CN110621697B - 拮抗性cd40单克隆抗体及其用途 - Google Patents

拮抗性cd40单克隆抗体及其用途 Download PDF

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CN110621697B
CN110621697B CN201880030151.8A CN201880030151A CN110621697B CN 110621697 B CN110621697 B CN 110621697B CN 201880030151 A CN201880030151 A CN 201880030151A CN 110621697 B CN110621697 B CN 110621697B
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A.亚梅钮克
M.斯特拉瑟斯
S.J.苏查德
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Abstract

本公开内容提供了结合CD40的抗体，包括具有不同Fc结构域的人源化抗体和嵌合抗体。抗体结合CD40，但不显示出CD40激动活性。抗体可以包含经修饰的IgG1Fc结构域，并显示出对未成熟树突状细胞的最小活化。提供了包含抗体的组合物，用于治疗涉及CD40活性的疾病的方法，以及在制备用于治疗涉及CD40活性的疾病的药物中的用途。

Description

拮抗性CD40单克隆抗体及其用途
序列表
本申请包含序列表，该序列表已经以ASCII格式电子提交，其全部内容通过引用在此并入。所述ASCII副本创建于2018年05月22日，名称为200896_0013_00_WO_ST25.txt，大小为187,153字节。
发明领域
本公开内容提供了结合CD40的抗体，包括具有不同Fc结构域的人源化抗体和嵌合抗体。所述抗体多肽结合CD40并且不显示出CD40激动活性。所述抗体可以包含经修饰的IgG1 Fc结构域，，并显示出对未成熟树突状细胞的最小活化。提供了包含抗体的组合物，用于治疗涉及CD40活性的疾病的方法，以及在制备用于治疗涉及CD40活性的疾病的药物中的用途。
背景技术
CD40共刺激分子属于肿瘤坏死因子(TNF)受体超家族，其存在于包括树突状细胞、B细胞和巨噬细胞在内的抗原提呈细胞(APC)上。当CD40与TH细胞上的其配体CD154(CD40L)结合时，APC被活化。CD40-介导的APC活化参与多种免疫应答，包括细胞因子产生、共刺激分子(如CD86)上调以及增强抗原提呈和B细胞增殖。CD40也可以由内皮细胞、平滑肌细胞、成纤维细胞和上皮细胞表达。
CD40激活也参与多种例如与自身免疫、移植排斥或变态反应相关的不需要的T细胞应答。控制不需要的T细胞应答的一种策略是使用拮抗性抗体靶向CD40。例如，单克隆抗体HCD122(鲁卡木单抗)，以前称为Chiron 1212，目前正在用于治疗某些CD40介导的炎性疾病的临床试验中。参见在因特网超文本传输协议clinicaltrialsfeeds.org/clinical-trials/show/NCT01275209上的临床试验摘要“HCD122(鲁卡木单抗)与苯达莫司汀联合疗法在CD40+利妥昔单-抗难治性滤泡性淋巴瘤中的研究”(最后更新于2011年01月11日)。但是，单克隆抗体可以显示出激动活性。例如，抗-CD40抗体Chi220的实用性受其较弱刺激潜能的限制。参见Adams等，“Development of a chimeric anti-CD40monoclonal antibodythat synergizes with LEA29Y to prolong islet allograft survival,”J.Immunol.174:542-50(2005)。
发明内容
在第一个实施方式中，本发明提供了一种分离的抗体或其抗原结合片段，其特异性结合人CD40，其中所述抗体包含重链和轻链，其中所述重链包含含有含有GYTFTDLSMHW(SEQ ID NO:1)的CDR1，含有YITPSSGYTAYNQKFKG(SEQ ID NO:2)的CDR2，含有LILQRGAY(SEQID NO:3)的CDR3；和所述轻链包含含有RASKNVDSYGNSFMHW(SEQ ID NO:4)的CDR1，含有RASNLES(SEQ ID NO:5)的CDR2，和含有QQSNEDPLT(SEQ ID NO:6)的CDR3。
在第二个实施方式中，本发明提供了一种分离的抗体或其抗原结合片段，其特异性结合人CD40，其中所述抗体包含重链和轻链，其中所述重链包含含有GYAFTNYLIE(SEQ IDNO:17)的CDR1，含有VINPGSGGTNYNEKFKG(SEQ ID NO:18)的CDR2，和含有SQLGRRFDY(SEQ IDNO:19)的CDR3；和所述轻链包含含有KASQDVRTGVA(SEQ ID NO:20)的CDR1，含有SASYRNT(SEQ ID NO:21)的CDR2，和含有QQHYSPPYT(SEQ ID NO:22)的CDR3。
分离的抗体或其抗原结合片段能够拮抗CD40的活性。分离的抗体或其抗原结合部分可以是嵌合抗体。分离的抗体或其抗原结合部分可以是人源化抗体。分离的抗体或其抗原结合部分可以包含人重链恒定区和人轻链恒定区。
本申请所述的分离的抗体或其抗原结合部分可以包含人IgG1 Fc结构域，其含有在Kabat位置238的突变，所述突变降低与Fc-γ-受体(FcgR)的结合，其中脯氨酸238(P238)突变为选自下组的残基之一：赖氨酸(K)、丝氨酸(S)、丙氨酸(A)、精氨酸(R)和色氨酸(W)，和其中所述抗体或抗原结合部分具有降低的FcgR结合。
在某些实施方式中，本申请所述的分离的抗体或其抗原结合部分可以在人IgG1Fc结构域中具有P238突变为赖氨酸(P238K)。本申请所述的分离的抗体或其抗原结合部分可以包含Fc结构域，其含有选自下述的氨基酸序列：
Figure SMS_1
Figure SMS_2
或者
Figure SMS_3
在某些特定的实施方式中，本申请所述分离的抗体或其抗原结合部分可以包含人IgG1 Fc结构域，其含有在Kabat位置297取代为丙氨酸。本申请所述分离的抗体或其抗原结合部分包含Fc结构域，其中所述Fc结构域包含选自下述的氨基酸序列：
Figure SMS_4
Figure SMS_5
或者
Figure SMS_6
在某些特定的实施方式中，分离的抗体或其抗原结合部分包含：(1)具有SEQ IDNO:7所示氨基酸序列的可变重链(VH)或(2)具有SEQ ID NO:8所示氨基酸序列的可变轻链(VL)。例如，分离的抗体或抗原结合部分包含：(1)具有SEQ ID NO:7所示氨基酸序列的可变重链(VH)和(2)具有SEQ ID NO:8所示氨基酸序列的可变轻链(VL)。在一个特定的实施方式中，分离的抗体或其抗原结合部分包含(1)具有SEQ ID NO:85所示氨基酸序列的重链，和(2)具有SEQ ID NO:88所示氨基酸序列的轻链。在其他特定的实施方式中，分离的抗体或其抗原结合部分包含：
(1)具有SEQ ID NO:106所示氨基酸序列的重链和(2)具有SEQ ID NO:88所示氨基酸序列的轻链；
(1)具有SEQ ID NO:107所示氨基酸序列的重链和(2)具有SEQ ID NO:88所示氨基酸序列的轻链；
(1)具有SEQ ID NO:108所示氨基酸序列的重链和(2)具有SEQ ID NO:88所示氨基酸序列的轻链；或者
(1)具有选自SEQ ID NO:84、109、110或111所示氨基酸序列的重链；和具有SEQ IDNO:88所示氨基酸序列的5F11-45轻链。
在其他特定的实施方式中，分离的抗体或其抗原结合部分包含(1)选自下组的可变重链(VH)：hz-HC1(SEQ ID NO:23)，hz-HC2(SEQ ID NO:24)，hz-HC3(SEQ ID NO:25)，hz-HC4(SEQ ID NO:26)，hz-HC5(SEQ ID NO:27)，hz-HC6(SEQ ID NO:28)，hz-HC7(SEQ ID NO:29)，hz-HC8(SEQ ID NO:30)，hz-HC9(SEQ ID NO:31)，hz-HC10(SEQ ID NO:32)和hz-HC11(SEQ ID NO:33)；和/或(2)选自下组的可变轻链(VL)：hz-LC1(SEQ ID NO:34)，hz-LC2(SEQID NO:35)，hz-LC3(SEQ ID NO:36)，hz-LC4(SEQ ID NO:37)和hz-LC5(SEQ ID NO:38)。
在某些特定的实施方式中，分离的抗体或其抗原结合部分包含(1)选自SEQ IDNO:39、SEQ ID NO:40、SEQ ID NO:42和SEQ ID NO:43的重链氨基酸序列，其中C-末端任选地还包含赖氨酸；和/或(2)选自SEQ ID NO:41和SEQ ID NO:44的可变轻链(VL)氨基酸序列。为了说明这一点，分离的抗体或其抗原结合部分选自下组：
a)具有SEQ ID NO:39所示重链和SEQ ID NO:41所示轻链的Y1238-hz1-P238K；
b)具有SEQ ID NO:40所示重链和SEQ ID NO:41所示轻链的Y1238-hz1-N297A；
c)具有SEQ ID NO:42所示重链和SEQ ID NO:44所示轻链的Y1238-hz2-P238K；和
d)具有SEQ ID NO:43所示重链和SEQ ID NO:44所示轻链的Y1238-hz2-N297A.
