CN114539403B - 靶向人bcma蛋白的兔重组单克隆抗体及应用 - Google Patents

靶向人bcma蛋白的兔重组单克隆抗体及应用 Download PDF

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CN114539403B
CN114539403B CN202011364095.7A CN202011364095A CN114539403B CN 114539403 B CN114539403 B CN 114539403B CN 202011364095 A CN202011364095 A CN 202011364095A CN 114539403 B CN114539403 B CN 114539403B
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马东晖
黄少梅
洪淑娟
蔡萌
李沛
梅芬
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Tecode Biotechnology Wuhan Co ltd
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Abstract

本发明涉及靶向人BCMA蛋白的特异性兔重组单克隆抗体及应用，靶向人BCMA蛋白的特异性兔重组单克隆抗体具有三个重链互补决定区和三个轻链互补决定区，亲和力高，具有良好的特异性，可用于ELISA和流式检测方法中，弥补市场上靶向BCMA蛋白的兔重组单克隆抗体的诊断与治疗相关的应用。此外，靶向人BCMA蛋白的特异性兔重组单克隆抗体的SCFV序列可以构建成CART或双特异性抗体或抗体偶联药，可用于后续的肿瘤疾病的治疗。

Description

靶向人BCMA蛋白的兔重组单克隆抗体及应用
技术领域
本发明属于生物技术领域，特别是涉及靶向人BCMA蛋白的兔重组单克隆抗体及应用。
背景技术
B细胞成熟抗原(BCMA)又称为CD269、TNFRSF17，属于TNF受体超家族，是一种由185个氨基酸残基组成的Ⅲ型跨膜蛋白，能够结合B淋巴细胞刺激因子(BAFF)和增殖诱导配体(APRIL)。
BCMA由人浆母细胞、扁桃体记忆B细胞、生发中心B细胞以及来自扁桃体、脾和骨髓的浆细胞表达而来，其具有穿膜蛋白活化物-B细胞激活因子受体(TACI-BAFFR)低表型，因此，BCMA通常限于表达在B细胞谱系，并且有报告在终末B细胞分化中增加。另外，BCMA并不存在于幼稚和记忆B细胞上。BCMA表达在细胞表面，但也有表达在高尔基体中。BCMA与其配体可以活化NF-κB的信号通路，而Jun氨基末端激酶(JNK)信号通路的活化也可以与BCMA的表达形成正反馈的回路，从而一起促进浆细胞和肿瘤细胞的生长。另外，B细胞表达BCMA于膜上后，γ分泌酶可以将BCMA胞外区域切断，从而导致其从膜上脱落下来形成可溶形式的BCMA，即sBCMA，并且可以进入到外周血进行循环。
由于BCMA高表达于浆细胞和多发性骨髓瘤细胞表面，其他正常组织细胞和造血干细胞中均不表达，因此可以作为多发性骨髓瘤(MM)的诊断和治疗用的理想靶点。
发明内容
本发明目的在于提供靶向人BCMA蛋白的特异性兔重组单克隆抗体，重要的是靶向人BCMA蛋白的特异性兔重组单克隆抗体亲和力高，具有良好的特异性，可以弥补市场上靶向人BCMA蛋白的兔重组单克隆抗体的诊断与治疗相关的应用。另外，该BCMA抗体序列可以构建成嵌合抗原受体T细胞免疫疗法(CART)或双特异性抗体或抗体偶联药，可用于后续的肿瘤疾病的治疗。
本发明提供的靶向人BCMA蛋白的兔重组单克隆抗体，选自以下兔重组单克隆抗体中的一种或多种：
命名为2B3的兔重组单克隆抗体：其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3所示的氨基酸序列；且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:4，SEQ ID NO:5、SEQ ID NO:6所示的氨基酸序列；
命名为2D3的兔重组单克隆抗体：其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15所示的氨基酸序列；
命名为4A12的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ IDNO:19、SEQ ID NO:20、SEQ ID NO:21所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:22、SEQ ID NO:23、SEQ ID NO:24所示的氨基酸序列；
命名为5C4的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:28、SEQ ID NO:29、SEQ ID NO:30所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33所示的氨基酸序列；
命名为5H7的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:40、SEQ ID NO:41及SEQ ID NO:42所示的氨基酸序列；
命名为6D4的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:46、SEQ ID NO:47、SEQ ID NO:48所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:49、SEQ ID NO:50、SEQ ID NO:51所示的氨基酸序列；
命名为1H2的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:55、SEQ ID NO:56、SEQ ID NO:57所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:58，SEQ ID NO:59、SEQ ID NO:60所示的氨基酸序列；
命名为2C11的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:64、SEQ ID NO:65、SEQ ID NO:66所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:67、SEQ ID NO:68、SEQ ID NO:69所示的氨基酸序列；
命名为2D7的兔重组单克隆抗体，其重链互补决定区的氨基酸序列分别为SEQ IDNO:73、SEQ ID NO:74、SEQ IDNO:75所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:76、SEQ ID NO:77、SEQ ID NO:78所示的氨基酸序列；
命名为4H3的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:82、SEQ ID NO:83、SEQ ID NO:84所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:85、SEQ ID NO:86、SEQ ID NO:87所示的氨基酸序列；
命名为5B3的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:91、SEQ ID NO:92、SEQ ID NO:93所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:94、SEQ ID NO:95、SEQ ID NO:96所示的氨基酸序列；
命名为6B9的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:100、SEQ ID NO:101、SEQ ID NO:102所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:103、SEQ ID NO:104、SEQ ID NO:105所示的氨基酸序列；
命名为4F1的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:109、SEQ ID NO:110、SEQ ID NO:111所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:112、SEQ ID NO:113、SEQ ID NO:114所示的氨基酸序列。
在上述技术方案的基础上，命名为2B3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:7所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:8所示的氨基酸序列；
命名为2D3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:16所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:17所示的氨基酸序列；
命名为4A12的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:25所示的氨基酸序列，且轻链可变区序列为SEQ IDNO:26所示的氨基酸序列；
命名为5C4的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO：34所示的氨基酸序列，且轻链可变区序列为SEQ IDNO:35所示的氨基酸序列；
命名为5H7的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:43所示的氨基酸序列，轻链可变区序列为SEQ ID NO:44所示的氨基酸序列；
命名为6D4的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:52所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:53所示的氨基酸序列；
命名为1H2的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:61所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:62所示的氨基酸序列；
命名为2C11的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:70所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:71所示的氨基酸序列；
命名为2D7的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:79所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:80所示的氨基酸序列；
命名为4H3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:88所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:89所示的氨基酸序列；
命名为5B3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:97所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:98所示的氨基酸序列；
命名为6B9的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:106所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:107所示的氨基酸序列；
命名为4F1的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:115所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:116所示的氨基酸序列。
在上述技术方案的基础上，命名为2B3的兔重组单克隆抗体，其SCFV序列为SEQ IDNO:9所示的氨基酸序列；
命名为2D3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:18所示的氨基酸序列；
命名为4A12的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:27所示的氨基酸序列；
命名为5C4的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:36所示的氨基酸序列；
命名为5H7的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:45所示的氨基酸序列；
命名为6D4的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:54所示的氨基酸序列；
