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CN113512116B - 一种抗igf-1r抗体及其应用 - Google Patents

一种抗igf-1r抗体及其应用 Download PDF

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CN113512116B
CN113512116B CN202010280882.7A CN202010280882A CN113512116B CN 113512116 B CN113512116 B CN 113512116B CN 202010280882 A CN202010280882 A CN 202010280882A CN 113512116 B CN113512116 B CN 113512116B
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郭树华
马琳
张鹏
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Suzhou Pharmaceutical Technology Co ltd
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Abstract

本发明提供一种可以结合IGF‑1R的抗体。该抗体由轻链和重链组成，所述轻链和重链可变区为SEQ ID NO：2和SEQ ID NO：3、SEQ ID NO：4和SEQ ID NO：5、SEQ ID NO：6和SEQ ID NO：7、SEQ ID NO：8和SEQ ID NO：9、SEQ ID NO：10和SEQ ID NO：11、SEQ ID NO：12和SEQ ID NO：13、SEQ ID NO：14和SEQ ID NO：15组合中的一种。本发明还提供了编码上述结合IGF‑1R的抗体的核酸分子、含有上述核酸的载体、上述载体转化的宿主细胞，以及这些核酸、载体、宿主细胞的应用。

Description

一种抗IGF-1R抗体及其应用
技术领域
本发明属于生物医药领域，具体涉及一种抗IGF-1R抗体及其应用。
背景技术
甲状腺相关性眼病(thyroid associated ophthalmopathy，TAO)又名内分泌突眼、浸润性突眼、甲状腺眼病，多并发于Graves病(弥漫性甲状腺肿伴甲状腺功能亢进症)，又称之为Graves眼病(Graves'Ophthalmopathy，GO)，约占眼眶疾病的20％。甲状腺相关性眼病可引起眼球突出、眼睑挛缩、眼外肌功能障碍、球结膜充血、眶周水肿等，严重时会导致暴露性角膜炎、复视和压迫性视神经病变，后者可导致失明，严重影响患者的生活质量。甲状腺相关性眼病是由甲状腺上皮细胞、眼眶前脂肪细胞及成纤维细胞一起表达的共同抗原引发的以细胞免疫为主的位点特异性自身免疫疾病。炎症反应、眼眶成纤维细胞增多和眼眶脂肪细胞的增多导致眼球突出，引发甲状腺相关性眼病的症状。胰岛素样生长因子1受体(insulin-like growth factor 1receptor，IGF-1R)和促甲状腺激素受体(thyroid-stimulating hormone receptor，TSHR)在甲状腺相关性眼病患者眼眶成纤维细胞中的水平异常偏高，且二者间存在着复杂的信号通路网络联系，与疾病的发生密切相关。
IGF-1R属于酪氨酸蛋白激酶受体家族，是一种细胞表面的跨膜蛋白，可被IGF-1及IGF-2(均为胰岛素增长因子)激活，其过度表达可能与多发性硬化、克罗恩病、肺纤维化等恶性肿瘤及自身免疫性疾病发病相关。大部分的Graves病患者体内可以检测到抗IGF-1R的抗体，而正常人体内少有发现。Graves病患者血清分离的IgG(Graves相关的免疫球蛋白G)可以替换IGF-1R与眼眶成纤维细胞表面上的位点结合，IGF-1R或Graves病相关的IgG可激活甲状腺相关性眼病患者来源的IGF-1R阳性的眼眶成纤维细胞，从而激活Akt/FRAP/mTOR/P70s6k通道诱导眼眶成纤维细胞上白细胞介素-16和调节活化正常T细胞表达和分泌的因子(regulated upon activation normal T cell expressed and secreted factor，RANTES)的表达，促进T细胞趋化因子的合成，引起T淋巴细胞的炎性浸润和透明质酸的产生，这些都表明IGF-1R可能作为第二抗原参与了Graves病的发展。另外，有报道显示，IGF-1R与TSHR的下游信号有大量重叠，且抗IGF-1R的单克隆抗体可以阻断IGF-1R的信号传导，及减弱TSHR通路的下游信号，减轻促甲状腺激素引起的炎症反应，提示IGF-1R参与了TSHR信号传导，并且IGF-1R与TSHR可能形成了一个复杂的功能复合体，在Graves病相关的异常信号传导方面起着协同作用。
抗IGF-1R抗体(参见专利WO2004087756、US20100158919、US20140193404)在治疗甲状腺眼病的临床实验中达到了主要和所有次要终点，可以显著改善眼球突出的症状。除了上述所列举的抗体外，目前国内外没有其他针对IGF-1R抗体用于治疗甲状腺眼病的应用，比如专利CN200880011970.4、CN200880114015.3、CN200780011979.0、CN200980137723.3、US20190040141、US20090175868、US20060018910中的抗IGF-1R抗体均是用于恶性肿瘤的治疗。在甲状腺相关眼病领域，开发新型的用于治疗该疾病的IGF-1R治疗性抗体十分的必要。