本申请公开的抗体或其抗原结合部分，其中所述抗原结合部分选自下组：Fv、Fab、F(ab’)2、Fab’、dsFv、scFv、sc(Fv)2、双抗体和scFv-Fc。
本申请公开的抗体或其抗原结合部分,其中所述抗体或其抗原结合部分与治疗剂连接。
本申请公开的抗体或其抗原结合部分,其中所述抗体或其抗原结合部分与第二功能性部分连接，所述第二功能性部分具有与所述抗体或其抗原结合部分不同的结合特异性。
本申请公开的抗体或其抗原结合部分还包含另外的部分。
本申请公开了编码分离的抗体或其抗原结合部分的核酸。本申请公开了包含核酸分子的表达载体。还涉及使用表达载体转化的细胞。
还公开了一种制备抗-人CD40抗体或其抗原结合部分的方法，包括：
a)在使用表达载体转化的细胞中表达所述抗体或其抗原结合部分，所述表达载体包含编码本申请公开的分离的抗体或其抗原结合部分的核酸分子；和
b)从所述细胞中分离所述抗体或其抗原结合部分。
还提供了一种药物组合物，所述药物组合物包含：a)本申请公开的抗体或其抗原结合部分；和b)药学上可接受的载体。
提供了一种在对象中治疗或预防免疫应答的方法，所述方法包括向所述对象施用本申请公开的所述抗体或其抗原结合部分。在此类在所述对象中治疗或预防免疫应答的方法中，所述对象具有选自下组的疾病：艾迪生氏病、变态反应、过敏反应、强制性脊柱炎、哮喘、动脉粥样硬化、特应性变态反应、耳部自身免疫性疾病、眼部自身免疫性疾病、自身免疫性肝炎、自身免疫性腮腺炎、支气管哮喘、冠心病、克罗恩病、糖尿病、附睾炎、肾小球肾炎、格雷夫斯病、格林-巴利综合征、桥本氏病、溶血性贫血、特发性血小板减少性紫癜、炎性肠病、对重组药品(例如，血友病中的因子VII)的免疫应答、系统性红斑狼疮、多发性硬化、重症肌无力、天疱疮、银屑病、风湿热、类风湿性关节炎、结节病、硬皮病、干燥综合征，脊柱关节炎、甲状腺炎、移植排斥、血管炎和溃疡性结肠炎。
还涉及本申请所公开的抗体或其抗原结合部分，或者包含其的药物用于治疗需要其的对象的用途。还涉及治疗有效量的本申请公开的抗体或其抗原结合部分用于治疗或预防免疫应答的用途，其中向需要其的患者施用所述抗体或其抗原结合部分。
附图说明
图1描述了在结合有1μM 5F11-45抗体的抗-His FAB表面上捕获7μg/ml hCD64-His的SPR传感图数据。
图2(包括图2A-2C)描述了在两个独立的供体中使用抗-CD40抗体5F11处理未成熟树突状细胞(iDC)的iDC活化数据。细胞培养基中的IL-6(白介素-6)(图2A)增加，以及细胞表面标志物的表达(CD54和CD86)如使用抗-CD86和抗-CD54抗体染色的流式细胞术的平均荧光所示。在图2B和图2C中在Y轴上测量了平均荧光强度(MFI)。抗体浓度以μg/ml表示。由“x-连接”表示在测定中包含CHO-CD32细胞。
图3(包括图3A-3C)描述了在两个独立的供体中使用嵌合Y1238抗-CD40抗体处理未成熟树突状细胞(iDC)的iDC活化数据。IL-6(图3A)增加，以及细胞表面标志物的表达(即，CD54和CD86)如使用抗-CD86(图3C)和抗-CD54(图3B)抗体染色的流式细胞术的平均荧光所示。抗体浓度以μg/ml表示。由“x-连接”表示在测定中包含CHO-CD32细胞。
图4(包括图4A-4C)描述了由抗-CD40抗体Y1238-P238K和Y1238-N297A处理未成熟树突状细胞(iDC)的iDC活化数据。测得细胞培养基中的IL-6增加(图4A)。如使用抗-CD86(图4B)和抗-CD54(图4C)抗体染色的流式细胞术平均荧光所示测量细胞表面标志物的表达(CD54和CD86)。抗体浓度以μg/ml表示。表明包含CHO-CD32细胞。
图5(包括图5A-5C)描述了在加入(x-连接)和不加入过表达CD32 CHO细胞的条件下使用Y1238抗体的人源化(hz)形式处理iDC的iDC活化数据。抗体的处理浓度为10、30或100μg/ml。图5A：细胞因子IL-6产生情况，以pg/ml计(上图)。图5B：针对CD54染色的平均荧光强度。图5C：CD86的平均荧光强度。
图6(包括图6A-6C)描述了在8个独立的供体中使用抗-CD40抗体5F11处理未成熟树突状细胞(iDC)的iDC活化数据。细胞培养基中的IL-6(白介素-6)(图6A)增加，以及细胞表面标志物的表达(CD54；图6B和CD86；图6C)如使用抗-CD86和抗-CD54抗体染色的流式细胞术的平均荧光所示。在图6B和图6C中在Y轴上测量了平均荧光强度(MFI)。抗体浓度以μg/ml表示(10、30或100μg/ml)。如所示的在测定中包含CHO-CD32细胞以介导FcgR介导的交联或聚簇。将CD40激动抗体#2141和抗-CD40-dAB-IgG4作为阳性对照。将IgG4-L6融合蛋白作为阴性对照。
具体实施方式
本公开内容涉及抗-CD40抗体，特别是拮抗性抗-CD40抗体。对于诸如CD40的治疗性靶点而言，抗-CD40抗体FcgR-介导的交联具有导致不需要的激动性信号传导的可能性以及导致毒性的可能性。本公开内容还描述了拮抗性抗-CD40抗体，其与“低亲和性”FcgRhCD32a/FcgRIIa、hCD32b/FcgRIIb、hCD16a/FcgRIIIa好hCD16b/FcgRIIIb具有降低的接合(engagement)。与低亲和性FcgR具有降低的接合预期降低不需要的激动性信号传导的可能性和不需要的毒性的可能性。
定义&缩写下面提供了进一步的缩写和定义。
APC 抗原提呈细胞
CD54 也称为ICAM-1
CDR 互补性决定区
CH或CH 恒定重链
CL或CL 恒定轻链
CHO细胞 中国仓鼠卵巢细胞
dAb 域抗体
FcgR 可与FcγR互换使用
FR 框架区
GM-CSF 粒细胞巨噬细胞集落刺激因子
HC 重链
iDC 未成熟树突状细胞
IFN 干扰素
IgG 免疫球蛋白G
IL-6 白介素-6
LC 轻链
mAb 单克隆抗体
mg 毫克
ml或mL 毫升
ng 纳克
nM 纳摩
pI 等电点
SPR 表面等离子体共振
TNF 肿瘤坏死因子
μg 微克
μM 微摩
VL或VL 可变轻链结构域
VH或VH 可变重链结构域
本申请提供了进一步的缩写和定义。
根据该详细描述，下述缩写和定义适用。必须注意的是，除非在上下文中另有明确规定，否则如在本申请中所使用的单数形式的“一个(a)”、“一种(an)”和“该(the)”包括复数指代。因而，例如，提及“一个抗体”时包括多个此类抗体和提及“该剂量”时包括一个或多个剂量以及本领域技术人员公知的等同物等等。
如本文中所使用的，术语“约”是本领域普通技术人员所理解的，并且将在使用其的上下文中在一定程度上变化。在通常情况下，除非在说明书中另外明示，否则“约”包括参照值正/负10％范围的值。
应当理解的是，本申请所述范围之间的任何和所有整数或部分整数都包括在内。
CD40也是已知的并且将其称为B-细胞表面抗原CD40、Bp50、CD40L受体、CDw40、CDW40、MGC9013、p50、TNFRSF5和肿瘤坏死因子受体超家族成员5。“人CD40”指包含下述氨基酸序列的CD40：
Figure SMS_7
如在本申请中所使用的，术语“可变结构域”指由Kabat等，Sequences ofImmunological Interest,5th ed.,U.S.Dept.Health&Human Services,Washington,D.C.(1991)定义的免疫球蛋白可变结构域。可变结构域内的CDR氨基酸残基的编号和位置是根据本领域熟知的Kabat编号确定的。VH“可变重链”和“可变重链结构域”指重链的可变结构域。VL“可变轻链”和“可变轻链结构域”指轻链的可变结构域。
当应用于抗体时，术语“人”指具有来源于人免疫球蛋白序列(例如，FR和/或CH结构域)的抗体。当序列(a)从人个体或来自人个体的细胞或细胞系分离；(b)从克隆的人抗体基因序列或人抗体可变结构域序列文库分离；或者(c)通过对一种或多种上述多肽突变和选择而多样化时，则序列“来自”人免疫球蛋白编码序列。
如在本申请中所使用的，“分离的”化合物指将所述化合物从与所述化合物天然结合的至少一种组分中除去。
可以向人类患者施用本公开内容的抗体，同时在很大程度上避免了通常因施用来自其他物种(例如，小鼠)的抗体而引起的抗-抗体免疫应答。例如，根据本领域公知的方法，可以通过将鼠CDR移植到人可变结构域FR上来“人源化”鼠抗体。然而，可以生产如本申请所公开的人抗体，而无需对鼠抗体序列进行基因操纵。
本公开内容中使用的抗-CD40抗体包含3个互补性决定区(CDR)和4个框架区(FR)，其从氨基末端到羧基末端按照以下顺序排布：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。这3个CDR含有与抗原特异性相互作用的大部分残基并且主要负责抗原识别。
在一个特定的实施方式中，本公开内容的抗-CD40抗体可以包含人源化抗体5F11-45的CDR。表1中提供了5F11-45的氨基酸序列。
表1：5F11-45的序列
Figure SMS_8
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Figure SMS_9
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Figure SMS_10
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Figure SMS_11
在一个实施方式中，本公开内容的抗体包含人源化5F11-45可变重链和轻链序列的CDR1、CDR2和CDR3区的氨基酸序列(参见例如，分别为SEQ ID NOS:7和8，作为实例)。单克隆抗体含有全部6个CDR(3个针对VH和3个针对VL)，例如，针对可变重链CDR 1-3分别为GYTFTDLSMHW(SEQ ID NO:1)，YITPSSGYTAYNQKFKG(SEQ ID NO:2)和LILQRGAY(SEQ ID NO:3)，以及针对可变轻链CDR 1-3分别为RASKNVDSYGNSFMHW(SEQ ID NO:4)，RASNLES(SEQ IDNO:5)和QQSNEDPLT(SEQ ID NO:6)。
在另一个实施方式中，本公开内容的抗体可以包括人源化Y1238可变重链和轻链序列的CDR1、CDR2和CDR3区的氨基酸序列(参见例如，分别为SEQ ID NOS:23和34，作为实例)。单克隆抗体含有全部6个CDR(3个针对VH和3个针对VL)，例如，针对可变重链CDR 1-3分别为GYAFTNYLIE(SEQ ID NO:17)，VINPGSGGTNYNEKFKG(SEQ ID NO:18)和
SQLGRRFDY(SEQ ID NO:19)，以及针对可变轻链CDR 1-3分别为
KASQDVRTGVA(SEQ ID NO:20)，SASYRNT(SEQ ID NO:21)和
QQHYSPPYT(SEQ ID NO:22)。
“抗体”(Ab)应包括但不限于与抗原特异性结合并且包含通过二硫键相互连接的至少2条重链(H)和2条轻链(L)的免疫球蛋白，或其抗原结合片段。每条H链包含重链可变区(在本申请中缩写为VH)和重链恒定区。重链恒定区包含3个恒定结构域CH1、CH2和CH3。每条轻链包含轻链可变区(在本申请中缩写为VL)和轻链恒定区。轻链恒定区包含一个恒定结构域CL。VH和VL区可以进一步细分为高变区，称为互补性决定区(CDR)，其间散布着更为保守的称为框架区(FR)的区域。每个VH和VL包含3个CDR和4个FR，其从氨基末端到羧基末端按照以下顺序排布：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。重链和轻链的可变区含有与抗原相互作用的结合结构域。
Ab的“抗原结合部分”(也称为“抗原结合片段”)或其抗原结合部分指保留与完整Ab结合的抗原特异性结合能力的Ab的一条或多条序列(全长或全长抗体的片段)。抗原结合片段的实例包括Fab、F(ab’)2、scFv(单链可变片段)、Fab’、dsFv、sc(Fv)2和scFv-Fc。
“人源化”抗体指其中非人Ab CDR结构域以外的一些、大部分或全部氨基酸被来源于人免疫球蛋白的相应氨基酸替代的Ab。在Ab人源化形式的一个实施方式中，CDR结构域以外的一些、大部分或全部氨基酸被来自人免疫球蛋白的氨基酸替代，而一个或多个CDR区内的一些、大部分或全部氨基酸未改变。氨基酸的少量加入、缺失、插入、取代或修饰是允许的，只要其不消除Ab与特定抗原结合的能力即可。“人源化”Ab保留与原始Ab相似的抗原特异性。
