包括上述所述的抗体或者抗原结合片段和药学可接受的载体。
本文提供的抗TrkA抗体可以掺入适合受试者施用的药物组合物中。通常，这些药物组合物包括本文提供的抗TrkA抗体以及药学上可接受的载体。“药学上可接受的载体”可以包括生理学上相容的任何和所有溶剂、分散介质、包衣、抗细菌剂和抗真菌剂、等渗剂和延迟吸收剂等等。具体实例可以是水、盐水、磷酸盐缓冲盐水、葡萄糖、甘油、乙醇等以及它们的组合物中的一种或多种。有许多情况下，药物组合物中包括等渗剂，例如糖类、多元醇(如甘露醇、山梨醇)或氯化钠等。当然药学上可接受的载体还可包括微量的辅助物质，例如润湿剂或乳化剂、防腐剂或缓冲剂，用来延长抗体的保存限期或效力。
例如，本发明的抗体可掺入适用于胃肠外施用(例如静脉内、皮下、腹膜内、肌肉内)的药物组合物中。这些药物组合物可以被制备成各种形式。例如液体、半固体和固体剂型等，包括但不限于液体溶液(例如，注射溶液和输注溶液)、分散剂或悬浮剂、片剂、丸剂、粉末、脂质体和栓剂。典型的药物组合物为注射溶液或输注溶液形式。所述抗体可通过静脉输注或注射或肌肉内或皮下注射来施用。
当然，本文中的抗TrkA抗体还可以根据需要被制成试剂盒或者其他诊断性试剂的一部分。根据本发明的实施例，本发明还提供了一种试剂盒，所述试剂盒包括上述TrkA抗体。应用本发明提供的试剂盒，例如可以用于免疫印迹、免疫沉淀等涉及到利用TrkA抗原和抗体特异性结合性能，来检测的试剂盒等。这些试剂盒可包含下列中的任意一种或多种：拮抗剂、抗TrkA抗体或者药物参照材料；蛋白纯化柱；免疫球蛋白亲和纯化缓冲剂；细胞的测定稀释剂；说明书或者文献等。抗TrkA抗体可被用于不同类型的诊断测试，例如可以在体外或者体内检测各种各样的疾病或者药物、毒素或者其他蛋白等的存在。例如可以通过对受试者的血清或者血液进行检测，用来测试相关疾病。这种相关疾病可包括TrkA相关疾病，例如疼痛、癌症、炎症或炎性疾病、神经变性疾病、舍格伦氏综合征、子宫内膜异位、糖尿病性周围神经病变、前列腺炎、盆腔疼痛综合征、与骨重塑调节失衡相关的疾病以及由结缔组织生长因子异常信号传导引起的疾病等等。当然本文提供的抗体也可以用于上述疾病的放射免疫检测和放射免疫治疗等等。
具体地，上述疼痛、炎症或炎性疾病、神经变性疾病、舍格伦氏综合征、子宫内膜异位、糖尿病性周围神经病变、前列腺炎、盆腔疼痛综合征、与骨重塑调节失衡相关的疾病以及由结缔组织生长因子异常信号传导引起的疾病包括神经性疼痛、炎性疼痛、与癌症有关的疼痛、与骨折有关的疼痛、与手术有关的疼痛、炎性肺病、间质性膀胱炎、痛性膀胱综合征、炎性肠疾病、炎性皮肤病、雷诺氏综合征、特发性肺纤维化、瘢痕(肥大型、瘢痕瘤型和其他形式)、硬化、心内膜心肌纤维化、心房纤维化、骨髓纤维化、进行性块状纤维化(肺)、肾源性系统性纤维化、硬皮病、系统性硬化、关节纤维化、眼部纤维化。
这些癌症或者肿瘤可以是任何不受调控的细胞生长。具体地，可以是非小细胞肺癌、乳头状甲状腺癌、多形性成胶质细胞瘤、结肠直肠癌、黑色素瘤、胆管癌或肉瘤、急性骨髓性白血病、大细胞神经内分泌癌、前列腺癌、成神经细胞瘤、胰腺癌、黑色素瘤、头颈鳞状细胞癌或胃癌等等。
在利用本发明所提供的抗TrkA抗体治疗上述疾病时，可以将本发明提供的抗TrkA抗体提供给受试者即可。为此，本发明提供了一种用于治疗上述疾病的方法，包括向有需要的受试者施用本发明所提供的的抗体或其抗原结合片段。
下面将结合实施例对本发明的方案进行解释。本领域技术人员将会理解，下面的实施例仅用于说明本发明，而不应视为限定本发明的范围。实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
实施例1杂交瘤技术筛选抗TrkA单克隆抗体
通过杂交瘤技术和ELISA方法筛选可以产生分子水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的杂交瘤细胞株。采用NCBI数据库设计出TrkA蛋白胞外区编码基因，构建到哺乳动物真核表达系统中，表达得到TrkA蛋白胞外区，用于抗TrkA单克隆抗体筛选。免疫原TrkA蛋白胞外区来源于北京义翘神州生物科技有限公司，Balb/C小鼠三次腹腔注射免疫后，ELISA方法测定免疫小鼠血清抗体滴度，达到105级。脾脏注射加强免疫后，与骨髓瘤细胞融合。采用HAT选择培养基和HT选择培养基选择性培养筛选融合的杂交瘤细胞株，采用ELISA方法筛选能产生抗TrkA抗体的阳性杂交瘤细胞株，采用有限稀释法进行阳性杂交瘤细胞的克隆化培养，筛选稳定的能产生抗TrkA单克隆抗体的杂交瘤细胞株。经三次克隆化培养，四次ELISA检测后，共筛选得到147株阳性单克隆杂交瘤细胞株。将NGF生物素化，利用NGF可以结合TrkA蛋白胞外区，抗TrkA单克隆抗体也可以结合TrkA蛋白胞外区，设计竞争性实验，通过ELISA方法，检测抗TrkA单克隆抗体作用下，NGF与TrkA蛋白胞外区结合情况，筛选分子水平阻断NGF和TrkA结合的抗TrkA单克隆抗体。图1是147株阳性单克隆杂交瘤细胞株中能产生分子水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的9株杂交瘤细胞株的阻断实验结果，即经筛选共得到9株能产生分子水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的单克隆杂交瘤细胞株，实验结果见图1。图中，OD450反映了与TrkA蛋白胞外区结合的NGF信号，读值越高，表明与TrkA蛋白胞外区结合的NGF信号越强，抗体阻断NGF与TrkA结合的效果越差；如图所示，与阴性克隆组相比，9个阳性克隆组的ELISA读值显著性的降低，与TrkA蛋白胞外区结合的NGF信号显著性降低；可见，这9个阳性克隆能通过产生抗TrkA单克隆抗体阻断NGF与TrkA蛋白胞外区结合，降低与TrkA蛋白胞外区结合的NGF信号。
实施例2 HEK-293T-TrkA细胞模型筛选抗TrkA单克隆抗体
利用慢病毒技术构建HEK-293T-TrkA细胞模型，通过流式细胞术，筛选可以产生细胞水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的杂交瘤细胞株。将NGF生物素化，利用NGF可以结合HEK-293T-TrkA细胞上的TrkA蛋白胞外区，抗TrkA单克隆抗体也可以结合HEK-293T-TrkA细胞上的TrkA蛋白胞外区，设计竞争性实验，通过流式细胞术，检测抗TrkA单克隆抗体作用下，NGF与HEK-293T-TrkA细胞上TrkA蛋白胞外区结合情况，筛选细胞水平阻断NGF和TrkA结合的抗TrkA单克隆抗体。经筛选共得到9株能产生细胞水平阻断NGF和TrkA结合的抗TrkA单克隆抗体的单克隆杂交瘤细胞株，与分子水平阻断实验结果一致，实验结果见图2，图中，％parent值反映了与HEK-293T-TrKA细胞上TrkA蛋白胞外区结合的NGF信号，读值越高，表明与HEK-293T-TrKA细胞上TrkA蛋白胞外区结合的NGF信号越强，抗体阻断NGF与TrkA结合的效果越差；如图所示，与阴性克隆组相比，9个阳性克隆组的％parent值显著性的降低，与TrkA蛋白胞外区结合的NGF信号显著性降低；可见，这9个阳性克隆能通过产生抗TrkA单克隆抗体阻断NGF与HEK-293T-TrkA细胞上TrkA蛋白胞外区结合，降低与HEK-293T-TrkA细胞上TrkA蛋白胞外区结合的NGF信号。
实施例3应用ELISA方法检测阳性克隆产生单克隆抗体的人鼠交叉反应
通过ELISA方法检测各阳性克隆上清产生的单克隆抗体与Mouse-TrKA受体的结合情况，检测阳性克隆产生单克隆抗体的人鼠交叉反应，结果见图3，图中，OD450值反映抗体与蛋白的结合强弱，读值越大表示抗体与蛋白的结合越强。如图所示，23E12、21E5、15D4、27H3组的OD450值在3.5左右，与阴性对照组相比有显著性差异；20C8、1B9、4H4、3A5、22D12组的OD450值几乎接近于0，与阴性克隆组相比没有显著性差异。可见，克隆23E12、21E5、15D4、27H3能产生与Human-TrKA和Mouse-TrKA都结合的单克隆抗体，存在人鼠交叉反应。
实施例4载体的构建
采用分子克隆的方法构建一系列(20C8、23E12、27H3、21E5、2A5、4H4、1B9、22D12、15D4)嵌合抗体表达载体，在CHO表达系统中，重组表达嵌合抗体。编码一系列(20C8、23E12、27H3、21E5、2A5、4H4、1B9、22D12、15D4)嵌合单克隆抗体轻重链的核苷酸序列是委托金唯智生物科技有限公司通过化学合成获得的，所获得的序列经双酶切后，插入到真核表达载体的相同酶切位点间，构建一系列(20C8、23E12、27H3、21E5、2A5、4H4、1B9、22D12、15D4)嵌合抗体表达载体。然后采用Invitrogen质粒提取试剂盒提取一系列经验证正确的表达载体，并用限制性内切酶进行线性化后纯化回收，-20℃保藏。
实施例5编码一系列嵌合抗体载体转染并在细胞中表达
将CHO宿主细胞用CD CHO培养基复苏培养后，当细胞密度约8*105cell/mL时收集细胞进行转染。转染细胞约1*107cell，载体约40μg，通过电击方法转染(Bio-Rad，Genepulser Xcell)。电击后细胞于20mL CD CHO培养基中培养。培养第二天，离心收集细胞，并在加入MSX至终浓度50μM的20mL CD CHO培养基中重悬培养。当细胞密度约0.6*106cell/mL时，对获得的混合克隆株用CD CHO培养基进行传代，传代细胞密度约0.2*106cell/mL。当细胞存活率约90％时，收集细胞培养液。
实施例6收集细胞发酵培养液纯化嵌合抗体
对一系列的嵌合抗体进行翻译水平上的检测。采用Protein A层析柱对收集的细胞培养液进行纯化，并收集吸收峰，进行质谱检测，质谱检测一系列嵌合抗体分子量150KD左右，与理论分子量一致。同时对收集的样品经还原与非还原后通过10％SDS-PAGE电泳检测，还原型SDS-PAGE电泳图谱显示二条带，分别在25KD和50KD左右，非还原型SDS-PAGE电泳图谱显示单一条带，在150KD左右，电泳图谱条带大小与理论一致。纯化后样品采用pH7.4的0.02M PBS缓冲液在4℃透析过夜。
在以下实施例7～16中，发明人评价了所构建纯化获得的嵌合抗体20C8、23E12、27H3、21E5、2A5、4H4、1B9、22D12、15D4的与TrkA的亲和力和结合TrkA、阻断NGF与TrkA蛋白胞外区结合的活性。
实施例7应用HEK-293T-TrkA细胞模型评价受试抗体的阻断活性
将NGF生物素化，利用NGF可以结合HEK-293T-TrkA细胞上的TrkA蛋白胞外区，抗TrkA单克隆抗体也可以结合HEK-293T-TrkA细胞上的TrkA蛋白胞外区，设计竞争性实验，通过流式细胞术，检测不同浓度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)受试抗体作用下，NGF与HEK-293T-TrkA细胞上TrkA蛋白胞外区结合情况，研究受试抗体对NGF和TrKA结合的抑制作用。实验结果见图4。图中，parent％值反映了与HEK-293T-TrkA细胞上TrKA蛋白胞外区结合的NGF信号，读值越低，与HEK-293T-TrkA细胞上TrkA蛋白胞外区结合的NGF信号越弱，抗体抑制NGF与TrKA结合的作用越大；如图所示，随着受试抗体浓度的增加，parent％值逐渐降低，直到趋近于零，即与TrkA蛋白胞外区结合的NGF信号逐渐降低，直到没有NGF结合TrkA蛋白胞外区，NGF与TrkA的结合全部抑制。可见，一定的浓度范围内，各受试抗体在细胞水平上能够剂量依赖性的抑制NGF与TrkA的结合。
实施例8应用NIH-3T3-TrkA细胞模型评价受试抗体的体外活性
在NGF刺激下，NIH-3T3-TrkA细胞膜上的TrkA蛋白酪氨酸磷酸化水平上调，TrkA下游信号通路激活。受试抗体可以结合NIH-3T3-TrkA细胞膜表面的TrkA蛋白，抑制NGF的刺激，下调TrkA蛋白酪氨酸磷酸化水平。试验中，以IgG4为阴性对照(该抗体不结合NGF及TrkA)，tanezumab(抗NGF单克隆抗体，轻链氨基酸序列如SEQ ID NO:149所示，重链氨基酸序列如SEQID NO:150所示，抑制NGF-TrkA通路药物)、MNAC13(抗TrKA单克隆抗体，轻链氨基酸序列如SEQ ID NO:151所示，重链氨基酸序列如SEQID NO:152所示，抑制NGF-TrkA通路药物)为阳性对照，其中，Tanezumab用于验证NIH-3T3-TrkA细胞模型是否可以用于评价抑制NGF-TrkA通路药物(受试药物)的体外活性，实验数据如图5所示。通过AlphaLISA的方法检测不同浓度受试抗体及阳性抗体MNAC13作用下，TrkA蛋白的酪氨酸磷酸化水平下调情况，衡量受试抗体的体外活性，p-TrkA检测结果见图6，实验结果显示，受试抗体及阳性抗体MNAC13均能抑制NGF-TrKA通路，剂量依赖性地下调TrkA蛋白酪氨酸磷酸化水平，其中，受试抗体23E12、20C8、27H3、21E5、1B9、4H4的半数抑制浓度IC50小于阳性抗体MNAC13。可见，受试抗体23E12、20C8、27H3、21E5、1B9、4H4的体外活性优于阳性抗体MNAC13。
DIQMTQSPSSLSASVGDRVTITCRASQSISNNLNWYQQKPGKAPKLLIYYTSRFHSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQEHTLPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:149)。
QVQLQESGPGLVKPSETLSLTCTVSGFSLIGYDLNWIRQPPGKGLEWIGIIWGDGTTDYNSAVKSRVTISKDTSKNQFSLKLSSVTAADTAVYYCARGGYWYATSYYFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO:150)。
DIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGQAPKLLIYTTSNLASGVPSRFSGSGSGTDYTLTISSLQPEDVATYYCHQWSSYPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:151)。
EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYTMSWARQAPGKGLEWVAYISKGGGSTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDSAVYYCARGAMFGNDFFFPMDRWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG(SEQ ID NO:152)。
实施例9应用Fortebio方法评价受试抗体与TrkA的亲和力
将受试抗体(23E12、20C8、21E5、27H3、1B9、4H4、2A5)样品分别用缓冲液(PBS缓冲液100ml，加入0.1gBSA搅拌至充分溶解，再加入20μL的吐温20，混合均匀)稀释至4个浓度梯度(80nM、26.67nM、8.89nM、2.93nM)，分别加入到样品板的各孔中，通过亲和力检测系统(OCTET RED 96 SYSTEM)，用带有Ni-NTA传感器(厂家：PALL，货号：18-5101)结合带有组氨酸标签的TRKA(50μg/ml，75kDa，HEC实验室提供)蛋白后，自动检测其与各受试抗体的结合情况，结果见表1。KD值是抗体与其抗原之间的平衡解离常数，KD值与亲和力成反比。KD值与抗体的浓度(特定实验所需的抗体量)有关，KD值越低(浓度越低)，抗体的亲和力越高。通常认为，高亲和力抗体处于低纳摩尔范围内(10-9)。表1结果显示，受试抗体(23E12、20C8、21E5、27H3、1B9、4H4、2A5)的KD值处于低纳摩尔范围内(10-9)，表明，受试抗体都具有很高的亲和力。
表1：
Figure BDA0002512640120000441
Figure BDA0002512640120000451
实施例10应用流式细胞术评价受试抗体的亲和力
将受试抗体(23E12、20C8、21E5、27H3、1B9、4H4、2A5)样品分别用PBS缓冲液稀释至11个浓度梯度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)，通过流式细胞术检测各浓度梯度受试抗体与HEK-293T-TrKA细胞表面TrKA受体的结合情况，从细胞水平评价受试抗体的亲和力，结果见图7。图中，EC50(半数结合浓度)值反映抗体的亲和力高低，EC50值越小，抗体的亲和力越高，通常认为，高亲和力抗体的EC50值低于1.5μg/mL。图中结果显示，受试抗体23E12、20C8、21E5、27H3、1B9、4H4、2A5的EC50值均低于1.5μg/mL，表明，受试抗体都具有很高的亲和力。
实施例11应用ELISA方法评价受试抗体与靶标TrKA结合的特异性
TrkA受体家族属于受体酪氨酸激酶(RTKs)，包括TrkA、TrkB和TrkC，它们具有很高的同源性。TrkA是神经生长因子(NGF)的受体酪氨酸激酶，选择性结合NGF，是NGF的功能性受体。除高亲和力受体TrkA外，NGF还可与其低亲和力受体p75结合。试验中，通过ELISA方法检测不同浓度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)受试抗体分别与TrKA、TrKB、TrKC、P75的结合情况，评价受试抗体与靶标TrKA结合的特异性，结果如图8A～8G所示。图中，一定的抗体浓度下，OD450值反映抗体与蛋白的结合强弱，读值越大抗体与蛋白的结合越强。实验结果显示，受试抗体23E12、20C8、27H3、21E5、1B9、4H4、2A5与TrKA受体都有很好的结合(受试抗体浓度由0μg/mL增加到20μg/mL，OD450值逐渐增加直至趋近稳定，接近3左右)，与TrKB、TrKC、P75基本不结合(各受试抗体浓度由0μg/mL增加到20μg/mL，OD450值基本没有变化，接近于0)。可见，受试抗体与靶标TrKA结合的特异性都很好。
实施例12应用ELISA方法检测受试抗体与Mouse-TrKA蛋白的结合能力
将受试抗体(23E12、21E5)样品分别用PBS缓冲液稀释至11个浓度梯度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)，通过ELISA方法检测各浓度梯度受试抗体与Mouse-TrKA受体的结合情况，检测受试抗体与Mouse-TrKA蛋白的结合能力，结果见图9，图中，一定的抗体浓度下，OD450值反映抗体与蛋白的结合强弱，读值越大抗体与蛋白的结合越强。实验结果显示，受试抗体的浓度由0μg/mL增加到20μg/mL，OD450值逐渐增加直至趋近稳定，接近3.5左右。可见，受试抗体23E12、21E5和Mouse-TrKA蛋白都有很好的结合能力。
实施例13应用流式细胞术检测受试抗体与Mouse-TrKA蛋白的结合能力
将受试抗体23E12样品分别用PBS缓冲液稀释至11个浓度梯度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)，通过流式细胞术检测各浓度梯度受试抗体与HEK293T-Mouse-TrKA细胞表面Mouse-TrKA受体的结合情况，检测受试抗体与Mouse-TrKA蛋白的结合能力，结果见图10。图中，EC50(半数结合浓度)值反映抗体的结合能力，EC50值越小，抗体的结合能力越强，实验结果显示，受试抗体的浓度由0μg/mL增加到20μg/mL，％Parent值逐渐增加直至趋近稳定，EC50值＝0.08012μg/mL。可见，受试抗体23E12与HEK293T-Mouse-TrKA细胞表面Mouse-TrKA受体有很好的结合能力。
实施例14应用ELISA方法检测受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用
试验中，以IgG4为阴性对照(该抗体不结合Mouse-NGF及Mouse-TrkA)，将Mouse-NGF生物素化，利用Mouse-NGF可以结合Mouse-TrkA蛋白，抗Mouse-TrkA单克隆抗体也可以结合Mouse-TrkA蛋白，设计竞争性实验，通过ELISA方法，检测不同浓度(2.5μg/mL，0.25μg/mL)受试抗体(23E12，21E5)作用下，Mouse-NGF与Mouse-TrkA蛋白结合情况，研究受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用。实验结果见图11。图中，OD450值反映了与Mouse-TrkA结合的Mouse-NGF信号。读值越低，与Mouse-TrkA结合的Mouse-NGF信号越弱，抗体抑制Mouse-NGF与Mouse-TrKA结合的作用越大；如图所示，与阴性对照组相比，不同浓度(2.5μg/mL，0.25μg/mL)受试抗体(23E12，21E5)作用下，与Mouse-TrkA结合的Mouse-NGF信号显著性降低。可见，受试抗体23E12、21E5能够抑制Mouse-NGF和Mouse-TrKA结合。
实施例15应用流式细胞术检测受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用
将Mouse-NGF生物素化，利用Mouse-NGF可以结合HEK293T-Mouse-TrkA细胞上的Mouse-TrkA蛋白胞外区，抗Mouse-TrkA单克隆抗体也可以结合HEK293T-Mouse-TrkA细胞上的Mouse-TrkA蛋白胞外区，设计竞争性实验，通过流式细胞术，检测不同浓度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)受试抗体作用下，Mouse-NGF与HEK-293T-Mouse-TrkA细胞上Mouse-TrkA蛋白胞外区结合情况，研究受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用。实验结果见图12。图中，parent％值反映了与HEK293T-Mouse-TrkA细胞上Mouse-TrKA蛋白胞外区结合的Mouse-NGF信号，读值越低，与HEK293T-Mouse-TrkA细胞上Mouse-TrkA蛋白胞外区结合的Mouse-NGF信号越弱，抗体抑制Mouse-NGF与Mouse-TrKA结合的作用越大；如图所示，随着受试抗体浓度的增加，％parent值逐渐降低，直到趋近于零，即与Mouse-TrkA蛋白胞外区结合的Mouse-NGF信号逐渐降低，直到没有Mouse-NGF结合Mouse-TrkA蛋白胞外区，Mouse-NGF与Mouse-TrkA的结合全部抑制，IC50＝0.05147μg/mL。可见，一定的浓度范围内，受试抗体23E12在细胞水平上能够剂量依赖性的抑制Mouse-NGF与Mouse-TrkA的结合。
实施例16应用ELISA方法检测受试抗体对Human-NGF和Human-TrKA结合的抑制作用
将Mouse-NGF生物素化，利用Mouse-NGF可以结合Mouse-TrkA蛋白，抗Mouse-TrkA单克隆抗体也可以结合Mouse-TrkA蛋白，设计竞争性实验，通过Elisa方法，检测不同浓度(20μg/mL，10μg/mL，5μg/mL，2.5μg/mL，1.25μg/mL，0.625μg/mL，0.313μg/mL，0.156μg/mL，0.078μg/mL，0.039μg/mL，0.019μg/mL)受试抗体作用下，Mouse-NGF与Mouse-TrkA蛋白结合情况，研究受试抗体对Mouse-NGF和Mouse-TrKA结合的抑制作用。实验结果见图13。图中，OD450值反映了与Mouse-TrkA结合的Mouse-NGF信号。读值越低，与Mouse-TrkA结合的Mouse-NGF信号越弱，抗体抑制Mouse-NGF与Mouse-TrKA结合的作用越大；如图所示，随着受试抗体浓度的增加，OD450值逐渐降低，直到趋近于零，即与Mouse-TrkA蛋白胞外区结合的Mouse-NGF信号逐渐降低，直到几乎没有Mouse-NGF结合Mouse-TrkA蛋白胞外区，Mouse-NGF与Mouse-TrkA的结合几乎完全抑制。可见，一定的浓度范围内，受试抗体23E12、20C8、21E5、27H3、4H4、2A5在分子水平上能够剂量依赖性的抑制NGF与TrkA的结合。
实施例17应用福尔马林致痛模型评价受试抗体的体内镇痛活性
福尔马林炎症痛模型是通过注射福尔马林来产生持续而非短暂的疼痛刺激及应答的验证痛模型。该模型产生两相疼痛，分别是第I相化学刺激疼痛和第II相炎症疼痛，造成的疼痛应答具有可重复性、可计量性，是临床前疼痛研究最佳的模型之一，且被广泛应用于评估不同药物的镇痛效果。试验中，选用6-8周雄性ICR小鼠，造模前根据体重随机分为6组：模型组(皮下注射生理盐水)、Tanezumab 60μg/mouse剂量组、20C8 60μg/mouse剂量组、21E5 60μg/mouse剂量组、23E12 15μg/mouse剂量组、23E12 60μg/mouse剂量组，每组10只。药物经皮下给药，18个小时后于小鼠右后爪足背皮下注射2.5％福尔马林溶液15μL，观察其后45分钟内小鼠抬脚次数，记1-15min为第I相疼痛，16-45min为第II相疼痛，一般而言，第I相疼痛反映的是急性疼痛，第II相疼痛反映的是慢性疼痛。结果见图14(图中数据展示为Mean±SEM,n＝10/组，*p<0.05与溶剂模型组比较，使用单因素方差分析附加LSD多重比较检验)，图中抬脚次数反映小鼠造模后的疼痛强度，抬脚次数越少，疼痛越弱。结果显示，与模型组相比，23E12 60μg/mouse剂量组能显著减少小鼠第II相疼痛的抬脚次数，其它剂量组未能降低小鼠造模后的抬脚次数。结论：在福尔马林致痛模型中，皮下注射60μg/mouse的受试抗体23E12对于慢性疼痛有效，而阳性药Tanezumab未体现镇痛效果，可能对福尔马林致痛模型不敏感。
实施例18应用完全弗氏佐剂诱导炎症疼痛模型评价受试抗体的体内镇痛活性
完全弗氏佐剂诱导炎症痛模型通过在小鼠掌心注射完全弗氏佐剂来产生类似骨关节炎症的慢性炎症疼痛刺激及应答的疼痛模型，通过机械痛测试对疼痛测量，机械刺激力度越大，说明动物对疼痛的耐受性越强。试验中，选用8周龄雄性ICR小鼠，在造模前根据疼痛敏感随机分为6组(15只/组)：对照组(小鼠左后肢足底注射生理盐水25μl+皮下注射生理盐水)，模型组(小鼠左后肢足底注射25μl CFA诱导炎性痛+皮下注射生理盐水)、Tanezumab 60μg/mouse剂量组，23E12 60μg/mouse剂量组，在造模后第4天皮下注射药物，给药后36hr进行机械痛测试。结果见图15(图中数据展示为Mean±SEM，n＝15/组，#p＜0.05，##p＜0.01与空白对照组比较，*p＜0.05，**p＜0.01与模型组比较，使用非参数统计分析附加独立样本T检验)，纵坐标表示机械刺激力度，小鼠爪子撤离的压力阈值越大，镇痛效果越好。结果显示：与模型组比较，皮下注射60μg/mouse 23E12、60μg/mouse tanezumab能够显著增加小鼠爪子撤离的压力阈值(P＜0.01；P＜0.05)，呈现镇痛作用，两者镇痛效果作用效果相当。结论：皮下注射60μg/mouse的受试抗体23E12，在完全弗氏佐剂诱导的炎症痛模型中呈现镇痛作用。
实施例19受试抗体毒理实验
在本实施例中，发明人所要进行的毒性测试如下表2所示。
表2：
Figure BDA0002512640120000481
Figure BDA0002512640120000491
其中，上述实验过程和实验结论如下所述：
毒理实验：
单次给药毒性
SD大鼠、食蟹猴，观察是否出现靶器官毒性。观察相关的症状，对体重、摄食、检眼镜检查、心电图、血液学、临床生化、尿液、脏器重量的影响，提供最大耐受剂量MTD，足够的安全系数。结果显示：无动物死亡，在器官和组织上观察到弥散型炎症浸润，但是无论临床病理还是组织学分析，都没有显示出靶器官毒性。对体重、摄食、检眼镜检查、心电图、血液学、临床生化、尿液、脏器重量无显著影响。大体解剖无显著异常。
重复给药毒性：SD大鼠、食蟹猴，一周一次静脉注射药物，连续4周，观察临床病理学和组织学确定是否存在靶器官毒性NOAEL，观察对体重、摄食、检眼镜检查、心电图、血液学、临床生化、尿液、脏器重量的影响，免疫原性(ADA(抗药抗体)分析方法)，免疫毒性研究(评价内容包括：血液学白细胞分类计数(包括巨噬细胞)、临床化学(球蛋白和白蛋白：球蛋白比值))，器官质量(胸腺、脾脏、淋巴结)和组织病理学(淋巴器官和组织)计算安全窗。结果显示：无动物死亡，在器官和组织上观察到弥散型炎症浸润，但是无论临床病理还是组织学分析，都没有显示出靶器官毒性。没有出现纳武单抗相关的症状，对体重、摄食、检眼镜检查、心电图、血液学、临床生化、尿液、脏器重量无显著影响。大体解剖无显著异常。
一般安全药理学：SD大鼠单次静脉注射药物，观察药物对中枢神经系统、心血管系统和呼吸系统的影响。结果显示：药物对中枢神经系统、心血管系统和呼吸系统的无影响。
局部给药安全性实验(血管刺激性实验和体外溶血实验)：新西兰大白兔静脉注射药物，检验药物对血管和血液是否有影响。结果显示：无影响。
高致敏性试验(豚鼠全身主动过敏实验)：豚鼠单次静脉注射药物，观察其是否有致敏作用。结果显示：无致敏作用。