发明内容
为了解决上述问题，本发明提供了一种结合IGF-1R的抗体及其在药品中的应用，该抗体及相关药品可用于甲状腺相关眼病的预防或者治疗。
一方面，本发明提供了一种结合IGF-1R的抗体。
所述的抗体由轻链和重链组成。
所述的抗体的轻链可变区选自SEQ ID NO：2、4、6、8、10、12或14所示的氨基酸序列中的一种。
所述的抗体的轻链恒定区的氨基酸序列如SEQ ID NO：16所示。
所述的抗体的轻链由轻链可变区与SEQ ID NO：16所示的轻链恒定区拼接形成。
所述的抗体的重链可变区选自SEQ ID NO：3、5、7、9、11、13、15所示的氨基酸序列中的一种。
所述的抗体的重链恒定区的氨基酸序列如SEQ ID NO：17所示。
所述的抗体的重链由重链可变区与SEQ ID NO：17所示的重链恒定区拼接形成。
具体地，所述的抗体的轻链和重链可变区为SEQ ID NO：2和SEQ ID NO：3所示的氨基酸序列；
或，SEQ ID NO：4和SEQ ID NO：5所示的氨基酸序列；
或，SEQ ID NO：6和SEQ ID NO：7所示的氨基酸序列；
或，SEQ ID NO：8和SEQ ID NO：9所示的氨基酸序列；
或，SEQ ID NO：10和SEQ ID NO：11所示的氨基酸序列；
或，SEQ ID NO：12和SEQ ID NO：13所示的氨基酸序列；
或，SEQ ID NO：14和SEQ ID NO：15所示的氨基酸序列。
具体地，所述的轻链和重链通过链间二硫键形成完整的抗IGF-1R抗体。
另一方面，本发明提供了编码上述任一技术方案得到的抗体的核苷酸序列。
再一方面，本发明提供了一种表达载体，所述的表达载体包含编码前述任一技术方案得到的抗体的核酸分子。
又一方面，本发明提供了一种宿主细胞，所述的宿主细胞包含前述表达载体。
又一方面，本发明提供了上述结合IGF-1R的抗体在制备甲状腺相关眼病的药物中的应用。
所述的应用包括但不限于利用上述结合IGF-1R的抗体、编码该抗体的核苷酸序列及其突变型、含有上述核苷酸序列的表达载体、含有上述载体的宿主细胞在制备甲状腺相关眼病药物中的应用。
具体地，所述的抗体经过分离纯化。
所述的核苷酸序列、表达载体和宿主细胞通过表达抗体实现所述应用。
所述的抗体、核苷酸序列、表达载体通过直接添加至药物中实现所述应用。
所述的药物可应用的甲状腺相关眼病包括但不限于：眼球突出、眼睑退缩、上睑迟落、眼外肌肥大、结膜充血、眶周组织水肿、眼睑闭合不全、畏光、流泪、异物感、视力下降或复视。
又一方面，本发明提供了上述结合IGF-1R的抗体的制备方法。
具体地，所述的制备方法为利用编码上述任一技术方案提供的抗体的核苷酸序列构建表达载体转染宿主细胞，通过宿主细胞的表达获得结合IGF-1R的抗体。
又一方面，本发明提供了一种药物组合物。
所述的药物组合物包含上述结合IGF-1R的抗体、编码该抗体的核苷酸序列及其突变体、含有上述核苷酸序列或其突变体的表达载体中的一种或多种。
所述的药物组合物还包括药学上可接受的载体和/或辅料。
进一步地，所述的药学上可接受的载体和/或辅料包括但不限于：增溶剂、稳定剂和赋形剂。
所述的药物组合物的给药方式包括但不限于：皮下注射、皮内注射、肌肉注射、静脉注射、静脉滴注、眼睑注射。
所述的药物组合物可应用于甲状腺相关眼病的预防和/或治疗，所述的甲状腺相关眼病包括但不限于眼球突出、眼睑退缩、上睑迟落、眼外肌肥大、结膜充血、眶周组织水肿、眼睑闭合不全、畏光、流泪、异物感、视力下降或复视。
附图说明
图1为PHP1003针对人IGF-1R的亲和力检测结果。
图2为PHP1003针对猴子IGF-1R的亲和力检测结果。
图3为PHP1003针对大鼠IGF-1R的亲和力检测结果。
图4为PHP1003蛋白的SEC-HPLC检测结果。
图5为对照抗体的SEC-HPLC检测结果。
图6为测试品PHP1003及对照品给药大鼠甲状腺功能亢进及甲状腺相关眼病模型4周后，眼外肌组织染色光镜下观察结果。
具体实施方式
下面结合具体实施例，对本发明作进一步详细的阐述，下述实施例不用于限制本发明，仅用于说明本发明。以下实施例中所使用的实验方法如无特殊说明，实施例中未注明具体条件的实验方法，通常按照常规条件，下述实施例中所使用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1抗IGF-1R抗体的序列合成与载体构建
轻链设计：将轻链可变区和轻链恒定区直接拼接形成轻链；
重链设计：将重链可变区和重链恒定区直接拼接形成重链；
针对轻链和重链的氨基酸序列，按照人宿主细胞进行密码子优化并常规合成基因，合成同时添加5'(EcoRI酶切位点)和5'UTR([SEQ ID NO：20])以及3'UTR([TGATGA])和3'(HindIII酶切位点)。
上述基因委托苏州金唯智生物科技有限公司合成。
将基因通过5'EcoRI和3'HindIII克隆至载体pTT5(氨苄抗性)。挑选克隆进行测序，选择测序正确的菌体进行保种并进行菌体的扩大培养，扩大的菌体用于质粒的提取。构建完成的轻链重组载体和重链重组载体组合命名为PHP1003。按照上述同样的基因合成与载体构建方法，获得可表达对照抗体的质粒，其中对照抗体的轻链序列如SEQ ID NO：18和重链序列如SEQ ID NO：19。
本实施例中轻、重链序列设计的序列组成如下：
序列组成
PHP1003轻链 轻链可变区SEQ ID NO：2、轻链恒定区SEQ ID NO：16
PHP1003重链 重链可变区SEQ ID NO：3、重链恒定区SEQ ID NO：17
