CN115836084A - 多特异性免疫调节剂 - Google Patents

多特异性免疫调节剂 Download PDF

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CN115836084A
CN115836084A CN202180049642.9A CN202180049642A CN115836084A CN 115836084 A CN115836084 A CN 115836084A CN 202180049642 A CN202180049642 A CN 202180049642A CN 115836084 A CN115836084 A CN 115836084A
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M·斯尔曼
O·黑尔
G·吉菲尔斯
K·比丽安-弗雷
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Abstract

本发明涉及免疫学和免疫肿瘤学领域。更具体地，本发明涉及多特异性和双特异性细胞因子和抗体衍生物，其能够靶向细胞和/或组织以局部增强免疫应答并降低全身毒性。

Description

多特异性免疫调节剂
技术领域
本发明涉及多特异性和双特异性TNF超家族融合蛋白组装体，其包含至少(i)一种包含单链TNF超家族受体结合结构域的蛋白质部分和(ii)能够特异性结合细胞表面抗原或免疫调节蛋白的蛋白质部分。在优选实施方案中，第一(i)和第二(ii)蛋白质部分通过杵入臼结构(knob-into-hole)Fc融合蛋白技术(US 5,731,168；US 7,695,936)连接。本发明进一步涉及核酸和转染的宿主细胞用于生产多特异性TNF超家族融合蛋白组装体。
背景技术
免疫系统的各种功能通过刺激和抑制信号的复杂且微妙的平衡相互作用来协调。免疫细胞功能的许多关键调节因子属于所谓的肿瘤坏死因子超家族(TNFSF)及其同源受体(所谓的TNF受体超家族)。TNFSF由19种结构相关的配体组成，每一种与TNF受体超家族的29个成员中的一个或多个结合。
TNFSF受体在抗肿瘤免疫应答和炎症过程的调节中非常重要。它们由多种免疫细胞表达，包括T细胞和抗原呈递细胞群体，例如树突状细胞和巨噬细胞，以及由肿瘤细胞其自身表达。这种不同的表达模式突出了TNFSF受体在身体的许多部分中以及在抗肿瘤免疫应答的不同阶段中发挥的关键作用。
WO 2010/010051公开了作为单个单链融合蛋白的三价蛋白质部分。所述融合蛋白包含由基于短(3-8个)氨基酸的接头连接的三个可溶的、柄缺失(stalk depleted)的TNF超家族(TNFSF)受体结合结构域。它们基本上是非聚集性的并且非常适合于治疗应用。在WO2015/164588、WO 2016/177771、WO 2017/068183、WO 2017/068180、WO 2017/068185、WO2017/072080和WO 2017/068192(所有前述专利申请的内容通过引用以其整体并入本文)中公开了具有降低的免疫原性和改变的稳定性的其它三价单链TNFSF受体结合结构域。
发明内容
尽管已经公开了三价TNFSF蛋白质部分，但仍需要其特异性靶向构建体。此类靶向构建体将允许局部增强或局部富集TNF受体超家族(TNFRSF)激动活性。因此，本发明的一个目的是提供多功能和双功能融合蛋白，其包含用于组织或细胞靶向或活性调节的至少两种不同的三价TNFSF蛋白质部分或三价TNFSF蛋白质部分和特异性抗原结合部分。
本发明进一步涉及编码如本文所述的融合蛋白的核酸分子，以及用如本文所述的核酸分子转化或转染的细胞或非人生物体。
本发明还涉及药物或诊断组合物，其包含如本文所述的多特异性融合蛋白、核酸分子或细胞作为活性剂。
本发明还涉及如本文所述的多特异性融合蛋白、核酸分子或细胞用于治疗的用途，例如，如本文所述的融合蛋白、核酸分子或细胞用于制备药物组合物的用途，所述药物组合物用于预防和/或治疗由TNFSF细胞因子功能障碍引起的、与之相关的和/或伴随的病症，特别是增殖性病症，诸如肿瘤，例如实体瘤或淋巴肿瘤；传染性疾病；炎性疾病；代谢性疾病；自身免疫性病症，例如类风湿性和/或关节炎疾病；退行性疾病，例如神经退行性疾病如多发性硬化；细胞凋亡相关疾病或移植排斥。
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图1双特异性Fab-Fc/scTNFSF-RBD-Fc异聚融合蛋白(所谓的单臂-双特异性或SAB蛋白)的示意图。三价scTNFSF-RBD-Fc和Fab-Fc的异二聚化是基于相应Fc-部分的野生型或特异性CH3-结构域变体。
图2双特异性、六价scTNFSF配体的示意图。两个三价scTNFSF-RBD-Fc融合蛋白的异二聚化是基于CH3结构域。这可以通过相应Fc-部分的野生型或特异性CH3-结构域变体实现。
图3双特异性、三价靶向构建体的示意图；构建体是基于Fab重链部分与三价scTNFSF-RBD的直接或接头介导的融合体。
图4GITR激动剂的细胞体外活性以GITR荧光素酶报告基因检测示出。aPDL1-scGITRL-SAB示出显著的GITR激动，其与scGITRL-Fc相比较低。
图5GITR激动剂的细胞体外活性以GITR荧光素酶报告基因检测示出。aPDL1-scGITRL-SAB示出显著的GITR激动，其与scGITRL-Fc相比较低，与aPDL1-scGITRL(三价)相比较高。
图6GITR激动剂的细胞体外活性以GITR荧光素酶报告基因检测示出。aPDL1-scGITRL-SAB示出显著的GITR激动，其与aPDL1-scGITRL(三价)相比较高。通过与抗人Fc(x-连接)交联，两种化合物的活性明显增强。
图7PD-L1抑制剂的细胞体外活性以PDL1荧光素酶报告基因检测示出。aPDL1-scGITRL-SAB示出显著的PDL1抑制，其与aPDL1-mAb相比在相似的范围内。
图8PD-L1抑制剂的细胞体外活性以PDL1荧光素酶报告基因检测示出。aPDL1-scCD27L-SAB示出显著的PDL1抑制，其与aPDL1-mAb相比在相似的范围内。
图9PDL1抑制剂的细胞体外活性以PDL1荧光素酶报告基因检测示出。aPDL1-scCD40L-SAB示出显著的PDL1抑制，其与aPDL1-mAb相比在相似的范围内。
图10PDL1抑制剂的细胞体外活性以PDL1荧光素酶报告基因检测示出。aPDL1-scCD40L和aPDL1-scCD40L-Fc示出显著的PDL1抑制，其与aPDL1-mAb相比在相似的范围内。
图11PDL1抑制剂的细胞体外活性以PDL1荧光素酶报告基因检测示出。aPDL1-scCD40L-Fc示出显著的PDL1抑制，其与aPDL1-mAb相比在相似的范围内。
图12PD-L1抑制剂的细胞体外活性以PDL1荧光素酶报告基因检测示出。aPDL1-scCD40L-SAB和aPDL1-scCD40L示出显著的PDL1抑制，其与aPDL1-mAb相比在相似的范围内。
图13CD95L抑制剂的体外活性通过评估CD95L-诱导的细胞凋亡的拮抗以JurkatA3细胞检测示出。细胞凋亡抑制与半胱天冬酶3/7活性的降低并行。aCD95L-scCD40L、aCD95L-mAb和aCD95L-scCD40L-Fc示出在相似的范围内的显著的细胞凋亡抑制。aCD95L-mAb和aCD95L-scCD40L-Fc比aCD95L-scCD40L活性高一点。
图14CD95L抑制剂的体外活性通过评估CD95L-诱导的细胞凋亡的拮抗以JurkatA3细胞检测示出。细胞凋亡抑制与半胱天冬酶3/7活性的降低并行。aCD95L-scCD40L和aCD95L-mAb示出在相似范围内的显著的细胞凋亡抑制。aCD95L-mAb比aCD95L-scCD40L活性高一点。
图15CD27激动剂的细胞体外活性以CD27荧光素酶报告基因检测示出。aPDL1-scCD27L-SAB示出轻微的CD27激动，其通过与抗人Fc(x-连接)交联明显增强。scCD27L-Fc活性较高并且可以通过与抗人Fc(x-连接)交联进一步增强。
图16CD40激动剂的细胞体外活性以CD40荧光素酶报告基因检测示出。三聚体CD40L和三聚体CD40L(stab)[stab＝稳定化]在该检测中示出基础活性。aPDL1-scCD40L-SAB和aPDL1-scCD40L示出明显较高的CD40激动，这表明PDL1-靶向对CD40激动活性的贡献。该发现由aCD95L-scCD40L(该化合物靶向CD95L而不是PD-L1)的较低活性证实。
图17CD40激动剂的细胞体外活性以CD40荧光素酶报告基因检测示出。三聚体CD40L在该检测中示出基础活性。相比之下，aCD95L-scCD40L示出较高的CD40激动。
图18CD40激动剂的细胞体外活性以CD40荧光素酶报告基因检测示出。scCD40L-Fc、aPDL1-scCD40L-SAB和aPDL1-scCD40L-Fc示出明显的CD40激动，其可通过与抗人Fc(x-连接)交联明显增强。六价CD40L-形式scCD40L-Fc和aPDL1-scCD40L-Fc显示比三价CD40L-形式aPDL1-scCD40L-SAB高的激动活性。