在本说明书的描述中，参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不必须针对的是相同的实施例或示例。而且，描述的具体特征、结构、材料或者特点可以在任一个或多个实施例或示例中以合适的方式结合。此外，在不相互矛盾的情况下，本领域的技术人员可以将本说明书中描述的不同实施例或示例以及不同实施例或示例的特征进行结合和组合。
尽管上面已经示出和描述了本发明的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本发明的限制，本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
SEQUENCE LISTING
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Ala Gln Asn Leu Glu Phe Pro Phe Thr
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<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 40
Ser Ser Val Ser Tyr
1 5
<210> 41
<211> 3
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 41
Leu Thr Ser
1
<210> 42
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 42
Gln Gln Trp Ser Ser Asn Pro Pro Thr
1 5
<210> 43
<211> 5
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 43
Ser Ser Val Ser Tyr
1 5
<210> 44
<211> 3
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 44
Asp Thr Ser
1
<210> 45
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 45
Gln Gln Trp Ser Ser Asn Pro Pro Thr
1 5
<210> 46
<211> 5
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 46
Ser Ser Val Ser Tyr
1 5
<210> 47
<211> 3
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 47
Ala Thr Ser
1
<210> 48
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 48
Gln Gln Trp Ser Ser Asn Pro Pro Thr
1 5
<210> 49
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 49
Lys Ser Val Ser Ser Ser Ala Tyr Ser Tyr
1 5 10
<210> 50
<211> 3
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 50
Leu Ala Ser
1
<210> 51
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 51
Gln His Ser Arg Glu Leu Pro Phe Thr
1 5
<210> 52
<211> 5
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 52
Ser Asn Ile Ser Tyr
1 5
<210> 53
<211> 3
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 53
Asp Thr Ser
1
<210> 54
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区CDR序列
<400> 54
Gln Gln Trp Ser Ser Val Pro Leu Thr
1 5
<210> 55
<211> 121
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 55
Glu Val Leu Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Val Tyr Ser Phe Thr Ala Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro His Asn Gly Gly Thr Arg Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Leu Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Asp Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ile Ser Arg Tyr Gly Ser Ser Ser Phe Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Ala Val Ser Ser
115 120
<210> 56
<211> 116
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 56
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Trp Leu Gly Trp Met Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Asp Phe Tyr Pro Arg Thr Gly Asn Thr Phe Tyr Asn Glu Asn Phe
50 55 60
Lys Gly Lys Val Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Leu Cys
85 90 95
Ala Arg Ala Gly Thr Gly Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu
100 105 110
Thr Val Ser Ser
115
<210> 57
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 57
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Thr Thr Gly Tyr Arg Phe Ser Ser Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu
35 40 45
Gly Glu Ile Leu Pro Gly Arg Gly Ile Ile Asn Tyr Asn Glu Asn Phe
50 55 60
Arg Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Val Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Thr Asp Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Thr
100 105 110
Val Thr Val Ser Ser
115
<210> 58
<211> 118
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 58
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Thr Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Glu Asn Leu Pro Gly Arg His Ile Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Arg Gly Thr Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Pro Leu Thr Val Ser Ser
115
<210> 59
<211> 121
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 59
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Phe Ser Tyr Tyr Gly Ser Arg Gly Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Thr Leu Thr Val Ser Ser
115 120
<210> 60
<211> 122
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 60
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Arg Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Ser Ser Tyr Arg Asn Asp Gly Asn Trp Tyr Phe Asp Val Trp
100 105 110
Gly Ala Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 61
<211> 121
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 61
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Ile Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Val Asn Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Lys Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ile Ser Arg Tyr Gly Ser Glu Ser Trp Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 62
<211> 118
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 62
Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Asn Ile Tyr
20 25 30
Trp Ile Asp Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Glu Ile Leu Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Asn Phe
50 55 60
Lys Gly Lys Ala Thr Met Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Thr Asp Gly Arg Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Thr Leu Thr Val Ser Ser
115
<210> 63
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链可变区
<400> 63
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu
35 40 45
Gly Glu Phe Leu Pro Arg Ser Gly Lys Thr Asn Tyr Asn Glu Glu Phe
50 55 60
Arg Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Thr Asp Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Met
100 105 110
Val Ala Val Ser Ser
115
<210> 64
<211> 106
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 64
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Ser
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Gly Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Asn Leu Glu Ile Lys
100 105
<210> 65
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 65
Ser Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met Ser Val Gly
1 5 10 15
Glu Arg Val Thr Leu Ser Cys Lys Ala Ser Glu Asn Val Gly Gly Tyr
20 25 30
Val Ser Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Ser Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Glu Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Ala Tyr His Cys Gly Gln Asn Tyr Ile Tyr Pro Phe
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 66
<211> 109
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 66
Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Ile Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly
35 40 45
Leu Ile Gly Gly Thr Asn Asn Arg Pro Pro Gly Val Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala
65 70 75 80
Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Val Leu Trp Tyr Ser Asn
85 90 95
His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 67
<211> 112
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 67
Asp Ile Val Met Thr Gln Ala Ala Phe Ser Thr Pro Val Thr Leu Gly
1 5 10 15
Thr Ser Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
20 25 30
Asn Gly Ile Thr Tyr Leu Tyr Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu Ala Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Ser Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Asn
85 90 95
Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110
<210> 68
<211> 106
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 68
Gln Ile Val Leu Thr Gln Ser Pro Ala Leu Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Thr Ser Ser Val Ser Tyr Ile
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Arg Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 69
<211> 106
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 69
Asp Ile Val Met Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr
35 40 45
Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Thr Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys
100 105
<210> 70
<211> 106
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 70
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Thr Ser Ser Val Ser Tyr Met
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Lys Lys
100 105
<210> 71
<211> 111
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 71
Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Val Val Ser Leu Gly
1 5 10 15
Gln Arg Ala Thr Ile Ser Cys Arg Thr Ser Lys Ser Val Ser Ser Ser
20 25 30
Ala Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Val Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His
65 70 75 80
Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110
<210> 72
<211> 106
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链可变区
<400> 72
Gln Ile Val Leu Thr Gln Ser Pro Pro Ile Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Asn Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr
35 40 45
Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Val Pro Leu Thr
85 90 95
Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 73
<211> 229
<212> PRT
<213> Artificial Sequence
<220>
<223> 人源IgG4野生型Fc序列
<400> 73
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe
1 5 10 15
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
20 25 30
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
35 40 45
Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val
50 55 60
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
65 70 75 80
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
85 90 95
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser
100 105 110
Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
115 120 125
Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln
130 135 140
Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
145 150 155 160
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
165 170 175
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
180 185 190
Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser
195 200 205
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
210 215 220
Leu Ser Leu Gly Lys
225
<210> 74
<211> 326
<212> PRT
<213> Artificial Sequence
<220>
<223> 抗体恒定区的全长序列
<400> 74
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
1 5 10 15
Ser Thr Ser Glu Ser 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 Lys Thr
65 70 75 80
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro
100 105 110
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
115 120 125
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
130 135 140
Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
145 150 155 160
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
180 185 190
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu
195 200 205
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
225 230 235 240
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
245 250 255
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
260 265 270
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
275 280 285
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
290 295 300
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
305 310 315 320
Leu Ser Leu Ser Leu Gly
325
<210> 75
<211> 107
<212> PRT
<213> Artificial Sequence
<220>
<223> 抗体恒定区的全长序列
<400> 75
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
<210> 76
<211> 447
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 76
Glu Val Leu Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Val Tyr Ser Phe Thr Ala Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro His Asn Gly Gly Thr Arg Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Leu Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Asp Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ile Ser Arg Tyr Gly Ser Ser Ser Phe Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Ala Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly
210 215 220
Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 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 Gln Glu Asp Pro
260 265 270
Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Phe 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 Gly Leu Pro Ser Ser 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 Gln 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 Glu 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 Leu Gly
435 440 445
<210> 77
<211> 442
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 77
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Trp Leu Gly Trp Met Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Asp Phe Tyr Pro Arg Thr Gly Asn Thr Phe Tyr Asn Glu Asn Phe
50 55 60
Lys Gly Lys Val Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Leu Cys
85 90 95
Ala Arg Ala Gly Thr Gly Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro
210 215 220
Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro
225 230 235 240
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
245 250 255
Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn
260 265 270
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
275 280 285
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
290 295 300
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