对照抗体 轻链序列SEQ ID NO：18、重链序列SEQ ID NO：19
实施例2抗IGF-1R抗体的瞬转表达及结合IGF-1R蛋白的检测
按照如下的方式对提取后的质粒进行细胞转染和蛋白的分离纯化：
1、测定细胞密度，活力应当大于95％，用(预热的)HEK293培养基将HEK293细胞密度调节成3×106细胞/mL，轻轻摇匀并分装细胞(转染量为转染体系的90％)，注意摇瓶中细胞体积不超过摇瓶规格的1/3，放入摇床待用。
2、根据转染细胞体积计算转染缓冲液opti-MEM的体积，该体积为转染体系的1/10；计算转染试剂PEI的量，其比例为3μL/mL转染细胞；计算转染DNA的总量，其比例为1μg/mL转染细胞。
具体转染的操作过程如下：
取1支50ml离心管，加入转染体系10％的MEM，加入质粒，混匀后过滤，静置5min，向DNA混悬液中加入PEI，轻柔混匀(轻轻颠倒混匀2-3次)后静置15-20min。随后将复合物轻柔加入分装好的细胞中，边加入边轻轻晃动摇瓶；转染完成的细胞放入37℃摇床进行培养。
培养3天后离心收集培养上清，测定表达上清(表达的轻、重链融合蛋白命名为PHP1003，具有抗IGF-1R抗性)与IGF-1R蛋白的亲和力。采用Octet RED96蛋白质相互作用工作站对上述表达的融合蛋白PHP1003进行与人IGF-1R(Human IGF-1R，ACRO，货号IGR-H5229)、猴子IGF-1R(Cyno IGF-1R，ACRO，货号IGR-C5225)和大鼠IGF-1R(Rat IGF-1R，ACRO，货号IGR-R5224)蛋白的亲和力检测，选用生物传感器AHC(货号18-5060)对样品进行捕获，之后将捕获的样品与人IGF-1R、猴子IGF-1R和大鼠IGF-1R蛋白进行结合、解离的动力学检测。动力学使用1：1结合模型进行拟合分析。其中简要的作用步骤为：蛋白加载200s，结合180s，解离300s，再生30s。
使用Fortebio仪器测定亲和力。结果如下表和图1-3所示。
Figure BDA0002446524100000061
实施例3抗IGF-1R抗体的分离纯化
用常规的亲和纯化方法分离纯化HEK293细胞瞬转表达的上清中的抗IGF-1R抗体，简要的步骤为用5-10倍体积的结合缓冲液平衡填料柱、上清样品上样、结合缓冲液洗脱杂蛋白、蛋白洗脱。将分离纯化后的蛋白进行SEC-HPLC纯度检测，检测结果如图4、5所示。其中PHP1003蛋白纯度为96.44％，对照抗体的纯度为99.76％。
实施例4抗IGF-1R抗体PHP1003和对照抗体在甲亢动物模型中的药效
本实施例通过腹腔注射牛甲状腺球蛋白诱导大鼠甲状腺功能亢进及甲状腺相关眼病模型，观察PHP1003对大鼠甲状腺相关眼病的影响，包括以下步骤：
(1)模型组大鼠每2周按150μg/只的剂量腹腔注射牛甲状腺球蛋白(Sigma-Aldrich，货号609310)进行造模，持续4周完成造模。
(2)分别用测试品及对照品给药，试验动物分组情况如下表所示：
Figure BDA0002446524100000062
组别设计：正常对照组(Sham Group)、阴性对照组(Vehicle Group)、供试品治疗组(L Group；H Group)、对照抗体组(P.Group)，共5组。
具体给药方案按照下表操作：
Figure BDA0002446524100000063
Figure BDA0002446524100000071
(3)给药4周后安乐死动物进行取材。将大鼠安乐死后眼外肌取材，进行组织染色，光镜下采图观察。
(4)结果如图6所示：
Sham组可见肌纤维均质红染，排列整齐，肌间毛细血管丰富，肌间距正常。Vehicle组可见各类眼外肌病变，主要包括肌纤维严重肿胀、变性、坏死、溶解，肌间距缩窄或增宽，肌间纤维结缔组织和血管增生。提示SD大鼠甲状腺眼病模型成功建立。
PHP1003-L组可见病变基本涵盖Vehicle组各类病变，肌纤维坏死溶解程度较Vehiclel组略轻微。PHP1003-H组病变轻微，肌纤维排列基本整齐，可见部分肌纤维轻微肿胀或变性，说明PHP1003-H组药物治疗SD大鼠甲状腺眼病有一定效果。P组眼外肌染色结果显示眼外肌肿胀肥大、肌纤维变性、坏死程度较Vehicle组减轻，结果说明PHP1003-H组和P组可减轻大鼠甲状腺眼病眼外肌病变程度。PHP1003-H组和P组相比，SD大鼠眼外肌病变程度更轻微，P组眼外肌肿胀程度更大，肌间距更窄，说明在同等剂量下PHP1003-H组对SD大鼠甲状腺眼病的治疗效果更优异。
序列表
<110> 苏州普乐康医药科技有限公司
<120> 一种IGF-1R抗体及其应用
<130> 20200409
<160> 20
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1337
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Glu Ile Cys Gly Pro Gly Ile Asp Ile Arg Asn Asp Tyr Gln Gln Leu
1 5 10 15
Lys Arg Leu Glu Asn Cys Thr Val Ile Glu Gly Tyr Leu His Ile Leu
20 25 30
Leu Ile Ser Lys Ala Glu Asp Tyr Arg Ser Tyr Arg Phe Pro Lys Leu
35 40 45
Thr Val Ile Thr Glu Tyr Leu Leu Leu Phe Arg Val Ala Gly Leu Glu
50 55 60