“嵌合抗体”指其中可变区来自一个物种和恒定区来自另一个物种的Ab，如其中可变区来自小鼠Ab和恒定区来自人Ab的Ab。
如在本申请中所使用的，“特异性结合”指例如通过表面等离子体共振(SPR)所测量的抗体以约1μM或更低的解离常数(Kd)与抗原的结合。适宜的测定系统包括BIAcoreTM(GEHealthcare Life Sciences,Marlborough,MA)表面等离子体共振系统和BIAcoreTM动力学评价软件(例如，2.1版)。
本申请所述抗体与CD40的结合拮抗CD40活性。“CD40活性”包括但不限于T细胞活化(例如，诱导T细胞增殖或细胞因子分泌)，巨噬细胞活化(例如，在巨噬细胞中诱导活性氧和一氧化氮)和B细胞活化(例如，B细胞增殖、抗体同种型转换或分化为浆细胞)。可以通过与其他分子相互作用介导CD40活性。“CD40活性”包括CD40与下述分子之间的功能性相互作用，括号中是由其Uniprot登录号标识的这些分子：
Figure SMS_12
Figure SMS_13
例如，CD40“活性”包括与TRAF2的相互作用。CD40/TRAF2相互作用活化NF-κB和JNK。参见Davies等，Mol.Cell Biol.25:9806-19(2005)。因此，这种CD40活性可以由相对于参照的CD40依赖性细胞NF-κB和JNK活化确定。
如在本申请中所使用的，术语“活化(activate)”、“活化(activates)”和“活化的(activated)”指与参照相比给定可测量的CD40活性增加至少10％，例如，至少10％、25％、50％、75％、或甚至100％或者更多。如果CD40活性降低至少10％，则CD40活性被“拮抗”，并且在一个示例性实施方式中，与不存在拮抗剂相比，降低至少约20％、30％、40％、50％、60％、70％、80％、90％、95％、97％或甚至100％(即，检测不到活性)。例如，抗体可以拮抗一些或全部CD40活性，但不活化CD40。例如，抗体可以不活化B细胞增殖。抗体可以不活化T细胞分泌细胞因子，其中所述细胞因子是选自下组的至少一种细胞因子：IL-2、IL-6、IL-10、IL-13、TNF-α、IFN-γ。
可变结构域可以包含一个或多个框架区(FR)，其具有与由人种系抗体基因区段编码的相应框架区相同的氨基酸序列。在本申请所述的抗体中使用的优选框架序列是与本申请所述的抗体使用的框架序列在结构上相似的那些。可以将VH CDR 1、2和3序列和VL CDR1、2和3序列移植到具有与框架序列所源自的种系免疫球蛋白基因中发现的序列相同序列的框架区上，或者可以将CDR序列移植到与种系序列相比含有多达20个(优选保守的)氨基酸取代的框架区上。例如，已经发现，在某些情况下，使框架区内的残基突变以维持或增强抗体的抗原结合能力是有益的(参见例如，Queen等的美国专利号5,530,101；5,585,089；5,693,762和6,180,370)。
示例性的框架区包括但不限于下表2和表3中所示的那些。序列为从氨基末端至羧基末端的格式。示例性的重链框架区如表2中所示。针对人源化5F11-45 VH重链框架序列优选的组为：QVQLVQSGAEVKKPGSSVKVSCKAS(FR1；SEQ ID NO:46)，WVRQAPGQGLEWMG(FR2；SEQID NO:49)，KTTLTADKSTSTAYMELSSLRSEDTAVYYCAR(FR3；SEQ ID NO:51)和WGQGTLVTVSS(FR4：SEQ ID NO:60)。针对人源化Y1238 VH重链框架序列优选的组为：QVQLVQSGAEVKKPGASVKVSCKAS(FR1；SEQ ID NO:47)，WVRQAPGQGLEWMG(FR2；SEQ ID NO:49)，RVTMTRDTSISTAYMELSRLRSDDTAVYYCAR(FR3；SEQ ID NO:52)和WGQGTLVTVSS(FW4；SEQ IDNO:60)。
表2
Figure SMS_14
Figure SMS_15
示例性的轻链框架区如表3中所示。人源化5F11-45 VL轻链框架区优选的组包括下述：DIVLTQSPDSLAVSLGERATINC(FR1；SEQ ID NO:61)，WYQQKPGQPPKLLIY(FR2；SEQ ID NO:63)，GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC(FR3；SEQ ID NO:66)和FGQGTKLEIK(FR4；SEQ IDNO:71)。人源化Y1238 VL轻链框架区优选的组包括下述：DIQMTQSPSSLSASVGDRVTITC(FR1；SEQ ID NO:62)，WYQQKPGKAPKLLIY(FR2；SEQ ID NO:64)，GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC(FR3；SEQ ID NO:67)，GVPSRFSGSRSGTDFTFTISSLQPEDIATYYC(FR3；SEQ ID NO:68)和FGGGTKVEIK(FR4；SEQ ID NO:72)。
表3
Figure SMS_16
Figure SMS_17
变体可变结构域可以与人源化5F11-45或Y1238序列的可变结构域相差至多10个氨基酸或其之间的任何整数值，其中所述变体可变结构域特异性结合CD40。或者，所述变体可变结构域可以分别相对于人源化5F11-45或Y1238序列的序列具有至少90％序列同一性(例如，至少92％、95％、98％或99％序列同一性)。在两条序列之间不同的氨基酸残基或氨基酸可以代表氨基酸取代、添加或缺失。当通过任意适宜的氨基酸序列比对算法如
Figure SMS_18
(美国国家医学图书馆的注册商标)对两条序列进行比对时，两条序列之间不同的残基显示为不同位置。
本发明的示例性CD40抗体可以包括分离的抗体或其抗原结合部分，其特异性结合人CD40，其中所述抗体包含重链和轻链，其中：
所述重链包含含有GYTFTDLSMHW(SEQ ID NO:1)的CDR1，含有YITPSSGYTAYNQKFKG(SEQ ID NO:2)的CDR2，含有LILQRGAY(SEQ ID NO:3)的CDR3；和
所述轻链包含含有RASKNVDSYGNSFMHW(SEQ ID NO:4)的CDR1，含有RASNLES(SEQID NO:5)的CDR2和含有QQSNEDPLT(SEQ ID NO:6)的CDR3；
或者
所述重链包含含有GYAFTNYLIE(SEQ ID NO:17)的CDR1，含有VINPGSGGTNYNEKFKG(SEQ ID NO:18)的CDR2，和含有SQLGRRFDY(SEQ ID NO:19)的CDR3；和
所述轻链包含含有KASQDVRTGVA(SEQ ID NO:20)的CDR1，含有SASYRNT(SEQ IDNO:21)的CDR2，和含有QQHYSPPYT(SEQ ID NO:22)的CDR3。
在一个特定的实施方式中，特异性结合人CD40的抗体或其抗原结合部分包含(1)重链，其包含：含有GYTFTDLSMHW(SEQ ID NO:1)的CDR1，含有YITPSSGYTAYNQKFKG(SEQ IDNO:2)的CDR2，含有LILQRGAY(SEQ ID NO:3)的CDR3；和/或(2)包含轻链，其包含：含有RASKNVDSYGNSFMHW(SEQ ID NO:4)的CDR1，含有RASNLES(SEQ ID NO:5)的CDR2和含有QQSNEDPLT(SEQ ID NO:6)的CDR3。任选地，此类抗体或其抗原结合部分包含(1)SEQ ID NO:7所示的可变重链；和/或(2)SEQ ID NO:8所示的可变轻链。为了说明这一点，此类抗体或其抗原结合部分包含(1)含有SEQ ID NO:7所示的可变重链的重链；和(2)含有SEQ ID NO:8所示的可变轻链的轻链。
特异性结合人CD40的抗体或其抗原结合部分可以包含含有SEQ ID NO:7所示的可变重链的重链和含有SEQ ID NO:8所示的可变轻链的轻链，其中所述重链包含含有SEQ IDNO:9或SEQ ID NO:98所示氨基酸序列的Fc结构域。特异性结合人CD40的抗体或其抗原结合部分可以包含含有SEQ ID NO:7所示的可变重链的重链和含有SEQ ID NO:8所示的可变轻链的轻链，其中所述重链包含含有SEQ ID NO:10或SEQ ID NO:99所示氨基酸序列的Fc结构域。
特异性结合人CD40的抗体或其抗原结合部分可以包含(1)含有SEQ ID NO:85，SEQID NO：106、107或108所示氨基酸序列的重链；和(2)含有SEQ ID NO:88所示氨基酸序列的轻链。为了说明这一点，此类抗体或其抗原结合部分包含(1)含有SEQ ID NO:85所示氨基酸序列的重链；和(2)含有SEQ ID NO:88所示氨基酸序列的轻链。
分离的抗体或其抗原结合部分能够拮抗CD40的活性。分离的抗体或其抗原结合部分可以是嵌合抗体。分离的抗体或其抗原结合部分可以是人源化抗体。分离的抗体或其抗原结合部分可以包含人重链恒定区和人轻链恒定区。
本申请所述分离的抗体或其抗原结合部分包含人IgG1 Fc结构域，其含有在Kabat位置238的突变，所述突变降低与Fc-γ-受体(FcgR)的结合，其中脯氨酸238(P238)突变为选自下组的残基之一：赖氨酸(K)、丝氨酸(S)、丙氨酸(A)、精氨酸(R)和色氨酸(W)，和其中所述抗体或抗原结合部分具有降低的FcgR结合。本申请所述分离的抗体或其抗原结合部分可以在人IgG1 Fc结构域具有P238突变为赖氨酸
分离的抗体或其抗原结合部分包含Fc结构域，其含有选自下述的氨基酸序列：
Figure SMS_19
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Figure SMS_20
或者
Figure SMS_21
本申请所述分离的抗体或其抗原结合部分可以包含人IgG1 Fc结构域，其含有在Kabat位置297取代为丙氨酸。例如，分离的抗体或其抗原结合部分包含Fc结构域，其中所述Fc结构域包含选自下述的氨基酸序列：
Figure SMS_22
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Figure SMS_23
或者
Figure SMS_24
本申请所述分离的抗体或抗原结合部分，其中所述抗体包含(1)选自下组的可变重链(VH)：hz-HC1(SEQ ID NO:23)，hz-HC2(SEQ ID NO:24)，hz-HC3(SEQ ID NO:25)，hz-HC4(SEQ ID NO:26)，hz-HC5(SEQ ID NO:27)，hz-HC6(SEQ ID NO:28)，hz-HC7(SEQ ID NO:29)，hz-HC8(SEQ ID NO:30)，hz-HC9(SEQ ID NO:31)，hz-HC10(SEQ ID NO:32)和hz-HC11(SEQ ID NO:33)；和/或(2)选自下组的可变轻链(VL)：hz-LC1(SEQ ID NO:34)，hz-LC2(SEQID NO:35)，hz-LC3(SEQ ID NO:36)，hz-LC4(SEQ ID NO:37)和hz-LC5(SEQ ID NO:38)。
本申请公开的分离的抗体或其抗原结合部分，其中所述抗体包含选自SEQ ID NO:39，SEQ ID NO:40，SEQ ID NO:42和SEQ ID NO:43的重链氨基酸序列。
本申请公开的分离的抗体或其抗原结合部分，其中所述抗体包含选自SEQ ID NO:41和SEQ ID NO:44的轻链氨基酸序列。
本申请公开的分离的抗体或其抗原结合部分，其中所述抗体选自下组：
a)具有SEQ ID NO:39所示重链和SEQ ID NO:41所示轻链的Y1238-hz1-P238K；
b)具有SEQ ID NO:40所示重链和SEQ ID NO:41所示轻链的Y1238-hz1-N297A；
c)具有SEQ ID NO:42所示重链和SEQ ID NO:44所示轻链的Y1238-hz2-P238K；和
d)具有SEQ ID NO:43所示重链和SEQ ID NO:44所示轻链的Y1238-hz2-N297A.