305 310 315 320
Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys
325 330 335
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu
340 345 350
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
355 360 365
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
370 375 380
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
385 390 395 400
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly
405 410 415
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
420 425 430
Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
435 440
<210> 78
<211> 443
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 78
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Thr Thr Gly Tyr Arg Phe Ser Ser Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu
35 40 45
Gly Glu Ile Leu Pro Gly Arg Gly Ile Ile Asn Tyr Asn Glu Asn Phe
50 55 60
Arg Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Val Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Thr Asp Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Thr
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro
210 215 220
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
245 250 255
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
260 265 270
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
275 280 285
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
290 295 300
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
305 310 315 320
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
325 330 335
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
340 345 350
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
355 360 365
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
370 375 380
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
385 390 395 400
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
405 410 415
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
420 425 430
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
435 440
<210> 79
<211> 444
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 79
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Thr Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Glu Asn Leu Pro Gly Arg His Ile Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Arg Gly Thr Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Pro Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys
210 215 220
Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu
225 230 235 240
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
245 250 255
Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln
260 265 270
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
275 280 285
Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu
290 295 300
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
305 310 315 320
Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys
325 330 335
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
340 345 350
Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
355 360 365
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
370 375 380
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
385 390 395 400
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
405 410 415
Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
420 425 430
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
435 440
<210> 80
<211> 447
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 80
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Phe Ser Tyr Tyr Gly Ser Arg Gly Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly
210 215 220
Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 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 Gln Glu Asp Pro
260 265 270
Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Phe 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 Gly Leu Pro Ser Ser 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 Gln 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 Glu 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 Leu Gly
435 440 445
<210> 81
<211> 448
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 81
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Arg Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Ser Ser Tyr Arg Asn Asp Gly Asn Trp Tyr Phe Asp Val Trp
100 105 110
Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr
210 215 220
Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 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 Gln Glu Asp
260 265 270
Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 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 Gly Leu Pro Ser Ser 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 Gln 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 Glu 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 Leu Gly
435 440 445
<210> 82
<211> 447
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 82
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Ile Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Val Asn Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Lys Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ile Ser Arg Tyr Gly Ser Glu Ser Trp Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly
210 215 220
Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 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 Gln Glu Asp Pro
260 265 270
Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Phe 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 Gly Leu Pro Ser Ser 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 Gln 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 Glu 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 Leu Gly
435 440 445
<210> 83
<211> 444
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 83
Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Asn Ile Tyr
20 25 30
Trp Ile Asp Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Glu Ile Leu Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Asn Phe
50 55 60
Lys Gly Lys Ala Thr Met Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Thr Asp Gly Arg Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys
210 215 220
Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu
225 230 235 240
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
245 250 255
Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln
260 265 270
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
275 280 285
Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu
290 295 300
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
305 310 315 320
Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys
325 330 335
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
340 345 350
Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
355 360 365
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
370 375 380
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
385 390 395 400
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
405 410 415
Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
420 425 430
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
435 440
<210> 84
<211> 443
<212> PRT
<213> Artificial Sequence
<220>
<223> 重链
<400> 84
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu
35 40 45
Gly Glu Phe Leu Pro Arg Ser Gly Lys Thr Asn Tyr Asn Glu Glu Phe
50 55 60
Arg Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Thr Asp Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Met
100 105 110
Val Ala Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro
210 215 220
Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
245 250 255
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
260 265 270
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
275 280 285
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
290 295 300
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
305 310 315 320
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
325 330 335
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
340 345 350
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
355 360 365
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
370 375 380
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
385 390 395 400
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
405 410 415
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
420 425 430
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
435 440
<210> 85
<211> 213
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 85
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Ser
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Gly Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Asn 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> 86
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 86
Ser Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met Ser Val Gly
1 5 10 15
Glu Arg Val Thr Leu Ser Cys Lys Ala Ser Glu Asn Val Gly Gly Tyr
20 25 30
Val Ser Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Ser Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Glu Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Ala Tyr His Cys Gly Gln Asn Tyr Ile Tyr Pro Phe
85 90 95
Thr Phe Gly Gly 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> 87
<211> 216
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 87
Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Ile Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly
35 40 45
Leu Ile Gly Gly Thr Asn Asn Arg Pro Pro Gly Val Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala
65 70 75 80
Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Val Leu Trp Tyr Ser Asn
85 90 95
His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr Val
100 105 110
Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys
115 120 125
Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg
130 135 140
Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
145 150 155 160
Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser
165 170 175
Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys
180 185 190
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
195 200 205
Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 88
<211> 219
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 88
Asp Ile Val Met Thr Gln Ala Ala Phe Ser Thr Pro Val Thr Leu Gly
1 5 10 15
Thr Ser Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
20 25 30
Asn Gly Ile Thr Tyr Leu Tyr Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu Ala Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Ser Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Asn
85 90 95
Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
130 135 140
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
145 150 155 160
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
165 170 175
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
180 185 190
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
195 200 205
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 89
<211> 213
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 89
Gln Ile Val Leu Thr Gln Ser Pro Ala Leu Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Thr Ser Ser Val Ser Tyr Ile
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Arg Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly 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> 90
<211> 213
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 90
Asp Ile Val Met Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr
35 40 45
Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Thr Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Leu 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> 91
<211> 213
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 91
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Thr Ser Ser Val Ser Tyr Met
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Lys 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> 92
<211> 218
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 92
Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Val Val Ser Leu Gly
1 5 10 15
Gln Arg Ala Thr Ile Ser Cys Arg Thr Ser Lys Ser Val Ser Ser Ser
20 25 30
Ala Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Val Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His
65 70 75 80
Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 93
<211> 213
<212> PRT
<213> Artificial Sequence
<220>
<223> 轻链
<400> 93
Gln Ile Val Leu Thr Gln Ser Pro Pro Ile Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Asn Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr
35 40 45
Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Val Pro Leu Thr
85 90 95
Phe Gly Ala 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> 94
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 94
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Ser
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Gly Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Asn Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Leu Leu Gln Gln Ser