图19CD40激动剂的细胞体外活性以CD40荧光素酶报告基因检测示出。scCD40L-Fc、aPDL1-scCD40L和三聚体CD40L(stab)示出明显的CD40激动，其可通过与StrepMAB Immo(x-连接)交联明显增强。六价CD40L-形式scCD40L-Fc示出比三价CD40L-形式aPDL1-scCD40L高的激动活性，两种形式的活性都明显高于三聚体CD40L(stab)[stab＝稳定化]。
图20CD40激动剂的细胞体外活性以CD40荧光素酶报告基因检测示出。将恒定浓度(如所示)的CD40激动剂与浓度增加直至100μg/ml的aPDL1-mAb一起孵育。如所预期的，scCD40L-Fc的CD40激动不受aPDL1-mAb竞争的影响，这是由于该分子不包含PDL1-靶向结构域。相比之下，aPDL1-scCD40L-SAB和aPDL1-scCD40L示出随着竞争性aPDL1-mAb的浓度的增加而明显降低的CD40激动。总之，aPDL1-scCD40L-SAB和aPDL1-scCD40L的PD-L1-靶向结构域明显有助于它们的CD40激动活性。
图21CD40激动剂的细胞体外活性以CD40荧光素酶报告基因检测示出。将恒定浓度(如所示)的CD40激动剂与浓度增加直至100μg/ml的aPDL1-mAb一起孵育。如所预期的，scCD40L-Fc的CD40激动不受aPDL1-mAb竞争的影响，这是由于该分子不包含a PDL1-靶向结构域。相比之下，aPDL1-scCD40L-SAB示出随着竞争性aPDL1-mAb的浓度的增加而明显降低的CD40激动。总之，aPDL1-scCD40L-SAB的PDL1-靶向结构域明显有助于其CD40激动活性。
图22ELISA阐释了CEA靶向构建体与CEA的结合。aCEA-Fab、aCEA-scCD40L和aCEA-scCD40L-Fc与人CEA以剂量依赖性方式结合。aCEA-Fab和aCEA-scCD40L-Fc的结合与aCEA-scCD40L的结合相当并且明显更强。
图23ELISA阐释了CEA靶向构建体与CEA的结合。aCEA-Fab和aCEA-scCD40L与人CEA以剂量依赖性方式结合。aCEA-Fab的结合比aCEA-scCD40L的结合明显更强。
图24描绘了非常适合于本发明的第一、第二和第三方面的多特异性和双特异性TNF超家族融合蛋白组装体的示例性scTNF RBD序列。
图25双特异性、三价靶向构建体的示意图；构建体是基于一个(A)或两个(B)单-结构域抗体部分(VHH)与三价scTNFSF-RBD的直接或接头介导的融合体。
图26双特异性VHH-Fc/scTNFSF-RBD-Fc异聚融合蛋白(所谓的单臂双特异性(SAB)蛋白，基于一个(A)或两个(B)单-结构域抗体部分(VHH))的示意图。三价scTNFSF-RBD-Fc和VHH-Fc的异二聚化是基于相应Fc-部分的野生型或特异性CH3-结构域变体。
图27双特异性scFv-Fc/scTNFSF-RBD-Fc异聚融合蛋白(所谓的单臂双特异性(SAB)蛋白，基于scFv抗体片段)的示意图。三价scTNFSF-RBD-Fc和scFv-Fc的异二聚化是基于相应Fc-部分的野生型或特异性CH3-结构域变体。
图28双特异性、六价靶向Fab-scTNFSF-RBD-Fc融合蛋白的示意图。构建体是基于Fab结构域与三价scTNFSF-RBD-Fc的直接融合，以及随后的Fab-scTNFSF-RBD-Fc经由Fc结构域如aPDL1-scCD40L-Fc的同二聚化。
图29CD137激动剂的细胞体外活性以CD137荧光素酶报告基因检测示出。scCD137L-Fc(A)和乌瑞芦单抗(Urelumab)(B)与HT1080细胞的细胞交联示出中度的活性增加，而非靶向对照aCD95L-scCD137L-SAB(C)未示出相关的活性增加。D aPDL1-scCD137L-SAB示出轻微的CD137激动，其通过与HT1080细胞(99％的HT1080细胞确实表达PD-L1)的交联而大幅增强。峰值活性比六价scCD137L-Fc所观察到的活性高约16倍(参见A)。
图30CD27激动剂的细胞体外活性以CD27荧光素酶报告基因检测示出。非靶向对照aCD95L-scCD27L-SAB与MDA-MB231细胞的细胞交联未示出活性增加。B aPDL1-scCD27L-SAB示出轻微的CD27激动，其通过与MDA-MB231细胞(99％的MDA-MB231细胞确实表达PD-L1)的交联而大幅增强。峰值活性比六价scCD27L-Fc所观察到的活性高约3倍(参见A)。
图31天然人T细胞活化检测。染色强度随着每个细胞分裂而降低，即，未分裂的细胞对于tag it violet染色是最阳性的。天然Pan T细胞分离自人PBMC。第0天用1μg/ml包被的抗-CD3(OKT3)刺激和添加培养基或100ng/ml aPDL1-scCD137L-SAB或100ng/ml aCD95L-scCD137L-SAB。第5天流式细胞术。双特异性分子aPDL1-scCD137L-SAB和aCD95L-scCD137L-SAB，在抗-CD3刺激(24.6和25.2％增殖vs.16.0％增殖(对于单独的CD3刺激))的存在下，导致相似的T细胞增殖。
图32采用试剂盒(Stem Cell)从来自健康人供体的血沉棕黄层(buffy coats)分离单核细胞。通过添加50ng/ml GM-CSF 3天、接着50ng/ml GM-CSF+50ng/ml IL-4另外3天，实现单核细胞的分化。然后用100ng/ml的指定化合物处理细胞24h，接着流式细胞术评估CD86和CD83表达。
三价scCD40L与抗PD-L1抗体片段结合的双特异性分子aPDL1-scCD40L-SAB是最有效的树突细胞激活剂，激活水平(CD86+/CD83+)为88.31％。因此，将两个部分结合在一个分子中远比将这些部分作为两个单独的分子即aPD-L1抗体+CD40L(三聚体)(示出仅39.48％的活化水平)添加更有效。aCD40和CD40L(三聚体)均示出中等水平的活性(40.59％和46.10％)，而aPD-L1具有与培养基对照相同的活性。
图33采用试剂盒(Stem Cell)从来自健康人供体的血沉棕黄层分离单核细胞。通过添加50ng/ml GM-CSF 3天、接着50ng/ml GM-CSF+50ng/ml IL-4另外3天，实现单核细胞的分化。然后用100ng/ml的指定化合物处理细胞24h，接着流式细胞术评估CD86和CD83表达。
三价scCD40L与抗PD-L1抗体片段结合的双特异性分子是非常有效的树突细胞激活剂，激活水平(CD86+/CD83+)与scCD40L-Fc相似：对于aPDL1-scCD40L-SAB为73.00％，对于aPDL1-scCD40L(三价)为87.18％和对于aPDL1-scCD40L-Fc为96.12％(图28中示出的形式)。相比之下，由于单核细胞上缺乏CD95L表达，靶向CD95L而不是PD-L1的双特异性分子aCD95L-scCD40L-SAB仅具有中等水平的活化。低活化水平还见于aCD40和CD40L(三聚体)。
图34GITR激动剂的细胞体外活性以GITR荧光素酶报告基因检测示出。aPDL1-scGITRL示出显著的GITR激动，其与scGITRL-Fc相比较低。三价GITR激动剂aPDL1-scGITRL的活性通过与抗人Fc(x-连接)的交联而明显增强至六价GITR激动剂scGITRL-Fc所观察到的水平。
具体实施方式
定义
二聚体形成：本文所用，二聚化是指折叠的肽链能够与折叠的第二条多肽链形成稳定结构，并且在多肽链中实现的某些二聚化结构域执行该过程。二聚体形成发生于两条多肽的每一条中存在的这些特定结构域之间。二聚化结构域的实例是本领域众所周知的。在天然人IgA-、IgD-和IgG抗体中，CH3结构域是重链的二聚化的驱动力。在天然IgE或IgM抗体中，CH4结构域在结构和功能上等同于IgG-CH3结构域，执行其重链二聚化。CH3结构域或它们的等同物仅对其自身具有选择性。这意味着，由于CH3/CH3二聚体形成，天然地或通过工程化方法包含功能性CH3结构域的任何多肽能够与包含功能性CH3结构域的第二多肽形成二聚体。
两条含CH3结构域的多肽异二聚化为功能性双特异性融合蛋白是通过在合适的宿主细胞中共表达两条多肽来实现的，确保在蛋白质折叠过程中同时存在两条链。宿主细胞中的蛋白质合成过程中，将形成本多肽链的任何CH3结构域组合：异二聚体以及同二聚体。所需的异二聚体蛋白产物需要通过合适的层析法程序在以后进行纯化。含CH3的多肽的共表达方法和异二聚体产物的后续纯化概念是本领域众所周知的。所用的CH3结构域可为野生型、或者它们可以包含稳定化某些组装体的点突变，例如如Carter等人所述(Merchant,A.,Zhu,Z.,Yuan,J.等人.An efficient route to human bispecific IgG.NatBiotechnol 16,677–681(1998).https://doi.org/10.1038/nbt0798-677)。为了产生本发明的多特异性免疫调节剂，高度优选使用CH3结构域衍生的二聚化技术。在优选实施方案中，在两条融合蛋白多肽中实现的CH3结构域是天然存在的序列。在优选实施方案中，CH3结构域包含点突变，这旨在稳定当前的二聚化产物。高度优选的是，稳定化突变导致两条多肽的共价连接，例如通过当前组装体的CH3-结构域之间的胱氨酸，从而抑制CH3-结构域解离。因此，减少了生产过程中纯化的异二聚体产物的链间交换反应和随后的多聚体和/或同二聚体形成。在优选实施方案中，CH3结构域包含在蛋白质表达过程中优先导致异二聚体形成的点突变，例如杵入臼结构(KiH)技术。除了KiH技术以外，还开发了其它更近期的产生基于CH3的异二聚化结构域的技术，采用静电转向(electrostatic steering)或免疫球蛋白结构域界面交换或两者的组合。本领域中存在的基础技术描述于Skegro等人.J BiolChem.2017Jun9；292(23):9745-9759)，Gunasekaran等人.J Biol Chem.2010Jun18；285(25):19637-46，Sampei等人.PLoS One.2013；8(2):e57479，Von Kreudenstein等人.MAbs.2013Sep-Oct；5(5):646-54，Davis等人.Protein Eng Des Sel.2010Apr；23(4):195-202。