Ser Leu Gly Asp Leu Phe Pro Asn Leu Thr Val Ile Arg Gly Trp Lys
65 70 75 80
Leu Phe Tyr Asn Tyr Ala Leu Val Ile Phe Glu Met Thr Asn Leu Lys
85 90 95
Asp Ile Gly Leu Tyr Asn Leu Arg Asn Ile Thr Arg Gly Ala Ile Arg
100 105 110
Ile Glu Lys Asn Ala Asp Leu Cys Tyr Leu Ser Thr Val Asp Trp Ser
115 120 125
Leu Ile Leu Asp Ala Val Ser Asn Asn Tyr Ile Val Gly Asn Lys Pro
130 135 140
Pro Lys Glu Cys Gly Asp Leu Cys Pro Gly Thr Met Glu Glu Lys Pro
145 150 155 160
Met Cys Glu Lys Thr Thr Ile Asn Asn Glu Tyr Asn Tyr Arg Cys Trp
165 170 175
Thr Thr Asn Arg Cys Gln Lys Met Cys Pro Ser Thr Cys Gly Lys Arg
180 185 190
Ala Cys Thr Glu Asn Asn Glu Cys Cys His Pro Glu Cys Leu Gly Ser
195 200 205
Cys Ser Ala Pro Asp Asn Asp Thr Ala Cys Val Ala Cys Arg His Tyr
210 215 220
Tyr Tyr Ala Gly Val Cys Val Pro Ala Cys Pro Pro Asn Thr Tyr Arg
225 230 235 240
Phe Glu Gly Trp Arg Cys Val Asp Arg Asp Phe Cys Ala Asn Ile Leu
245 250 255
Ser Ala Glu Ser Ser Asp Ser Glu Gly Phe Val Ile His Asp Gly Glu
260 265 270
Cys Met Gln Glu Cys Pro Ser Gly Phe Ile Arg Asn Gly Ser Gln Ser
275 280 285
Met Tyr Cys Ile Pro Cys Glu Gly Pro Cys Pro Lys Val Cys Glu Glu
290 295 300
Glu Lys Lys Thr Lys Thr Ile Asp Ser Val Thr Ser Ala Gln Met Leu
305 310 315 320
Gln Gly Cys Thr Ile Phe Lys Gly Asn Leu Leu Ile Asn Ile Arg Arg
325 330 335
Gly Asn Asn Ile Ala Ser Glu Leu Glu Asn Phe Met Gly Leu Ile Glu
340 345 350
Val Val Thr Gly Tyr Val Lys Ile Arg His Ser His Ala Leu Val Ser
355 360 365
Leu Ser Phe Leu Lys Asn Leu Arg Leu Ile Leu Gly Glu Glu Gln Leu
370 375 380
Glu Gly Asn Tyr Ser Phe Tyr Val Leu Asp Asn Gln Asn Leu Gln Gln
385 390 395 400
Leu Trp Asp Trp Asp His Arg Asn Leu Thr Ile Lys Ala Gly Lys Met
405 410 415
Tyr Phe Ala Phe Asn Pro Lys Leu Cys Val Ser Glu Ile Tyr Arg Met
420 425 430
Glu Glu Val Thr Gly Thr Lys Gly Arg Gln Ser Lys Gly Asp Ile Asn
435 440 445
Thr Arg Asn Asn Gly Glu Arg Ala Ser Cys Glu Ser Asp Val Leu His
450 455 460
Phe Thr Ser Thr Thr Thr Ser Lys Asn Arg Ile Ile Ile Thr Trp His
465 470 475 480
Arg Tyr Arg Pro Pro Asp Tyr Arg Asp Leu Ile Ser Phe Thr Val Tyr
485 490 495
Tyr Lys Glu Ala Pro Phe Lys Asn Val Thr Glu Tyr Asp Gly Gln Asp
500 505 510
Ala Cys Gly Ser Asn Ser Trp Asn Met Val Asp Val Asp Leu Pro Pro
515 520 525
Asn Lys Asp Val Glu Pro Gly Ile Leu Leu His Gly Leu Lys Pro Trp
530 535 540
Thr Gln Tyr Ala Val Tyr Val Lys Ala Val Thr Leu Thr Met Val Glu
545 550 555 560
Asn Asp His Ile Arg Gly Ala Lys Ser Glu Ile Leu Tyr Ile Arg Thr