115 120 125
Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Met Lys Ile Ser Cys Lys
130 135 140
Ala Ser Val Tyr Ser Phe Thr Ala Tyr Thr Met Asn Trp Val Lys Gln
145 150 155 160
Ser His Gly Lys Asn Leu Glu Trp Ile Gly Leu Ile Asn Pro His Asn
165 170 175
Gly Gly Thr Arg Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Leu Asp Lys Ser Ser Ser Thr Ala Tyr Met Asp Leu Leu Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Ile Ser Arg Tyr Gly Ser
210 215 220
Ser Ser Phe Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Ala Val
225 230 235 240
Ser Ser
<210> 95
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 95
Glu Val Leu Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Val Tyr Ser Phe Thr Ala Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro His Asn Gly Gly Thr Arg Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Leu Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Asp Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ile Ser Arg Tyr Gly Ser Ser Ser Phe Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Ala Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Ser Ser Ser Ile Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly
165 170 175
Ser Ser Pro Lys Pro Trp Ile Ser Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Gly Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Ser Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Asn Leu Glu
225 230 235 240
Ile Lys
<210> 96
<211> 238
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 96
Ser Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met Ser Val Gly
1 5 10 15
Glu Arg Val Thr Leu Ser Cys Lys Ala Ser Glu Asn Val Gly Gly Tyr
20 25 30
Val Ser Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Ser Ser Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Glu Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Ala Tyr His Cys Gly Gln Asn Tyr Ile Tyr Pro Phe
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln
115 120 125
Ser Gly Ala Glu Leu Val Arg Pro Gly Thr Ser Val Lys Ile Ser Cys
130 135 140
Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr Trp Leu Gly Trp Met Lys
145 150 155 160
Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly Asp Phe Tyr Pro Arg
165 170 175
Thr Gly Asn Thr Phe Tyr Asn Glu Asn Phe Lys Gly Lys Val Thr Leu
180 185 190
Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr Met Gln Leu Ser Ser Leu
195 200 205
Thr Ser Glu Asp Ser Ala Val Tyr Leu Cys Ala Arg Ala Gly Thr Gly
210 215 220
Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser
225 230 235
<210> 97
<211> 238
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 97
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Trp Leu Gly Trp Met Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Asp Phe Tyr Pro Arg Thr Gly Asn Thr Phe Tyr Asn Glu Asn Phe
50 55 60
Lys Gly Lys Val Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Leu Cys
85 90 95
Ala Arg Ala Gly Thr Gly Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Ser Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met
130 135 140
Ser Val Gly Glu Arg Val Thr Leu Ser Cys Lys Ala Ser Glu Asn Val
145 150 155 160
Gly Gly Tyr Val Ser Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys
165 170 175
Leu Leu Ile Tyr Gly Ala Ser Ser Arg His Thr Gly Val Pro Asp Arg
180 185 190
Phe Thr Gly Ser Gly Ser Glu Thr Asp Phe Thr Leu Thr Ile Ser Ser
195 200 205
Val Gln Ala Glu Asp Leu Ala Ala Tyr His Cys Gly Gln Asn Tyr Ile
210 215 220
Tyr Pro Phe Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
225 230 235
<210> 98
<211> 241
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 98
Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu
1 5 10 15
Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Ile Ser
20 25 30
Asn Tyr Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly
35 40 45
Leu Ile Gly Gly Thr Asn Asn Arg Pro Pro Gly Val Pro Ala Arg Phe
50 55 60
Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala
65 70 75 80
Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Val Leu Trp Tyr Ser Asn
85 90 95
His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly
100 105 110
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu
115 120 125
Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala Ser Val Lys Ile
130 135 140
Ser Cys Lys Thr Thr Gly Tyr Arg Phe Ser Ser Tyr Trp Ile Glu Trp
145 150 155 160
Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu Gly Glu Ile Leu
165 170 175
Pro Gly Arg Gly Ile Ile Asn Tyr Asn Glu Asn Phe Arg Gly Lys Ala
180 185 190
Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr Val Gln Leu Ser
195 200 205
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala Arg Thr Asp
210 215 220
Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser
225 230 235 240
Ser
<210> 99
<211> 241
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 99
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Thr Thr Gly Tyr Arg Phe Ser Ser Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu
35 40 45
Gly Glu Ile Leu Pro Gly Arg Gly Ile Ile Asn Tyr Asn Glu Asn Phe
50 55 60
Arg Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Val Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Thr Asp Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Thr
100 105 110
Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
115 120 125
Gly Gly Gly Ser Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr
130 135 140
Ser Pro Gly Glu Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala
145 150 155 160
Val Thr Ile Ser Asn Tyr Ala Asn Trp Val Gln Glu Lys Pro Asp His
165 170 175
Leu Phe Thr Gly Leu Ile Gly Gly Thr Asn Asn Arg Pro Pro Gly Val
180 185 190
Pro Ala Arg Phe Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr
195 200 205
Ile Thr Gly Ala Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Val Leu
210 215 220
Trp Tyr Ser Asn His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 100
<211> 245
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 100
Asp Ile Val Met Thr Gln Ala Ala Phe Ser Thr Pro Val Thr Leu Gly
1 5 10 15
Thr Ser Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
20 25 30
Asn Gly Ile Thr Tyr Leu Tyr Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu Ala Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Ser Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Asn
85 90 95
Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
115 120 125
Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala Ser
130 135 140
Met Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Thr Tyr Trp
145 150 155 160
Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly
165 170 175
Glu Asn Leu Pro Gly Arg His Ile Thr Asn Tyr Asn Glu Lys Phe Lys
180 185 190
Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr Met
195 200 205
Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala
210 215 220
Arg Gly Arg Gly Thr Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Pro
225 230 235 240
Leu Thr Val Ser Ser
245
<210> 101
<211> 245
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 101
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Thr Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Glu Asn Leu Pro Gly Arg His Ile Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Arg Gly Thr Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Pro Leu Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Ala Ala Phe Ser Thr
130 135 140
Pro Val Thr Leu Gly Thr Ser Ala Ser Ile Ser Cys Arg Ser Ser Gln
145 150 155 160
Ser Leu Leu His Ser Asn Gly Ile Thr Tyr Leu Tyr Trp Tyr Leu Gln
165 170 175
Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu
180 185 190
Ala Ser Gly Val Pro Asp Arg Phe Ser Ser Ser Gly Ser Gly Thr Asp
195 200 205
Phe Thr Leu Arg Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr
210 215 220
Tyr Cys Ala Gln Asn Leu Glu Phe Pro Phe Thr Phe Gly Ser Gly Thr
225 230 235 240
Lys Leu Glu Ile Lys
245
<210> 102
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 102
Gln Ile Val Leu Thr Gln Ser Pro Ala Leu Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Thr Ser Ser Val Ser Tyr Ile
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Arg Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Leu Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Gln Gln Ser
115 120 125
Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Met Lys Ile Ser Cys Lys
130 135 140
Ala Ser Gly Tyr Ser Phe Thr Gly Tyr Thr Met Asn Trp Val Lys Gln
145 150 155 160
Ser His Gly Lys Asn Leu Glu Trp Ile Gly Leu Ile Asn Pro Tyr Asn
165 170 175
Gly Gly Thr Asn Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Val Asp Lys Ser Ser Ser Thr Ala Tyr Met Glu Leu Leu Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Phe Ser Tyr Tyr Gly Ser
210 215 220
Arg Gly Phe Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val
225 230 235 240
Ser Ser
<210> 103
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 103
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Phe Ser Tyr Tyr Gly Ser Arg Gly Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Thr Leu Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Thr Gln Ser Pro
130 135 140
Ala Leu Met Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Ser
145 150 155 160
Ala Thr Ser Ser Val Ser Tyr Ile Tyr Trp Tyr Gln Gln Lys Pro Arg
165 170 175
Ser Ser Pro Lys Pro Trp Ile Tyr Leu Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Ser Met Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Ser Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Ile Lys
<210> 104
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 104
Asp Ile Val Met Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr
35 40 45
Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Thr Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Gln Gln Ser
115 120 125
Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Met Lys Ile Ser Cys Lys
130 135 140
Ala Ser Gly Tyr Ser Phe Thr Gly Tyr Thr Met Asn Trp Val Lys Gln
145 150 155 160
Ser His Gly Lys Asn Leu Glu Trp Ile Gly Leu Ile Asn Pro Tyr Asn
165 170 175
Gly Gly Thr Arg Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Val Asp Lys Ser Ser Ser Thr Ala Tyr Met Glu Leu Leu Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Ser Ser Ser Tyr Arg Asn
210 215 220
Asp Gly Asn Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr
225 230 235 240
Val Ser Ser
<210> 105
<211> 243
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 105
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Ile Asn Pro Tyr Asn Gly Gly Thr Arg Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Ser Ser Tyr Arg Asn Asp Gly Asn Trp Tyr Phe Asp Val Trp
100 105 110
Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln Ser
130 135 140
Pro Ala Ile Met Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys
145 150 155 160
Ser Ala Ser Ser Ser Val Ser Tyr Met His Trp Tyr Gln Gln Lys Ser
165 170 175
Gly Thr Ser Pro Lys Arg Trp Ile Tyr Asp Thr Ser Lys Leu Ala Ser
180 185 190
Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser
195 200 205
Leu Thr Ile Ser Ser Met Glu Thr Glu Asp Ala Ala Thr Tyr Tyr Cys
210 215 220
Gln Gln Trp Ser Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu
225 230 235 240
Glu Leu Lys
<210> 106
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 106
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Thr Ser Ser Val Ser Tyr Met
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Lys Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Gln Gln Ser
115 120 125
Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Met Lys Ile Ser Cys Lys
130 135 140
Ala Ser Gly Tyr Ser Ile Thr Gly Tyr Thr Met Asn Trp Val Lys Gln
145 150 155 160
Ser His Gly Lys Asn Leu Glu Trp Ile Gly Leu Val Asn Pro Tyr Asn
165 170 175
Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys Ala Thr Leu Thr
180 185 190
Val Asp Lys Ser Ser Ser Thr Ala Tyr Met Glu Leu Leu Ser Leu Lys
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Ile Ser Arg Tyr Gly Ser
210 215 220
Glu Ser Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val
225 230 235 240
Ser Ser
<210> 107
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 107
Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Ile Thr Gly Tyr
20 25 30
Thr Met Asn Trp Val Lys Gln Ser His Gly Lys Asn Leu Glu Trp Ile
35 40 45
Gly Leu Val Asn Pro Tyr Asn Gly Gly Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Leu Ser Leu Lys Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Ile Ser Arg Tyr Gly Ser Glu Ser Trp Tyr Phe Asp Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Ile Val Leu Ser Gln Ser Pro
130 135 140
Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Arg
145 150 155 160
Ala Thr Ser Ser Val Ser Tyr Met Tyr Trp Tyr Gln Gln Lys Pro Gly
165 170 175
Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser Gly
180 185 190
Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu
195 200 205
Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln
210 215 220
Gln Trp Ser Ser Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu
225 230 235 240
Lys Lys
<210> 108
<211> 244