命名为1H2的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:63所示的氨基酸序列；
命名为2C11的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:72所示的氨基酸序列；
命名为2D7的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:81所示的氨基酸序列；
命名为4H3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:90所示的氨基酸序列；
命名为5B3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:99所示的氨基酸序列；
命名为6B9的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:108所示的氨基酸序列；
命名为4F1的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:117所示的氨基酸序列。
在上述技术方案的基础上，兔重组单克隆抗体的轻链恒定区为κ链，重链恒定区为IgG型。
本发明还提供一种核酸分子，其包含能够编码靶向人BCMA蛋白的兔重组单克隆抗体的重链互补决定区或轻链互补决定区的核酸序列。
本发明还提供一种载体，其含有上述核酸分子。
本发明还提供一种宿主细胞，该宿主细胞含有上述靶向人BCMA蛋白的兔重组单克隆抗体、上述核酸分子或上述载体。
本发明还提供一种偶联物，含有上述抗体。
本发明还提供一种药物组合物，含有主成分和辅成分，其中：主成分上述靶向人BCMA蛋白的兔重组单克隆抗体、上述核酸分子、上述载体、上述宿主细胞、上述偶联物中的一种或多种，辅成分选自药学上可接受的载体或赋形剂，以及任选的其它生物活性物质。
本发明还提供上述靶向人BCMA蛋白的兔重组单克隆抗体、上述酸分子上述载体、上述宿主细胞、上述偶联物在制备治疗疾病的药物或检测试剂中的应用。
本发明还提供一种试剂盒，该试剂盒包含上述靶向人BCMA蛋白的兔重组单克隆抗体。
与现有技术相比，本发明具有以下优点和有益效果：
本发明利用B细胞克隆技术提供靶向人BCMA蛋白的兔重组单克隆抗体，重要的是靶向人BCMA蛋白的特异性兔重组单克隆抗体亲和力高，具有良好的特异性，可以弥补市场上靶向人BCMA蛋白的兔重组单克隆抗体的诊断与治疗相关的应用。另外，该BCMA抗体序列可以构建成CART或双特异性抗体或抗体偶联药，可用于后续的肿瘤疾病的治疗。
附图说明
为了更清楚地说明本发明实施例中的技术方案，下面将对实施例描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1显示了BCMA胞外区蛋白的SDS电泳图。
图2～图5显示了FACs检测的BCMA兔重组单克隆抗体与K562-BCMA和NCI-H929结合力。
图6和图7显示了靶向人BCMA蛋白的兔重组单克隆抗体的表位分析结果。
图8显示了靶向人BCMA蛋白的兔重组单克隆抗体构建成双特异性抗体后的体外肿瘤细胞杀伤结果。
具体实施方式
以下对本发明做进一步描述：在本发明中，除非另有说明，否则本文中使用的科学和技术名词具有本领域技术人员所通常理解的含义。并且，本文中所用的蛋白质和核酸化学、分子生物学、细胞和组织培养、微生物学、免疫学相关术语和实验室操作步骤均为相应领域内广泛使用的术语和常规步骤。同时，为了更好地理解本发明，下面提供相关术语的定义和解释。
这里提到的术语“抗体”包括完整抗体及其任何抗原结合片段(即“抗原结合部分”)或单链。“抗体”是指包含通过二硫键互相连接在一起的至少两条重(H)链和两条轻(L)链的糖蛋白，或其抗原结合部分。每条重链由重链可变区和重链恒定区组成。本发明中涉及的蛋白或其片段可以是天然纯化的产物，或是化学合成的产物，或使用重组技术从真核宿主(例如哺乳动物细胞)中产生。本发明所用原料及试剂均可由市场购得。
实施例：
(1)靶向人BCMA的兔重组单克隆抗体的制备
1)动物免疫
为了获得靶向人BCMA蛋白的兔重组单克隆抗体，本发明构建表达了BCMA胞外区的哺乳动物细胞表达的重组蛋白作为免疫原，利用新西兰大白兔进行免疫。首免时每只兔子免疫300μg免疫原，将免疫原与等体积的弗氏完全佐剂混合后制成乳化剂，通过背部皮下进行多点注射；间隔3周后，取150μg免疫原与等体积的弗氏不完全佐剂混合制成乳化剂，背部皮下多点注射，加强免疫两次。三免后耳部静脉采血，采用间接酶联免疫吸附实验(ELISA)测试血清效价，取效价高的兔子，用150μg免疫原与等体积的弗氏不完全佐剂混合皮下多点注射加强一次，一周后无菌取外周血。
2)分离外周血单个核细胞(PBMC)
无菌采集兔外周抗凝血，用Ficoll密度梯度离心法分离PBMC，用无菌的PBS洗细胞2次，悬浮细胞计数，PBMC放置液氮中长期保存。
3)免疫磁珠分离B淋巴细胞
PBMC用抗原包被的磁珠进行筛选，即将人BCMA蛋白按照磁珠包被的方法包被在磁珠上，将PBMC细胞与磁珠室温充分孵育1h，将混合物放入专用的磁极中静置5min，弃掉上清，用无菌PBS洗细胞两次，经分离后可获得抗原特异性的B淋巴细胞。
4)B淋巴细胞的培养
将分离的抗原特异性的B淋巴细胞稀释后用完全1640培养基(含10％胎牛血清(FBS)，1％青霉素-链霉素，1％谷氨酰胺，2μg/mL的人IL2)培养于96孔细胞培养板，37℃5％CO2条件下培养6天。
5)特异性兔单克隆抗体的鉴定
采用ELISA方法鉴定，先利用包被人BCMA蛋白浓度1μg/mL，4℃作用酶标板15h；然后用封闭液(含10wt.％血清的PBS)封闭酶标板，150μL/孔，37℃温育一小时；再加入B细胞上清，37℃反应1h，用磷酸盐吐温缓冲液(PBST)洗酶标板5次，加入辣根过氧化物酶标记羊抗兔IgG(羊抗兔IgG-HRP，1：5000)，37℃反应1h，用PBST洗酶标板5次，加入底物进行显色。
流式方法鉴定：即收集稳转的K562-BCMA细胞与K562细胞，用PBS洗细胞两次，加入FC受体封闭液4℃孵育30min；用PBS洗细胞两次，加入B细胞培养上清，4℃孵育30min，用PBS洗细胞两次，加入羊抗兔IgG-488二抗，4℃孵育30min，用PBS洗细胞两次，最后用200μLPBS重悬细胞，流式机器上机测试。
6)特异性B细胞的基因克隆
收集阳性抗体对应的B细胞，采用常规方法提取RNA并反转录成cDNA。利用引物即重链基因引物序列为：5’AAGCTTGCCACCATGGAGACTGGGCTGCGCTGGCTTC-3’和5’CCATTGGTGAGGGTGCCCGAG-3’，轻链基因扩增引物为：5’AAGCTTGCCACCATG-GACAYGAGGGCCCCCACTC3’和5’CAGAGTRCTGCTGAGGTTGTAGGTAC-3’，进行重轻链基因的扩增，其PCR反应体系为：95℃3min，(95℃1min，58℃30s，72℃1min)25个循环，最后72℃5min，将PCR产物进行回收纯化。将兔重组单克隆抗体的重轻链基因克隆至表达载体，即将含兔单克隆抗体重链恒定区和轻链恒定区哺乳动物细胞表达载体进行线性化处理；然后利用同源重组的方式，将重轻链可变区分别构建到相应的哺乳动物表达载体中，将连接后的载体进行转化，涂板，最后进行菌落PCR验证阳性克隆，将阳性克隆进行基因测序，即可获得兔单克隆抗体的基因序列。
7)兔重组单克隆抗体的生产和纯化
将测序正确的兔单克隆抗体的重、轻链基因的表达质粒共转染入293细胞中；转染72h后，离心去除细胞，将培养基上清用proteinA树脂进行纯化。
(2)BCMA兔重组单克隆抗体的鉴定
对纯化后的抗体进行初步鉴定，包括抗体的ELISA效价、流式检测抗体的特异性和结合力以及表位分析，具体如下：
1)ELISA效价测定
采用间接ELISA方法测试，即包被BCMA蛋白浓度1μg/mL，4℃作用15h；用封闭液即10％血清的PBS封闭酶标板，150μL/孔，37℃温育一小时。加入倍比稀释的纯化抗体，37℃反应1h，用PBST洗酶标板5次，加入羊抗兔IgG-HRP(1：5000)，37℃反应1h，用PBST洗酶标板5次，加入底物进行显色。以抗体检测孔与对照孔的2倍所对应的抗体的稀释浓度为抗体的最高ELISA效价。
表1靶向人BCMA的兔重组单克隆抗体的ELISA测定结果
抗体稀释倍数 6D4 2B3 2D3 4A12 5C4 5H7 6B9
1K Overflow Overflow Overflow Overflow Overflow Overflow Overflow
5K Overflow Overflow Overflow Overflow Overflow Overflow Overflow
25K 3.7916 3.6667 3.7459 3.4906 3.6319 3.5698 Overflow
125K 2.5632 1.9329 1.9184 1.6425 1.9827 1.5056 2.2786
625K 0.7269 0.4949 0.5441 0.4295 0.5288 0.4227 0.7859
3215K 0.2211 0.1669 0.1719 0.1374 0.172 0.1495 0.3902
15625K 0.123 0.0877 0.1081 0.0899 0.0989 0.0869 0.294
抗体稀释倍数 5B3 4H3 2C11 2D7 1H2 4F1 -
1K Overflow Overflow Overflow Overflow Overflow Overflow -
5K Overflow Overflow Overflow Overflow Overflow Overflow -
25K 3.9679 Overflow Overflow Overflow Overflow 3.0739 -
125K 1.513 1.7141 2.4182 2.0023 2.1694 0.8561 -
625K 0.5324 0.6325 0.7943 0.6878 1.0279 0.4758 -
3215K 0.3755 0.3824 0.4153 0.3244 0.4265 0.3343 -
15625K 0.2599 0.2428 0.2747 0.2481 0.2754 0.2813 -
2)流式检测抗体的结合力
用表达人BCMA蛋白的天然细胞系NCI-H929和稳转BCMA细胞系K562-BCMA细胞测试抗体的结合力。即收集细胞，用PBS洗细胞两次，加入FC受体封闭液4℃孵育30min；用PBS洗细胞两次，加入倍比稀释的纯化抗体，4℃孵育30min，用PBS洗细胞两次，加入羊抗兔IgG-488二抗，4℃孵育30min，用PBS洗细胞两次，最后用200μL PBS重悬细胞，流式机器上机测试。
3)表位分析
采用竞争ELISA方法测试BCMA兔重组单克隆抗体与Bluebird(克隆号为huC11D5.3)的BCMA抗体进行表位分析。即包被人BCMA蛋白浓度1μg/mL，4℃作用15h；用封闭液即10％血清的PBS封闭酶标板，150μL/孔，37℃温育一小时。加入bluebird抗体5μg/mL，100μL/孔37℃反应1h，用PBST洗酶标板5次，加入兔重组单克隆抗体抗体1ng/mL，100μL/孔37℃反应1h，加入羊抗兔IgG-HRP(1：5000)，37℃反应1h，用PBST洗酶标板5次，加入底物进行显色。
4)靶向人BCMA的兔重组单克隆抗体SCFV功能测试
将靶向人BCMA的兔重组单克隆抗体的重轻链序列构建成双特异性抗体，利用稳转荧光素酶的NCI-H929细胞(NCI-H929-Luc)与人的PBCM细胞共培养于96孔避光的培养板中，加入不同浓度的双特异性抗体，孵育48h后，测试细胞荧光值。靶向人BCMA的兔重组单克隆抗体的SCFV序列信息见SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:16、SEQ ID NO:17、SEQ IDNO:25、SEQ ID NO:26、SEQ ID NO:34、SEQ ID NO:35、SEQ ID NO:43、SEQ ID NO:44、SEQ IDNO:52、SEQ ID NO:53、SEQ ID NO:61、SEQ ID NO:62、SEQ ID NO:70、SEQ ID NO:71、SEQ IDNO:79、SEQ ID NO:80、SEQ ID NO:88、SEQ ID NO:89、SEQ ID NO:97、SEQ ID NO:98、SEQ IDNO:106、SEQ ID NO:107、SEQ ID NO:115、SEQ ID NO:116。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，可轻易想到的变化或替换，都应涵盖在本发明的保护范围内。
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Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Arg Ile Asp Leu Asn Tyr Tyr Gly
20 25 30
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Ile Ile Ala Gly Asp Arg Asp Thr Tyr Tyr Ala Thr Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Ser
85 90 95