抗体：术语“全长抗体”、“完整抗体”、“全抗体”和“天然抗体”在本文中可交换地使用，是指具有与天然抗体结构基本相似的结构的抗体。“天然抗体”是指具有不同结构的天然存在的免疫球蛋白分子。例如，天然IgG类抗体是约150,000道尔顿的异源四聚糖蛋白，由二硫键连接的两条轻链和两条重链构成。从N末端至C末端，每条重链具有可变区(VH)，也称为可变重结构域或重链可变结构域，接着是三个恒定结构域(CH1、CH2和CH3)，也称为重链恒定区。如本文所用，本发明的上下文中所用的典型IgG衍生的恒定重链结构域是SEQ-ID:28、SEQ-ID:29、SEQ-ID:30、SEQ-ID:31、SEQ-ID:111、SEQ-ID:112，它们都被定义为根据EU编号以Ala118开始。如本文所用，本发明的上下文中所用的典型IgG衍生的CH1结构域是SEQ-ID:27，CLκ是SEQ-ID:26。CH1和CH2结构域通过铰链区连接，所述铰链区通过半胱氨酸桥来稳定抗体。类似地，从N末端至C末端，每条轻链具有可变区(VL)，也称为可变轻结构域或轻链可变结构域，然后是轻链恒定结构域(CL)，也称为轻链恒定区。抗体的重链可属于五种类型之一，称为α(IgA)、δ(IgD)、ε(IgE)、γ(IgG)、或μ(IgM)，其中一些可进一步分为亚型，例如γ1(IgG1)、γ2(IgG2)、γ3(IgG3)、γ4(IgG4)、α1(IgA1)和α2(IgA2)。抗体的轻链可属于两种类型之一，称为卡帕(κ)和拉姆达(λ)。此外，可以工程化包含λVL和κCL的杂合轻链形式，反之亦然。在优选的实施方案中，轻链是基于κLC或由VLλ/CLκ构成的杂合LC，以改善溶解度和更快的折叠动力学。如本文所用，本发明的上下文中所用的典型CLκ结构域是SEQ-ID:26。
抗体片段：“抗体片段”是指除了完整抗体以外的分子，其包含与完整抗体所结合的抗原结合的一部分完整抗体。抗体片段的实例包括但不限于Fv、Fab、Fab′、Fab′-SH、F(ab′)2；双抗体、三抗体(triabodies)、四抗体(tetrabodies)、交叉Fab片段；线性抗体；单链抗体分子(例如scFv)；和单结构域抗体(例如VHH)。对于某些抗体片段的综述，参见Hudson等人,Nat Med 9,129-134(2003)。对于scFv片段的综述，参见，例如Plückthun,inThe Pharmacology of Monoclonal Antibodies,vol.113,Rosenburg and Moore eds.,Springer-Verlag,New York,pp.269-315(1994)；也参见WO 93/16185；和美国专利号5,571,894和5,587,458。双抗体是具有两个抗原结合位点的抗体片段，可为二价或双特异性的，参见，例如，EP 404,097；WO 1993/01161；Hudson等人,Nat Med 9,129-134(2003)；和Hollinger等人,Proc Natl Acad Sci USA 90,6444-6448(1993)。三抗体和四抗体也描述于Hudson等人,Nat Med 9,129-134(2003)。对于基于双特异性抗体片段的构建体的综述，参见，Brinkmann U,Kontermann RE.MAbs.2017Feb/Mar；9(2):182-212。单结构域抗体是包含抗体的全部或一部分的重链可变结构域或全部或一部分的轻链可变结构域的抗体片段。第一单结构域抗体源自来自骆驼科动物的抗体重链的可变结构域(纳米抗体或VHH片段)。此外，术语单结构域抗体包括源自鲨鱼的自主人重链可变结构域(aVH)或VNAR片段。在某些实施方案中，单结构域抗体是人单结构域抗体(Domantis,Inc.,Waltham,Mass.；参见，例如美国专利号6,248,516B1)。制备抗体片段的方法为本领域技术人员所熟悉的。广泛使用的方法包括蛋白水解消化或在宿主细胞中重组生产。抗体和抗体片段的制备方法的非限制性综述示于US20160200833A1。
Fab片段和scFv片段：术语“Fab片段”指包含由轻链的VL结构域和恒定结构域(CL)构成的轻链片段、和重链的VH结构域和第一恒定结构域(CH1)的抗体片段。CH1和CL结构域可以包含野生型序列或用于改善连接的点突变(CH1：L128F，EU编号)。
“单链可变片段(scFv)”是抗体重链(VH)和轻链(CL)的可变区的融合蛋白，由10至约25个氨基酸的短接头肽连接。所述接头通常富含甘氨酸以获得柔性，富含丝氨酸或苏氨酸以获得溶解性，并且可使VH的N末端与VL的C末端连接，反之亦然。尽管去除了恒定区并引入了接头，这种蛋白质仍保留了原始抗体的特异性。scFv抗体例如描述于Houston,J.S.,Methods in Enzymol.203(1991)46-96)。
TNF-SF：术语“TNF配体家族成员”或“TNF家族配体”或“TNF超家族”(TNF-SF)指促炎细胞因子。通常，细胞因子，特别是TNF配体超家族的成员，在免疫系统的刺激和协调中起到至关重要的作用。目前，基于序列、功能和结构相似性，已将19种细胞因子鉴定为TNF(肿瘤坏死因子)配体超家族的成员。所有这些配体是II型跨膜蛋白，具有C末端胞外结构域(胞外域)、N末端胞内结构域和单个跨膜结构域。TNF-SF胞外域包括柄区和位于C末端的序列，称为TNF同源结构域(THD)，其在超家族成员之间具有20-30％的氨基酸同一性。TNF胞外域的C末端部分也负责TNF配体以形成被其特异性受体识别的三聚体复合物。这些三聚体复合物是有结合能力的结构(binding competent structure)，因为受体结合发生在所谓的TNF-SF受体结合结构域(RBD)的原聚体(protomer)界面处。换言之：三个单独的TNF-SF多肽的C末端区形成功能单元，并且三聚体形成是人TNF-SF成员的适当受体募集的结构前提。
Fc结构域：本文中的术语“Fc结构域”或“Fc区”用于定义包含至少一部分的恒定区的抗体重链的C末端区。该术语包括天然序列Fc区和变体Fc区。IgG Fc区包含IgG CH2和IgGCH3结构域。然而，如本文经常使用的，Fc从氨基酸残基P230延伸至氨基酸K447(CH2：230-340，CH3：341-447)。人IgG Fc区的“CH2结构域”通常从约位置231处的氨基酸残基延伸至约位置340处的氨基酸残基。在一个实施方案中，糖链附接至CH2结构域。本文中的CH2结构域可以是天然序列CH2结构域或变体CH2结构域。CH2结构域的位置N297位在天然序列中被糖基化，并且是Fc受体结合所需的。在一个实施方案中，N297处的突变消除了Fc受体结合。“CH3结构域”包含Fc区中C末端至CH2结构域的残基的延伸(即从IgG的约位置341处的氨基酸残基至约位置447处的氨基酸残基)。本文中的CH3区可以是天然序列CH3结构域或变体CH3结构域(例如在其一条链中具有引入的“突起”(“杵”)并在其另一条链中具有相应地引入的“空腔”(“臼”)的CH3结构域；参见美国专利号5,821,333，通过引用明确地并入本文)。此类变体CH3结构域可用于促进如本文所述的两条不同的抗体重链的异二聚化。在一个实施方案中，人IgG重链Fc区从重链的Cys226或从Pro230延伸至羧基末端。然而，可能存在或也可能不存在Fc区的C末端赖氨酸(Lys447)。除非本文另有说明，否则Fc区或恒定区中的氨基酸残基的编号是根据EU编号系统，也称为EU索引，如Edelman,G.M.等人,Proc.Natl.Acad.USA,63,78-85(1969)中所述。
“杵臼结构”技术描述于例如美国专利号5,731,168；美国专利号7,695,936。通常，所述方法涉及在第一条多肽的界面处引入一个突起(“杵”)并在第二条多肽的界面中引入相应的空腔(“臼”)，使得突起可以置于空腔中，从而促进异二聚体形成并阻碍同二聚体形成。通过用较大的侧链(例如酪氨酸或色氨酸)替代来自第一条多肽的界面的小氨基酸侧链来构建突起。通过用较小的氨基酸侧链(例如丙氨酸或苏氨酸)替代大的氨基酸侧链而在第二条多肽的界面中产生与突起相同或类似大小的补偿性空腔。通过改变编码多肽的核酸，例如通过定点诱变，或通过肽合成，可以形成突起和空腔。在一个具体实施方案中，杵修饰包括Fc结构域的两个亚基之一中的氨基酸取代T366W，臼修饰包括Fc结构域的两个亚基中的另一者中的氨基酸取代T366S、L368A和Y407V。在其它具体实施方案中，包含杵修饰的Fc结构域的亚基额外地包含氨基酸取代S354C，包含臼修饰的Fc结构域的亚基额外地包含氨基酸取代Y349C。这两个半胱氨酸残基的引入导致Fc区的两个亚基之间形成二硫键，从而进一步稳定化二聚体(Carter,J Immunol Methods 248,7-15(2001))。编号是根据EU编号。如本文所用，本发明的上下文中所用的典型IgG衍生的Fc结构域为SEQ-ID:20、SEQ-ID:21、SEQ-ID:22、SEQ-ID:23、SEQ-ID:24、SEQ-ID:25、SEQ-ID:20、SEQ-ID:109和SEQ-ID:110，它们都被定义为根据EU编号以Pro230开始。