565 570 575
Asn Ala Ser Val Pro Ser Ile Pro Leu Asp Val Leu Ser Ala Ser Asn
580 585 590
Ser Ser Ser Gln Leu Ile Val Lys Trp Asn Pro Pro Ser Leu Pro Asn
595 600 605
Gly Asn Leu Ser Tyr Tyr Ile Val Arg Trp Gln Arg Gln Pro Gln Asp
610 615 620
Gly Tyr Leu Tyr Arg His Asn Tyr Cys Ser Lys Asp Lys Ile Pro Ile
625 630 635 640
Arg Lys Tyr Ala Asp Gly Thr Ile Asp Ile Glu Glu Val Thr Glu Asn
645 650 655
Pro Lys Thr Glu Val Cys Gly Gly Glu Lys Gly Pro Cys Cys Ala Cys
660 665 670
Pro Lys Thr Glu Ala Glu Lys Gln Ala Glu Lys Glu Glu Ala Glu Tyr
675 680 685
Arg Lys Val Phe Glu Asn Phe Leu His Asn Ser Ile Phe Val Pro Arg
690 695 700
Pro Glu Arg Lys Arg Arg Asp Val Met Gln Val Ala Asn Thr Thr Met
705 710 715 720
Ser Ser Arg Ser Arg Asn Thr Thr Ala Ala Asp Thr Tyr Asn Ile Thr
725 730 735
Asp Pro Glu Glu Leu Glu Thr Glu Tyr Pro Phe Phe Glu Ser Arg Val
740 745 750
Asp Asn Lys Glu Arg Thr Val Ile Ser Asn Leu Arg Pro Phe Thr Leu
755 760 765
Tyr Arg Ile Asp Ile His Ser Cys Asn His Glu Ala Glu Lys Leu Gly
770 775 780
Cys Ser Ala Ser Asn Phe Val Phe Ala Arg Thr Met Pro Ala Glu Gly
785 790 795 800
Ala Asp Asp Ile Pro Gly Pro Val Thr Trp Glu Pro Arg Pro Glu Asn
805 810 815
Ser Ile Phe Leu Lys Trp Pro Glu Pro Glu Asn Pro Asn Gly Leu Ile
820 825 830
Leu Met Tyr Glu Ile Lys Tyr Gly Ser Gln Val Glu Asp Gln Arg Glu
835 840 845
Cys Val Ser Arg Gln Glu Tyr Arg Lys Tyr Gly Gly Ala Lys Leu Asn
850 855 860
Arg Leu Asn Pro Gly Asn Tyr Thr Ala Arg Ile Gln Ala Thr Ser Leu
865 870 875 880
Ser Gly Asn Gly Ser Trp Thr Asp Pro Val Phe Phe Tyr Val Gln Ala
885 890 895
Lys Thr Gly Tyr Glu Asn Phe Ile His Leu Ile Ile Ala Leu Pro Val
900 905 910
Ala Val Leu Leu Ile Val Gly Gly Leu Val Ile Met Leu Tyr Val Phe
915 920 925
His Arg Lys Arg Asn Asn Ser Arg Leu Gly Asn Gly Val Leu Tyr Ala
930 935 940
Ser Val Asn Pro Glu Tyr Phe Ser Ala Ala Asp Val Tyr Val Pro Asp
945 950 955 960
Glu Trp Glu Val Ala Arg Glu Lys Ile Thr Met Ser Arg Glu Leu Gly
965 970 975
Gln Gly Ser Phe Gly Met Val Tyr Glu Gly Val Ala Lys Gly Val Val
980 985 990
Lys Asp Glu Pro Glu Thr Arg Val Ala Ile Lys Thr Val Asn Glu Ala
995 1000 1005
Ala Ser Met Arg Glu Arg Ile Glu Phe Leu Asn Glu Ala Ser Val Met
1010 1015 1020
Lys Glu Phe Asn Cys His His Val Val Arg Leu Leu Gly Val Val Ser
1025 1030 1035 1040
Gln Gly Gln Pro Thr Leu Val Ile Met Glu Leu Met Thr Arg Gly Asp
1045 1050 1055
Leu Lys Ser Tyr Leu Arg Ser Leu Arg Pro Glu Met Glu Asn Asn Pro