本申请公开的抗体或其抗原结合部分，其中所述抗原结合部分选自下组：Fv、Fab、F(ab’)2、Fab’、dsFv、scFv、sc(Fv)2、双抗体和scFv-Fc。
本申请公开的抗体或其抗原结合部分可以是免疫缀合物，其中所述抗体或其抗原结合部分与治疗剂连接。
本申请公开的抗体或其抗原结合部分可以是双特异性抗体，其中所述抗体或其抗原结合部分与第二功能性部分连接，所述第二功能性部分具有与所述抗体或其抗原结合部分不同的结合特异性。
本申请公开的抗体或其抗原结合部分还包含另外的部分。
Fc结构域
使用重链羧基末端“一半”定义恒定区(FC)并且其主要负责效应功能。如在本申请中所使用的，术语“Fc结构域”指包含根据Kabat等，Sequences of ImmunologicalInterest,5th ed.,U.S.Dept.Health&Human Services,Washington,D.C.(1991)所限定的CH2和CH3恒定结构域的恒定区抗体序列。Fc区可以来自人IgG。例如，Fc区可以来自人IgG1或人IgG4 Fc区。重链可变结构域可以与Fc结构域融合。可变结构域的羧基末端可以与FcCH2结构域的氨基末端连接或融合。或者，可变结构域的羧基末端可以与接头氨基酸序列的氨基末端连接或融合，所述接头氨基酸序列本身与Fc结构域的氨基末端融合。或者，可变结构域的羧基末端可以与CH1结构域的氨基末端连接或融合，所述CH1结构域本身与Fc CH2结构域融合。任选地，所述蛋白可以全部或部分地在CH1结构域之后包含铰链区。任选地，氨基酸接头序列存在于可变结构域与Fc结构域之间。轻链可变结构域的羧基末端可以与CL结构域的氨基末端连接或融合。
重链CH1的示例性序列是SEQ ID NO:85的氨基酸118-215。轻链CL的示例性序列是SEQ ID NO:88的氨基酸112-218。
抗体可以是融合抗体，其包含特异性结合人CD40的第一可变结构域和包含Fc结构域的第二结构域。
在融合蛋白中使用的示例性Fc结构域可以包含人IgG结构域。示例性人IgG Fc结构域包括IgG4 Fc结构域和IgG1 Fc结构域。尽管人IgG重链基因编码C-末端赖氨酸，但是由于血液循环的裂解导致内源性抗体通常不存在赖氨酸。具有包含C-末端赖氨酸的IgG重链的抗体，当在哺乳动物细胞培养物中表达时，也可能具有可变水平的C-末端赖氨酸存在(Cai等，2011,Biotechnol Bioeng.108(2):404-12)。因此，可以将本申请公开的任意IgG重链Fc结构域的C-末端赖氨酸省去。
本申请所述分离的抗体或其抗原结合部分可以包含Fc结构域，其含有选自下述的氨基酸序列：
Figure SMS_25
Figure SMS_26
Fc结构域可以包含选自表4中的序列的氨基酸序列。SEQ ID NOS：10、12、14、16、99、101、103和105在N-末端包含CH1结构域(残基1-98)。
表4
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Figure SMS_28
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Figure SMS_29
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Figure SMS_30
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Figure SMS_31
本申请抗体的可变区可以通过“氨基酸接头”或“接头”任选地与Fc结构域连接。例如，可变重链结构域的C-末端可以与氨基酸接头的N-末端融合，和Fc结构域可以与接头的C-末端融合。尽管氨基酸接头可以是任何长度并且可以由氨基酸的任何组合构成，但是接头的长度可以相对较短(例如，5个或更少的氨基酸)，以减少所连接结构域之间的相互作用。还可以对接头的氨基酸组成进行调整以减少具有大侧链的氨基酸或可能引入二级结构的氨基酸的数量。适宜的氨基酸接头包括但不限于长度可达3、4、5、6、7、10、15、20或25个氨基酸的那些。代表性的氨基酸接头序列包含GGGGS(SEQ ID NO:73)，以及接头包含GGGGS的2、3、4或5个拷贝(分别为SEQ ID NO：74-77)。表5列出了用于本公开内容的适宜的接头序列。
表5
代表性的接头序列
Figure SMS_32
Figure SMS_33
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抗体的制备
可以使用常规技术在适宜哺乳动物宿主细胞系(如CHO、293、COS、NSO等)中生产和纯化抗体，随后使用一种或多种方法进行纯化，包括蛋白A亲和层析、离子交换、反相技术等。
如本领域所熟知的，多个密码子能够编码相同氨基酸。因此，编码蛋白序列的核酸包括具有密码子简并性的核酸。本申请公开的多肽序列可以由多种核酸编码。遗传密码是通用的和众所周知的。基于本领域的常规知识能够容易地构思出编码本申请公开的任何多肽序列的核酸，并且可以对其进行优化以进行生产。尽管编码给定多肽的核酸序列可能数目很大，但是给定遗传密码的标准表，并在计算机的辅助下，本领域普通技术人员能够容易地做出编码给定多肽的核酸序列的每种可能的组合。
编码5F11-45重链可变结构域的代表性核酸序列是：
Figure SMS_34
Figure SMS_35
在该序列中，核苷酸76-105编码5F11-45重链可变结构域的CDR1，核苷酸148-198编码CDR2和核苷酸295-318编码CDR3。
编码5F11-45轻链可变结构域的代表性核酸序列是：
Figure SMS_36
Figure SMS_37
在该序列中，核苷酸70-117编码5F11-45轻链可变结构域的CDR1，核苷酸160-180编码CDR2和核苷酸277-303编码CDR3。
编码包含P238K Fc结构域的5F11-45重链的代表性核酸序列是：
Figure SMS_38
Figure SMS_39
在该序列中，核苷酸1-351编码重链的可变结构域。核苷酸352-645编码CH1结构域，和核苷酸646-1338编码Fc结构域。任选地，可以将赖氨酸的密码子加入Fc结构域序列的3’末端。
编码5F11-45轻链的代表性核酸序列是：
Figure SMS_40
Figure SMS_41
Figure SMS_42
在该序列中，核苷酸1-333编码轻链的可变结构域，和核苷酸334-654编码CL。
重链和/或轻链的编码序列可以编码在编码序列5’末端的信号肽，如MRAWIFFLLCLAGRALA(SEQ ID NO:119)。5F11-45重链和轻链的示例性编码序列(其均包含信号肽编码序列)为：
Figure SMS_43
Figure SMS_44
和
Figure SMS_45
Figure SMS_46
因此，还涉及编码本申请公开的抗体的核酸。可以将此类核酸插入载体，如适宜的表达载体，例如pHEN-1(Hoogenboom等，(1991)Nucleic Acids Res.19:4133-4137)。还提供了包含载体和/或核酸的分离的宿主细胞。
可以仅使用常规技术在任意适宜哺乳动物宿主细胞系(如CHO(中国仓鼠卵巢细胞)、293(人胚肾293细胞)、COS细胞、NSO细胞等)中生产和纯化本公开内容的抗体，随后使用一种或多种方法进行纯化，包括蛋白A亲和层析、离子交换、反相技术等。
药物组合物和治疗方法
药物组合物包含治疗有效量的一种或多种抗体和任选地药学上可接受的运载体。药学上可接受的运载体包括例如水、盐水、磷酸盐缓冲液、右旋糖、甘油、乙醇等及其组合。药学上可接受的运载体可以进一步包含少量辅助物质，如润湿剂或乳化剂、防腐剂或者增加融合蛋白半衰期或有效性的缓冲剂。可以将组合物制剂，以便在施用后提供活性成分的快速、持续或延迟释放。适宜的药物组合物以及用于制备其的方法是本领域公知的。参见例如，Remington,The Science and Practice of Pharmacy,A.Gennaro等编著，21st ed.,Mack Publishing Co.(2005)。
药物组合物可以进一步包含免疫抑制剂/免疫调节剂和/或抗炎剂。
一种在需要此类治疗的患者中治疗免疫疾病的方法可以包括向患者施用治疗有效量的抗体。拮抗CD40-介导的T细胞活化能够抑制例如在自身免疫、移植排斥或变态反应期间出现的不利T细胞应答。抑制CD40-介导的T细胞活化能够调节这些疾病的进展和/或严重程度。
还提供了使用本公开内容的抗体或其药学上可接受的盐在制备用于在需要此类治疗的患者中治疗免疫疾病的药物中的用途。例如，可以将所述药物与免疫抑制剂/免疫调节剂和/或抗炎剂联合施用。
如在本申请中所使用的，“患者”指动物，例如哺乳动物，包括人。患者可以被诊断为患有免疫疾病。“治疗(treatment)”或“治疗(treat)”或“治疗(treating)”指涉及减轻症状、病症、病况或疾病的进展或严重程度的方法。“免疫疾病”指与个体中的免疫反应进展相关的任何疾病，包括细胞和/或体液免疫反应。免疫疾病的实例包括但不限于炎症、变态反应、自身免疫性疾病或移植相关疾病。“自身免疫性疾病”指与个体中的自身免疫反应进展相关的任何疾病，包括细胞和/或体液免疫反应。自身免疫性疾病的实例是炎性肠病(IBD)，包括但不限于溃疡性结肠炎和克罗恩病。其他自身免疫性疾病包括系统性红斑狼疮、多发性硬化症、类风湿性关节炎、糖尿病、银屑病、硬皮病和动脉粥样硬化。移植相关疾病包括移植物抗宿主病(GVHD)、急性移植排斥和慢性移植排斥。
能够通过施用本公开内容的抗体治疗的疾病可以选自下组：艾迪生氏病、变态反应、过敏反应、强制性脊柱炎、哮喘、动脉粥样硬化、特应性变态反应、耳部自身免疫性疾病、眼部自身免疫性疾病、自身免疫性肝炎、自身免疫性腮腺炎、支气管哮喘、冠心病、克罗恩病、糖尿病、附睾炎、肾小球肾炎、格雷夫斯病、格林-巴利综合征、桥本氏病、溶血性贫血、特发性血小板减少性紫癜、炎性肠病、对重组药品(例如，血友病中的因子VII)的免疫应答、系统性红斑狼疮、多发性硬化、重症肌无力、天疱疮、银屑病、风湿热、类风湿性关节炎、结节病、硬皮病、干燥综合征，脊柱关节炎、甲状腺炎、移植排斥、血管炎和溃疡性结肠炎。
药物组合物可以单独施用，或者作为与免疫抑制剂/免疫调节剂和/或抗炎剂的联合疗法(即，同时或顺序)施用。不同免疫疾病可能需要使用用于治疗免疫疾病的特定辅助化合物，其取决于患者的具体情况。例如，可以将药物组合物与一种或多种适宜的佐剂(例如，细胞因子(例如，IL-10和IL-13))或他免疫刺激剂(例如，趋化因子、肿瘤相关抗原和肽)联合施用。适宜的佐剂是本领域公知的。
可以将任何适宜的方法或途径用于施用抗体或药物组合物。施用途径包括例如静脉内、腹膜内、皮下或肌内施用。所施用抗体的治疗有效剂量取决于很多因素，例如包括所治疗的免疫疾病的类型和严重程度，联合疗法的使用，抗体或药物组合物的施用途径以及患者的体重。相对于患者的体重，域抗体治疗有效量的非限制性范围是0.1-20毫克/千克(mg/kg)，以及在一个方面中是1-10mg/kg。
试剂盒
提供了用于在人类患者中治疗免疫疾病的试剂盒。试剂盒可以包含(a)一剂本公开内容的抗体和(b)在治疗患者免疫疾病的方法中使用所述抗体的说明性材料。
作为在本申请中使用的术语，“说明性材料”包括出版物、记录、图表或任何其他表达媒介，可以将其用于传达试剂盒中本发明组合物和/或化合物的有用性。例如，可以将试剂盒的说明性材料固定在含有本发明化合物和/或组合物的容器上，或者与装有所述化合物和/或组合物的容器一起运输。或者，可以将说明性材料与容器分开运输，其目的是使接收者协同使用说明性材料和化合物。例如，可以通过出版物的实际交付或传达试剂盒有用性的其他表达媒介，或者可以通过电子传输(例如，通过计算机，如通过电子邮件)或从网站下载来实现传递说明性材料。
实施例
实施例1：抗-CD40抗体的Fc形式影响未成熟树突状细胞.的活化
可以通过突变重链恒定区(如在铰链和Fc结构域中)对抗体进行工程化改造以使其针对FcgR(FcγR)具有不同亲和性和选择性。可以引入突变以增强或减弱FcgR结合。这些突变能够增加或减少FcgR-介导的交联和/或信号传导。对于诸如CD40的治疗性靶点而言，抗-CD40抗体FcgR-介导的交联具有导致不需要的激动性信号传导的可能性以及导致毒性的可能性。重要的是要鉴定出抗-CD40拮抗剂抗体，这些抗体具有降低的FcgR结合以及交联和/或信号传导的可能性，特别是降低与“低亲和性”FcgR hCD32a/FcgRIIa、hCD32b/FcgRIIb、hCD16a/FcgRIIIa和hCD16b/FcgRIIIb的接合。通常对“高亲和性”受体CD64/FcgRI的接合给予较少关注，因为该受体被血清IgG饱和。
在美国专利公开号20160376371(通过引用并入本申请)中公开了抗体5F11-45，其是人源化抗体，对人CD40具有较高亲和性，该抗体被工程化改造以具有激动剂活性，并且能够优先与CD32a-R131和CD32b结合。下表6提供了5F11-45重链和轻链可变区的氨基酸序列。在重链可变区中，CDR1是氨基酸26-35，CDR2是氨基酸50-66和CDR3是氨基酸99-106。在轻链可变区中，CDR1是氨基酸24-38，CDR2是氨基酸54-60和CDR3是氨基酸93-101。在每个可变区中的CDR用下划线表示。
为了探索Fc形式对未成熟树突状细胞(iDC)活化的影响，使用野生型人IgG1f同种型形成人源化抗-CD40抗体5F11-45(5F11-45-IgG1f)，以及设计4个带有突变的同种型以减少FcgR结合(即，5F11-45-IgG1.3f、5F11-45-CTza、5F11-45-P238K、5F11-45-N297A)。见表6中的SEQ ID NOS:81-85。表6还列出了每种5F11-45抗体的重链和轻链恒定区。Fc结构域在表6中用下划线表示。重链可变区(氨基酸1-117)和轻链可变区(氨基酸1-111)以粗体显示。在重链中可变区(粗体显示)和Fc结构域(用下划线表示)之间的序列(重链的氨基酸118-215)包含CH1结构域。在轻链中未加粗的序列(氨基酸112-218)包含CL结构域。
表6
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Figure SMS_50
作为对照，对相同5F11-45抗体进行工程化改造以增强与人CD32a和CD32b蛋白的结合(5F11-45-SE；重链(HC)和轻链(LC)分别如SEQ ID NOS:86和88所示)或者增强对hCD32b相对于hCD32a的选择性(5F11-45-V11；重链和轻链分别如SEQ ID NOS:87和88所示)。
通过表面等离子体共振(SPR)表7证明，所有5G11-45 Fc变体显示出与CD40靶点具有较高的亲和性。
表7：与在蛋白A传感器芯片表面上捕获的5F11-45抗体结合的人-CD40单体的SPR动力学和亲和性数据。
Figure SMS_51
通过SPR对Fc-工程化改造5F11-45抗体的FcgR结合性质进行表征以确定是否Fc突变对FcgR的结合和选择性具有所需影响。针对FcgR结合的SPR数据如表8中所示。
表8：与抗-his Fab捕获的hFcgR-his蛋白结合的1μM或10μM 5F11-45抗体的％Rmax数据。
Figure SMS_52
如表8中所示，5F11-45-IgG1f与所有FcgR结合，与CD64、CD16a-V158和CD32a-H131具有特别强的结合。S267E变体(5F11-45-SE)(对照)显示出与CD32a和CD32b蛋白预期增强的结合。在5F11-45-V11的IgG1 V11同种型中存在的5个突变提供了对CD32b增强的选择性。
其他4个5F11-45 Fc变体显示出降低的FcgR结合。5F11-45-IgG1.3f显示出与所有FcgR降低的结合，包括与CD64的结合最弱。在非常高的抗体浓度(即，10μM)下，观察到5F11-45-IgG1.3f与CD32a-R131一些较弱的结合。而相反的是，即使在10μM抗体浓度下，其他3种变体(5F11-45-CTza、5F11-45-P238K和5F11-45-N297A)也几乎无法检测到与任何低亲和性FcgR的结合。参见上表7。还观察到了CD64结合的差异，特别是在解离速率方面，与5F11-45-CTza或5F11-45-IgG1f相比，5F11-45-P238K和5F11-45-N297A的解离速率要快得多，图1。
未成熟树突状细胞(iDC)对通过成簇/交联的CD40间接活化敏感，其能够导致未成熟DC活化，从而引起细胞因子释放(例如，白介素-6，IL-6)和细胞表面活化信号上调(例如，CD86和CD54)。使用高度过表达CD32a并对FcgR具有低亲和性的中国仓鼠卵巢(CHO)细胞证明FcgR介导的成簇/交联的可能性。CHO细胞与iDC的比率为1:6，表示具有极高的成簇/交联水平。
BMS-986090是与IgG4 Fc结构域融合的抗-CD40激动剂结构域抗体，并且其具有氨基酸序列：
Figure SMS_53
Figure SMS_54
(其是在WO2012/145673中的SEQ ID NO:1287)。BMS-986090序列的实线下划线部分对应于BMS3h-56-269域抗体(其是在WO2012/145673中的SEQ ID NO:417)。虚线下划线“AST”(SEQ ID NO:78)是接头，其余是IgG4.1的Fc结构域。2141(mAb 134-2141)是CD40激动剂抗体(参见RobertVonderheide等，“Clinical Activity and Immune Modulation in Cancer PatientsTreated with CP-870,893,a Novel CD40 Agonist Monoclonal Antibody,”25(7):876-883(2007))。将BMS-986090和2141作为交联介导的抗-CD40 iDC活化的阳性对照。将L6-IgG4(不具有CD40结合能力的融合蛋白)作为阴性对照。如图2中所示，当是可溶性的或与细胞表面CD32交联/成簇时，如通过细胞因子增加(白介素-6；IL-6；图2A)或活化标记物增加(CD86 MFI；图2C和CD54MFI；图2B)所测量的，5F11-45-CTza、5F11-45-P238K和5F11-45-N297A未显示出交联介导的抗-CD40 iDC活化。如CD54和CD86细胞表面染色增加所示，仅当与CD32 CHO细胞共同孵育时，5F11-45-IgG1.3f才显示出适度iDC活化。表现出与低亲和性FcgR结合的所有变体(即，5F11-45-IgG1f、5F11-45-SE和5F11-45-V11)显示出iDC活化，其在加入CD32 CHO细胞后显著增强，图2。具有CTza、P238K和N297A的5F11-45抗体未显示出任何可检测的iDC活化。这些数据支持了具有CTza、P238K和N297A的5F11-45抗体拮抗CD40活性。
为了在其他抗体中进一步评估这些观察结果，制备强效小鼠抗-人-CD40抗体Y1238，作为与这些同种型的嵌合抗体并且作为IgG1.3f(重链为SEQ ID NOS:90-93和轻链为SEQ ID NO:94)。见表9。重链可变区是氨基酸1-118(粗体表示)。在重链可变区中，CDR1是氨基酸26-36，CDR2是氨基酸50-66和CDR3是氨基酸99-107。Fc结构域是氨基酸217-448。在重链中，可变区(粗体表示)与Fc结构域(下划线表示)之间的序列(重链的氨基酸119-216)包含CH1结构域。轻链可变区是氨基酸1-107。在轻链可变区中，CDR1是氨基酸24-34，CDR2是氨基酸50-56和CDR3是氨基酸89-97。在轻链中的氨基酸108-214包含CL结构域。
表9
Figure SMS_55
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Figure SMS_56
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Figure SMS_57
通过SPR针对CD40和FcgR结合以及针对iDC活化对这些嵌合抗体进行检测。所有Fc变体Y1238嵌合抗体均显示出针对人-CD40较高的亲和性，表10。
表10：与在蛋白A传感器芯片表面上捕获的嵌合Y1238抗体结合的人-CD40单体的SPR动力学和亲和性数据。
样品 ka(1/Ms) kd(1/s) KD(nM)
Y1238-hIgG1.3f 1.0E+05 2.1E-04 2.1
Y1238-CTza 1.0E+05 2.3E-04 2.3
Y1238-N297A 9.7E+04 2.4E-04 2.5
Y1238-P238K 9.7E+04 2.6E-04 2.6
与5F11-45类似，与IgG1f同种型对照相比，具有经工程化改造Fc结构域的嵌合Y1238抗体显示出更弱的FcgR结合。Y1238-IgG1.3f分子对低亲和性FcgR(特别是CD32a-R131和CD32b)显示出有些弱，但能够测量的结合应答。而相反的是，即使在10μM的抗体浓度下，CTza、P238K和N297A变体也显示出与这些FcgR非常弱或无法检测的结合，表11。
表11：与抗-his Fab捕获的hFcgR-his蛋白结合的1μM或10μM嵌合Y1238抗体的％Rmax数据。
Figure SMS_58
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Figure SMS_59
嵌合Y1238抗体的CD64结合与5F11-45变体类似，IgG1.3f表现出最弱的CD64结合。无论是Y1238嵌合抗体形式还是5F11-45，P238K和N297A变体具有比CTza显著更快的解离速率。
与5F11-45类似，当形成CTza、P238K或N297A同种型时，单独或加入过表达FcgR(CD32)的CHO细胞，嵌合Y1238均显示出无iDC活化作用，图3。在具有IgG1.3f Fc序列的嵌合Y1238中观察到最小活化。然而，通过与表达CD32的CHO细胞共同孵育，增加了iDC细胞因子的释放(图3A)和活化标志物的上调(图3B和3C)。
在来自其他共同的iDC中对嵌合Y1238-P238K和Y1238-N297A进行了进一步检测，证实了单独或加入CD32 CHO细胞时，这些分子均不活化未成熟树突状细胞，图4。在这些研究中的阳性对照是BMS-986090和2141；当在溶液中使用时，这两种抗体在来自供体子集的细胞中均导致iDC活化(图3和4)。然而，随着CD32 CHO细胞的成簇/交联，iDC活化显著增加，在这些条件下在来自每种测试供体的细胞中均观察到了这一现象。
实施例2：当制成含有P238K或N297A突变的同种型时，人源化CD40拮抗性抗体Y1238显示出对CD40信号传导的高效抑制以及无激动剂信号。
人源化背景/程序如在WO2017004006的章节“II经工程化改造和修饰的抗体”中所讨论的，其全部内容通过引用并入本申请。基于该分析，选择11个11条VH序列(SEQ ID NOS:23-33)和5条Vκ序列(SEQ ID NOS:34-38)进行检测。取代用粗体和虚线下划线表示。每条可变重链和可变轻链中的CDR(按照CDR1-CDR2-CDR3方向)用下划线表示，即可变重链CDR分别为GYAFTNYLIE(SEQ ID NO:17)，VINPGSGGTNYNEKFKG(SEQ ID NO:18)和SQLGRRFDY(SEQ IDNO:19)(表12)和可变轻链CDR分别为KASQDVRTGVA(SEQ ID NO:20)，SASYRNT(SEQ ID NO:21)和QQHYSPPYT(SEQ ID NO:22)(表13)。