“相当于免疫球蛋白的Fc区的区域”旨在包括免疫球蛋白的Fc区的天然存在的等位基因变体(例如D356E/L358M)以及具有产生取代、添加、或缺失但不会显著降低免疫球蛋白介导效应子功能的能力(例如抗体依赖性细胞毒性)的变化的变体。例如，可以从免疫球蛋白的Fc区的N末端或C末端删除一个或多个氨基酸，而基本上不丧失生物学功能。可以根据本领域已知的一般规则选择此类变体，以便对活性具有最小的影响(参见，例如，Bowie,J.U.等人,Science 247:1306-10(1990))。
如本文所用，对于上述提及的三价非聚集TNF-SF受体结合结构域，同义地使用术语单链TNF-SF受体结合结构域、单链TNFSF受体结合结构域以及TNF-SF RBD和TNFSF RBD。此外，当提及所述受体结合结构域时，词语‘单链’通常缩写为‘sc’，例如scTNFSF-RBD。
如本文所用，具有抗PD-L1特异性的抗PD-L1抗体或抗体片段通常被称为“aPDL1”或“aPD-L1”抗体或相应的抗体片段。对于其它抗体特异性也是如此；例如，对于抗CD95L，也使用aCD95L，并且对于抗CEA，也使用aCEA。
在本说明书中，本发明的第一方面的蛋白组装体被称为“单臂双特异性”或SABs。
此外，特别是在命名本发明的分子或蛋白组装体时，术语抗体通常缩写为“AB”或“Ab”。
此外，术语“异聚融合蛋白”和“异聚蛋白组装体”或“蛋白组装体”可交换地使用。
实施方案
根据本发明，多特异性TNF超家族融合蛋白组装体至少包含(i)一种包含单链TNF超家族受体结合结构域的蛋白质部分和(ii)能够特异性结合细胞表面抗原或活性调节效应子的蛋白质部分。
在本发明的第一方面中，双特异性TNF超家族融合蛋白组装体至少包含
(a)单链TNF-SF受体结合结构域，其融合至
(b)第一肽接头，其融合至
(c)第一(异)二聚化结构域，和
(d)抗原结合或相互作用蛋白质部分，其融合至
(e)第二肽接头，其融合至
(f)第二(异)二聚化结构域。
图1给出了本发明的第一方面的多特异性TNF超家族融合蛋白组装体的总体概述。
如图1所示，本发明的典型多特异性免疫调节剂是一种蛋白质单元，其一侧包含IgG抗体衍生的重链和轻链组装体，另一侧包含三价单链TNFSF-RBD-Fc融合多肽。蛋白质单元的两半的异二聚化由CH3结构域执行，并由铰链链间半胱氨酸额外稳定化。三条多肽链的共表达和正确组装是形成这种功能性双特异性蛋白质单元所必需的。这种融合蛋白形式称为Ab-scTNFSF-SAB(SAB＝单臂双特异性)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗PD-L1(aPDL1)靶向与CD40激动相结合。这种特异性组装体称为aPDL1-scCD40L-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:33(scCD40L-Fc-杵_b)或SEQ-ID:84(scCD40L-Fc-杵_c)与SEQ-ID:55(aPD-L1-LC)和SEQ-ID:54(aPD-L1-HC-RF-臼)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗PD-L1(aPDL1)靶向与CD27激动相结合。这种特异性组装体称为aPDL1-scCD27L-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:39(scCD27L-Fc-杵_b)、SEQ-ID:55(aPD-L1-LC)和SEQ-ID:54(aPD-L1-HC-RF-臼)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:119(scCD27L-V2-RBD)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗PD-L1靶向与GITR激动相结合。这种特异性组装体称为aPDL1-scGITRL-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:41(scGITRL-Fc-杵_b)、SEQ-ID:55(aPD-L1-LC)和SEQ-ID:54(aPD-L1-HC-RF-臼)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗PD-L1靶向与CD137激动相结合。这种特异性组装体称为aPDL1-scCD137L-SAB。非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:86(scCD137L-V1-Fc-杵_b)或SEQ-ID:90(scCD137L-V2-Fc-杵_b)或SEQ-ID:94(scCD137L-V3-Fc-杵_b)结合有SEQ-ID:55(aPD-L1-LC)和SEQ-ID:54(aPD-L1-HC-RF-臼)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:107(scCD137L-V4-RBD)或SEQ-ID:108(scCD137L-V5-RBD)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗PD-L1靶向与HVEM/LTbR激动相结合。这种特异性组装体称为aPDL1-scLIGHT-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:98(scLIGHT-Fc-杵_b)、SEQ-ID:55(aPD-L1-LC)和SEQ-ID:54(aPD-L1-HC-RF-臼)。
在一个优选实施方案中，上述PD-L1特异性Ab-scTNFSF-SAB(SAB＝单臂双特异性)多特异性免疫调节剂(aPDL1-scCD40L-SAB、aPDL1-scCD27L-SAB、aPDL1-scGITRL-SAB、aPDL1-scCD137L-SAB，aPDL1-scLIGHT-SAB)包含相同的抗原特异性序列(aPD-L1抗体的VHCH和VLCL)和相同的三价scTNFSF模块，但在其Fc部分包含不同的CH3结构域序列。CH3结构域可以是突变的或可以是野生型的，但仍然能够与其对应物形成二聚体，引导结构组装，如图1所示。IgG衍生的CH3结构域的非限制性实例表示为SEQ-ID:44、SEQ-ID:45、SEQ-ID:48、SEQ-ID:49、SEQ-ID:52、SEQ-ID:53、SEQ-ID:28、SEQ-ID:29、SEQ-ID:30、SEQ-ID:31、SEQ-ID:111、SEQ-ID:112、SEQ-ID:20、SEQ-ID:21、SEQ-ID:22、SEQ-ID:23、SEQ-ID:24、SEQ-ID:25、SEQ-ID:109、SEQ-ID:110的部分。
使用上述aPDL1特异性、单臂双特异性免疫调节剂的实例，对于本领域技术人员显而易见的是，这种设计可以容易地与如图24中所示的scTNFSF RBD、或其变体结合，以进一步构建具有CD40L、GITRL、OX40L、LIGHT、TL1A、CD137L、CD27L或TRAIL作为第二特异性结合靶标的aPDL1-SAB。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CD95L(aCD95L)靶向与CD40激动相结合。这种特异性组装体称为aCD95L-scCD40L-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:33(scCD40L-Fc-杵_b)、SEQ-ID:47(aCD95L-LC)和SEQ-ID:46(aCD95L-HC-RF-臼)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CD95L(aCD95L)靶向与CD27激动相结合。这种特异性组装体称为aCD95L-scCD27L-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:39(scCD27L-Fc-杵_b)、SEQ-ID:47(aCD95L-LC)和SEQ-ID:46(aCD95L-HC-RF-臼)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:119(scCD27L-V2-RBD)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CD95L靶向与GITR激动相结合。这种特异性组装体称为aCD95L-scGITRL-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:41(scGITRL-Fc-杵_b)、SEQ-ID:47(aCD95L-LC)和SEQ-ID:46(aCD95L-HC-RF-臼)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CD95L靶向与CD137激动相结合。这种特异性组装体称为aCD95L-scCD137L-SAB。非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:86(scCD137L-V1-Fc-杵_b)或SEQ-ID:90(scCD137L-V2-Fc-杵_b)或SEQ-ID:94(scCD137L-V3-Fc-杵_b)结合有SEQ-ID:47(aCD95L-LC)和SEQ-ID:46(aCD95L-HC-RF-臼)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:107(scCD137L-V4-RBD)或SEQ-ID:108(scCD137L-V5-RBD)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CD95L靶向与HVEM/LTbR激动相结合。这种特异性组装体称为aCD95L-scLIGHT-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:98(scLIGHT-Fc-杵_b)、SEQ-ID:47(aCD95L-LC)和SEQ-ID:46(aCD95L-HC-RF-臼)。