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 108
Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Val Val Ser Leu Gly
1 5 10 15
Gln Arg Ala Thr Ile Ser Cys Arg Thr Ser Lys Ser Val Ser Ser Ser
20 25 30
Ala Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Val Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His
65 70 75 80
Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val
115 120 125
His Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala Ser Val
130 135 140
Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Asn Ile Tyr Trp Ile
145 150 155 160
Asp Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly Glu
165 170 175
Ile Leu Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Asn Phe Lys Gly
180 185 190
Lys Ala Thr Met Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr Met Gln
195 200 205
Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg
210 215 220
Thr Asp Gly Arg Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu
225 230 235 240
Thr Val Ser Ser
<210> 109
<211> 244
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 109
Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Asn Ile Tyr
20 25 30
Trp Ile Asp Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile
35 40 45
Gly Glu Ile Leu Pro Gly Ser Gly Asn Thr His Tyr Asn Glu Asn Phe
50 55 60
Lys Gly Lys Ala Thr Met Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Thr Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Thr Asp Gly Arg Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Thr Leu Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu
130 135 140
Val Val Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys Arg Thr Ser Lys
145 150 155 160
Ser Val Ser Ser Ser Ala Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys
165 170 175
Pro Gly Gln Pro Pro Lys Val Leu Ile Tyr Leu Ala Ser Asn Leu Glu
180 185 190
Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
195 200 205
Thr Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr
210 215 220
Cys Gln His Ser Arg Glu Leu Pro Phe Thr Phe Gly Ser Gly Thr Lys
225 230 235 240
Leu Glu Ile Lys
<210> 110
<211> 238
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 110
Gln Ile Val Leu Thr Gln Ser Pro Pro Ile Met Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Asn Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr
35 40 45
Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Val Pro Leu Thr
85 90 95
Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
100 105 110
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Gln Gln Ser
115 120 125
Gly Ala Glu Leu Met Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys
130 135 140
Ala Thr Gly Tyr Thr Phe Ser Ser Tyr Trp Ile Glu Trp Val Lys Gln
145 150 155 160
Arg Pro Gly His Gly Leu Glu Trp Leu Gly Glu Phe Leu Pro Arg Ser
165 170 175
Gly Lys Thr Asn Tyr Asn Glu Glu Phe Arg Gly Lys Ala Thr Phe Thr
180 185 190
Ala Asp Thr Ser Ser Asn Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr
195 200 205
Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Thr Asp Pro Pro Tyr
210 215 220
Phe Gly Val Trp Gly Ala Gly Thr Met Val Ala Val Ser Ser
225 230 235
<210> 111
<211> 238
<212> PRT
<213> Artificial Sequence
<220>
<223> 单链抗体
<400> 111
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Tyr
20 25 30
Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Leu
35 40 45
Gly Glu Phe Leu Pro Arg Ser Gly Lys Thr Asn Tyr Asn Glu Glu Phe
50 55 60
Arg Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Thr Asp Pro Pro Tyr Phe Gly Val Trp Gly Ala Gly Thr Met
100 105 110
Val Ala Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
115 120 125
Gly Gly Gly Ser Gln Ile Val Leu Thr Gln Ser Pro Pro Ile Met Ser
130 135 140
Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Asn
145 150 155 160
Ile Ser Tyr Met His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys
165 170 175
Arg Trp Ile Tyr Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg
180 185 190
Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser
195 200 205
Met Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser
210 215 220
Val Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys
225 230 235
<210> 112
<211> 1353
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 112
gaagtgctgc tgcagcagag cggaccagag ctggtcaagc ccggcgcctc catgaagatc 60
agctgtaagg cctccgtgta cagcttcaca gcctacacaa tgaactgggt gaagcagagc 120
cacggcaaga atctggagtg gatcggactg atcaacccac acaatggcgg cactaggtac 180
aaccagaagt tcaagggcaa ggccacactg actctggata agtccagcag cactgcctac 240
atggatctgc tgtctctgac aagcgaggac agcgccgtct actattgcgc catctctagg 300
tacggcagca gcagcttcta cttcgatgtg tggggcgccg gcacaacagt ggctgtgagc 360
agcgccagca ccaagggccc cagcgtgttc cctctggctc cttgtagccg gtccacctcc 420
gagtccacag ctgctctggg ctgcctcgtg aaggactact ttcccgaacc cgttaccgtg 480
agctggaata gcggcgcttt aacctccgga gtgcacacct tccccgctgt gctccagtcc 540
tccggtttat actctttatc ctccgtggtg accgtgcctt cctccagcct cggcaccaag 600
acctacactt gtaacgtgga ccacaagccc agcaacacca aggtggacaa gagggtggag 660
tccaagtacg gacctccttg tcccccttgc cccgcccccg aggccgctgg cggaccctcc 720
gtgttcctct tcccccccaa acccaaggac actttaatga tctcccggac ccccgaagtg 780
acttgtgtgg tggtggacgt gtcccaagaa gaccccgagg tgcagtttaa ctggtacgtg 840
gatggcgtgg aggtgcacaa cgccaagacc aagcctaggg aggaacagtt caactccacc 900
taccgggtgg tgtccgtgct caccgtgctg catcaagatt ggctgaacgg caaggagtac 960
aagtgcaagg tgagcaacaa gggactgccc agctccatcg agaagaccat cagcaaggcc 1020
aaaggccagc cccgggaacc tcaagtttat acactgcccc ccagccaaga agagatgacc 1080
aagaaccaag tttctttaac ttgtttagtg aagggcttct accctagcga catcgctgtg 1140
gagtgggagt ccaatggcca gcccgaaaac aattataaga ccaccccccc cgtgctggac 1200
tccgatggtt cttttttttt atactccaag ctgacagtgg acaagtctcg ttggcaagaa 1260
ggcaacgtgt tctcttgtag cgtgatgcac gaggctttac acaaccacta cacccagaag 1320
tctttatctc tgtctttagg ctgatgagaa ttc 1353
<210> 113
<211> 1338
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 113
caagtgcagc tccagcagag cggagctgag ctggtgagac ccggcactag cgtgaagatc 60
agctgtaagg ccagcggcta cgccttcact aattactggc tgggctggat gaagcagaga 120
cccggccatg gactggagtg gatcggcgac ttctacccaa ggactggcaa cactttctac 180
aacgagaact ttaagggcaa ggtgactctg actgccgata agtccagcaa cactgcctac 240
atgcagctgt cctctctgac tagcgaagat agcgccgtgt atctgtgtgc tagggctggc 300
actggcttcg attactgggg ccaaggcaca acactgacag tgagcagcgc cagcaccaag 360
ggccccagcg tgttccctct ggctccttgt agccggtcca cctccgagtc cacagctgct 420
ctgggctgcc tcgtgaagga ctactttccc gaacccgtta ccgtgagctg gaatagcggc 480
gctttaacct ccggagtgca caccttcccc gctgtgctcc agtcctccgg tttatactct 540
ttatcctccg tggtgaccgt gccttcctcc agcctcggca ccaagaccta cacttgtaac 600
gtggaccaca agcccagcaa caccaaggtg gacaagaggg tggagtccaa gtacggacct 660
ccttgtcccc cttgccccgc ccccgaggcc gctggcggac cctccgtgtt cctcttcccc 720
cccaaaccca aggacacttt aatgatctcc cggacccccg aagtgacttg tgtggtggtg 780
gacgtgtccc aagaagaccc cgaggtgcag tttaactggt acgtggatgg cgtggaggtg 840
cacaacgcca agaccaagcc tagggaggaa cagttcaact ccacctaccg ggtggtgtcc 900
gtgctcaccg tgctgcatca agattggctg aacggcaagg agtacaagtg caaggtgagc 960
aacaagggac tgcccagctc catcgagaag accatcagca aggccaaagg ccagccccgg 1020
gaacctcaag tttatacact gccccccagc caagaagaga tgaccaagaa ccaagtttct 1080
ttaacttgtt tagtgaaggg cttctaccct agcgacatcg ctgtggagtg ggagtccaat 1140
ggccagcccg aaaacaatta taagaccacc ccccccgtgc tggactccga tggttctttt 1200
tttttatact ccaagctgac agtggacaag tctcgttggc aagaaggcaa cgtgttctct 1260
tgtagcgtga tgcacgaggc tttacacaac cactacaccc agaagtcttt atctctgtct 1320
ttaggctgat gagaattc 1338
<210> 114
<211> 1341
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 114
caagtgcagc tgcagcagag cggcgctgag ctgatgaagc ccggcgcttc cgtcaagatc 60
agctgcaaga ctactggcta taggttcagc agctactgga tcgagtgggt gaagcagaga 120
cccggccacg gactggaatg gctgggcgaa atcctccccg gaaggggcat catcaactac 180
aatgagaact ttaggggcaa ggccacattc acagccgaca caagcagcaa cactgcctac 240
gtgcagctga gctctctgac ttccgaggac agcgccgtgt acttctgcgc cagaacagac 300
cctccttatt tcggcgtgtg gggcgccgga actactgtga ctgtgtccag cgccagcacc 360
aagggcccca gcgtgttccc tctggctcct tgtagccggt ccacctccga gtccacagct 420
gctctgggct gcctcgtgaa ggactacttt cccgaacccg ttaccgtgag ctggaatagc 480
ggcgctttaa cctccggagt gcacaccttc cccgctgtgc tccagtcctc cggtttatac 540
tctttatcct ccgtggtgac cgtgccttcc tccagcctcg gcaccaagac ctacacttgt 600
aacgtggacc acaagcccag caacaccaag gtggacaaga gggtggagtc caagtacgga 660
cctccttgtc ccccttgccc cgcccccgag gccgctggcg gaccctccgt gttcctcttc 720
ccccccaaac ccaaggacac tttaatgatc tcccggaccc ccgaagtgac ttgtgtggtg 780
gtggacgtgt cccaagaaga ccccgaggtg cagtttaact ggtacgtgga tggcgtggag 840
gtgcacaacg ccaagaccaa gcctagggag gaacagttca actccaccta ccgggtggtg 900
tccgtgctca ccgtgctgca tcaagattgg ctgaacggca aggagtacaa gtgcaaggtg 960
agcaacaagg gactgcccag ctccatcgag aagaccatca gcaaggccaa aggccagccc 1020
cgggaacctc aagtttatac actgcccccc agccaagaag agatgaccaa gaaccaagtt 1080
tctttaactt gtttagtgaa gggcttctac cctagcgaca tcgctgtgga gtgggagtcc 1140
aatggccagc ccgaaaacaa ttataagacc accccccccg tgctggactc cgatggttct 1200
ttttttttat actccaagct gacagtggac aagtctcgtt ggcaagaagg caacgtgttc 1260
tcttgtagcg tgatgcacga ggctttacac aaccactaca cccagaagtc tttatctctg 1320
tctttaggct gatgagaatt c 1341
<210> 115
<211> 1344
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 115
caagtgcagc tgcagcagag cggagccgag ctgatgaagc ccggcgcctc catgaagatc 60
agctgcaagg ccactggcta cacattcagc acatactgga tcgagtgggt caagcagaga 120
cccggccacg gactggagtg gatcggagag aatctgcccg gaaggcacat cactaactac 180
aacgagaagt tcaagggcaa ggccactttc acagccgaca ctagcagcaa cactgcctac 240
atgcagctca gctctctgac aagcgaagat agcgccgtgt actactgtgc taggggaagg 300
ggcacttact acttcgatta ctggggccaa ggcactccac tgactgtgtc cagcgccagc 360
accaagggcc ccagcgtgtt ccctctggct ccttgtagcc ggtccacctc cgagtccaca 420
gctgctctgg gctgcctcgt gaaggactac tttcccgaac ccgttaccgt gagctggaat 480
agcggcgctt taacctccgg agtgcacacc ttccccgctg tgctccagtc ctccggttta 540
tactctttat cctccgtggt gaccgtgcct tcctccagcc tcggcaccaa gacctacact 600
tgtaacgtgg accacaagcc cagcaacacc aaggtggaca agagggtgga gtccaagtac 660
ggacctcctt gtcccccttg ccccgccccc gaggccgctg gcggaccctc cgtgttcctc 720
ttccccccca aacccaagga cactttaatg atctcccgga cccccgaagt gacttgtgtg 780
gtggtggacg tgtcccaaga agaccccgag gtgcagttta actggtacgt ggatggcgtg 840
gaggtgcaca acgccaagac caagcctagg gaggaacagt tcaactccac ctaccgggtg 900
gtgtccgtgc tcaccgtgct gcatcaagat tggctgaacg gcaaggagta caagtgcaag 960
gtgagcaaca agggactgcc cagctccatc gagaagacca tcagcaaggc caaaggccag 1020
ccccgggaac ctcaagttta tacactgccc cccagccaag aagagatgac caagaaccaa 1080
gtttctttaa cttgtttagt gaagggcttc taccctagcg acatcgctgt ggagtgggag 1140
tccaatggcc agcccgaaaa caattataag accacccccc ccgtgctgga ctccgatggt 1200
tctttttttt tatactccaa gctgacagtg gacaagtctc gttggcaaga aggcaacgtg 1260
ttctcttgta gcgtgatgca cgaggcttta cacaaccact acacccagaa gtctttatct 1320
ctgtctttag gctgatgaga attc 1344
<210> 116
<211> 1353
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 116
gaggtgcagc tccaacagag cggaccagag ctggtgaaac ccggcgccag catgaagatc 60
agctgtaagg cctccggcta cagcttcact ggctacacta tgaactgggt gaagcagtcc 120
cacggcaaga atctggagtg gatcggactg atcaacccat acaacggcgg cacaaactac 180
aaccagaagt tcaagggcaa ggccactctg acagtcgata agagctccag cactgcctac 240
atggagctgc tgagcctcac tagcgaggac agcgctgtgt actactgtgc cttctcctac 300
tacggctcta ggggcttcta cttcgattac tggggccaag gcacaacact gacagtgtcc 360
agcgccagca ccaagggccc cagcgtgttc cctctggctc cttgtagccg gtccacctcc 420
gagtccacag ctgctctggg ctgcctcgtg aaggactact ttcccgaacc cgttaccgtg 480
agctggaata gcggcgcttt aacctccgga gtgcacacct tccccgctgt gctccagtcc 540
tccggtttat actctttatc ctccgtggtg accgtgcctt cctccagcct cggcaccaag 600
acctacactt gtaacgtgga ccacaagccc agcaacacca aggtggacaa gagggtggag 660
tccaagtacg gacctccttg tcccccttgc cccgcccccg aggccgctgg cggaccctcc 720
gtgttcctct tcccccccaa acccaaggac actttaatga tctcccggac ccccgaagtg 780
acttgtgtgg tggtggacgt gtcccaagaa gaccccgagg tgcagtttaa ctggtacgtg 840
gatggcgtgg aggtgcacaa cgccaagacc aagcctaggg aggaacagtt caactccacc 900
taccgggtgg tgtccgtgct caccgtgctg catcaagatt ggctgaacgg caaggagtac 960
aagtgcaagg tgagcaacaa gggactgccc agctccatcg agaagaccat cagcaaggcc 1020
aaaggccagc cccgggaacc tcaagtttat acactgcccc ccagccaaga agagatgacc 1080
aagaaccaag tttctttaac ttgtttagtg aagggcttct accctagcga catcgctgtg 1140
gagtgggagt ccaatggcca gcccgaaaac aattataaga ccaccccccc cgtgctggac 1200
tccgatggtt cttttttttt atactccaag ctgacagtgg acaagtctcg ttggcaagaa 1260
ggcaacgtgt tctcttgtag cgtgatgcac gaggctttac acaaccacta cacccagaag 1320
tctttatctc tgtctttagg ctgatgagaa ttc 1353
<210> 117
<211> 1356
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 117
gaagtgcagc tgcaacagag cggcccagag ctcgtgaagc ccggcgccag catgaagatc 60
agctgcaagg ccagcggcta cagcttcact ggctacacaa tgaactgggt caagcagagc 120
cacggaaaaa atctggagtg gatcggactg atcaaccctt acaacggcgg cacaaggtac 180
aatcagaagt tcaagggcaa ggccactctc actgtggata agagcagcag cactgcctac 240
atggagctgc tgtctctgac aagcgaggat agcgccgtgt actactgcgc cagctcctcc 300
tataggaacg acggcaactg gtacttcgat gtgtggggcg ccggcactac tgtgacagtg 360