Tyr Ser Ser Thr Trp Asp Gly Ala Phe Asp His Trp Gly Pro Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 26
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Ala Arg Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asp Cys Gln Ala Ser Gln Ser Val Phe Ser Asn
20 25 30
Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Lys Leu
35 40 45
Leu Ile Tyr Arg Ala Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe
50 55 60
Ser Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val
65 70 75 80
Gln Cys Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Pro Thr Tyr Leu Ser
85 90 95
Gly Asp Trp Tyr Val Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 27
<211> 230
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Arg Ile Asp Leu Asn Tyr Tyr Gly
20 25 30
Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Ile Ile Ala Gly Asp Arg Asp Thr Tyr Tyr Ala Thr Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Ser
85 90 95
Tyr Ser Ser Thr Trp Asp Gly Ala Phe Asp His Trp Gly Pro Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Arg Val Leu Thr Gln Thr Pro Ser Pro
115 120 125
Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asp Cys Gln Ala Ser
130 135 140
Gln Ser Val Phe Ser Asn Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
145 150 155 160
Gly Gln Ala Pro Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Ala Ser
165 170 175
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gln Phe Thr
180 185 190
Leu Thr Ile Ser Gly Val Gln Cys Ala Asp Ala Ala Thr Tyr Tyr Cys
195 200 205
Gln Pro Thr Tyr Leu Ser Gly Asp Trp Tyr Val Ala Phe Gly Gly Gly
210 215 220
Thr Glu Val Val Val Lys
225 230
<210> 28
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Gly Phe Ser Leu Ser Ser Tyr Val
1 5
<210> 29
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Ile Asp Pro Ser Asn Gly Ala
1 5
<210> 30
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Val Arg Glu Arg Tyr Pro Ser Asn Asp Asn Leu
1 5 10
<210> 31
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Gln Asn Val Tyr Ser Asn Asn Tyr
1 5
<210> 32
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Glu Ala Ser
1
<210> 33
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Gln Gly Gly His Leu Ser Gly Asp Trp Tyr Leu Leu
1 5 10
<210> 34
<211> 114
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Gln Ser Val Glu Glu Ser Gly Gly Gly Leu Val Thr Pro Gly Thr Ser
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Ser Tyr Val
20 25 30
Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Tyr Ile Asp Pro Ser Asn Gly Ala Tyr Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Ser Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Glu Arg
85 90 95
Tyr Pro Ser Asn Asp Asn Leu Trp Gly Gln Gly Thr Leu Val Thr Val
100 105 110
Ser Ser
<210> 35
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Asp Pro Val Met Thr Gln Thr Pro Ser Ser Thr Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Asn Val Tyr Ser Asn
20 25 30
Asn Tyr Leu Ser Trp Phe Gln Lys Lys Pro Gly Gln Pro Pro Lys Leu
35 40 45
Leu Ile Tyr Glu Ala Ser Lys Leu Ala Ser Gly Val Pro Pro Arg Phe
50 55 60
Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val
65 70 75 80
Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Gly His Leu Ser
85 90 95
Gly Asp Trp Tyr Leu Leu Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 36
<211> 226
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Gln Ser Val Glu Glu Ser Gly Gly Gly Leu Val Thr Pro Gly Thr Ser
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Ser Tyr Val
20 25 30
Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Tyr Ile Asp Pro Ser Asn Gly Ala Tyr Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Ser Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Glu Arg
85 90 95
Tyr Pro Ser Asn Asp Asn Leu Trp Gly Gln Gly Thr Leu Val Thr Val
100 105 110
Ser Ser Asp Pro Val Met Thr Gln Thr Pro Ser Ser Thr Ser Ala Ala
115 120 125
Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Asn Val Tyr
130 135 140
Ser Asn Asn Tyr Leu Ser Trp Phe Gln Lys Lys Pro Gly Gln Pro Pro
145 150 155 160
Lys Leu Leu Ile Tyr Glu Ala Ser Lys Leu Ala Ser Gly Val Pro Pro
165 170 175
Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser
180 185 190
Asp Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Gly His
195 200 205
Leu Ser Gly Asp Trp Tyr Leu Leu Phe Gly Gly Gly Thr Glu Val Val
210 215 220
Val Lys
225
<210> 37
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Gly Phe Ser Leu Ser Ser Tyr Ala
1 5
<210> 38
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Ile Ser Ser Ser Gly Ser Thr
1 5
<210> 39
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Ala Arg Asp Phe Tyr Val Tyr Ser Ser Gly Thr Tyr Leu Glu Tyr Gly
1 5 10 15
Met Asp Leu
<210> 40
<211> 6
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Glu Asn Ile Tyr Arg Asn
1 5
<210> 41
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Ala Ala Ser
1
<210> 42
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Gln Asn Ala Phe Tyr Ser Ser Ser Ala Asp Asn Pro
1 5 10
<210> 43
<211> 119
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr Ala
20 25 30
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Met Ile Ser Ser Ser Gly Ser Thr His Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Phe Phe Cys Ala Arg Asp Phe
85 90 95
Tyr Val Tyr Ser Ser Gly Thr Tyr Leu Glu Tyr Gly Met Asp Leu Trp
100 105 110
Gly Pro Gly Thr Leu Val Thr
115
<210> 44
<211> 110
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Ala Ile Glu Met Thr Gln Thr Pro Phe Ser Val Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asn Cys Gln Ala Ser Glu Asn Ile Tyr Arg Asn
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Leu Leu Ala Ser Gly Val Pro Ser Arg Phe Lys Gly
50 55 60
Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Gly Val Gln Cys
65 70 75 80
Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Asn Ala Phe Tyr Ser Ser Ser
85 90 95