表12：人源化Y1238可变重链序列：
Figure SMS_60
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Figure SMS_61
表13：人源化Y1238可变轻链序列：
Figure SMS_62
在IgG1.3f同种型背景下形成VH序列。作为具有共同CL序列的完整轻链形成Vκ序列。将这些人源化HC和LC构建体的所有55种组合，以嵌合Y1238分子在3mL上清液中表达，以通过SPR进行结合分析。在蛋白A传感器芯片上捕获56种上清液以及纯化Y1238嵌合IgG1抗体的每一个，并且在筛选测定中使用BIAcoreTM SPR测定其与两种浓度(即，200nM和1000nM)hCD40-单体的结合。使用100倍稀释的表达上清液，除了含有hz-HC3的那些以外，所有抗体均以较高水平(>300RU)捕获，表明在除了hz-HC3样品以外的上清液中具有较高的抗体滴度。将两种浓度hCD40-单体的动力学数据拟合为1:1Langmuir模型，以得出这些相互作用的动力学和亲和性值的估算值，并对不同分子进行比较。这些数据表明捕获的抗体以较高亲和性与CD40结合；估算的KD值在2.65nM至18.2nM之间，见表14。针对给定的轻链，具有重链hzHC1、hz-HC2、hz-HC4、hz-HC5、hz-HC6、hz-HC7、hz-HC8和hz-HC9的抗体与具有hz-HC10或hz-HC11的抗体相比具有更高的CD40亲和性。而且，针对给定的重链，具有轻链hz-LC1或hz-LC2的抗体与具有hz-LC3、hz-LC4或hz-LC5的抗体相比具有更高的CD40亲和性。对代表性上清液和纯化嵌合Y1238进行的CD40结合的全动力学滴定系列还表明，当与如表14中所示的对分析物的全动力学滴定系列比较时，基于2点(200nM和1000nM)筛选数据估算的KD值是对上清液样品结合动力学和亲和性的合理估算值。
表14：人-CD40单体与在蛋白A传感器芯片表面上捕获的人源化Y1238抗体结合的SPR动力学和亲和性数据。显示了2点筛选测定的数据，其中对人-CD40单体200nM和1000nM样品进行检测，以及“全动力学”测定的数据，其中对人-CD40单体的全浓度系列进行检测。
Figure SMS_63
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Figure SMS_64
基于上清液筛选SPR数据以及倾向于选择具有更少框架回复突变的构建体，选择了用于表达、纯化和进一步表征的人源化Y1238抗体子集。使用这些抗体中的两种(即，Y1238-hz1和Y1238-hz2)制备P238K和N297A同种型(即，重链为SEQ ID NOS:39、40、42和43；轻链见表15)并通过SPR检测CD40和FcgR结合。CD40结合的SPR数据表明每种抗体均以较高亲和性结合CD40靶点，表16。将该数据与另一种抗-CD40抗体抗体BI-mAb-B(即，在美国专利号9,090,696中公开的SEQ ID NOS:95和96，且在本申请中称为“抗体B”)进行比较，其显示出比人源化Y1238分子低得多的结合亲和性，表15。Fc结构域用下划线表示。
表15
Figure SMS_65
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Figure SMS_66
对表15中Hz1和Hz2的两条轻链序列(SEQ ID NOS:41和44)的比对如下所示，在两条序列之间显示出一个突变差异(在位置66，用下划线表示)：
Figure SMS_67
Figure SMS_68
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表16：人-CD40单体与在蛋白A传感器芯片表面上捕获的人源化Y1238抗体或BI-mAb-B结合的SPR动力学和亲和性数据。
样品 ka(1/Ms) kd(1/s) KD(nM)
Y1238-hz1-P238K(n＝3) 7.8±0.8E+04 3.9±0.7E-04 5.0±0.7
Y1238-hz1-N297A(n＝2) 8.0±1.4E+04 3.9±0.7E-04 5.0±1.8
Y1238-hz2-P238K(n＝2) 7.0±0.9E+04 4.3±0.7E-04 6.2±0.2
Y1238-hz2-N297A 6.6E+04 4.0E-04 6.0
BI-mAb-B(n＝2) 6.1±0.3E+04 2.1±1.6E-03 35±1
所有Y1238抗体均显示出对由可溶性CD40L三聚体或来自表达CD40L的CHO细胞的细胞CD40L所驱动的B细胞增殖的高效拮抗作用(见表17)。而相反的是，竞争抗体BI-mAb-B和5F11-45-P238K对由可溶性CD40L信号驱动的B细胞增殖显示出高效抑制作用，但是在抑制由细胞CD40L(CHO细胞过表达的CD40L)驱动的B细胞增殖方面几乎没有作用。当使用细胞CD40L刺激B细胞时，Y1238嵌合和人源化抗体的效能仅显示出约10倍变化。
表17：对由可溶性CD40L三聚体或来自CD40L过表达CHO细胞的细胞相关CD40L驱动的扁桃体人B细胞的抑制作用。抑制B细胞增殖的IC50以ng/mL表示。显示了标准偏差(STDEV)和供体数量(n)。*BI mAb-B和5F11-45-P238K不能完全抑制B细胞增殖，平均抑制作用为1μg/ml。
Figure SMS_69
Figure SMS_70
人源化Y1238抗体(即，Y1238-hz1-N297A、Y1238-hz1-P238K、Y1238-hz2-P238K和Y1238-hz1-N297A)的FcgR结合与嵌合Y1238抗体和5F11-45抗体此前的数据一致，其对所有低亲和性FcgR均显示出极弱的结合应答，表18。
表18：与抗-his Fab捕获的hFcgR-his蛋白结合的1μM或10μM人源化Y1238抗体的％Rmax数据。
Figure SMS_71
此外，当单独或加入表达FcgR CD32的CHO细胞进行检测时，Y1238抗体的人源化形式与阴性对照蛋白L6-IgG4相比显示出最小的iDC活化。该观察结果与人源化Y1238抗体不能活化CD40一致，即使是在由过表达CD32的CHO细胞提供了非常强的交联的情况下。而相反的是，单独的2141和BMS-986090对照均显示出供体依赖性适度的iDC活化，当由CD32 CHO细胞交联或成簇时，该活化显著增强，图5和图6。类似地，当在更大范围的iDC供体上进行检测时，单独或使用CD32介导的成簇或交联时，5F11-45-P238K均未显示出活化作用，图6。
实施例的材料和方法
FCGR结合SPR：可以使用纯化的FcgR利用诸如BIAcoreTM表面等离子体共振(SPR)的方法在体外测量FcgR结合。一种方法检测纯化抗体与His-标记的FcgR蛋白(FcgR-His)的结合，所述蛋白被捕获在固化的抗-His抗体的Fab片段上。这些实验在25℃下在BIAcoreTMT100或BIAcoreTM T200仪器(GE Healthcare Life Sciences,Marlborough,MA)上进行。使用标准的乙基(二甲基氨基丙基)碳二亚胺(EDC)/N-羟基琥珀酰亚胺(NHS)化学试剂和乙醇胺阻断剂，将鼠抗-6xHis抗体(内部制备)的Fab片段固化在CM5传感器芯片上，以使其在10mM HEPES pH 7.4、150mM NaCl、3mM EDTA、0.05％表面活性剂p20(HBS-EP+)的运行缓冲液中的密度为～3000RU。使用运行缓冲液10mM NaPO4、130mM NaCl、0.05％p20(PBS-T)pH7.1进行其余所有研究。使用30sec接触时间以10μl/min将含有C-末端6x多聚组氨酸标签的各种FcγR蛋白(内部制备)捕获在该表面上(通常使用的FcgR-his蛋白浓度为～7μg/ml)。例如，使用120秒结合时间以30μl/min以及120秒解离时间以30μl/min对各种浓度纯化抗体的结合情况进行检测。在这些研究中检测的FcgR蛋白包括“高亲和性”FcgR CD64(hFcgRI)，以及“低亲和性”FcgR CD32a-H131(FcgRIIa-H131)、CD32a-R131(FcgRIIa-R131)、CD32b(FcgRIIb)、CD16a-V158(FcgRIIIa-V158)和CD16b-NA2(FcgRIIIb-NA2)。
为了对结合应答进行定量分析并对不同分子的FcgR结合进行比较，可以通过使用下述公式计算最大结合应答(以占理论最大结合应答的百分比(％Rmax)表示)对SPR结合数据进行分析：
公式1：％Rmax＝(分析物的结合应答)/[((分析物Mw)/(配体Mw))x(配体应答)x(分析物：配体的化学计量)]
其中“分析物”是抗体和配体是捕获的FcgR蛋白。
该分析未考虑抗体或FcgR糖基化的质量，并假设捕获配体的部分活性为100％。由于FcgR是糖基化的，因而即使在饱和条件下其％Rmax值通常也小于100％。对于评价“低亲和性”FcgR CD32a-H131、CD32a-R131、CD32b、CD16a-V158和CD16b-NA2的结合，％Rmax分析是特别有用的，其具有相对迅速的结合和解离速率并且亲和性接近或低于所检测的分析物浓度(1uM)，这样在这些条件下通常无法达到表面的饱和。而相反的是，与特异性针对IgG1和IgG4的其他FcgR相比，“高亲和性”FcgR CD64以更高的亲和性和更缓慢的解离动力学结合。因而，这些同种型通常确实会在微摩尔分析物浓度下使CD64表面饱和，并且使用％Rmax很难区分结合亲和性。对于这些相互作用，可以通过比较传感图数据中的解离速率容易地观察到抗体之间的差异。
CD40结合动力学和亲和性：在25℃下通过在BIAcoreTM T100或T200仪器(GEHealthcare)上进行SPR测量抗体分子的单价CD40结合亲和性，其在固化的蛋白A传感器芯片表面上捕获抗体，然后结合人-CD40-单体蛋白(内部制备)，该测定使用的结合时间为180秒，解离时间为360秒，以30μl/min在pH 7.1的PBS-T(含有
Figure SMS_72
20的磷酸盐缓冲液)中进行。
原代细胞的分离和培养：通过Ficoll密度梯度分离法从肝素化的人血液中分离PBMC(外周血单核细胞)。按照Manual EasySep方案(STEMCELL,Vancouver,Canada)从PBMC中分离单核细胞。在6孔板的每个孔中接种一百万个分离的单核细胞，每孔含6ml完全培养基(RPMI-1640，10％热灭活胎牛血清，100单位/ml青霉素-链霉素)，其中含有人IL-4(白介素-4，100ng/ml)和人GM-CSF(粒细胞巨噬细胞集落刺激因子，100ng/ml)，在37℃/5％CO2下孵育6天，隔日更换培养基并更换为含有相同浓度细胞因子的新鲜培养基。在第6天收集iDC，彻底洗涤，并在完全培养基中重悬。
在存在或不存在FcγR成簇/交联的条件下使用抗-CD40抗体处理iDC：在完全培养基中制备各种生物制剂的滴定物，并加入重复的96孔板中。在交联的情况下，将抗体加入细胞30min，随后以1:6的比例加入表达CD32a的CHO细胞。在37℃，5％CO2条件下孵育细胞约18-20小时。从每孔中取出150μL上清液，按照1:5稀释，并根据生产厂商的说明书使用市售ELISA试剂盒(R&D Systems,Minneapolis,MN)确定IL-6、TNFα和IL-12的蛋白浓度。将每个重复处理收集上清液后残留在培养板中的细胞合成1个样品，并转移至新的96孔圆底(RB)培养板上中，并置于4℃下。使用不含Ca++和Mg++的D-PBS(Dulbecco磷酸盐缓冲液)洗涤细胞，并使用
Figure SMS_73
可固定的近红外死细胞染色试剂盒(Invitrogen,Carlsbad,CA)在冰上染色30min，以测定细胞活力。洗涤细胞并将其重悬于不含Ca++和Mg++，含2％胎牛血清(FBS)，0.1％NaN3(染色缓冲液)的D-PBS中，并且使用含人TruStain FcXTM(Fc受体封闭溶液，Biolegend,San Diego,CA)的染色缓冲液进行封闭(5μl/孔)。
使用下述对DC进行免疫染色：PerCpCy5.5-缀合的αCD3、αCD19、αCD14(Lin-)，BUV395-缀合的αCD11c(BD Biosciences,San Diego,CA)，APC-缀合的αCD86(Biolegend,San Diego,CA)，PE-缀合的αCD83(eBioscience,San Diego,CA)，FITC-缀合的αCD54(Biolegend,San Diego,CA)，并在4℃下孵育45分钟。在染色缓冲液中洗涤细胞两次，并通过加入100μl BD Cytofix固定缓冲液(在室温下15min，避光)固定。使用LSRII-Fortessa流式细胞仪(BD Biosciences,San Diego,CA)和FlowJo分析软件(Treestar,Ashland,OR)针对CD86,ICAM-1and CD83的表达情况对DC进行评价。
抑制CD40L诱导人B细胞增殖：人扁桃体B细胞来自儿科患者的常规扁桃体切除术。通过切碎和轻轻捣碎组织，将细胞过筛并使用人
Figure SMS_74
分离培养基(CedarlaneLabs,Burlington,ON)通过密度梯度分离分离单核细胞分离所述细胞。从界面收集单核细胞，洗涤，并用绵羊红细胞(SRBC,Colorado Serum Company；Denver,CO)在4℃下孵育1小时，然后进行密度梯度分离以除去T细胞。再次洗涤细胞并重悬于含10％FBS的RPMI(完全培养基)中。在完全培养基中制备抗体的滴定物，并加入三次重复的96孔圆底(RB)培养板中。加入1X105个扁桃体人B细胞，并使用可溶性IZ-hCD40L(2μg/mL)或使用经10,000Rad辐照的使用人CD40L稳定转染的中国仓鼠卵巢细胞(CHO-hCD40L)进行刺激；然后以2X103个细胞/孔接种细胞，每孔的终体积为200μL。将板在37℃，5％CO2条件下孵育72小时，最后6小时每孔用0.5μCi3[H]-胸腺嘧啶标记，收集并通过液闪仪计数。根据胸腺嘧啶的掺入情况对B细胞增殖进行定量。/>
序列表
<110> 百时美施贵宝公司
<120> 拮抗性CD40单克隆抗体及其用途
<130> 200896-0013-00-WO-577479
<150> 62/511,079
<151> 2017-05-25
<160> 119
<170> PatentIn 3.5版
<210> 1
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-45-VH_CDR1"
<400> 1
Gly Tyr Thr Phe Thr Asp Leu Ser Met His Trp
1 5 10
<210> 2
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-45-VH_CDR2"
<400> 2
Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe Lys
1 5 10 15
Gly
<210> 3
<211> 8
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-45-VH_CDR3"
<400> 3
Leu Ile Leu Gln Arg Gly Ala Tyr
1 5
<210> 4
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-45-VL_CDR1"
<400> 4
Arg Ala Ser Lys Asn Val Asp Ser Tyr Gly Asn Ser Phe Met His Trp
1 5 10 15
<210> 5
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-45-VL_CDR2"
<400> 5
Arg Ala Ser Asn Leu Glu Ser
1 5
<210> 6
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-45-VL_CDR3"
<400> 6
Gln Gln Ser Asn Glu Asp Pro Leu Thr
1 5
<210> 7
<211> 117
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_VH_ 5F11-45"
<400> 7
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser
115
<210> 8
<211> 111
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：LC_VL_ 5F11-45"
<400> 8
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Lys Asn Val Asp Ser Tyr
20 25 30
Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Asp
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110
<210> 9
<211> 232
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1a-P238K"
<400> 9
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 10
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1a-P238K"
<400> 10
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 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> 11
<211> 232
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1f-P238K"
<400> 11
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 12
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1f-P238K"
<400> 12
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
225 230 235 240
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 13
<211> 232
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1a-N297A"
<400> 13
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 14
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1a-N297A"
<400> 14
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 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> 15
<211> 232
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1f-N297A"
<400> 15
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 16
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1f-N297A"
<400> 16
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
225 230 235 240
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 17
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VH_CDR1"
<400> 17
Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu
1 5 10
<210> 18
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VH_CDR2"
<400> 18
Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe Lys
1 5 10 15
Gly
<210> 19
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VH_CDR3"
<400> 19
Ser Gln Leu Gly Arg Arg Phe Asp Tyr
1 5
<210> 20
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_CDR1"
<400> 20
Lys Ala Ser Gln Asp Val Arg Thr Gly Val Ala
1 5 10
<210> 21
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_CDR2"
<400> 21
Ser Ala Ser Tyr Arg Asn Thr
1 5
<210> 22
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_CDR3"
<400> 22
Gln Gln His Tyr Ser Pro Pro Tyr Thr
1 5
<210> 23
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC1 (h-Y1238_VH)"
<400> 23
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 24
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC2"
<400> 24
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Ala Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 25
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC3"
<400> 25
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 26
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC4"
<400> 26