agctccgcca gcaccaaggg ccccagcgtg ttccctctgg ctccttgtag ccggtccacc 420
tccgagtcca cagctgctct gggctgcctc gtgaaggact actttcccga acccgttacc 480
gtgagctgga atagcggcgc tttaacctcc ggagtgcaca ccttccccgc tgtgctccag 540
tcctccggtt tatactcttt atcctccgtg gtgaccgtgc cttcctccag cctcggcacc 600
aagacctaca cttgtaacgt ggaccacaag cccagcaaca ccaaggtgga caagagggtg 660
gagtccaagt acggacctcc ttgtccccct tgccccgccc ccgaggccgc tggcggaccc 720
tccgtgttcc tcttcccccc caaacccaag gacactttaa tgatctcccg gacccccgaa 780
gtgacttgtg tggtggtgga cgtgtcccaa gaagaccccg aggtgcagtt taactggtac 840
gtggatggcg tggaggtgca caacgccaag accaagccta gggaggaaca gttcaactcc 900
acctaccggg tggtgtccgt gctcaccgtg ctgcatcaag attggctgaa cggcaaggag 960
tacaagtgca aggtgagcaa caagggactg cccagctcca tcgagaagac catcagcaag 1020
gccaaaggcc agccccggga acctcaagtt tatacactgc cccccagcca agaagagatg 1080
accaagaacc aagtttcttt aacttgttta gtgaagggct tctaccctag cgacatcgct 1140
gtggagtggg agtccaatgg ccagcccgaa aacaattata agaccacccc ccccgtgctg 1200
gactccgatg gttctttttt tttatactcc aagctgacag tggacaagtc tcgttggcaa 1260
gaaggcaacg tgttctcttg tagcgtgatg cacgaggctt tacacaacca ctacacccag 1320
aagtctttat ctctgtcttt aggctgatga gaattc 1356
<210> 118
<211> 1353
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 118
gaagtgcagc tgcagcagag cggaccagag ctggtcaagc ccggcgccag catgaagatc 60
agctgtaagg ccagcggcta cagcatcact ggctacacta tgaactgggt gaagcagagc 120
cacggcaaga acctcgagtg gattggcctc gtgaacccat acaacggcgg cacttcctac 180
aaccagaagt tcaaaggcaa ggccacactc acagtcgata agtccagctc cacagcctac 240
atggagctgc tgtctctgaa gagcgaggat agcgctgtct actactgtgc catcagcaga 300
tacggcagcg agagctggta cttcgacgtg tggggcgccg gcacaacagt gacagtgagc 360
agcgccagca caaagggccc cagcgtgttc cctctggctc cttgtagccg gtccacctcc 420
gagtccacag ctgctctggg ctgcctcgtg aaggactact ttcccgaacc cgttaccgtg 480
agctggaata gcggcgcttt aacctccgga gtgcacacct tccccgctgt gctccagtcc 540
tccggtttat actctttatc ctccgtggtg accgtgcctt cctccagcct cggcaccaag 600
acctacactt gtaacgtgga ccacaagccc agcaacacca aggtggacaa gagggtggag 660
tccaagtacg gacctccttg tcccccttgc cccgcccccg aggccgctgg cggaccctcc 720
gtgttcctct tcccccccaa acccaaggac actttaatga tctcccggac ccccgaagtg 780
acttgtgtgg tggtggacgt gtcccaagaa gaccccgagg tgcagtttaa ctggtacgtg 840
gatggcgtgg aggtgcacaa cgccaagacc aagcctaggg aggaacagtt caactccacc 900
taccgggtgg tgtccgtgct caccgtgctg catcaagatt ggctgaacgg caaggagtac 960
aagtgcaagg tgagcaacaa gggactgccc agctccatcg agaagaccat cagcaaggcc 1020
aaaggccagc cccgggaacc tcaagtttat acactgcccc ccagccaaga agagatgacc 1080
aagaaccaag tttctttaac ttgtttagtg aagggcttct accctagcga catcgctgtg 1140
gagtgggagt ccaatggcca gcccgaaaac aattataaga ccaccccccc cgtgctggac 1200
tccgatggtt cttttttttt atactccaag ctgacagtgg acaagtctcg ttggcaagaa 1260
ggcaacgtgt tctcttgtag cgtgatgcac gaggctttac acaaccacta cacccagaag 1320
tctttatctc tgtctttagg ctgatgagaa ttc 1353
<210> 119
<211> 1344
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 119
caagtgcatc tgcagcagag cggcgctgag ctgatgaagc ccggcgccag cgtgaagatt 60
agctgcaagg ccactggcta cacattcaac atctactgga tcgactgggt gaagcagagg 120
cccggccacg gactggaatg gatcggcgaa attctgcccg gcagcggcaa cactcactac 180
aacgagaact tcaagggcaa ggccacaatg acagccgaca caagctccaa cactgcttac 240
atgcagctga cttctctgac tagcgaggac agcgccgtgt actattgcgc taggacagac 300
ggaaggggct acttcgatta ctggggccaa ggcactacac tcacagtgag cagcgccagc 360
actaagggcc ccagcgtgtt ccctctggct ccttgtagcc ggtccacctc cgagtccaca 420
gctgctctgg gctgcctcgt gaaggactac tttcccgaac ccgttaccgt gagctggaat 480
agcggcgctt taacctccgg agtgcacacc ttccccgctg tgctccagtc ctccggttta 540
tactctttat cctccgtggt gaccgtgcct tcctccagcc tcggcaccaa gacctacact 600
tgtaacgtgg accacaagcc cagcaacacc aaggtggaca agagggtgga gtccaagtac 660
ggacctcctt gtcccccttg ccccgccccc gaggccgctg gcggaccctc cgtgttcctc 720
ttccccccca aacccaagga cactttaatg atctcccgga cccccgaagt gacttgtgtg 780
gtggtggacg tgtcccaaga agaccccgag gtgcagttta actggtacgt ggatggcgtg 840
gaggtgcaca acgccaagac caagcctagg gaggaacagt tcaactccac ctaccgggtg 900
gtgtccgtgc tcaccgtgct gcatcaagat tggctgaacg gcaaggagta caagtgcaag 960
gtgagcaaca agggactgcc cagctccatc gagaagacca tcagcaaggc caaaggccag 1020
ccccgggaac ctcaagttta tacactgccc cccagccaag aagagatgac caagaaccaa 1080
gtttctttaa cttgtttagt gaagggcttc taccctagcg acatcgctgt ggagtgggag 1140
tccaatggcc agcccgaaaa caattataag accacccccc ccgtgctgga ctccgatggt 1200
tctttttttt tatactccaa gctgacagtg gacaagtctc gttggcaaga aggcaacgtg 1260
ttctcttgta gcgtgatgca cgaggcttta cacaaccact acacccagaa gtctttatct 1320
ctgtctttag gctgatgaga attc 1344
<210> 120
<211> 1341
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 120
caggtgcagc tgcagcagtc cggcgctgag ctcatgaagc ccggcgccag cgtgaagatc 60
agctgcaagg ccactggcta cacattcagc agctactgga tcgagtgggt gaagcagaga 120
cccggccacg gactggaatg gctgggagag tttctgccta gaagcggcaa gacaaactac 180
aacgaggagt ttaggggcaa ggctacattc actgccgaca catccagcaa cactgcctac 240
atgcagctga gcagcctcac aagcgaggat tccgccgtct actactgtgc taggactgat 300
cctccttact tcggagtgtg gggcgctggc acaatggtgg ctgtgagcag cgcctccact 360
aagggcccca gcgtgttccc tctggctcct tgtagccggt ccacctccga gtccacagct 420
gctctgggct gcctcgtgaa ggactacttt cccgaacccg ttaccgtgag ctggaatagc 480
ggcgctttaa cctccggagt gcacaccttc cccgctgtgc tccagtcctc cggtttatac 540
tctttatcct ccgtggtgac cgtgccttcc tccagcctcg gcaccaagac ctacacttgt 600
aacgtggacc acaagcccag caacaccaag gtggacaaga gggtggagtc caagtacgga 660
cctccttgtc ccccttgccc cgcccccgag gccgctggcg gaccctccgt gttcctcttc 720
ccccccaaac ccaaggacac tttaatgatc tcccggaccc ccgaagtgac ttgtgtggtg 780
gtggacgtgt cccaagaaga ccccgaggtg cagtttaact ggtacgtgga tggcgtggag 840
gtgcacaacg ccaagaccaa gcctagggag gaacagttca actccaccta ccgggtggtg 900
tccgtgctca ccgtgctgca tcaagattgg ctgaacggca aggagtacaa gtgcaaggtg 960
agcaacaagg gactgcccag ctccatcgag aagaccatca gcaaggccaa aggccagccc 1020
cgggaacctc aagtttatac actgcccccc agccaagaag agatgaccaa gaaccaagtt 1080
tctttaactt gtttagtgaa gggcttctac cctagcgaca tcgctgtgga gtgggagtcc 1140
aatggccagc ccgaaaacaa ttataagacc accccccccg tgctggactc cgatggttct 1200
ttttttttat actccaagct gacagtggac aagtctcgtt ggcaagaagg caacgtgttc 1260
tcttgtagcg tgatgcacga ggctttacac aaccactaca cccagaagtc tttatctctg 1320
tctttaggct gatgagaatt c 1341
<210> 121
<211> 651
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 121
cagatcgtgc tgagccagag cccagctatt ctgtccgcca gccccggcga gaaggtgact 60
atgacttgta gggccagctc cagcatcagc tacatgcact ggtaccagca gaagcccggc 120
tcctccccaa agccttggat cagcgccact agcaatctgg ccagcggcgt gccagccaga 180
ttcagcggaa gcggcagcgg cactagctac tctctgacta tctccggcgt ggaagctgag 240
gatgccgcca cttactactg ccagcagtgg tccagcaacc ctcctacttt cggcggcggc 300
acaaatctgg agatcaagcg gaccgtggct gccccctccg tgttcatctt ccccccttcc 360
gacgagcagc tgaagtccgg caccgctagc gtggtgtgtt tactgaacaa cttctaccct 420
cgtgaggcca aggtgcagtg gaaggtggac aacgctttac agtccggcaa ctcccaagaa 480
tccgtgaccg agcaagattc caaggactcc acctactctt tatcctccac tttaacttta 540
tccaaggccg actacgagaa gcacaaggtg tacgcttgtg aggtgaccca tcaaggttta 600
tcctcccccg tgaccaagtc cttcaatcgt ggcgagtgct gatgagaatt c 651
<210> 122
<211> 654
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 122
agcatcgtca tgacacaaag ccctaagagc atgagcatga gcgtgggcga gagagtgact 60
ctgagctgta aggccagcga gaatgtgggc ggctacgtga gctggtatca gcagaagcca 120
gatcagagcc caaagctgct gatctacggc gccagcagca gacacactgg cgtgccagat 180
aggttcactg gcagcggctc cgagacagac ttcactctga ctatcagcag cgtccaagcc 240
gaagatctgg ccgcctatca ctgcggccag aactacatct acccattcac attcggcggc 300
ggcacaaagc tggagatcaa gcggaccgtg gctgccccct ccgtgttcat cttcccccct 360
tccgacgagc agctgaagtc cggcaccgct agcgtggtgt gtttactgaa caacttctac 420
cctcgtgagg ccaaggtgca gtggaaggtg gacaacgctt tacagtccgg caactcccaa 480
gaatccgtga ccgagcaaga ttccaaggac tccacctact ctttatcctc cactttaact 540
ttatccaagg ccgactacga gaagcacaag gtgtacgctt gtgaggtgac ccatcaaggt 600
ttatcctccc ccgtgaccaa gtccttcaat cgtggcgagt gctgatgaga attc 654
<210> 123
<211> 660
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 123
caagccgtcg tgacacaaga gtccgctctg acaacttccc ccggcgagac tgtgacactg 60
acttgtagga gcagcactgg cgccgtgact atcagcaact acgccaactg ggtccaagag 120
aagccagatc atctgttcac tggactgatc ggcggcacaa ataataggcc tcccggcgtg 180
ccagccagat ttagcggctc tctgattggc gataaggctg ctctgacaat cactggcgcc 240
cagactgagg acgaggccat ctacttctgc gtcctctggt acagcaacca ctgggtgttc 300
ggcggcggca ctaagctgac tgtgctgcgg accgtggctg ccccctccgt gttcatcttc 360
cccccttccg acgagcagct gaagtccggc accgctagcg tggtgtgttt actgaacaac 420
ttctaccctc gtgaggccaa ggtgcagtgg aaggtggaca acgctttaca gtccggcaac 480
tcccaagaat ccgtgaccga gcaagattcc aaggactcca cctactcttt atcctccact 540
ttaactttat ccaaggccga ctacgagaag cacaaggtgt acgcttgtga ggtgacccat 600
caaggtttat cctcccccgt gaccaagtcc ttcaatcgtg gcgagtgctg atgagaattc 660
<210> 124
<211> 669
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 124
gatatcgtca tgactcaagc cgccttcagc actccagtca ctctcggcac aagcgccagc 60
atcagctgta ggtccagcca gtctctgctg cacagcaacg gcatcactta tctgtactgg 120
tatctgcaga agcccggcca aagcccacag ctgctgatct accagatgag caatctggcc 180
agcggcgtgc cagatagatt cagcagcagc ggaagcggaa cagacttcac actgaggatc 240
tctagggtgg aagccgaaga tgtgggcgtc tattactgcg cccagaacct cgagttccct 300
ttcacattcg gcagcggcac taagctggag atcaagcgga ccgtggctgc cccctccgtg 360
ttcatcttcc ccccttccga cgagcagctg aagtccggca ccgctagcgt ggtgtgttta 420
ctgaacaact tctaccctcg tgaggccaag gtgcagtgga aggtggacaa cgctttacag 480
tccggcaact cccaagaatc cgtgaccgag caagattcca aggactccac ctactcttta 540
tcctccactt taactttatc caaggccgac tacgagaagc acaaggtgta cgcttgtgag 600
gtgacccatc aaggtttatc ctcccccgtg accaagtcct tcaatcgtgg cgagtgctga 660
tgagaattc 669
<210> 125
<211> 651
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 125
cagatcgtgc tgacacagag cccagctctg atgagcgcca gccccggcga gaaggtcaca 60
atgacttgca gcgccacatc cagcgtgagc tacatctact ggtaccagca gaagcctagg 120
agcagcccta agccttggat ctacctcaca agcaatctgg ccagcggagt gccagctagg 180
ttcagcggaa gcggcagcgg cacaagctac tctctgacaa tctccagcat ggaagccgaa 240
gatgccgcca cttactactg ccagcagtgg agcagcaatc cacctacatt cggaggcggc 300
actaagctgg agatcaagcg gaccgtggct gccccctccg tgttcatctt ccccccttcc 360
gacgagcagc tgaagtccgg caccgctagc gtggtgtgtt tactgaacaa cttctaccct 420
cgtgaggcca aggtgcagtg gaaggtggac aacgctttac agtccggcaa ctcccaagaa 480
tccgtgaccg agcaagattc caaggactcc acctactctt tatcctccac tttaacttta 540
tccaaggccg actacgagaa gcacaaggtg tacgcttgtg aggtgaccca tcaaggttta 600
tcctcccccg tgaccaagtc cttcaatcgt ggcgagtgct gatgagaatt c 651
<210> 126
<211> 651
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 126
gatatcgtga tgactcagtc cccagccatc atgtccgcca gccccggcga gaaggtgact 60
atgacttgct ccgccagcag cagcgtgagc tacatgcact ggtaccagca gaagagcggc 120
acatccccaa agaggtggat ctacgacaca agcaagctgg ccagcggcgt gccagccaga 180
ttcagcggct ccggcagcgg aacaagctac tctctgacta tcagcagcat ggagacagag 240
gacgctgcca cttactactg ccagcagtgg agcagcaatc ctccaacttt cggcggaggc 300
actaagctgg agctgaagcg gaccgtggct gccccctccg tgttcatctt ccccccttcc 360
gacgagcagc tgaagtccgg caccgctagc gtggtgtgtt tactgaacaa cttctaccct 420
cgtgaggcca aggtgcagtg gaaggtggac aacgctttac agtccggcaa ctcccaagaa 480
tccgtgaccg agcaagattc caaggactcc acctactctt tatcctccac tttaacttta 540
tccaaggccg actacgagaa gcacaaggtg tacgcttgtg aggtgaccca tcaaggttta 600
tcctcccccg tgaccaagtc cttcaatcgt ggcgagtgct gatgagaatt c 651
<210> 127
<211> 651
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 127
cagattgtgc tgtcccagtc cccagccatt ctgagcgcca gccccggcga gaaggtgact 60
atgacttgta gggccacaag cagcgtgagc tacatgtact ggtaccagca gaagcccggc 120
agcagcccta agccttggat ctacgccaca agcaatctgg ccagcggcgt cccagctaga 180
tttagcggca gcggatccgg cactagctat tctctgacta tctctagggt cgaggccgaa 240
gatgccgcca catactactg ccagcagtgg tcctccaatc ctccaacatt cggcggcgga 300
actaagctgg agaagaagag gacagtggct gccccttccg tgttcatctt ccctccaagc 360
gacgagcagc tgaagtccgg cactgctagc gtggtgtgtt tactgaacaa cttctaccct 420
cgtgaggcca aggtgcagtg gaaggtggac aacgctttac agtccggcaa ctcccaagaa 480
tccgtgaccg agcaagattc caaggactcc acctactctt tatcctccac tttaacttta 540
tccaaggccg actacgagaa gcacaaggtg tacgcttgtg aggtgaccca tcaaggttta 600
tcctcccccg tgaccaagtc cttcaatcgt ggcgagtgct gatgagaatt c 651
<210> 128
<211> 666
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 128
gatatcgtgc tgactcagag cccagcctct ctggtggtgt ctctgggaca gagggccaca 60
atcagctgta ggacttccaa gagcgtgagc agctccgcct acagctacat gcactggtac 120
cagcagaagc ccggccagcc tcctaaggtg ctgatctatc tggccagcaa tctggagagc 180
ggcgtcccag ctagattcag cggctccgga agcggcactg acttcactct gaacatccac 240
ccagtggaag aggaggatgc cgccacatac tactgccagc actctaggga gctgcctttc 300
acatttggca gcggaactaa gctggagatc aagaggactg tcgccgcccc tagcgtgttc 360
atcttccctc caagcgatga gcagctgaag agcggcactg ctagcgtggt gtgtttactg 420
aacaacttct accctcgtga ggccaaggtg cagtggaagg tggacaacgc tttacagtcc 480
ggcaactccc aagaatccgt gaccgagcaa gattccaagg actccaccta ctctttatcc 540
tccactttaa ctttatccaa ggccgactac gagaagcaca aggtgtacgc ttgtgaggtg 600
acccatcaag gtttatcctc ccccgtgacc aagtccttca atcgtggcga gtgctgatga 660