Ala Asp Asn Pro Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 45
<211> 229
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr Ala
20 25 30
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Met Ile Ser Ser Ser Gly Ser Thr His Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Phe Phe Cys Ala Arg Asp Phe
85 90 95
Tyr Val Tyr Ser Ser Gly Thr Tyr Leu Glu Tyr Gly Met Asp Leu Trp
100 105 110
Gly Pro Gly Thr Leu Val Thr Ala Ile Glu Met Thr Gln Thr Pro Phe
115 120 125
Ser Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ala
130 135 140
Ser Glu Asn Ile Tyr Arg Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly
145 150 155 160
Gln Pro Pro Lys Leu Leu Ile Tyr Ala Ala Ser Leu Leu Ala Ser Gly
165 170 175
Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Glu Tyr Thr Leu
180 185 190
Thr Ile Ser Gly Val Gln Cys Ala Asp Ala Ala Thr Tyr Tyr Cys Gln
195 200 205
Asn Ala Phe Tyr Ser Ser Ser Ala Asp Asn Pro Phe Gly Gly Gly Thr
210 215 220
Glu Val Val Val Lys
225
<210> 46
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Gly Phe Ser Leu Ser Ser Tyr Gly
1 5
<210> 47
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Ile Glu Val Gly Gly Ser Ser
1 5
<210> 48
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Ala Arg Asn Leu Phe Ser Gly Gly Asp Asp Thr Arg Asn Leu
1 5 10
<210> 49
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
Pro Ser Val Tyr Ser Asn Tyr
1 5
<210> 50
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Ser Ala Ser
1
<210> 51
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Leu Gly Gly Tyr Ile Ile Asn Ser Asp Ser Ala
1 5 10
<210> 52
<211> 117
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr Gly
20 25 30
Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Phe Ile Glu Val Gly Gly Ser Ser Asp Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Ile Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asn Leu
85 90 95
Phe Ser Gly Gly Asp Asp Thr Arg Asn Leu Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 53
<211> 110
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Ala Ala Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asn Cys Gln Ala Ser Pro Ser Val Tyr Ser Asn
20 25 30
Tyr Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Gln Leu
35 40 45
Ile Tyr Ser Ala Ser Ser Leu Ala Ser Gly Val Pro Ser Arg Phe Lys
50 55 60
Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val Gln
65 70 75 80
Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Gly Tyr Ile Ile Asn
85 90 95
Ser Asp Ser Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 54
<211> 227
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr Gly
20 25 30
Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Phe Ile Glu Val Gly Gly Ser Ser Asp Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Ile Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asn Leu
85 90 95
Phe Ser Gly Gly Asp Asp Thr Arg Asn Leu Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ala Val Leu Thr Gln Thr Pro Ser Pro Val
115 120 125
Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ala Ser Pro
130 135 140
Ser Val Tyr Ser Asn Tyr Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln
145 150 155 160
Pro Pro Lys Gln Leu Ile Tyr Ser Ala Ser Ser Leu Ala Ser Gly Val
165 170 175
Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr
180 185 190
Ile Ser Asp Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Leu Gly
195 200 205
Gly Tyr Ile Ile Asn Ser Asp Ser Ala Phe Gly Gly Gly Thr Glu Val
210 215 220
Val Val Lys
225
<210> 55
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
Gly Phe Ser Leu Ser Ser Asn Ala
1 5
<210> 56
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Ile Asp Pro Tyr Asn Gly Ala
1 5
<210> 57
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Val Arg Glu Arg Tyr Pro Gly Asn Asp Asn Leu
1 5 10
<210> 58
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
Gln Asn Val Tyr Ser Asn Asn Tyr
1 5
<210> 59
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Gly Ala Ser
1
<210> 60
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Gln Gly Gly His Leu Ser Gly Asp Trp Tyr Leu Pro
1 5 10
<210> 61
<211> 114
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Ser Asn Ala
20 25 30
Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Tyr Ile Asp Pro Tyr Asn Gly Ala Tyr Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Ser Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Glu Arg
85 90 95
Tyr Pro Gly Asn Asp Asn Leu Trp Gly Gln Gly Thr Leu Val Thr Val
100 105 110
Ser Ser
<210> 62
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Asp Pro Val Met Thr Gln Thr Pro Ser Ser Thr Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Asn Val Tyr Ser Asn
20 25 30
Asn Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly His Ser Pro Lys Leu
35 40 45
Leu Ile Tyr Gly Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe
50 55 60
Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Val
65 70 75 80
Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Gly His Leu Ser
85 90 95
Gly Asp Trp Tyr Leu Pro Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 63
<211> 226
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 63
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Ser Asn Ala
20 25 30
Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Tyr Ile Asp Pro Tyr Asn Gly Ala Tyr Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Ser Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Glu Arg
85 90 95
Tyr Pro Gly Asn Asp Asn Leu Trp Gly Gln Gly Thr Leu Val Thr Val
100 105 110
Ser Ser Asp Pro Val Met Thr Gln Thr Pro Ser Ser Thr Ser Ala Ala
115 120 125
Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Asn Val Tyr
130 135 140
Ser Asn Asn Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly His Ser Pro
145 150 155 160
Lys Leu Leu Ile Tyr Gly Ala Ser Thr Leu Ala Ser Gly Val Pro Ser
165 170 175
Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser
180 185 190