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Ala Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 27
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC5"
<400> 27
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Ala Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 28
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC6"
<400> 28
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 29
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC7"
<400> 29
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 30
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC8"
<400> 30
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 31
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC9"
<400> 31
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Thr Val Thr Val Ser Ser
115
<210> 32
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC10"
<400> 32
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 33
<211> 118
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-HC11"
<400> 33
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 34
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-LC1"
<400> 34
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 Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn 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 Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 35
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-LC2"
<400> 35
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 Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg 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 His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 36
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-LC3"
<400> 36
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Ile Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 37
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-LC4"
<400> 37
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 Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Ala
65 70 75 80
Glu Asp Ile Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 38
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：hz-LC5"
<400> 38
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Ile Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 39
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_Y1238-hz1-P238K"
<400> 39
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg 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 Lys 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
435 440 445
<210> 40
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ Y1238-hz1-N297A"
<400> 40
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg 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 Ala Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 41
<211> 214
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：LC_Y1238-hz1"
<400> 41
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 Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn 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 Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 42
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_Y1238-hz2-P238K"
<400> 42
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg 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 Lys 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
435 440 445
<210> 43
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ Y1238-hz2-N297A"
<400> 43
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg 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 Ala Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 44
<211> 214
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：LC_Y1238-hz2"
<400> 44
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn Thr Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg 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 His Tyr Ser Pro Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 45
<211> 277
<212> PRT
<213> 智人
<400> 45
Met Val Arg Leu Pro Leu Gln Cys Val Leu Trp Gly Cys Leu Leu Thr
1 5 10 15
Ala Val His Pro Glu Pro Pro Thr Ala Cys Arg Glu Lys Gln Tyr Leu
20 25 30
Ile Asn Ser Gln Cys Cys Ser Leu Cys Gln Pro Gly Gln Lys Leu Val
35 40 45
Ser Asp Cys Thr Glu Phe Thr Glu Thr Glu Cys Leu Pro Cys Gly Glu
50 55 60
Ser Glu Phe Leu Asp Thr Trp Asn Arg Glu Thr His Cys His Gln His
65 70 75 80
Lys Tyr Cys Asp Pro Asn Leu Gly Leu Arg Val Gln Gln Lys Gly Thr
85 90 95
Ser Glu Thr Asp Thr Ile Cys Thr Cys Glu Glu Gly Trp His Cys Thr
100 105 110
Ser Glu Ala Cys Glu Ser Cys Val Leu His Arg Ser Cys Ser Pro Gly
115 120 125
Phe Gly Val Lys Gln Ile Ala Thr Gly Val Ser Asp Thr Ile Cys Glu
130 135 140
Pro Cys Pro Val Gly Phe Phe Ser Asn Val Ser Ser Ala Phe Glu Lys
145 150 155 160
Cys His Pro Trp Thr Ser Cys Glu Thr Lys Asp Leu Val Val Gln Gln
165 170 175
Ala Gly Thr Asn Lys Thr Asp Val Val Cys Gly Pro Gln Asp Arg Leu
180 185 190
Arg Ala Leu Val Val Ile Pro Ile Ile Phe Gly Ile Leu Phe Ala Ile
195 200 205
Leu Leu Val Leu Val Phe Ile Lys Lys Val Ala Lys Lys Pro Thr Asn
210 215 220
Lys Ala Pro His Pro Lys Gln Glu Pro Gln Glu Ile Asn Phe Pro Asp
225 230 235 240
Asp Leu Pro Gly Ser Asn Thr Ala Ala Pro Val Gln Glu Thr Leu His
245 250 255
Gly Cys Gln Pro Val Thr Gln Glu Asp Gly Lys Glu Ser Arg Ile Ser
260 265 270
Val Gln Glu Arg Gln
275
<210> 46
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-VH_FR1"
<400> 46
Gln 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
20 25
<210> 47
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VH_FR1"
<400> 47
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser
20 25
<210> 48
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VH_FR1"
<400> 48
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser
20 25
<210> 49
<211> 14
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11- and Y1238-VH_FR2"
<400> 49
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly
1 5 10
<210> 50
<211> 14
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VH_FR2"
<400> 50
Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met Gly
1 5 10
<210> 51
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-VH_FR3"
<400> 51
Lys Thr Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 52
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 52
Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 53
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 53
Arg Val Thr Met Thr Ala Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 54
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 54
Arg Val Thr Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 55
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 55
Arg Val Thr Met Thr Ala Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 56
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 56
Arg Val Thr Leu Thr Ala Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 57
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 57
Arg Val Thr Met Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys Ala Arg
20 25 30
<210> 58
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 58
Arg Val Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys Ala Arg
20 25 30
<210> 59
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VH_FR3"
<400> 59
Arg Val Thr Met Thr Arg Asp Lys Ser Ser Ser Thr Ala Tyr Met Glu
1 5 10 15
Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys Ala Arg
20 25 30
<210> 60
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-和Y1238-VH_FR4"
<400> 60
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
1 5 10
<210> 61
<211> 23
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-VL_FR1"
<400> 61
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys
20
<210> 62
<211> 23
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_FR1"
<400> 62
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys
20
<210> 63
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-VL_FR2"
<400> 63
Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr
1 5 10 15
<210> 64
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_FR2"
<400> 64
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr
1 5 10 15
<210> 65
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_FR2"
<400> 65
Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr
1 5 10 15
<210> 66
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-VL_FR3"
<400> 66
Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys
20 25 30
<210> 67
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VL_FR3"
<400> 67
Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys
20 25 30
<210> 68
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VL_FR3"
<400> 68
Gly Val Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr
1 5 10 15
Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys
20 25 30
<210> 69
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VL_FR3"
<400> 69
Gly Val Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr
1 5 10 15
Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Val Tyr Tyr Cys
20 25 30
<210> 70
<211> 32
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Y1238-VL_FR3"
<400> 70
Gly Val Pro Ser Arg Phe Ser Gly Ser Arg Ser Gly Thr Asp Phe Thr
1 5 10 15
Phe Thr Ile Ser Ser Leu Gln Ala Glu Asp Ile Ala Val Tyr Tyr Cys
20 25 30
<210> 71
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：5F11-VL_FR4"
<400> 71
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
1 5 10
<210> 72
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：Y1238-VL_FR4"
<400> 72
Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
1 5 10
<210> 73
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头GGGGS"
<400> 73
Gly Gly Gly Gly Ser
1 5
<210> 74
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头(GGGGS)2"
<400> 74
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10
<210> 75
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头(GGGGS)3"
<400> 75
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 76
<211> 20
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头(GGGGS)4"
<400> 76
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1 5 10 15
Gly Gly Gly Ser
20
<210> 77
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头(GGGGS)5"
<400> 77
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1 5 10 15
Gly Gly Gly Ser Gly Gly Gly Gly Ser
20 25