在一个优选实施方案中，上述CD95L特异性Ab-scTNFSF-SAB(SAB＝单臂双特异性)多特异性免疫调节剂(aCD95L-scCD40L-SAB、aCD95L-scCD27L-SAB、aCD95L-scGITRL-SAB、aCD95L-scCD137L-SAB、aCD95L-scLIGHT-SAB)包含相同的抗原特异性序列(aCD95L抗体的VHCH和VLCL)和相同的三价scTNFSF模块，但在其Fc部分包含不同的CH3结构域序列。CH3结构域可以是突变的或可以是野生型的，但仍然能够与其对应物形成二聚体，引导结构组装，如图1所示。IgG衍生的CH3结构域的非限制性实例表示为SEQ-ID:44、SEQ-ID:45、SEQ-ID:48、SEQ-ID:49、SEQ-ID:52、SEQ-ID:53、SEQ-ID:28、SEQ-ID:29、SEQ-ID:30、SEQ-ID:31、SEQ-ID:111、SEQ-ID:112、SEQ-ID:20、SEQ-ID:21、SEQ-ID:22、SEQ-ID:23、SEQ-ID:24、SEQ-ID:25、SEQ-ID:109、SEQ-ID:110的部分。
使用上述aCD95L特异性、单臂双特异性免疫调节剂的实例，对于本领域技术人员显而易见的是，这种设计可以容易地与如图24所示的scTNFSF RBD、或其变体结合，以进一步构建具有CD40L、GITRL、OX40L、LIGHT、TL1A、CD137L、CD27L或TRAIL作为第二特异性结合靶标的aCD95L-SAB。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CEA(aCEA)靶向与CD40激动相结合。这种特异性组装体称为aCEA-scCD40L-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:33(scCD40L-Fc-杵_b)、SEQ-ID:51(aCEA-LC)和SEQ-ID:50(aCEA-HC-RF-臼)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CEA(aCEA)靶向与CD27激动相结合。这种特异性组装体称为aCEA-scCD27L-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:39(scCD27L-Fc-杵_b)、SEQ-ID:51(aCEA-LC)和SEQ-ID:50(aCEA-HC-RF-臼)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:119(scCD27L-V2-RBD)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CEA靶向与GITR激动相结合。这种特异性组装体称为aCEA-scGITRL-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:41(scGITRL-Fc-杵_b)、SEQ-ID:51(aCEA-LC)和SEQ-ID:50(aCEA-HC-RF-臼)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CEA靶向与CD137激动相结合。这种特异性组装体称为aCEA-scCD137L-SAB。非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:86(scCD137L-V1-Fc-杵_b)或SEQ-ID:90(scCD137L-V2-Fc-杵_b)或SEQ-ID:94(scCD137L-V3-Fc-杵_b)结合有SEQ-ID:51(aCEA-LC)和SEQ-ID:50(aCEA-HC-RF-臼)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:107(scCD137L-V4-RBD)或SEQ-ID:108(scCD137L-V5-RBD)。
在一个优选实施方案中，Ab-scTNFSF-SAB多特异性免疫调节剂将抗CEA靶向与HVEM/LTbR激动相结合。这种特异性组装体称为aCEA-scLIGHT-SAB。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:98(scLIGHT-Fc-杵_b)、SEQ-ID:51(aCEA-LC)和SEQ-ID:50(aCEA-HC-RF-臼)。
在一个优选实施方案中，上述CEA特异性Ab-scTNFSF-SAB(SAB＝单臂双特异性)多特异性免疫调节剂(aCEA-scCD40L-SAB、aCEA-scCD27L-SAB、aCEA-scGITRL-SAB、aCEA-scCD137L-SAB、aCEA-scLIGHT-SAB)包含相同的抗原特异性序列(aCEA抗体的VHCH和VLCL)和相同的三价scTNFSF模块，但在其Fc部分包含不同的CH3结构域序列。CH3结构域可以是突变的或可以是野生型的，但仍然能够与其对应物形成二聚体，引导结构组装，如图1所示。IgG衍生的CH3结构域的非限制性实例表示为SEQ-ID:44、SEQ-ID:45、SEQ-ID:48、SEQ-ID:49、SEQ-ID:52、SEQ-ID:53、SEQ-ID:28、SEQ-ID:29、SEQ-ID:30、SEQ-ID:31、SEQ-ID:111、SEQ-ID:112、SEQ-ID:20、SEQ-ID:21、SEQ-ID:22、SEQ-ID:23、SEQ-ID:24、SEQ-ID:25、SEQ-ID:109、SEQ-ID:110的部分。
使用上述CEA特异性、单臂双特异性免疫调节剂的实例，对于本领域技术人员显而易见的是，这种设计可以容易地与如图24中所示的scTNFSF RBD、或其变体结合，以进一步构建具有CD40L、GITRL、OX40L、LIGHT、TL1A、CD137L CD27L或TRAIL作为第二特异性结合靶标的aCEA-SAB。
scCD27L RBD的一种特异性变体包含Seq-ID:36(scCD27L-Fc-杵_a)、Seq-ID:37(scCD27L-Fc-杵_b)、Seq-ID:38(scCD27L-Fc-臼_a)、Seq-ID:39(scCD27L-Fc-臼_b)、Seq-ID:70(scCD27L-RBD)的N-末端谷氨酰胺到谷氨酸的交换。
在进一步优选的实施方案中，本领域技术人员可以通过将来自aPD-L1的VH和VL结构域交换为其他抗体特异性来修饰本发明第一方面中描述的Ab-scTNFSF-SAB的实例，包括但不仅限于
-抗CD137(SEQ-ID:59aCD137-VH，SEQ-ID:60aCD137-VL)
-抗间皮素(SEQ-ID:61aMeso-VH，SEQ-ID:62aMeso-VL)
-抗CD25(SEQ-ID:63aCD25-VH，SEQ-ID:64aCD25-VL)
-抗PD-1(SEQ-ID:65aPD1-a-VH，SEQ-ID:66aPD1-a-VL，SEQ-ID:67aPD1-b-VH，SEQ-ID:68aPD1-b-VL)
-抗CEA(SEQ-ID:113aCEA-a-VH，SEQ-ID:114aCEA-a-VL，SEQ-ID:115aCEA-b-VH，SEQ-ID:116aCEA-b-VL)
-抗CD95L(SEQ-ID:117aCD95L-VH，SEQ-ID:118aCD95L-VL)，
从而产生aCD137-scTNFSF-SAB、aMeso-scTNFSF-SAB、aCD25-scTNFSF-SAB、aPD1-scTNFSF-SAB、aCEA-scTNFSF-SAB或aCD95L-scTNFSF-SAB。
在本发明的第二方面中，多特异性TNF超家族融合蛋白组装体至少包含
(a)单链TNF-SF受体结合结构域，其融合至
(b)第一肽接头，其融合至
(c)第一(异)二聚化结构域，和
(d)第二单链TNF-SF受体结合结构域，其融合至
(e)第二肽接头，其融合至
(f)第二(异)二聚化结构域。
图2给出了本发明的第二方面的多特异性TNF超家族融合蛋白组装体的总体概述。