1060 1065 1070
Val Leu Ala Pro Pro Ser Leu Ser Lys Met Ile Gln Met Ala Gly Glu
1075 1080 1085
Ile Ala Asp Gly Met Ala Tyr Leu Asn Ala Asn Lys Phe Val His Arg
1090 1095 1100
Asp Leu Ala Ala Arg Asn Cys Met Val Ala Glu Asp Phe Thr Val Lys
1105 1110 1115 1120
Ile Gly Asp Phe Gly Met Thr Arg Asp Ile Tyr Glu Thr Asp Tyr Tyr
1125 1130 1135
Arg Lys Gly Gly Lys Gly Leu Leu Pro Val Arg Trp Met Ser Pro Glu
1140 1145 1150
Ser Leu Lys Asp Gly Val Phe Thr Thr Tyr Ser Asp Val Trp Ser Phe
1155 1160 1165
Gly Val Val Leu Trp Glu Ile Ala Thr Leu Ala Glu Gln Pro Tyr Gln
1170 1175 1180
Gly Leu Ser Asn Glu Gln Val Leu Arg Phe Val Met Glu Gly Gly Leu
1185 1190 1195 1200
Leu Asp Lys Pro Asp Asn Cys Pro Asp Met Leu Phe Glu Leu Met Arg
1205 1210 1215
Met Cys Trp Gln Tyr Asn Pro Lys Met Arg Pro Ser Phe Leu Glu Ile
1220 1225 1230
Ile Ser Ser Ile Lys Glu Glu Met Glu Pro Gly Phe Arg Glu Val Ser
1235 1240 1245
Phe Tyr Tyr Ser Glu Glu Asn Lys Leu Pro Glu Pro Glu Glu Leu Asp
1250 1255 1260
Leu Glu Pro Glu Asn Met Glu Ser Val Pro Leu Asp Pro Ser Ala Ser
1265 1270 1275 1280
Ser Ser Ser Leu Pro Leu Pro Asp Arg His Ser Gly His Lys Ala Glu
1285 1290 1295
Asn Gly Pro Gly Pro Gly Val Leu Val Leu Arg Ala Ser Phe Asp Glu
1300 1305 1310
Arg Gln Pro Tyr Ala His Met Asn Gly Gly Arg Lys Asn Glu Arg Ala
1315 1320 1325
Leu Pro Leu Pro Gln Ser Ser Thr Cys
1330 1335
<210> 2
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
20 25 30
Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly
85 90 95
Thr His Trp Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
<210> 3
<211> 119
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Ile Tyr His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Thr Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Trp Thr Gly Arg Thr Asp Ala Phe Asp Ile Trp Gly Gln Gly
100 105 110
Thr Met Val Thr Val Ser Ser
115
<210> 4
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Gly Ser Ser
20 25 30
Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg Leu Pro His
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 5
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr
100 105 110
Thr Val Thr Val Ser Ser
115
<210> 6
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser
20 25 30
Asn Gly Asn Thr Trp Leu Gln Trp Trp Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Trp Lys Val Ser Asn Arg Leu Trp Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Gln Ala Gln Asp Val Gly Val Trp Trp Cys Phe Gln Gly