<210> 78
<211> 3
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头"
<400> 78
Ala Ser Thr
1
<210> 79
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头"
<400> 79
Thr Val Ala Ala Pro Ser
1 5
<210> 80
<211> 3
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头"
<400> 80
Thr Val Ala
1
<210> 81
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ 5F11-45-IgG1f"
<400> 81
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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> 82
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-IgG1.3f (也称为
HC_5F11-45-hIgG1.3f)"
<400> 82
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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> 83
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-CTza"
<400> 83
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Ser 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu 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 Val 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> 84
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-N297A"
<400> 84
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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> 85
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-P238K"
<400> 85
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Lys 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu 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 Val 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> 86
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-SE"
<400> 86
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 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 Glu His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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> 87
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-V11"
<400> 87
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Asp Asp 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 Asp Glu Asp Gly Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Arg Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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> 88
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：LC_5F11-45"
<400> 88
Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Lys Asn Val Asp Ser Tyr
20 25 30
Gly Asn Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Leu Leu Ile Tyr Arg Ala Ser Asn Leu Glu Ser Gly Val Pro Asp
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ser Asn
85 90 95
Glu Asp Pro Leu Thr Phe Gly Gln 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> 89
<211> 348
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：BMS-986090"
<400> 89
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 Arg Asp Tyr
20 25 30
Glu Met Trp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Arg Val
35 40 45
Ser Ala Ile Asn Pro Gln Gly Thr Arg Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Leu Pro Phe Arg Phe Ser Asp Arg Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Glu Ser Lys Tyr Gly Pro Pro Cys Pro
115 120 125
Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe
130 135 140
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
145 150 155 160
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
165 170 175
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
180 185 190
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
195 200 205
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
210 215 220
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
225 230 235 240
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
245 250 255
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
260 265 270
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
275 280 285
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
290 295 300
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
305 310 315 320
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
325 330 335
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
340 345
<210> 90
<211> 448
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ Y1238-IgG1.3f"
<400> 90
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg 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 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 Arg Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 91
<211> 448
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ Y1238-CTza"
<400> 91
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg 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 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 Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu Gly Gly Ser 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> 92
<211> 448
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ Y1238-N297A"
<400> 92
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg 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 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 Arg 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 Ala Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 93
<211> 448
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_ Y1238-P238K"
<400> 93
Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Thr
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr
20 25 30
Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Asn Pro Gly Ser Gly Gly Thr Asn Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Ser Gln Leu Gly Arg Arg 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 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 Arg 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 Lys 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> 94
<211> 214
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：LC_Y1238"
<400> 94
Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly
1 5 10 15
Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Arg Thr Gly
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Asn Thr Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Arg Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala
65 70 75 80
Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr
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> 95
<211> 449
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_BI-mAb-B"
<400> 95
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Tyr Ile Ser Ser Gly Asn Arg Ile Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gln Asp Gly Tyr Arg Tyr Ala Met Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr 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 His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Lys
<210> 96
<211> 220
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：LC_BI-mAb-B"
<400> 96
Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Glu Lys Val Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30
Gly Asn Gln Lys Asn Tyr Leu Thr Trp His Gln Gln Lys Pro Gly Gln
35 40 45
Pro Pro Lys Leu Leu Ile Tyr Trp Thr Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Asn
85 90 95
Asp Tyr Thr Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile
100 105 110
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
115 120 125
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
130 135 140
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
145 150 155 160
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
165 170 175
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
180 185 190
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
195 200 205
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215 220
<210> 97
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：接头"
<400> 97
Ala Ser Thr Ser Gly Pro Ser
1 5
<210> 98
<211> 231
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1a-P238K- (-C-末端Lys)"
<400> 98
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly
225 230
<210> 99
<211> 329
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：CH1- IgG1a-P238K (-C-末端Lys)"
<400> 99
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 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
325
<210> 100
<211> 231
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1f-P238K (-C-末端Lys)"
<400> 100
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly
225 230
<210> 101
<211> 329
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1f-P238K (-C-末端Lys)"
<400> 101
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Lys Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
225 230 235 240
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly
325
<210> 102
<211> 231
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1a-N297A (-C-末端Lys)"
<400> 102
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly
225 230
<210> 103
<211> 329
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1a-N297A (-C-末端Lys)"
<400> 103
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 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
325
<210> 104
<211> 231
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域IgG1f-N297A (-C-末端Lys)"
<400> 104
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly
225 230
<210> 105
<211> 329
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：5F11-CH1- IgG1f-N297A (-C-末端Lys)"
<400> 105
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
Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu
225 230 235 240
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
245 250 255
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
275 280 285
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
290 295 300
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly
325
<210> 106
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-P238K (-C-末端LYS)"
<400> 106
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Lys 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu 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 Val 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
435 440 445
<210> 107
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-P238K (-C-末端LYS)"
<400> 107
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Lys 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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
435 440 445
<210> 108
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-P238K"
<400> 108