如图2所示，本发明的典型多特异性免疫调节剂可通过将本发明的两种scTNFSF-Fc融合多肽相结合来实现。在一个优选实施方案中，多特异性免疫调节剂包含作为成熟蛋白质的多肽SEQ-ID:32和SEQ-ID:36。在一个优选实施方案中，多特异性免疫调节剂包含作为成熟蛋白质的多肽SEQ-ID:33和SEQ-ID:37。两种结构对CD40和CD27为双特异性的，对于两种TNFRSF成员各有三个结合位点。
在本发明的第二方面中，多特异性TNF超家族融合蛋白组装体包含部分a)(列)的至少一个单链TNF-SF受体结合结构域和部分e)(行)的一个单链结构域。这允许在蛋白组装体内自由组合所有公开的单链TNF-SF受体结合结构域。作为非限制性列表，部分a)(列)和部分e)(行)的可能组合在随后的表中用‘X’标记。
Figure BDA0004048583020000211
在本发明的第三方面中，多特异性TNF超家族融合蛋白组装体至少包含
(a)抗体的功能性Fab结构域，其融合至
(b)单链TNF-SF受体结合结构域，
其中Fab片段(a)的恒定重链结构域的C-末端(C-terminal end)通过肽接头(Seq-ID:13–Seq-ID:19)融合至单链TNF-SF受体结合。
图3给出了本发明的第三方面的多特异性TNF超家族融合蛋白组装体的总体概述。这种融合蛋白形式称为Ab-scTNFSF。
在一个优选实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗PD-L1(aPDL1)靶向与CD40激动相结合。这种特异性组装体称为aPDL1-scCD40L。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:58(aPDL1-hc-scCD40L-RBD)和SEQ-ID:55(aPD-L1-LC)。
在一个优选实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗PD-L1(aPDL1)靶向与CD27激动相结合。这种特异性组装体称为aPDL1-scCD27L。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:102(aPDL1-hc-scCD27L-RBD)和SEQ-ID:55(aPD-L1-LC)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:119(scCD27L-V2-RBD)。
在一个优选实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗PD-L1靶向与GITR激动相结合。这种特异性组装体称为aPDL1-scGITRL。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:104(aPDL1-hc-scGITRL-RBD)和SEQ-ID:55(aPD-L1-LC)。
在一个优选实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗CD95L(aCD95L)靶向与CD40激动相结合。这种特异性组装体称为aCD95L-scCD40L。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:57(aCD95L-hc-scCD40L-RBD)和SEQ-ID:47(aCD95L-LC)。
在一个优选实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗CD95L(aCD95L)靶向与CD27激动相结合。这种特异性组装体称为aCD95L-scCD27L。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:101(aCD95L-hc-scCD27L-RBD)和SEQ-ID:47(aCD95L-LC)。一个进一步优选的实施方案使用TNFSF模块SEQ-ID:119(scCD27L-V2-RBD)。
在一个优选实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗CD95L靶向与GITR激动相结合。这种特异性组装体称为aCD95L-scGITRL。一个非限制性实例包含作为成熟蛋白质的多肽SEQ-ID:103(aCD95L-hc-scGITRL-RBD)和SEQ-ID:47(aCD95L-LC)。
在进一步优选的实施方案中，Ab-scTNFSF多特异性免疫调节剂将抗CD95L靶向与CD137激动或与HVEM/LTbR激动相结合。特异性组装体称为aCD95L-scCD137L和aCD95L-scLIGHT。本领域技术人员可以容易地将上述aPDL1靶向或aCD95L靶向的Ab-scTNFSF实例中的TNFSF模块交换为SEQ-ID:72(scCD137L-RBD)、SEQ-ID:105(scCD137L-V2-RBD)、SEQ-ID:106(scCD137L-V3-RBD)、SEQ-ID:107(scCD137L-V4-RBD)、SEQ-ID:108(scCD137L-V5-RBD)或SEQ-ID:73(scLIGHT-RBD)。
在进一步优选的实施方案中，本领域技术人员可以通过将来自aPD-L1或aCD95L的VH和VL结构域交换为其他抗体特异性来容易地修饰本发明的第三方面中描述的Ab-scTNFSF的实例，包括但不仅限于
-抗CD137(SEQ-ID:59aCD137-VH，SEQ-ID:60aCD137-VL)
-抗间皮素(SEQ-ID:61aMeso-VH，SEQ-ID:62aMeso-VL)
-抗CD25(SEQ-ID:63aCD25-VH，SEQ-ID:64aCD25-VL)
-抗PD-1(SEQ-ID:65aPD1-a-VH，SEQ-ID:66aPD1-a-VL，SEQ-ID:67aPD1-b-VH，SEQ-ID:68aPD1-b-VL)
-抗CEA(SEQ-ID:113aCEA-a-VH，SEQ-ID:114aCEA-a-VL，SEQ-ID:115aCEA-b-VH，SEQ-ID:116aCEA-b-VL；SEQ-ID:56aCEA-hc-scCD40L-RBD/SEQ-ID:51aCEA-LC)
从而产生aCD137-scTNFSF、aMeso-scTNFSF、aCD25-scTNFSF、aPD1-scTNFSF或aCEA-scTNFSF。
使用上述aCD95L-scTNFSF双特异性免疫调节剂的实例，对于本领域技术人员显而易见的是，这种设计可以容易地与如图24中所示的scTNFSF RBD、或其变体结合，以进一步构建具有CD40L、GITRL、OX40L、LIGHT、TL1A、CD137L、CD27L或TRAIL作为第二特异性结合靶标的aCD95L-scTNFSF双特异性免疫调节剂。
在本发明的进一步的方面中，多特异性TNF超家族融合蛋白组装体至少包含
(a)抗体的功能性单VH(可变重链)结构域，其融合至
(b)单链TNF-SF受体结合结构域，
其中VH结构域的c-末端通过肽接头融合至单链TNF-SF受体结合(图25)。
功能性单VH结构域的实例是所谓的VH衍生的单结构域抗体(VHH)。
从WO2010/010051，技术人员已知构建适用于本发明的任何上述方面的单链TNF-SF受体结合结构域的方法。一般而言，合适的非聚集性TNF-SF受体结合结构域由三个可溶性、柄缺失的受体结合结构域组成，它们通过短的、优选3-8个氨基酸长的接头连接。
在一个具体实施方案中，受体结合结构域可以通过较短的接头连接或甚至在没有额外氨基酸的情况下融合。
如上所述，特别合适的三价的、非聚集性TNF-SF受体结合结构域公开于WO2015/164588、WO2016/177771、WO2017/068183、WO2017/068180、WO2017/068185、WO2017/072080和WO2017/068192。作为一个非限制性实例，有利的单链TNF-SF受体结合结构域可以选自图24的序列。
本发明的第一和/或第三方面的抗原结合或相互作用部分可以是抗体片段，例如单特异性抗体片段或其功能性片段。进一步合适的结合和相互作用部分是本领域已知的。非限制性实例是：单链抗体或其功能性片段、单结构域抗体、功能性scFv片段。这些形式的实例在图1、图26、图27以及图3和图25中所示。
在本发明的第一和/或第三方面的具体实施方案中，功能性抗体片段针对细胞表面标志物或活性调节靶标。作为非限制性实例，抗体或抗体片段针对：酪氨酸激酶受体(EGFR、HER2、HER3、HER4)、VEGFR、异聚整合素a或β受体家族包括VLA-4和LFA-1、E-选择素、L-选择素、P-选择素、肿瘤基质标志物如成纤维细胞活化蛋白(FAP)、endoglyx-1、MCSP或内皮唾液酸蛋白(endosialin)、半乳糖凝集素(galectin)、N-CAM(髓鞘蛋白零)、ICAM1-ICAM5、VCAM-1、PE-CAM、L1-CAM、粘连蛋白(Nectin)(PVRL1、PVRL2、PVRL3)、EpCAM、肿瘤抗原包括NY-ESO-1、MAGE1、MAGE2、CA-125、癌胚抗原(CEA)、CAMPATH-1(CD52)、CD44及其肿瘤特异性变体和其他肿瘤选择性细胞表面标志物，CD2、CD5、CD7、CD19、CD20、CD21、CD22、CD24、CD25、CD30、CD33、CD38、CD40、CD52、CD56、CD71、CD72、CD73、CD105、CD117、CD123、CD133、c-Met、PDGFR、IGF1-R、HMW-MAA、TAG-72、GD2、GD3、GM2、叶酸受体、Lgr5、Ley、Muc-1、Muc-2、PSMA、PSCA和uPAR。更优选地，靶标分子是FAP、EGFR、HER2或HER、黑色素瘤相关的硫酸软骨素蛋白多糖(MCSP)。