Asp Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Gly His
195 200 205
Leu Ser Gly Asp Trp Tyr Leu Pro Phe Gly Gly Gly Thr Glu Val Val
210 215 220
Val Lys
225
<210> 64
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Gly Phe Ser Leu Ser Thr Tyr His
1 5
<210> 65
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Ile Ser Ser Ser Gly Ser Thr
1 5
<210> 66
<211> 10
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ala Arg Asp Leu Asp Tyr Val Ile Asp Leu
1 5 10
<210> 67
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Pro Ser Val Tyr Asn Asn Tyr
1 5
<210> 68
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Glu Thr Ser
1
<210> 69
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Ala Gly Thr Tyr Val Ser Gly Asp Arg Arg Ala
1 5 10
<210> 70
<211> 114
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 70
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Thr Tyr His
20 25 30
Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Val Ile Ser Ser Ser Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys Met
65 70 75 80
Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp
85 90 95
Leu Asp Tyr Val Ile Asp Leu Trp Gly Gln Gly Thr Leu Val Thr Val
100 105 110
Ser Ser
<210> 71
<211> 110
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 71
Ala Gln Val Leu Thr Gln Thr Pro Ser Ser Val Ser Ser Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Pro Ser Val Tyr Asn Asn
20 25 30
Tyr Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu
35 40 45
Ile Tyr Glu Thr Ser Thr Leu Pro Ser Gly Val Pro Ser Arg Phe Ser
50 55 60
Ala Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val Gln
65 70 75 80
Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Thr Tyr Val Ser Gly
85 90 95
Asp Arg Arg Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 72
<211> 224
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 72
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser Thr Tyr His
20 25 30
Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Val Ile Ser Ser Ser Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys Met
65 70 75 80
Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp
85 90 95
Leu Asp Tyr Val Ile Asp Leu Trp Gly Gln Gly Thr Leu Val Thr Val
100 105 110
Ser Ser Ala Gln Val Leu Thr Gln Thr Pro Ser Ser Val Ser Ser Ala
115 120 125
Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Pro Ser Val Tyr
130 135 140
Asn Asn Tyr Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys
145 150 155 160
Leu Leu Ile Tyr Glu Thr Ser Thr Leu Pro Ser Gly Val Pro Ser Arg
165 170 175
Phe Ser Ala Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly
180 185 190
Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Thr Tyr Val
195 200 205
Ser Gly Asp Arg Arg Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys
210 215 220
<210> 73
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 73
Gly Phe Ser Leu Asn Ala Tyr Ala
1 5
<210> 74
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 74
Ile Asp Ala Gly Asp Thr Thr
1 5
<210> 75
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 75
Ala Arg Asp Leu Phe Tyr Asn Ile
1 5
<210> 76
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 76
Glu Ser Val Tyr Ser Asn Trp
1 5
<210> 77
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 77
Gly Ala Ser
1
<210> 78
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 78
Ala Gly Val Tyr Asn Gly Asp Asp Gly Arg Ala
1 5 10
<210> 79
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 79
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Asn Ala Tyr Ala
20 25 30
Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly
35 40 45
Met Ile Asp Ala Gly Asp Thr Thr Tyr Tyr Ala Ser Trp Ala Arg Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys Ile
65 70 75 80
Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp
85 90 95
Leu Phe Tyr Asn Ile Trp Gly Pro Gly Thr Leu Val Thr Val Ser Ser
100 105 110
<210> 80
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 80
Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly Gly Thr
1 5 10 15
Val Ser Ile Ser Cys Gln Ser Ser Glu Ser Val Tyr Ser Asn Trp Leu
20 25 30
Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Phe
35 40 45
Gly Ala Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Lys Gly Ser
50 55 60
Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Asp Leu Glu Cys Ala
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Val Tyr Asn Gly Asp Asp Gly
85 90 95
Arg Ala Phe Gly Gly Gly Thr Glu Val Val Val
100 105
<210> 81
<211> 219
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 81
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Asn Ala Tyr Ala
20 25 30
Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly
35 40 45
Met Ile Asp Ala Gly Asp Thr Thr Tyr Tyr Ala Ser Trp Ala Arg Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Asp Leu Lys Ile
65 70 75 80
Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp
85 90 95
Leu Phe Tyr Asn Ile Trp Gly Pro Gly Thr Leu Val Thr Val Ser Ser
100 105 110
Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly Gly Thr
115 120 125
Val Ser Ile Ser Cys Gln Ser Ser Glu Ser Val Tyr Ser Asn Trp Leu
130 135 140
Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Phe
145 150 155 160
Gly Ala Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Lys Gly Ser
165 170 175
Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Asp Leu Glu Cys Ala
180 185 190
Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Val Tyr Asn Gly Asp Asp Gly
195 200 205
Arg Ala Phe Gly Gly Gly Thr Glu Val Val Val
210 215
<210> 82
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 82
Gly Phe Ser Pro Ser Ser Thr Tyr
1 5
<210> 83
<211> 9
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 83
Ile Tyr Val Gly Gly Ala Gly Ser Thr
1 5
<210> 84
<211> 16
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 84
Ala Arg Pro Tyr Pro Asn Trp Asn Ile Asp Thr Phe Arg Trp Asp Leu
1 5 10 15