gaattc 666
<210> 129
<211> 651
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 129
caaattgttc tcacacagag ccctcctatc atgagcgcca gccccggcga gaaggtgact 60
atgacttgtt ccgccagcag caacatcagc tacatgcact ggtaccagca gaagtccggc 120
acaagcccaa agaggtggat ctacgacaca agcaagctgg ccagcggcgt gccagccaga 180
tttagcggct ccggaagcgg cactagctac tctctgacaa tcagcagcat ggaagccgag 240
gacgccgcta catactactg ccagcagtgg agctccgtcc cactgacttt cggcgctggc 300
actaagctgg agatcaagag gactgtggcc gccccttccg tgttcatctt ccctcctagc 360
gacgaacagc tcaagagcgg cactgctagc gtggtgtgtt tactgaacaa cttctaccct 420
cgtgaggcca aggtgcagtg gaaggtggac aacgctttac agtccggcaa ctcccaagaa 480
tccgtgaccg agcaagattc caaggactcc acctactctt tatcctccac tttaacttta 540
tccaaggccg actacgagaa gcacaaggtg tacgcttgtg aggtgaccca tcaaggttta 600
tcctcccccg tgaccaagtc cttcaatcgt ggcgagtgct gatgagaatt c 651
<210> 130
<211> 726
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 130
cagatcgtgc tgagccagag cccagctatt ctgtccgcca gccccggcga gaaggtgact 60
atgacttgta gggccagctc cagcatcagc tacatgcact ggtaccagca gaagcccggc 120
tcctccccaa agccttggat cagcgccact agcaatctgg ccagcggcgt gccagccaga 180
ttcagcggaa gcggcagcgg cactagctac tctctgacta tctccggcgt ggaagctgag 240
gatgccgcca cttactactg ccagcagtgg tccagcaacc ctcctacttt cggcggcggc 300
acaaatctgg agatcaaggg tggcggtggc tcgggcggtg gtgggtcggg tggcggcgga 360
tctgaagtgc tgctgcagca gagcggacca gagctggtca agcccggcgc ctccatgaag 420
atcagctgta aggcctccgt gtacagcttc acagcctaca caatgaactg ggtgaagcag 480
agccacggca agaatctgga gtggatcgga ctgatcaacc cacacaatgg cggcactagg 540
tacaaccaga agttcaaggg caaggccaca ctgactctgg ataagtccag cagcactgcc 600
tacatggatc tgctgtctct gacaagcgag gacagcgccg tctactattg cgccatctct 660
aggtacggca gcagcagctt ctacttcgat gtgtggggcg ccggcacaac agtggctgtg 720
agcagc 726
<210> 131
<211> 726
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 131
gaagtgctgc tgcagcagag cggaccagag ctggtcaagc ccggcgcctc catgaagatc 60
agctgtaagg cctccgtgta cagcttcaca gcctacacaa tgaactgggt gaagcagagc 120
cacggcaaga atctggagtg gatcggactg atcaacccac acaatggcgg cactaggtac 180
aaccagaagt tcaagggcaa ggccacactg actctggata agtccagcag cactgcctac 240
atggatctgc tgtctctgac aagcgaggac agcgccgtct actattgcgc catctctagg 300
tacggcagca gcagcttcta cttcgatgtg tggggcgccg gcacaacagt ggctgtgagc 360
agcggtggcg gtggctcggg cggtggtggg tcgggtggcg gcggatctca gatcgtgctg 420
agccagagcc cagctattct gtccgccagc cccggcgaga aggtgactat gacttgtagg 480
gccagctcca gcatcagcta catgcactgg taccagcaga agcccggctc ctccccaaag 540
ccttggatca gcgccactag caatctggcc agcggcgtgc cagccagatt cagcggaagc 600
ggcagcggca ctagctactc tctgactatc tccggcgtgg aagctgagga tgccgccact 660
tactactgcc agcagtggtc cagcaaccct cctactttcg gcggcggcac aaatctggag 720
atcaag 726
<210> 132
<211> 714
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 132
agcatcgtca tgacacaaag ccctaagagc atgagcatga gcgtgggcga gagagtgact 60
ctgagctgta aggccagcga gaatgtgggc ggctacgtga gctggtatca gcagaagcca 120
gatcagagcc caaagctgct gatctacggc gccagcagca gacacactgg cgtgccagat 180
aggttcactg gcagcggctc cgagacagac ttcactctga ctatcagcag cgtccaagcc 240
gaagatctgg ccgcctatca ctgcggccag aactacatct acccattcac attcggcggc 300
ggcacaaagc tggagatcaa gggtggcggt ggctcgggcg gtggtgggtc gggtggcggc 360
ggatctcaag tgcagctcca gcagagcgga gctgagctgg tgagacccgg cactagcgtg 420
aagatcagct gtaaggccag cggctacgcc ttcactaatt actggctggg ctggatgaag 480
cagagacccg gccatggact ggagtggatc ggcgacttct acccaaggac tggcaacact 540
ttctacaacg agaactttaa gggcaaggtg actctgactg ccgataagtc cagcaacact 600
gcctacatgc agctgtcctc tctgactagc gaagatagcg ccgtgtatct gtgtgctagg 660
gctggcactg gcttcgatta ctggggccaa ggcacaacac tgacagtgag cagc 714
<210> 133
<211> 714
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 133
caagtgcagc tccagcagag cggagctgag ctggtgagac ccggcactag cgtgaagatc 60
agctgtaagg ccagcggcta cgccttcact aattactggc tgggctggat gaagcagaga 120
cccggccatg gactggagtg gatcggcgac ttctacccaa ggactggcaa cactttctac 180
aacgagaact ttaagggcaa ggtgactctg actgccgata agtccagcaa cactgcctac 240
atgcagctgt cctctctgac tagcgaagat agcgccgtgt atctgtgtgc tagggctggc 300
actggcttcg attactgggg ccaaggcaca acactgacag tgagcagcgg tggcggtggc 360
tcgggcggtg gtgggtcggg tggcggcgga tctagcatcg tcatgacaca aagccctaag 420
agcatgagca tgagcgtggg cgagagagtg actctgagct gtaaggccag cgagaatgtg 480
ggcggctacg tgagctggta tcagcagaag ccagatcaga gcccaaagct gctgatctac 540
ggcgccagca gcagacacac tggcgtgcca gataggttca ctggcagcgg ctccgagaca 600
gacttcactc tgactatcag cagcgtccaa gccgaagatc tggccgccta tcactgcggc 660
cagaactaca tctacccatt cacattcggc ggcggcacaa agctggagat caag 714
<210> 134
<211> 723
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 134
caagccgtcg tgacacaaga gtccgctctg acaacttccc ccggcgagac tgtgacactg 60
acttgtagga gcagcactgg cgccgtgact atcagcaact acgccaactg ggtccaagag 120
aagccagatc atctgttcac tggactgatc ggcggcacaa ataataggcc tcccggcgtg 180
ccagccagat ttagcggctc tctgattggc gataaggctg ctctgacaat cactggcgcc 240
cagactgagg acgaggccat ctacttctgc gtcctctggt acagcaacca ctgggtgttc 300
ggcggcggca ctaagctgac tgtgctgggt ggcggtggct cgggcggtgg tgggtcgggt 360
ggcggcggat ctcaagtgca gctgcagcag agcggcgctg agctgatgaa gcccggcgct 420
tccgtcaaga tcagctgcaa gactactggc tataggttca gcagctactg gatcgagtgg 480
gtgaagcaga gacccggcca cggactggaa tggctgggcg aaatcctccc cggaaggggc 540
atcatcaact acaatgagaa ctttaggggc aaggccacat tcacagccga cacaagcagc 600
aacactgcct acgtgcagct gagctctctg acttccgagg acagcgccgt gtacttctgc 660
gccagaacag accctcctta tttcggcgtg tggggcgccg gaactactgt gactgtgtcc 720
agc 723
<210> 135
<211> 723
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 135
caagtgcagc tgcagcagag cggcgctgag ctgatgaagc ccggcgcttc cgtcaagatc 60
agctgcaaga ctactggcta taggttcagc agctactgga tcgagtgggt gaagcagaga 120
cccggccacg gactggaatg gctgggcgaa atcctccccg gaaggggcat catcaactac 180
aatgagaact ttaggggcaa ggccacattc acagccgaca caagcagcaa cactgcctac 240
gtgcagctga gctctctgac ttccgaggac agcgccgtgt acttctgcgc cagaacagac 300
cctccttatt tcggcgtgtg gggcgccgga actactgtga ctgtgtccag cggtggcggt 360
ggctcgggcg gtggtgggtc gggtggcggc ggatctcaag ccgtcgtgac acaagagtcc 420
gctctgacaa cttcccccgg cgagactgtg acactgactt gtaggagcag cactggcgcc 480
gtgactatca gcaactacgc caactgggtc caagagaagc cagatcatct gttcactgga 540
ctgatcggcg gcacaaataa taggcctccc ggcgtgccag ccagatttag cggctctctg 600
attggcgata aggctgctct gacaatcact ggcgcccaga ctgaggacga ggccatctac 660
ttctgcgtcc tctggtacag caaccactgg gtgttcggcg gcggcactaa gctgactgtg 720
ctg 723
<210> 136
<211> 735
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 136
gatatcgtca tgactcaagc cgccttcagc actccagtca ctctcggcac aagcgccagc 60
atcagctgta ggtccagcca gtctctgctg cacagcaacg gcatcactta tctgtactgg 120
tatctgcaga agcccggcca aagcccacag ctgctgatct accagatgag caatctggcc 180
agcggcgtgc cagatagatt cagcagcagc ggaagcggaa cagacttcac actgaggatc 240
tctagggtgg aagccgaaga tgtgggcgtc tattactgcg cccagaacct cgagttccct 300
ttcacattcg gcagcggcac taagctggag atcaagggtg gcggtggctc gggcggtggt 360
gggtcgggtg gcggcggatc tcaagtgcag ctgcagcaga gcggagccga gctgatgaag 420
cccggcgcct ccatgaagat cagctgcaag gccactggct acacattcag cacatactgg 480
atcgagtggg tcaagcagag acccggccac ggactggagt ggatcggaga gaatctgccc 540
ggaaggcaca tcactaacta caacgagaag ttcaagggca aggccacttt cacagccgac 600
actagcagca acactgccta catgcagctc agctctctga caagcgaaga tagcgccgtg 660
tactactgtg ctaggggaag gggcacttac tacttcgatt actggggcca aggcactcca 720
ctgactgtgt ccagc 735
<210> 137
<211> 735
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 137
caagtgcagc tgcagcagag cggagccgag ctgatgaagc ccggcgcctc catgaagatc 60
agctgcaagg ccactggcta cacattcagc acatactgga tcgagtgggt caagcagaga 120
cccggccacg gactggagtg gatcggagag aatctgcccg gaaggcacat cactaactac 180
aacgagaagt tcaagggcaa ggccactttc acagccgaca ctagcagcaa cactgcctac 240
atgcagctca gctctctgac aagcgaagat agcgccgtgt actactgtgc taggggaagg 300
ggcacttact acttcgatta ctggggccaa ggcactccac tgactgtgtc cagcggtggc 360
ggtggctcgg gcggtggtgg gtcgggtggc ggcggatctg atatcgtcat gactcaagcc 420
gccttcagca ctccagtcac tctcggcaca agcgccagca tcagctgtag gtccagccag 480
tctctgctgc acagcaacgg catcacttat ctgtactggt atctgcagaa gcccggccaa 540
agcccacagc tgctgatcta ccagatgagc aatctggcca gcggcgtgcc agatagattc 600
agcagcagcg gaagcggaac agacttcaca ctgaggatct ctagggtgga agccgaagat 660
gtgggcgtct attactgcgc ccagaacctc gagttccctt tcacattcgg cagcggcact 720
aagctggaga tcaag 735
<210> 138
<211> 726
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 138
cagatcgtgc tgacacagag cccagctctg atgagcgcca gccccggcga gaaggtcaca 60
atgacttgca gcgccacatc cagcgtgagc tacatctact ggtaccagca gaagcctagg 120
agcagcccta agccttggat ctacctcaca agcaatctgg ccagcggagt gccagctagg 180
ttcagcggaa gcggcagcgg cacaagctac tctctgacaa tctccagcat ggaagccgaa 240
gatgccgcca cttactactg ccagcagtgg agcagcaatc cacctacatt cggaggcggc 300
actaagctgg agatcaaggg tggcggtggc tcgggcggtg gtgggtcggg tggcggcgga 360
tctgaggtgc agctccaaca gagcggacca gagctggtga aacccggcgc cagcatgaag 420
atcagctgta aggcctccgg ctacagcttc actggctaca ctatgaactg ggtgaagcag 480
tcccacggca agaatctgga gtggatcgga ctgatcaacc catacaacgg cggcacaaac 540
tacaaccaga agttcaaggg caaggccact ctgacagtcg ataagagctc cagcactgcc 600
tacatggagc tgctgagcct cactagcgag gacagcgctg tgtactactg tgccttctcc 660
tactacggct ctaggggctt ctacttcgat tactggggcc aaggcacaac actgacagtg 720
tccagc 726
<210> 139
<211> 726
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 139
gaggtgcagc tccaacagag cggaccagag ctggtgaaac ccggcgccag catgaagatc 60
agctgtaagg cctccggcta cagcttcact ggctacacta tgaactgggt gaagcagtcc 120
cacggcaaga atctggagtg gatcggactg atcaacccat acaacggcgg cacaaactac 180
aaccagaagt tcaagggcaa ggccactctg acagtcgata agagctccag cactgcctac 240
atggagctgc tgagcctcac tagcgaggac agcgctgtgt actactgtgc cttctcctac 300
tacggctcta ggggcttcta cttcgattac tggggccaag gcacaacact gacagtgtcc 360
agcggtggcg gtggctcggg cggtggtggg tcgggtggcg gcggatctca gatcgtgctg 420
acacagagcc cagctctgat gagcgccagc cccggcgaga aggtcacaat gacttgcagc 480
gccacatcca gcgtgagcta catctactgg taccagcaga agcctaggag cagccctaag 540
ccttggatct acctcacaag caatctggcc agcggagtgc cagctaggtt cagcggaagc 600
ggcagcggca caagctactc tctgacaatc tccagcatgg aagccgaaga tgccgccact 660
tactactgcc agcagtggag cagcaatcca cctacattcg gaggcggcac taagctggag 720
atcaag 726
<210> 140
<211> 729
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 140
gatatcgtga tgactcagtc cccagccatc atgtccgcca gccccggcga gaaggtgact 60
atgacttgct ccgccagcag cagcgtgagc tacatgcact ggtaccagca gaagagcggc 120
acatccccaa agaggtggat ctacgacaca agcaagctgg ccagcggcgt gccagccaga 180
ttcagcggct ccggcagcgg aacaagctac tctctgacta tcagcagcat ggagacagag 240
gacgctgcca cttactactg ccagcagtgg agcagcaatc ctccaacttt cggcggaggc 300
actaagctgg agctgaaggg tggcggtggc tcgggcggtg gtgggtcggg tggcggcgga 360
tctgaagtgc agctgcaaca gagcggccca gagctcgtga agcccggcgc cagcatgaag 420
atcagctgca aggccagcgg ctacagcttc actggctaca caatgaactg ggtcaagcag 480
agccacggaa aaaatctgga gtggatcgga ctgatcaacc cttacaacgg cggcacaagg 540
tacaatcaga agttcaaggg caaggccact ctcactgtgg ataagagcag cagcactgcc 600
tacatggagc tgctgtctct gacaagcgag gatagcgccg tgtactactg cgccagctcc 660
tcctatagga acgacggcaa ctggtacttc gatgtgtggg gcgccggcac tactgtgaca 720
gtgagctcc 729
<210> 141
<211> 729
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 141
gaagtgcagc tgcaacagag cggcccagag ctcgtgaagc ccggcgccag catgaagatc 60
agctgcaagg ccagcggcta cagcttcact ggctacacaa tgaactgggt caagcagagc 120
cacggaaaaa atctggagtg gatcggactg atcaaccctt acaacggcgg cacaaggtac 180
aatcagaagt tcaagggcaa ggccactctc actgtggata agagcagcag cactgcctac 240
atggagctgc tgtctctgac aagcgaggat agcgccgtgt actactgcgc cagctcctcc 300
tataggaacg acggcaactg gtacttcgat gtgtggggcg ccggcactac tgtgacagtg 360
agctccggtg gcggtggctc gggcggtggt gggtcgggtg gcggcggatc tgatatcgtg 420
atgactcagt ccccagccat catgtccgcc agccccggcg agaaggtgac tatgacttgc 480
tccgccagca gcagcgtgag ctacatgcac tggtaccagc agaagagcgg cacatcccca 540
aagaggtgga tctacgacac aagcaagctg gccagcggcg tgccagccag attcagcggc 600
tccggcagcg gaacaagcta ctctctgact atcagcagca tggagacaga ggacgctgcc 660
acttactact gccagcagtg gagcagcaat cctccaactt tcggcggagg cactaagctg 720
gagctgaag 729
<210> 142
<211> 726
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 142
cagattgtgc tgtcccagtc cccagccatt ctgagcgcca gccccggcga gaaggtgact 60
atgacttgta gggccacaag cagcgtgagc tacatgtact ggtaccagca gaagcccggc 120
agcagcccta agccttggat ctacgccaca agcaatctgg ccagcggcgt cccagctaga 180
tttagcggca gcggatccgg cactagctat tctctgacta tctctagggt cgaggccgaa 240
gatgccgcca catactactg ccagcagtgg tcctccaatc ctccaacatt cggcggcgga 300
actaagctgg agaagaaggg tggcggtggc tcgggcggtg gtgggtcggg tggcggcgga 360
tctgaagtgc agctgcagca gagcggacca gagctggtca agcccggcgc cagcatgaag 420
atcagctgta aggccagcgg ctacagcatc actggctaca ctatgaactg ggtgaagcag 480
agccacggca agaacctcga gtggattggc ctcgtgaacc catacaacgg cggcacttcc 540
tacaaccaga agttcaaagg caaggccaca ctcacagtcg ataagtccag ctccacagcc 600
tacatggagc tgctgtctct gaagagcgag gatagcgctg tctactactg tgccatcagc 660