抗体或抗体片段也可能针对B7家族的成员，包括B7-1(CD80)、B7-2(CD86)、B7-DC(PDCD1LG2、PD-L2、CD273)、B7-H1(PD-L1、CD274)、B7-H2(ICOSLG、B7RP1、CD275)、B7-H3(CD276)、B7-H4(VTCN1)、B7-H5(VISTA、血小板受体Gi24、SISP1)、B7-H6(NCR3LG1)和B7-H7(HHLA2)。
在一个进一步的实施方案中，抗体或抗体片段也可以针对活性调节靶标，包括但不限于CTLA-4、PD1、CD3、CD4、CD8、CD28、HLA I类和II类、LAG3(CD223)，ICOS(CD278)、CD39、CD73、TIGIT、CD96、PTA1(CD226)、TIM-3、TIM-1、CD47、SIRP-α(SIRP-alpha)、DNAM-1和白细胞介素(抗炎)，包括但不限于IL4、IL6、IL9、IL10、IL11、IL13、IL18、IL21和IL22。
应注意，TNF-SF和TNFR-SF的所有胞外域都是特别适合本发明的第一方面的抗体片段的靶标。优选但非限制性列表包含TNF-SF配体结构域如CD95L、TNF-α(TNF-alpha)、CD40L、CD27L、LIGHT、TL1A和TWEAK以及TNF-受体结构域如CD40、CD27、4-1BB、OX40、GITR、HVEM、BCMA、LTBR和TWEAKR的胞外域。
与TNFR-SF的胞外域结合的抗体的实例是抗CD137 mAb乌瑞芦单抗和乌托鲁单抗(Utomilumab)。与TNFR-SF的胞外域结合的单克隆抗体的进一步实例是伐立鲁单抗(Varlilumab)(抗CD27)、塞鲁单抗(Selicrelumab)(抗CD40)、APX005M(抗CD40)和TRX518(抗GITR)。
从科学和商业的角度来看，TNFSF配体与抗体的组合是特别有吸引力的，所述抗体结合已经评价的癌细胞的表面标志物，如CEA或HER2，或者干预检查点调节剂(PD-1、CTLA4、CD95)的信号级联。因此，实施例和图中所示的具有抗PDL1(aPDL1)和抗CD95L(aCD95L)或抗CEA(aCEA)活性的肽代表了本发明的进一步特别优选的实施方案。
本发明的一个进一步的方面涉及编码如本文所述的多特异性融合蛋白的蛋白质部分的核酸分子。核酸分子可以是DNA分子，例如双链或单链DNA分子，或RNA分子。核酸分子可编码融合蛋白或其前体，例如融合蛋白的原形式(pro-proform)或前形式(pre-proform)，其可包含用于分泌或纯化的信号序列或其他异源氨基酸部分，优选位于融合蛋白的N-末端和/或C-末端。异源氨基酸部分可以通过蛋白酶切割位点例如因子Xa、凝血酶或IgA蛋白酶切割位点连接至第一和/或第二结构域。
核酸分子可以可操作地连接至表达控制序列，例如允许核酸分子在所需宿主细胞中表达的表达控制序列。核酸分子可以位于载体上，例如质粒、噬菌体、病毒载体、染色体整合载体等。合适的表达控制序列和载体的实例例如由Sambrook等(1989)MolecularCloning,A Laboratory Manual,Cold Spring Harbor出版社和Ausubel等(1989),CurrentProtocols in Molecular Biology,John Wiley&Sons或其更新版本描述。
各种表达载体/宿主细胞系统可用于表达编码本发明的融合蛋白的核酸序列。合适的宿主细胞包括但不限于原核细胞如细菌、例如大肠杆菌(E.coli)，真核宿主细胞如酵母细胞、昆虫细胞、植物细胞或动物细胞，优选哺乳动物细胞，更优选人细胞。
进一步，本发明涉及用如上所述的核酸分子转化或转染的非人生物体。这种转基因生物体可以通过已知的遗传转移方法产生，包括同源重组。
本发明的一个进一步的方面涉及药物或诊断组合物，其包含作为活性剂的至少一种融合蛋白、编码融合蛋白的相应核酸、或转化或转染的细胞，全部如本文所述。
本发明的融合蛋白、编码所述融合蛋白的相应核酸、用于产生所述融合蛋白的转化或转染的细胞可用于治疗，例如，用于预防和/或治疗由TNF-SF细胞因子功能障碍引起的、与之相关的和/或伴随的病症，特别是增殖性病症，诸如肿瘤，例如实体瘤或淋巴肿瘤；传染性疾病；炎性疾病；代谢性疾病；自身免疫性病症，例如类风湿和/或关节炎疾病；退行性疾病，例如神经退行性疾病如多发性硬化症；细胞凋亡相关疾病或移植排斥。
实施例
实施例1：重组多特异性/双特异性TNF超家族融合蛋白组装体的大规模表达和纯化方法
对于前述本发明的多特异性免疫调节剂的大规模表达，将编码必要多肽(例如，scTNFSF-Fc，抗体-HC，抗体-LC，VH-CH1-scTNFSF)的合成DNA盒插入至真核表达载体中，所述真核表达载体包含适当的选择标志物(例如，包含杀稻瘟素、嘌呤霉素、潮霉素、或博莱霉素抗性基因的功能性表达盒)和适于提高宿主细胞基因组内转录活性插入位点数量的遗传元件，例如，人β-珠蛋白核基质附着区(humanβ-globin matrix attachment region,MAR)。通过电穿孔将经序列验证的表达载体引入至适应悬浮的中国仓鼠卵巢细胞(CHO-S,Invitrogen)中。在转染后三天向转染的细胞施加适当的选择压力。通过随后在选择压力下培养来回收携带源自载体的抗性基因的幸存细胞。在所选细胞汇集物(cell pool)在轨道式振荡培养箱(100rpm，50mm摇距(shaking throw))中在37℃和7％ CO2气氛下在化学成分确定的培养基(PowerCHO-2CD,Lonza,补充有4mM谷氨酰胺(glutamine/glutamax))中稳定生长后，通过检测上述蛋白质的ELISA检测来分析各个上清液。在摇瓶中扩增具有最高单位生产率(specific productivity)的细胞汇集物(轨道式摇床，100rpm，摇距50mm)用于蛋白质生产。
对于实验室规模的生产，将各个细胞汇聚物在轨道式振荡(100rpm，55mm摇距)的摇瓶中或在Wave生物反应器20/50EHT(GE Healthcare/Cytiva)中、在37℃和7％ CO2气氛下在化学成分确定的培养基(PowerCHO-2CD,Lonza,补充有4mM glutamax)中培养7-12天。Wave培养以0.3x10e6个细胞/ml的活细胞浓度和以下设置(对于五或十升)开始：摇动频率18rpm，摇动角度7°，气流0.2-0.3L/分钟，7％CO2，36.5℃。Wave运行期间，用PowerFeed A(Lonza)和Lipids给细胞培养物进料两次，通常在第3天(20％进料)和第6天(30％进料)。在第二次进料后，将摇动频率增加至22rpm并将摇动角度增加至8°。在第7天至第10天之间当细胞活力降至低于80％时收获wave生物反应器。使用深度过滤系统(Millipore MillistakPod MC0HC 0.054m2)澄清含有双特异性TNFSF激动剂的培养物上清液，然后使用0.22μm瓶顶过滤器(PES,Corning)对澄清的收获物进行灭菌过滤并保存在2-8℃直至进一步加工。
对于本发明的第一和第二方面的多特异性免疫调节剂的亲和纯化，在
Figure BDA0004048583020000271
层析系统(GE Healthcare/Cytiva)上进行纯化过程，利用了通过所用的两种Fc支架的每一者中的特定突变而引入的前述双特异性TNFSF Fc融合蛋白的不同性质。首先，使用作为固相亲和配体的MabSelect SuReTM ProteinA(GE Healthcare/Cytiva)，其以高结合能力与双特异性TNFSF激动剂Fc融合蛋白的Fc结构域结合。简言之，将无菌过滤的澄清的细胞培养物上清液/收获物加载在HiTrap MabSelect SuRe柱(CV＝5ml)上，所述柱在洗涤缓冲液1(20mMPi,95mM NaCl,pH 7.2)中平衡，不超过每ml柱体积10mg融合蛋白的载量。用10个柱体积(10CV)的洗涤缓冲液1洗涤柱，接着用4个柱体积(4CV)的洗涤缓冲液2(20mM Pi,95mMNaCl,pH 8.0)洗涤柱，以耗竭宿主细胞蛋白和宿主细胞DNA。也去除了缺少蛋白A结合位点的同二聚体污染物，因为它保留在柱流穿液(flowthrough)中并且不与柱结合。在一系列的洗涤步骤后，用两倍柱体积洗脱缓冲液(20mM Pi,95mM NaCl,pH 3.5)从柱洗脱蛋白质。收集洗脱液级分，并立即用1M Tris-HCl pH 8.0中和至中性pH。在上述亲和层析方法过程中，将线速度设定为150cm/h并保持恒定。
在纯化本发明的第二方面的多特异性免疫调节剂的情况下，存在于洗脱液中的异二聚体融合蛋白通过SEC和离子交换层析法的组合进行精制。