<210> 85
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 85
Lys Ser Val Tyr Asn Asn Asn Trp
1 5
<210> 86
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 86
Lys Ala Ser
1
<210> 87
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 87
Ala Gly Gly Phe Asn Asn Asp Ser Asp Arg Gly Thr
1 5 10
<210> 88
<211> 119
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 88
Gly Gly Ala Glu Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Glu Leu
1 5 10 15
Cys Cys Lys Ala Ser Gly Phe Ser Pro Ser Ser Thr Tyr Trp Ile Cys
20 25 30
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Cys Ile
35 40 45
Tyr Val Gly Gly Ala Gly Ser Thr Tyr Tyr Ala Thr Trp Val Asn Gly
50 55 60
Arg Phe Thr Leu Ser Arg Asp Ile Asp Gln Ser Thr Ser Cys Leu Gln
65 70 75 80
Leu Asn Ser Leu Thr Ala Ala Asp Thr Ala Met Tyr Tyr Cys Ala Arg
85 90 95
Pro Tyr Pro Asn Trp Asn Ile Asp Thr Phe Arg Trp Asp Leu Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val
115
<210> 89
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 89
Ala Ala Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Ser Ile Ser Cys Gln Ser Ser Lys Ser Val Tyr Asn Asn
20 25 30
Asn Trp Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu
35 40 45
Leu Ile Tyr Lys Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe
50 55 60
Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val
65 70 75 80
Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Phe Asn Asn
85 90 95
Asp Ser Asp Arg Gly Thr Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 90
<211> 231
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 90
Gly Gly Ala Glu Gly Gly Leu Val Lys Pro Gly Gly Ser Leu Glu Leu
1 5 10 15
Cys Cys Lys Ala Ser Gly Phe Ser Pro Ser Ser Thr Tyr Trp Ile Cys
20 25 30
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Cys Ile
35 40 45
Tyr Val Gly Gly Ala Gly Ser Thr Tyr Tyr Ala Thr Trp Val Asn Gly
50 55 60
Arg Phe Thr Leu Ser Arg Asp Ile Asp Gln Ser Thr Ser Cys Leu Gln
65 70 75 80
Leu Asn Ser Leu Thr Ala Ala Asp Thr Ala Met Tyr Tyr Cys Ala Arg
85 90 95
Pro Tyr Pro Asn Trp Asn Ile Asp Thr Phe Arg Trp Asp Leu Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ala Ala Val Leu Thr Gln Thr Pro Ser
115 120 125
Pro Val Ser Ala Ala Val Gly Gly Thr Val Ser Ile Ser Cys Gln Ser
130 135 140
Ser Lys Ser Val Tyr Asn Asn Asn Trp Leu Ser Trp Tyr Gln Gln Lys
145 150 155 160
Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Lys Ala Ser Thr Leu Ala
165 170 175
Ser Gly Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe
180 185 190
Thr Leu Thr Ile Ser Gly Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr
195 200 205
Cys Ala Gly Gly Phe Asn Asn Asp Ser Asp Arg Gly Thr Phe Gly Gly
210 215 220
Gly Thr Glu Val Val Val Lys
225 230
<210> 91
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 91
Gly Phe Ser Leu Ser Ser Tyr Asp
1 5
<210> 92
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 92
Ile Asn Gly Tyr Gly Gly Thr
1 5
<210> 93
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 93
Val Arg Gly Ile Val Gly Val Tyr Ser Thr Ala Phe Asn Leu
1 5 10
<210> 94
<211> 6
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 94
Glu Ser Ile Ser Ser Trp
1 5
<210> 95
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 95
Asp Ala Ser
1
<210> 96
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 96
Gln Thr Tyr Trp Tyr Ser Ser Tyr Ser Thr Thr Gly Phe Tyr Ala
1 5 10 15
<210> 97
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 97
Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Gly Ser
1 5 10 15
Leu Thr Leu Thr Cys Thr Gly Ser Gly Phe Ser Leu Ser Ser Tyr Asp
20 25 30
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Ile Ile Asn Gly Tyr Gly Gly Thr Tyr Tyr Thr Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Glu Leu Lys Met
65 70 75 80
Ala Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Gly
85 90 95
Ile Val Gly Val Tyr Ser Thr Ala Phe Asn Leu Trp Gly Pro Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 98
<211> 111
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 98
Glu Leu Thr Gln Thr Pro Ala Ser Met Glu Ala Ala Val Gly Gly Thr
1 5 10 15
Val Thr Ile Lys Cys Gln Ala Ser Glu Ser Ile Ser Ser Trp Leu Ala
20 25 30
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Asp
35 40 45
Ala Ser Thr Leu Ala Ser Gly Val Ser Ser Arg Phe Lys Gly Ser Gly
50 55 60
Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Asp Leu Glu Cys Ala Asp
65 70 75 80
Ala Ala Thr Tyr Tyr Cys Gln Thr Tyr Trp Tyr Ser Ser Tyr Ser Thr
85 90 95
Thr Gly Phe Tyr Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 99
<211> 229
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 99
Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Gly Ser
1 5 10 15
Leu Thr Leu Thr Cys Thr Gly Ser Gly Phe Ser Leu Ser Ser Tyr Asp
20 25 30
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly
35 40 45
Ile Ile Asn Gly Tyr Gly Gly Thr Tyr Tyr Thr Ser Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Glu Leu Lys Met
65 70 75 80
Ala Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Gly
85 90 95
Ile Val Gly Val Tyr Ser Thr Ala Phe Asn Leu Trp Gly Pro Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Glu Leu Thr Gln Thr Pro Ala Ser Met Glu
115 120 125
Ala Ala Val Gly Gly Thr Val Thr Ile Lys Cys Gln Ala Ser Glu Ser
130 135 140
Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
145 150 155 160
Lys Leu Leu Ile Tyr Asp Ala Ser Thr Leu Ala Ser Gly Val Ser Ser
165 170 175
Arg Phe Lys Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser
180 185 190
Asp Leu Glu Cys Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Thr Tyr Trp
195 200 205
Tyr Ser Ser Tyr Ser Thr Thr Gly Phe Tyr Ala Phe Gly Gly Gly Thr
210 215 220
Glu Val Val Val Lys
225
<210> 100
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 100
Arg Phe Ser Leu Ser Ser Tyr Gly
1 5
<210> 101
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 101
Ile Trp Thr Asp Tyr Ser Thr
1 5