agatacggca gcgagagctg gtacttcgac gtgtggggcg ccggcacaac agtgacagtg 720
agcagc 726
<210> 143
<211> 726
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 143
gaagtgcagc tgcagcagag cggaccagag ctggtcaagc ccggcgccag catgaagatc 60
agctgtaagg ccagcggcta cagcatcact ggctacacta tgaactgggt gaagcagagc 120
cacggcaaga acctcgagtg gattggcctc gtgaacccat acaacggcgg cacttcctac 180
aaccagaagt tcaaaggcaa ggccacactc acagtcgata agtccagctc cacagcctac 240
atggagctgc tgtctctgaa gagcgaggat agcgctgtct actactgtgc catcagcaga 300
tacggcagcg agagctggta cttcgacgtg tggggcgccg gcacaacagt gacagtgagc 360
agcggtggcg gtggctcggg cggtggtggg tcgggtggcg gcggatctca gattgtgctg 420
tcccagtccc cagccattct gagcgccagc cccggcgaga aggtgactat gacttgtagg 480
gccacaagca gcgtgagcta catgtactgg taccagcaga agcccggcag cagccctaag 540
ccttggatct acgccacaag caatctggcc agcggcgtcc cagctagatt tagcggcagc 600
ggatccggca ctagctattc tctgactatc tctagggtcg aggccgaaga tgccgccaca 660
tactactgcc agcagtggtc ctccaatcct ccaacattcg gcggcggaac taagctggag 720
aagaag 726
<210> 144
<211> 732
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 144
gatatcgtgc tgactcagag cccagcctct ctggtggtgt ctctgggaca gagggccaca 60
atcagctgta ggacttccaa gagcgtgagc agctccgcct acagctacat gcactggtac 120
cagcagaagc ccggccagcc tcctaaggtg ctgatctatc tggccagcaa tctggagagc 180
ggcgtcccag ctagattcag cggctccgga agcggcactg acttcactct gaacatccac 240
ccagtggaag aggaggatgc cgccacatac tactgccagc actctaggga gctgcctttc 300
acatttggca gcggaactaa gctggagatc aagggtggcg gtggctcggg cggtggtggg 360
tcgggtggcg gcggatctca agtgcatctg cagcagagcg gcgctgagct gatgaagccc 420
ggcgccagcg tgaagattag ctgcaaggcc actggctaca cattcaacat ctactggatc 480
gactgggtga agcagaggcc cggccacgga ctggaatgga tcggcgaaat tctgcccggc 540
agcggcaaca ctcactacaa cgagaacttc aagggcaagg ccacaatgac agccgacaca 600
agctccaaca ctgcttacat gcagctgact tctctgacta gcgaggacag cgccgtgtac 660
tattgcgcta ggacagacgg aaggggctac ttcgattact ggggccaagg cactacactc 720
acagtgagca gc 732
<210> 145
<211> 732
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 145
caagtgcatc tgcagcagag cggcgctgag ctgatgaagc ccggcgccag cgtgaagatt 60
agctgcaagg ccactggcta cacattcaac atctactgga tcgactgggt gaagcagagg 120
cccggccacg gactggaatg gatcggcgaa attctgcccg gcagcggcaa cactcactac 180
aacgagaact tcaagggcaa ggccacaatg acagccgaca caagctccaa cactgcttac 240
atgcagctga cttctctgac tagcgaggac agcgccgtgt actattgcgc taggacagac 300
ggaaggggct acttcgatta ctggggccaa ggcactacac tcacagtgag cagcggtggc 360
ggtggctcgg gcggtggtgg gtcgggtggc ggcggatctg atatcgtgct gactcagagc 420
ccagcctctc tggtggtgtc tctgggacag agggccacaa tcagctgtag gacttccaag 480
agcgtgagca gctccgccta cagctacatg cactggtacc agcagaagcc cggccagcct 540
cctaaggtgc tgatctatct ggccagcaat ctggagagcg gcgtcccagc tagattcagc 600
ggctccggaa gcggcactga cttcactctg aacatccacc cagtggaaga ggaggatgcc 660
gccacatact actgccagca ctctagggag ctgcctttca catttggcag cggaactaag 720
ctggagatca ag 732
<210> 146
<211> 714
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 146
caaattgttc tcacacagag ccctcctatc atgagcgcca gccccggcga gaaggtgact 60
atgacttgtt ccgccagcag caacatcagc tacatgcact ggtaccagca gaagtccggc 120
acaagcccaa agaggtggat ctacgacaca agcaagctgg ccagcggcgt gccagccaga 180
tttagcggct ccggaagcgg cactagctac tctctgacaa tcagcagcat ggaagccgag 240
gacgccgcta catactactg ccagcagtgg agctccgtcc cactgacttt cggcgctggc 300
actaagctgg agatcaaggg tggcggtggc tcgggcggtg gtgggtcggg tggcggcgga 360
tctcaggttc agctgcagca gtccggcgct gagctcatga agcccggcgc cagcgtgaag 420
atcagctgca aggccactgg ctacacattc agcagctact ggatcgagtg ggtgaagcag 480
agacccggcc acggactgga atggctggga gagtttctgc ctagaagcgg caagacaaac 540
tacaacgagg agtttagggg caaggctaca ttcactgccg acacatccag caacactgcc 600
tacatgcagc tgagcagcct cacaagcgag gattccgccg tctactactg tgctaggact 660
gatcctcctt acttcggagt gtggggcgct ggcacaatgg tggctgtgag cagc 714
<210> 147
<211> 714
<212> DNA
<213> Artificial Sequence
<220>
<223> 核酸分子
<400> 147
caggttcagc tgcagcagtc cggcgctgag ctcatgaagc ccggcgccag cgtgaagatc 60
agctgcaagg ccactggcta cacattcagc agctactgga tcgagtgggt gaagcagaga 120
cccggccacg gactggaatg gctgggagag tttctgccta gaagcggcaa gacaaactac 180
aacgaggagt ttaggggcaa ggctacattc actgccgaca catccagcaa cactgcctac 240
atgcagctga gcagcctcac aagcgaggat tccgccgtct actactgtgc taggactgat 300
cctccttact tcggagtgtg gggcgctggc acaatggtgg ctgtgagcag cggtggcggt 360
ggctcgggcg gtggtgggtc gggtggcggc ggatctcaaa ttgttctcac acagagccct 420
cctatcatga gcgccagccc cggcgagaag gtgactatga cttgttccgc cagcagcaac 480
atcagctaca tgcactggta ccagcagaag tccggcacaa gcccaaagag gtggatctac 540
gacacaagca agctggccag cggcgtgcca gccagattta gcggctccgg aagcggcact 600
agctactctc tgacaatcag cagcatggaa gccgaggacg ccgctacata ctactgccag 660
cagtggagct ccgtcccact gactttcggc gctggcacta agctggagat caag 714
<210> 148
<211> 327
<212> PRT
<213> Artificial Sequence
<220>
<223> 人IgG4的恒定区
<400> 148
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
1 5 10 15
Ser Thr Ser Glu Ser 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 Lys Thr
65 70 75 80
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro
100 105 110
Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
115 120 125
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
130 135 140
Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
145 150 155 160
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
180 185 190
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu
195 200 205
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
225 230 235 240
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
245 250 255
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
260 265 270
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
275 280 285
Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
290 295 300
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
305 310 315 320
Leu Ser Leu Ser Leu Gly Lys
325
<210> 149
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> tanezumab轻链氨基酸序列
<400> 149
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Phe His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Glu His Thr Leu 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> 150
<211> 447
<212> PRT
<213> Artificial Sequence
<220>
<223> tanezumab重链氨基酸序列
<400> 150
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ile Gly Tyr
20 25 30
Asp Leu Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Gly Ile Ile Trp Gly Asp Gly Thr Thr Asp Tyr Asn Ser Ala Val Lys
50 55 60
Ser Arg Val Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Phe Ser Leu
65 70 75 80
Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Gly Tyr Trp Tyr Ala Thr Ser Tyr Tyr Phe Asp Tyr 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 Cys Ser Arg Ser Thr Ser Glu Ser 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 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys
210 215 220
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser
290 295 300
Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 151
<211> 213
<212> PRT
<213> Artificial Sequence
<220>
<223> MNAC13轻链氨基酸序列
<400> 151
Asp Ile Val Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Lys Leu Leu Ile Tyr
35 40 45
Thr Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu
65 70 75 80
Asp Val Ala Thr Tyr Tyr Cys His Gln Trp Ser Ser Tyr Pro Trp Thr
85 90 95
Phe Gly Gly Gly Thr Lys Val 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> 152
<211> 449
<212> PRT
<213> Artificial Sequence
<220>
<223> MNAC13重链氨基酸序列
<400> 152
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30
Thr Met Ser Trp Ala Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Tyr Ile Ser Lys Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr 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 Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Ala Met Phe Gly Asn Asp Phe Phe Phe Pro Met Asp Arg
100 105 110
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125
Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
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 Lys Thr Tyr Thr Cys Asn Val
195 200 205
Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys
210 215 220
Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 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 Gln Glu
260 265 270
Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe 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 Gly Leu Pro Ser Ser 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 Gln 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 Glu 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 Leu
435 440 445
Gly

Claims (33)

1.一种能够特异性识别TrkA的抗体或其抗原结合片段，其特征在于，所述抗体含有选自下列至少之一的CDR序列或与其具有至少95％同一性的氨基酸序列：
重链可变区CDR序列：SEQ ID NO:1～27，
轻链可变区CDR序列：SEQ IN NO:28～54。
2.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体包括：
分别如SEQ ID NO:1、2和3或者与SEQ ID NO:1、2和3具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:4、5和6或者与SEQ ID NO:4、5和6具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:7、8和9或者与SEQ ID NO:7、8和9具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:10、11和12或者与SEQ ID NO:10、11和12具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:13、14和15或者与SEQ ID NO:13、14和15具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:16、17和18或者与SEQ ID NO:16、17和18具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO19、20和21或者与SEQ ID NO:19、20和21具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:22、23和24或者与SEQ ID NO:22、23和24具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:25、26和27或者与SEQ ID NO:25、26和27具有至少95％同一性的氨基酸序列所示的重链可变区CDR1、CDR2、CDR3序列。
3.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体包括：
分别如SEQ ID NO:28、29和30或者与SEQ ID NO:28、39和30具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:31、32和33或者与SEQ ID NO:31、32和33具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:34、35和36或者与SEQ ID NO:34、35和36具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:37、38和39或者与SEQ ID NO:37、38和39具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:40、41和42或者与SEQ ID NO:40、41和42具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:43、44和45或者与SEQ ID NO:43、44和45具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:46、47和48或者与SEQ ID NO:46、47和48具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:49、50和51或者与SEQ ID NO:49、50和51具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列；或者
分别如SEQ ID NO:52、53和54或者与SEQ ID NO:52、53和54具有至少95％同一性的氨基酸序列所示的轻链可变区CDR1、CDR2、CDR3序列。
4.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体或其抗原结合片段特异性识别TrkA的胞外区。
5.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体含有重链框架区序列和轻链框架区序列的至少之一，所述重链框架区序列和轻链框架区序列的至少之一的至少一部分来自于鼠源抗体、人源抗体、灵长目源抗体或其突变体的至少之一。
6.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体具有如SEQ IDNO：55～63任一项所示氨基酸序列的重链可变区。
7.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体具有如SEQ IDNO：64～72任一项所示氨基酸序列的轻链可变区。
8.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体含有重链恒定区和轻链恒定区的至少之一，所述重链恒定区和轻链恒定区的至少之一的至少一部分来自于鼠源抗体、人源抗体、灵长目源抗体或其突变体的至少之一。
9.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体的轻链恒定区和重链恒定区均来自于人源IgG抗体或其突变体。
10.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体的轻链恒定区和重链恒定区均来自于人源IgG4。
11.根据权利要求10所述的抗体或其抗原结合片段，其特征在于，所述抗体的Fc区域与野生型相比具有S10P，F16A，L17A，R191K突变以及229K缺失突变。
12.根据权利要求10所述的抗体或其抗原结合片段，其特征在于，所述抗体恒定区的全长序列如SEQ ID NO:74或75所示。
13.根据权利要求10所述的抗体或其抗原结合片段，其特征在于，所述抗体具有SEQ IDNO:76～84任一项所示氨基酸序列的重链和具有SEQ ID NO:85～93任一项所示氨基酸序列的轻链。
14.根据权利要求1所述的抗体或其抗原结合片段，其特征在于，所述抗体为单链抗体、多聚体抗体、CDR移植抗体或小分子抗体。
15.根据权利要求14所述的抗体或其抗原结合片段，其特征在于，所述抗体为单链抗体，所述抗体具有SEQ ID NO:94～111所示的氨基酸序列。
16.根据权利要求14所述的抗体或其抗原结合片段，其特征在于，所述小分子抗体包括Fab抗体、Fv抗体、单域抗体以及最小识别单位的至少之一。
17.一种核酸分子，其特征在于，所述核酸分子编码权利要求1～16任一项所述的抗体或其抗原结合片段。
18.根据权利要求17所述的核酸分子，其特征在于，所述核酸分子为DNA。
19.根据权利要求18所述的核酸分子，其特征在于，所述核酸分子具有如SEQ ID NO:112～120任一项所示核苷酸序列或具有SEQ ID NO:121～129任一项所示核苷酸序列或具有SEQ ID NO:130～147任一项所示核苷酸序列。
20.一种表达载体，其特征在于，携带权利要求17～19任一项所述的核酸分子。
21.根据权利要求20所述的表达载体，其特征在于，所述表达载体为真核表达载体。
22.一种重组细胞，其特征在于，所述重组细胞携带权利要求17～19任一项所述的核酸分子，或者表达权利要求1～16任一项所述的抗体或其抗原结合片段。
23.根据权利要求22所述的重组细胞，其特征在于，所述重组细胞是通过将权利要求20或21所述的表达载体引入至宿主细胞中而获得的。
24.根据权利要求23所述的重组细胞，其特征在于，通过电转导的方法将所述表达载体引入所述宿主细胞中。
25.根据权利要求22所述的重组细胞，其特征在于，所述重组细胞为真核细胞。
26.根据权利要求22所述的重组细胞，其特征在于，所述重组细胞为哺乳动物细胞。
27.一种药物组合物，其特征在于，含有权利要求1～16任一项所述的抗体，权利要求17～19任一项所述的核酸分子，权利要求20～21任一项所述的表达载体或权利要求22～26任一项所述的重组细胞。
28.权利要求1～16任一项所述的抗体、权利要求17～19任一项所述的核酸分子、权利要求20～21任一项所述的表达载体或权利要求22～26任一项所述的重组细胞、权利要求27所述的药物组合物在制备药物中的用途，所述药物用于治疗或者预防疼痛、癌症、炎症或炎性疾病、神经变性疾病、舍格伦氏综合征、子宫内膜异位、糖尿病性周围神经病变、前列腺炎、盆腔疼痛综合征、与骨重塑调节失衡相关的疾病以及由结缔组织生长因子异常信号传导引起的疾病。
29.根据权利要求28所述的用途，其特征在于，所述药物用于治疗或预防神经性疼痛、炎性疼痛、与癌症有关的疼痛、与骨折有关的疼痛、与手术有关的疼痛、炎性肺病、间质性膀胱炎、痛性膀胱综合征、炎性肠疾病、炎性皮肤病、雷诺氏综合征、特发性肺纤维化、瘢痕(肥大型、瘢痕瘤型和其他形式)、硬化、心内膜心肌纤维化、心房纤维化、骨髓纤维化、进行性块状纤维化(肺)、肾源性系统性纤维化、硬皮病、系统性硬化、关节纤维化、眼部纤维化、非小细胞肺癌、乳头状甲状腺癌、多形性成胶质细胞瘤、结肠直肠癌、黑色素瘤、胆管癌或肉瘤、急性骨髓性白血病、大细胞神经内分泌癌、成神经细胞瘤、前列腺癌、成神经细胞瘤、胰腺癌、黑色素瘤、头颈鳞状细胞癌或胃癌。
30.一种检测TrkA的试剂盒，其特征在于，包括权利要求利要求1～16任一项所述的抗体。
31.权利要求1～16任一项所述的抗体、权利要求17～19任一项所述的核酸分子、权利要求20～21任一项所述的表达载体或权利要求22～26任一项所述的重组细胞在制备试剂盒中的用途，所述试剂盒用于检测TrkA或者诊断TrkA相关的疾病。
32.一种小鼠B细胞，其特征在于，所述B细胞的基因组中携带编码恒定区的序列，所述恒定区序列与人IgG4的恒定区相比，存在S108P，F114A，L115A，R289K突变以及327K缺失突变。
33.权利要求32所述的小鼠B细胞在制备单克隆抗体中的用途。
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