85 90 95
Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Gln Ile Lys
100 105 110
<210> 7
<211> 117
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Trp Ser Ile Ser Gly Gly
20 25 30
Trp Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Gln Trp
35 40 45
Ile Gly Trp Ile Ser Trp Asp Gly Thr Asn Asn Trp Lys Pro Ser Leu
50 55 60
Lys Asp Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Trp Trp Cys
85 90 95
Ala Arg Trp Gly Arg Val Phe Phe Asp Trp Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 8
<211> 108
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln
1 5 10 15
Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala
20 25 30
Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ile Leu Val Ile Tyr
35 40 45
Gly Glu Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser
50 55 60
Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu
65 70 75 80
Asp Glu Ala Asp Tyr Tyr Cys Lys Ser Arg Asp Gly Ser Gly Gln His
85 90 95
Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105
<210> 9
<211> 130
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Pro Leu Arg Phe Leu Glu Trp Ser Thr Gln Asp His Tyr
100 105 110
Tyr Tyr Tyr Tyr Met Asp Val Trp Gly Lys Gly Thr Thr Val Thr Val
115 120 125
Ser Ser
130
<210> 10
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 10
Asp Ile Gln Met Thr Gln Phe Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile
35 40 45
Tyr Ala Ala Ser Arg Leu His Arg Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Cys
85 90 95
Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 11
<211> 125
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 11
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 Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Gly Ser Gly Gly Thr Thr Phe Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Thr Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Asp Leu Gly Trp Ser Asp Ser Tyr Tyr Tyr Tyr Tyr Gly Met
100 105 110
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120 125
<210> 12
<211> 108
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Glu Val Gly Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr 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 Phe Cys Gln Gln Tyr Ser Asn Tyr Pro Leu
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105
<210> 13
<211> 121
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Tyr Ile Thr Asn Tyr Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Phe