<210> 102
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 102
Val Arg Gly Gly Gly Leu Glu Trp Leu Asp Leu
1 5 10
<210> 103
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 103
Lys Ser Val Tyr Ser Asn Tyr
1 5
<210> 104
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 104
Gly Ala Ser
1
<210> 105
<211> 11
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 105
Ala Gly Ala Tyr Val Thr Thr Ser Asp Ser Gly
1 5 10
<210> 106
<211> 114
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 106
Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Arg Phe Ser Leu Ser Ser Tyr Gly
20 25 30
Val Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ala
35 40 45
Tyr Ile Trp Thr Asp Tyr Ser Thr His Tyr Ala Arg Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Arg Thr Thr Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Gly Gly
85 90 95
Gly Leu Glu Trp Leu Asp Leu Trp Gly Gln Gly Thr Leu Val Ser Val
100 105 110
Ser Ser
<210> 107
<211> 109
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 107
Ala Gln Val Leu Thr Gln Thr Pro Ser Ser Val Ser Ala Ala Val Gly
1 5 10 15
Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Lys Ser Val Tyr Ser Asn
20 25 30
Tyr Leu Phe Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Arg Leu
35 40 45
Ile Tyr Gly Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Lys
50 55 60
Gly Ser Gly Phe Gly Ser Gln Phe Thr Leu Thr Ile Asn Asp Val Gln
65 70 75 80
Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Ala Tyr Val Thr Thr
85 90 95
Ser Asp Ser Gly Phe Gly Gly Gly Thr Glu Val Val Val
100 105
<210> 108
<211> 223
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 108
Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Ala Ser Arg Phe Ser Leu Ser Ser Tyr Gly
20 25 30
Val Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ala
35 40 45
Tyr Ile Trp Thr Asp Tyr Ser Thr His Tyr Ala Arg Trp Ala Lys Gly
50 55 60
Arg Phe Thr Ile Ser Arg Thr Thr Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Gly Gly
85 90 95
Gly Leu Glu Trp Leu Asp Leu Trp Gly Gln Gly Thr Leu Val Ser Val
100 105 110
Ser Ser Ala Gln Val Leu Thr Gln Thr Pro Ser Ser Val Ser Ala Ala
115 120 125
Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Lys Ser Val Tyr
130 135 140
Ser Asn Tyr Leu Phe Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys
145 150 155 160
Arg Leu Ile Tyr Gly Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg
165 170 175
Phe Lys Gly Ser Gly Phe Gly Ser Gln Phe Thr Leu Thr Ile Asn Asp
180 185 190
Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Ala Tyr Val
195 200 205
Thr Thr Ser Asp Ser Gly Phe Gly Gly Gly Thr Glu Val Val Val
210 215 220
<210> 109
<211> 8
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 109
Gly Phe Ser Leu Ser Ile Tyr Asp
1 5
<210> 110
<211> 7
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 110
Ile Ile Tyr Gly Gly Ser Ala
1 5
<210> 111
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 111
Ala Arg Gly Ala Ser Ser Tyr Ser Tyr Ser Tyr Gly Phe Ser Leu
1 5 10 15
<210> 112
<211> 6
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 112
Gln Ser Ile Asp Ser Trp
1 5
<210> 113
<211> 3
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 113
Leu Ala Ser
1
<210> 114
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 114
Gln Ser Asn Asp Glu Ser Ser Ile Tyr Gly Ala Ala
1 5 10
<210> 115
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 115
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ile Tyr Asp
20 25 30
Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly
35 40 45
Ile Ile Ile Tyr Gly Gly Ser Ala Ser Tyr Ala Asn Trp Ala Asn Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Ala
85 90 95
Ser Ser Tyr Ser Tyr Ser Tyr Gly Phe Ser Leu Trp Gly Pro Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 116
<211> 110
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 116
Asp Val Val Met Thr Gln Thr Pro Ser Ser Val Ser Glu Pro Val Gly
1 5 10 15
Gly Thr Val Thr Ile Lys Cys Gln Ala Ser Gln Ser Ile Asp Ser Trp
20 25 30
Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile
35 40 45
Tyr Leu Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Arg Gly
50 55 60
Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Asp Leu Glu Cys
65 70 75 80
Ala Asp Ala Ala Thr Tyr Ser Cys Gln Ser Asn Asp Glu Ser Ser Ile
85 90 95
Tyr Gly Ala Ala Phe Gly Gly Gly Thr Glu Val Val Val Lys
100 105 110
<210> 117
<211> 228
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 117
Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro
1 5 10 15
Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ile Tyr Asp
20 25 30
Met Ile Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Tyr Ile Gly
35 40 45
Ile Ile Ile Tyr Gly Gly Ser Ala Ser Tyr Ala Asn Trp Ala Asn Gly
50 55 60
Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr
65 70 75 80
Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Ala
85 90 95
Ser Ser Tyr Ser Tyr Ser Tyr Gly Phe Ser Leu Trp Gly Pro Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Asp Val Val Met Thr Gln Thr Pro Ser Ser
115 120 125
Val Ser Glu Pro Val Gly Gly Thr Val Thr Ile Lys Cys Gln Ala Ser
130 135 140
Gln Ser Ile Asp Ser Trp Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln
145 150 155 160
Pro Pro Lys Leu Leu Ile Tyr Leu Ala Ser Thr Leu Ala Ser Gly Val
165 170 175
Pro Ser Arg Phe Arg Gly Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr
180 185 190
Ile Ser Asp Leu Glu Cys Ala Asp Ala Ala Thr Tyr Ser Cys Gln Ser
195 200 205
Asn Asp Glu Ser Ser Ile Tyr Gly Ala Ala Phe Gly Gly Gly Thr Glu
210 215 220
Val Val Val Lys
225
<210> 118
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
aagcttgcca ccatggagac tgggctgcgc tggcttc 37
<210> 119
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
ccattggtga gggtgcccga g 21
<210> 120
<211> 34
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
aagcttgcca ccatggacay gagggccccc actc 34
<210> 121