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Lys 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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> 109
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-N297A (-C-末端LYS)"
<400> 109
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala 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 Val 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
435 440 445
<210> 110
<211> 447
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-N297A"
<400> 110
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu 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 Val 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> 111
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：HC_5F11-45-N297A (-C-末端LYS)"
<400> 111
Gln 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 Tyr Thr Phe Thr Asp Leu
20 25 30
Ser Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Tyr Ile Thr Pro Ser Ser Gly Tyr Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Thr Thr Leu 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 Leu Ile Leu Gln Arg Gly Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 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 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu 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 Val 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
435 440 445
<210> 112
<211> 232
<212> PRT
<213> 人工序列
<220>
<223> "合成多肽：Fc结构域共有序列"
<220>
<221> MISC_FEATURE
<222> (1)..(232)
<223> "序列中给出的变体残基相对于注释中的变体位置没有偏好"
<220>
<221> 变体
<222> (23)..(23)
<223> Pro或Lys
<220>
<221> 变体
<222> (82)..(82)
<223> Asn或Ala
<220>
<221> 变体
<222> (141)..(141)
<223> Asp或Glu
<220>
<221> 变体
<222> (143)..(143)
<223> Leu或Met
<220>
<221> 变体
<222> (232)..(232)
<223> Lys或不存在
<400> 112
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Xaa Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Xaa Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Xaa Glu Xaa Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Xaa
225 230
<210> 113
<211> 351
<212> DNA
<213> 人工序列
<220>
<223> "合成多核苷酸：5F11-45 VH"
<400> 113
caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60
tcctgcaagg cttctggata caccttcact gacttatcga tgcactgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatac attactccta gcagtggata tactgcgtac 180
aatcagaagt tcaagggcaa gaccacgttg accgcggaca aatccacgag cacagcctac 240
atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagattgatc 300
ttacaacggg gagcttactg gggccaggga accctggtca ccgtctcctc a 351
<210> 114
<211> 333
<212> DNA
<213> 人工序列
<220>
<223> "合成多核苷酸：5F11-45 VL"
<400> 114
gacatcgtgc tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 60
atcaactgca gagccagtaa aaatgttgat agttatggca atagttttat gcactggtac 120
cagcagaaac caggacagcc tcctaagctg ctcatttacc gtgcatccaa cctagaatct 180
ggggtccctg accgattcag tggcagcggg tctgggacag atttcactct caccatcagc 240
agcctgcagg ctgaagatgt ggcagtttat tactgtcagc aaagtaatga ggatcctctc 300
acgtttggcc aggggaccaa gctggagatc aaa 333
<210> 115
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> "合成多核苷酸：5F11-45-HC (无C-末端赖氨酸) 具有
P238K"
<400> 115
caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60
tcctgcaagg cttctggata caccttcact gacttatcga tgcactgggt gcgacaggcc 120
cctggacaag ggcttgagtg gatgggatac attactccta gcagtggata tactgcgtac 180
aatcagaagt tcaagggcaa gaccacgttg accgcggaca aatccacgag cacagcctac 240
atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagattgatc 300
ttacaacggg gagcttactg gggccaggga accctggtca ccgtctcctc agctagcacc 360
aagggcccat cggtcttccc cctggcaccc tcctccaaga gcacctctgg gggcacagcg 420
gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480
ggcgccctga ccagcggcgt gcacaccttc ccggccgtcc tacagtcctc aggactctac 540
tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc 600
aacgtgaatc acaagcccag caacaccaag gtggacaaga gagttgagcc caaatcttgt 660
gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg aaagtcagtc 720
ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 780
tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 840
ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 900
cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 960
tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 1020
gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag 1080
aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 1140
tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 1200
gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg 1260
aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 1320
ctctccctgt ctccgggt 1338
<210> 116
<211> 654
<212> DNA
<213> 人工序列
<220>
<223> "合成多核苷酸：5F11-45-LC"
<400> 116
gacatcgtgc tgacccagtc tccagactcc ctggctgtgt ctctgggcga gagggccacc 60
atcaactgca gagccagtaa aaatgttgat agttatggca atagttttat gcactggtac 120
cagcagaaac caggacagcc tcctaagctg ctcatttacc gtgcatccaa cctagaatct 180
ggggtccctg accgattcag tggcagcggg tctgggacag atttcactct caccatcagc 240
agcctgcagg ctgaagatgt ggcagtttat tactgtcagc aaagtaatga ggatcctctc 300
acgtttggcc aggggaccaa gctggagatc aaacgtacgg tggctgcacc atctgtcttc 360
atcttcccgc catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg 420
aataacttct atcccagaga ggccaaagta cagtggaagg tggataacgc cctccaatcg 480
ggtaactccc aggagagtgt cacagagcag gacagcaagg acagcaccta cagcctcagc 540
agcaccctga cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc 600
acccatcagg gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgt 654
<210> 117
<211> 1389
<212> DNA
<213> 人工序列
<220>
<223> "合成多核苷酸：5F11-45-HC具有信号肽（无C-末端赖氨酸）具有P238K"
<400> 117
atgagggctt ggatcttctt tctgctctgc ctggccggga gagcgctcgc acaggtgcag 60
ctggtgcagt ctggggctga ggtgaagaag cctgggtcct cggtgaaggt ctcctgcaag 120
gcttctggat acaccttcac tgacttatcg atgcactggg tgcgacaggc ccctggacaa 180
gggcttgagt ggatgggata cattactcct agcagtggat atactgcgta caatcagaag 240
ttcaagggca agaccacgtt gaccgcggac aaatccacga gcacagccta catggagctg 300
agcagcctga gatctgagga cacggccgtg tattactgtg cgagattgat cttacaacgg 360
ggagcttact ggggccaggg aaccctggtc accgtctcct cagctagcac caagggccca 420
tcggtcttcc ccctggcacc ctcctccaag agcacctctg ggggcacagc ggccctgggc 480
tgcctggtca aggactactt ccccgaaccg gtgacggtgt cgtggaactc aggcgccctg 540
accagcggcg tgcacacctt cccggccgtc ctacagtcct caggactcta ctccctcagc 600
agcgtggtga ccgtgccctc cagcagcttg ggcacccaga cctacatctg caacgtgaat 660
cacaagccca gcaacaccaa ggtggacaag agagttgagc ccaaatcttg tgacaaaact 720
cacacatgcc caccgtgccc agcacctgaa ctcctggggg gaaagtcagt cttcctcttc 780
cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg 840
gtggacgtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga cggcgtggag 900
gtgcataatg ccaagacaaa gccgcgggag gagcagtaca acagcacgta ccgtgtggtc 960
agcgtcctca ccgtcctgca ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc 1020
tccaacaaag ccctcccagc ccccatcgag aaaaccatct ccaaagccaa agggcagccc 1080
cgagaaccac aggtgtacac cctgccccca tcccgggatg agctgaccaa gaaccaggtc 1140
agcctgacct gcctggtcaa aggcttctat cccagcgaca tcgccgtgga gtgggagagc 1200
aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1260
ttcttcctct acagcaagct caccgtggac aagagcaggt ggcagcaggg gaacgtcttc 1320
tcatgctccg tgatgcatga ggctctgcac aaccactaca cgcagaagag cctctccctg 1380
tctccgggt 1389
<210> 118
<211> 705
<212> DNA
<213> 人工序列
<220>
<223> "合成多核苷酸：5F11-45-LC具有信号肽（无C-末端赖氨酸）"
<400> 118
atgagggctt ggatcttctt tctgctctgc ctggccgggc gcgccttggc cgacatcgtg 60
ctgacccagt ctccagactc cctggctgtg tctctgggcg agagggccac catcaactgc 120
agagccagta aaaatgttga tagttatggc aatagtttta tgcactggta ccagcagaaa 180
ccaggacagc ctcctaagct gctcatttac cgtgcatcca acctagaatc tggggtccct 240
gaccgattca gtggcagcgg gtctgggaca gatttcactc tcaccatcag cagcctgcag 300
gctgaagatg tggcagttta ttactgtcag caaagtaatg aggatcctct cacgtttggc 360
caggggacca agctggagat caaacgtacg gtggctgcac catctgtctt catcttcccg 420
ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 480
tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 540
caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 600
acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 660
ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 705
<210> 119
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> "合成肽：信号肽"
<400> 119
Met Arg Ala Trp Ile Phe Phe Leu Leu Cys Leu Ala Gly Arg Ala Leu
1 5 10 15
Ala

Claims (20)

1.一种分离的抗体或其抗原结合部分，其特异性结合人CD40，其中所述抗体包含重链和轻链，其中：
所述重链包含人重链恒定区和重链可变区，所述重链可变区包含GYTFTDLSMHW（SEQ IDNO: 1）的CDR1，YITPSSGYTAYNQKFKG（SEQ ID NO: 2）的CDR2和LILQRGAY（SEQ ID NO: 3）的CDR3；和
所述轻链包含人轻链恒定区和轻链可变区，所述轻链可变区包含RASKNVDSYGNSFMHW（SEQ ID NO: 4）的CDR1，RASNLES（SEQ ID NO: 5）的CDR2和QQSNEDPLT（SEQ ID NO: 6）的CDR3，和
所述人重链恒定区是在Kabat位置238上发生脯氨酸到赖氨酸突变的人IgG1 Fc结构域，所述突变降低与Fc-γ-受体（FcgR）的结合，并且其中所述抗体或抗原结合部分具有降低的FcgR结合。
2.根据权利要求1所述的分离的抗体或其抗原结合部分，其中所述抗体或其抗原结合部分拮抗CD40的活性。
3.根据权利要求1所述的分离的抗体或其抗原结合部分，其中所述重链可变区包含SEQID NO: 7所示的氨基酸序列，并且所述轻链可变区包含SEQ ID NO: 8所示的氨基酸序列。
4.根据权利要求1至3中任一项所述的分离的抗体或其抗原结合部分，其中所述Fc结构域包含选自下述的氨基酸序列：SEQ ID NO: 98，SEQ ID NO: 9，SEQ ID NO: 99，SEQ IDNO: 10，SEQ ID NO: 100，SEQ ID NO: 11，SEQ ID NO: 101或SEQ ID No: 12。
5.根据权利要求3所述的分离的抗体或其抗原结合部分，其中所述人IgG1 Fc结构域包含SEQ ID NO: 98或SEQ ID NO: 100所示的氨基酸序列。
6.根据权利要求1所述的分离的抗体或其抗原结合部分，其中所述重链包含选自下组的氨基酸序列或由选自下组的氨基酸序列组成：SEQ ID NO: 85、SEQ ID NO: 106、SEQ IDNO: 107和SEQ ID NO: 108；和所述轻链包含SEQ ID NO: 88所示的氨基酸序列或由SEQ IDNO: 88所示的氨基酸序列组成。
7.根据权利要求1至3、5和6中任一项所述的分离的抗体或其抗原结合部分，其中所述分离的抗体或其抗原结合部分是人源化的。
8.根据权利要求1所述的抗体或其抗原结合部分，其中所述抗原结合部分是scFv-Fc。
9.根据权利要求1至3、5、6和8中任一项所述的分离的抗体或其抗原结合部分，其中所述抗体或其抗原结合部分具有与至少一个选自以下的Fcg受体降低的结合：hCD32a-H131(FcgRIIa-H131)、hCD32a-R131 (FcgRIIa-R131)和/或hCD16b-NA2 (FcgRIIIb-NA2)。
10.一种编码根据权利要求1至9中任一项所述的分离的抗体或其抗原结合部分的核酸。
11.包含权利要求10所述的核酸的表达载体。
12.使用权利要求11所述的表达载体转化的细胞。
13.制备抗-人CD40抗体或其抗原结合部分的方法，包括：
a）在权利要求12所述的细胞中表达所述抗体或其抗原结合部分；和
b）从所述细胞中分离所述抗体或其抗原结合部分。
14.药物组合物，包含：a）权利要求1至9中任一项所述的抗体或其抗原结合部分；和b）药学上可接受的载体。
15.权利要求1至9中任一项所述的抗体或其抗原结合部分在制备用于在对象中治疗选自以下疾病的药物中的用途：艾迪生氏病（Addison’s disease）、强直性脊柱炎、哮喘、动脉粥样硬化、特应性变态反应、耳部自身免疫性疾病、眼部自身免疫性疾病、自身免疫性肝炎、自身免疫性腮腺炎、冠心病、糖尿病、附睾炎、肾小球肾炎、格雷夫斯病(Graves’ disease)、格林-巴利综合征、桥本氏病、溶血性贫血、特发性血小板减少性紫癜、炎性肠病、对重组药品的免疫应答、系统性红斑狼疮、多发性硬化、重症肌无力、天疱疮、银屑病、风湿热、类风湿性关节炎、结节病、硬皮病、干燥综合征，脊柱关节病、甲状腺炎、移植排斥和血管炎。
16.根据权利要求15所述的用途，其中所述疾病选自以下：炎性肠病、系统性红斑狼疮、多发性硬化、类风湿性关节炎、干燥综合征和移植排斥。
17.根据权利要求15所述的用途，其中所述疾病是变态反应（allergy）。
18.根据权利要求15所述的用途，其中所述疾病是过敏反应（anaphylaxis）。
19.根据权利要求15所述的用途，其中所述炎性肠病是克罗恩病或溃疡性结肠炎。
20.根据权利要求15所述的用途，其中所述哮喘是支气管哮喘。
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