65 70 75 80
Leu Gln Met Asp Ser Leu Arg Pro Glu Asp Thr Gly Val Tyr Phe Cys
85 90 95
Ala Arg Gln Ser Asn Tyr Asp Tyr Asp Gly Trp Phe Ala Tyr Trp Gly
100 105 110
Gln Gly Thr Pro Val Thr Val Ser Ser
115 120
<210> 14
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu Gln Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly
85 90 95
Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
<210> 15
<211> 117
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
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 Tyr Ser Ile Thr Gly Gly
20 25 30
Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
35 40 45
Ile Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu
50 55 60
Lys Asp Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Arg Val Phe Phe Asp Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 16
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
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> 17
<211> 330
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 18
<211> 215
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Lys Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Lys Trp Pro Pro
85 90 95
Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ser Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 19
<211> 448
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Gln Val Glu Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Gln Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Ile Ile Trp Phe Asp Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Leu Gly Arg Arg Tyr Phe Asp Leu Trp Gly Arg Gly Thr
100 105 110
Leu Val Ser Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 20
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
gccaccatgg agacagatac cctgctgctg tgggtgctgc tgctgtgggt ccctggcagc 60
accgga 66

Claims (8)

1.一种结合IGF-1R的抗体，其特征在于，所述的抗体的轻链可变区为SEQ ID NO：2；所述的抗体的重链可变区为SEQ ID NO：3；所述的抗体的轻链由轻链可变区与SEQ ID NO：16所示的轻链恒定区拼接形成；所述的抗体的重链由重链可变区与SEQ ID NO：17所示的重链恒定区拼接形成。
2.一种核酸分子，其特征在于，所述的核酸分子编码权利要求1中的所述的抗体序列。
3.一种表达载体，其特征在于，所述的载体包含权利要求2中所述的核酸分子。
4.一种宿主细胞，其特征在于，所述的宿主细胞包含权利要求3所述的表达载体。
5.权利要求1所述的抗体在制备治疗或辅助治疗甲状腺相关眼病的药物中的应用。
6.权利要求2所述的核酸分子、权利要求3所述的表达载体和权利要求4所述的宿主细胞在制备治疗或辅助治疗甲状腺相关眼病的药物中的应用。
7.根据权利要求5或6所述的应用，其特征在于，所述的甲状腺相关眼病为眼球突出、眼睑退缩、上睑迟落、眼外肌肥大、结膜充血、眶周组织水肿、眼睑闭合不全、畏光、流泪、异物感、视力下降或复视。
8.一种药物组合物，其特征在于，所述的药物组合物包含权利要求1所述的抗体、权利要求2所述的核酸分子、权利要求3所述的表达载体和/或权利要求4所述的宿主细胞。
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