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 121
cagagtrctg ctgaggttgt aggtac 26

Claims (10)

1.靶向人BCMA蛋白的兔重组单克隆抗体，其特征在于：选自以下兔重组单克隆抗体中的一种或多种：
命名为2B3的兔重组单克隆抗体：其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3所示的氨基酸序列；且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:4，SEQ ID NO:5、SEQ ID NO:6所示的氨基酸序列；
命名为2D3的兔重组单克隆抗体：其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15所示的氨基酸序列；
命名为4A12的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:22、SEQ ID NO:23、SEQ ID NO:24所示的氨基酸序列；
命名为5C4的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:28、SEQ ID NO:29、SEQ ID NO:30所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33所示的氨基酸序列；
命名为5H7的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:40、SEQ ID NO:41及SEQ ID NO:42所示的氨基酸序列；
命名为6D4的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:46、SEQ ID NO:47、SEQ ID NO:48所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:49、SEQ ID NO:50、SEQ ID NO:51所示的氨基酸序列；
命名为1H2的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:55、SEQ ID NO:56、SEQ ID NO:57所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:58，SEQ ID NO:59、SEQ ID NO:60所示的氨基酸序列；
命名为2C11的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:64、SEQ ID NO:65、SEQ ID NO:66所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:67、SEQ ID NO:68、SEQ ID NO:69所示的氨基酸序列；
命名为2D7的兔重组单克隆抗体，其重链互补决定区的氨基酸序列分别为SEQ ID NO:73、SEQ ID NO:74、SEQ ID NO:75所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:76、SEQ ID NO:77、SEQ ID NO:78所示的氨基酸序列；
命名为4H3的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:82、SEQ ID NO:83、SEQ ID NO:84所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:85、SEQ ID NO:86、SEQ ID NO:87所示的氨基酸序列；
命名为5B3的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:91、SEQ ID NO:92、SEQ ID NO:93所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:94、SEQ ID NO:95、SEQ ID NO:96所示的氨基酸序列；
命名为6B9的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:100、SEQ ID NO:101、SEQ ID NO:102所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:103、SEQ ID NO:104、SEQ ID NO:105所示的氨基酸序列；
命名为4F1的兔重组单克隆抗体，其重链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:109、SEQ ID NO:110、SEQ ID NO:111所示的氨基酸序列，且轻链互补决定区CDR1、CDR2、CDR3的氨基酸序列分别为SEQ ID NO:112、SEQ ID NO:113、SEQ ID NO:114所示的氨基酸序列。
2.根据权利要求1所述的靶向人BCMA蛋白的兔重组单克隆抗体，其特征在于：
所述命名为2B3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:7所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:8所示的氨基酸序列；
所述命名为2D3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:16所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:17所示的氨基酸序列；
所述命名为4A12的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:25所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:26所示的氨基酸序列；
所述命名为5C4的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO：34所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:35所示的氨基酸序列；
所述命名为5H7的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:43所示的氨基酸序列，轻链可变区序列为SEQ ID NO:44所示的氨基酸序列；
所述命名为6D4的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:52所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:53所示的氨基酸序列；
所述命名为1H2的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:61所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:62所示的氨基酸序列；
所述命名为2C11的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:70所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:71所示的氨基酸序列；
所述命名为2D7的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:79所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:80所示的氨基酸序列；
所述命名为4H3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:88所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:89所示的氨基酸序列；
所述命名为5B3的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:97所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:98所示的氨基酸序列；
所述命名为6B9的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:106所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:107所示的氨基酸序列；
所述命名为4F1的兔重组单克隆抗体，其重链可变区序列为SEQ ID NO:115所示的氨基酸序列，且轻链可变区序列为SEQ ID NO:116所示的氨基酸序列。
3.根据权利要求1或2所述的靶向人BCMA蛋白的兔重组单克隆抗体，其特征在于：
所述命名为2B3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:9所示的氨基酸序列；
所述命名为2D3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:18所示的氨基酸序列；
所述命名为4A12的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:27所示的氨基酸序列；
所述命名为5C4的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:36所示的氨基酸序列；
所述命名为5H7的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:45所示的氨基酸序列；
所述命名为6D4的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:54所示的氨基酸序列；
所述命名为1H2的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:63所示的氨基酸序列；
所述命名为2C11的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:72所示的氨基酸序列；
所述命名为2D7的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:81所示的氨基酸序列；
所述命名为4H3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:90所示的氨基酸序列；
所述命名为5B3的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:99所示的氨基酸序列；
所述命名为6B9的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:108所示的氨基酸序列；
所述命名为4F1的兔重组单克隆抗体，其SCFV序列为SEQ ID NO:117所示的氨基酸序列。
4.根据权利要求1所述的靶向人BCMA蛋白的兔重组单克隆抗体，其特征在于：所述兔重组单克隆抗体的轻链恒定区为κ链，且重链恒定区为IgG型。
5.一种核酸分子，其特征在于：其包含能够编码权利要求1-4任一项所述的靶向人BCMA蛋白的兔重组单克隆抗体的重链互补决定区或轻链互补决定区的核酸序列。
6.一种载体，其特征在于：其含有权利要求5所述的核酸分子。
7.一种宿主细胞，其特征在于：所述宿主细胞含有权利要求1至4任一项所述的靶向人BCMA蛋白的兔重组单克隆抗体、权利要求5所述的核酸分子或含有权利要求6所述的载体。
8.一种偶联物，其特征在于：其含有权利要求1至4任一项所述的靶向人BCMA蛋白的兔重组单克隆抗体。
9.一种药物组合物，其特征在于：含有主成分和辅成分，所述主成分含有权利要求1至4任一项所述的靶向人BCMA蛋白的兔重组单克隆抗体、权利要求5所述的核酸分子、权利要求6所述的载体、权利要求7所述的宿主细胞、权利要求8所述的偶联物中的一种或多种，所述辅成分选自药学上可接受的载体或赋形剂，以及任选的其它生物活性物质。
10.权利要求1至4任一项所述的靶向人BCMA蛋白的兔重组单克隆抗体、权利要求5所述的核酸分子、权利要求6所述的载体、权利要求7所述的宿主细胞或者权利要求8所述的偶联物在制备治疗疾病的药物或检测试剂中的应用。
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CN107759691A (zh) * 2017-12-04 2018-03-06 杭州尚健生物技术有限公司 一种特异性结合axl的单克隆抗体
CN109293772A (zh) * 2018-09-25 2019-02-01 上海邦耀生物科技有限公司 靶向bcma蛋白的抗体、嵌合抗原受体和药物
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CN107759691A (zh) * 2017-12-04 2018-03-06 杭州尚健生物技术有限公司 一种特异性结合axl的单克隆抗体
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