用于本发明第一方面的多特异性免疫调节剂的纯化的第二亲和步骤使用KappaSelectTM树脂(GE Healthcare/Cytiva)，其与双特异性TNFSF激动剂的Fab结构域的CL-κ结构域结合并耗竭同二聚体激动剂Fc融合蛋白。可选地，第二亲和步骤使用CaptureSelectTM IgG-CH1树脂(Thermo Scientific)，其以高亲和力与Fab结构域的CH1结构域结合。这也导致耗竭同二聚体激动剂Fc融合蛋白。将第一基于MabSelect SuReTM ProteinA的亲和层析法的洗脱液加载在Capture Select IgG-CH1(Thermo Scientific)上或加载在用洗涤缓冲液(PBS pH 7.4＝10mM Pi,2.7mM KCl,140mM NaCl)进行平衡的KappaSelect树脂(GE Healthcare/Cytiva)(CV＝5ml)上，不超过每ml柱体积10mg Fab。在用洗涤缓冲液(6CV)进行的洗涤步骤后，用2CV洗脱缓冲液(0.1M甘氨酸,pH 3.5)洗脱前述双特异性TNFSF激动剂，并立即用1M Tris-HCl pH 8.0中和至中性pH(0.4CV)。通过OD 280测量来定量洗脱液级分的蛋白质量并通过超滤来浓缩用于随后的尺寸排阻层析法(SEC)。
对于本发明的第三方面的多特异性免疫调节剂的亲和纯化，仅采用前述基于CH1的亲和纯化，并且通过随后的尺寸排阻层析法对蛋白质进行精制。
使用
Figure BDA0004048583020000291
层析系统在HiLoad 26/600Superdex 200pg或Superdex 200Increase10/300GL柱(GE Healthcare/Cytiva)上进行尺寸排阻层析法(SEC)。用磷酸盐缓冲盐水或中性pH(pH 7.4)的等效的基于Tris的缓冲系统平衡柱。
将浓缩的、亲和纯化的蛋白加载在SEC柱上，其中样品体积不超过柱体积的2％(v/v)。应用每分钟2.5ml的流速(HiLoad 26/600Superdex 200pg)或每分钟0.5ml的流速(Superdex 200Increase 10/300GL)，并通过280nm处的吸光度来监测洗脱特性。对于确定纯化的蛋白在天然条件下的表观分子量，用具有已知分子量的标准蛋白质加载SEC柱。基于标准蛋白质的洗脱体积，绘制校准曲线并确定纯化的蛋白的表观分子量。来自本发明的第一方面的双特异性TNFRSF激动剂融合蛋白(SAB形式)和来自本发明的第二方面的双特异性TNFSF-配体融合蛋白从Superdex SEC柱洗脱，表观分子量为约150kDa，而本发明的第三方面的双特异性基于Fab的融合蛋白具有约100kDa的表观分子量。使用HPLC、具有两种靶标的基于ELISA的夹心检测法和基于TNFRSF报告基因细胞(reporter-cell)的活性检测用于确定上述双特异性TNFSF激动剂的双特异性性质。
实施例2：材料和方法
CD40激动化合物的细胞活性
使用来自Promega的CD40荧光素酶报告基因检测(产品号JA2155)评价CD40激动剂的细胞活性。将表达NFκB-luc2的U2OS细胞(其在细胞膜上组成型表达CD40)铺板在96孔板中，并在添加CD40激动剂之前在37℃下培养16-20小时。通过用激动化合物处理来诱导的生产性(Productive)CD40信号传导驱动NFκB-luc2 U2OS细胞中萤火虫荧光素酶的表达。在37℃下诱导4小时后，添加荧光素酶测定试剂并测量发光(RLU)(Tecan Infinite F500)。
CD27激动化合物的细胞活性
使用来自Promega的CD27荧光素酶报告基因检测(产品号CS1979A25)评价CD27激动剂的细胞活性。将NFκB-luc2/CD27 Jurkat细胞(其在细胞膜上表达CD27)铺板在96孔板中，并在添加CD27激动剂之前在37℃下培养16-20小时。通过用激动化合物处理来诱导的生产性CD27信号传导驱动NFκB-luc2/CD27 Jurkat细胞中萤火虫荧光素酶的表达。在37℃下诱导6小时后，添加荧光素酶测定试剂并测量发光(RLU)(Tecan Infinite F500)。
GITR激动化合物的细胞活性
使用来自Promega的GITR荧光素酶报告基因检测(产品号CS184009)评价GITR激动剂的细胞活性。将NFκB-luc2/GITR Jurkat细胞(其在细胞膜上表达GITR)铺板在96孔板中，并在添加GITR激动剂之前在37℃下短暂培养。通过用激动化合物处理来诱导的生产性GITR信号传导驱动NFκB-luc2/GITR Jurkat细胞中萤火虫荧光素酶的表达。在37℃下诱导5小时后，添加荧光素酶测定试剂并测量发光(RLU)(Tecan Infinite F500)。
PD-L1靶向化合物的细胞活性
使用来自Promega的PD-1/PD-L1荧光素酶报告基因检测(产品号J1250)评价PD-L1靶向化合物的细胞活性。将PD-L1 aAPC/CHO-K1细胞(表达人PD-L1和设计成以抗原非依赖性方式激活同源TCR的工程化细胞表面蛋白的细胞)在添加PD-L1靶向化合物和PD-1效应细胞之前在37℃下培养16-20小时。PD-1效应细胞是表达人PD-1和由NFAT应答元件(NFAT-RE)驱动的荧光素酶报告的Jurkat T细胞。在将两种细胞类型共培养时，PD-1/PD-L1相互作用抑制TCR信号传导和NFAT-RE介导的发光。添加阻断PD-1/PD-L1相互作用的抗PD-1或抗PD-L1抗体中的任一者释放抑制信号并导致TCR激活和NFAT-RE介导的发光。
在37℃下诱导6小时后，添加荧光素酶测定试剂并测量发光(RLU)(TecanInfinite F500)。
T细胞激活(流式细胞术)
为了测试CD137激动剂对原代人T细胞的活性，使用基于间接磁珠的分离试剂盒(目录编号130-094-131,Miltenyi)从PBMC分离天然pan T细胞。用Tag-it VioletTM增殖和细胞示踪染料(Biolegend)标记纯化的T细胞，重悬浮于培养基(AIM-V w/5％人血清,Gibco)中，并在37℃下用预包被的抗CD3抗体，克隆OKT3，1μg/mL)刺激4h、或用培养基对照刺激。立即添加CD137激动剂(100ng/ml)。在第五天，收获T细胞并通过流式细胞术来检测。
未成熟树突状细胞的刺激(流式细胞术)
使用标准试剂盒(Stem Cell)从健康人供体的血沉棕黄层中分离单核细胞。通过添加50ng/ml GM-CSF 3天、然后添加50ng/ml GM-CSF+50ng/ml IL-4另外的3天来实现单核细胞的分化。然后用100ng/ml的指示的CD40激动剂处理细胞24小时，随后通过流式细胞术评价CD86和CD83表达。
实施例3：多特异性免疫调节剂的序列
对于所有基于Fc结构域的异聚体构建体，使用了杵臼结构异二聚化技术，其中杵链的CH3结构域中具有S354C/T366W突变，臼链的CH3结构域中具有相应的Y349C/T366S/L368A/Y407V突变(Carter,J Immunol Methods 248,7-15(2001))。
为了消除与Fcγ受体的结合，将N297S突变引入至杵和臼重链的CH2结构域(SEQ-ID 28-31中的“CH2”。在另一实施方案中，根据国际专利申请公开号WO 2012/130831 A1中所述的方法，可将Pro329Gly、Leu234Ala和Leu235Ala突变引入至杵和臼重链的恒定区。
表1：示例性铰链-接头序列
Figure BDA0004048583020000311
Figure BDA0004048583020000321
铰链接头1-5和17可用于构建本发明的第二方面的蛋白质部分。铰链接头6-16可用于构建本发明的第一和第二方面的蛋白质部分。
表2：本发明的序列
下表2和图24中给出了本发明的关键序列的概述。
Figure BDA0004048583020000322
Figure BDA0004048583020000331
Figure BDA0004048583020000341
Figure BDA0004048583020000351
Figure BDA0004048583020000361
Figure BDA0004048583020000371
Figure BDA0004048583020000381
Figure BDA0004048583020000391
Figure BDA0004048583020000401
Figure BDA0004048583020000411
Figure BDA0004048583020000421
Figure BDA0004048583020000431
Figure BDA0004048583020000441
Figure BDA0004048583020000451
Figure BDA0004048583020000461
Figure BDA0004048583020000471
Figure BDA0004048583020000481
Figure BDA0004048583020000491
Figure BDA0004048583020000501
Figure BDA0004048583020000511
Figure BDA0004048583020000521
Figure BDA0004048583020000531
Figure BDA0004048583020000541
Figure BDA0004048583020000551
Figure BDA0004048583020000561
Figure BDA0004048583020000571
Figure BDA0004048583020000581
Figure BDA0004048583020000591
Figure BDA0004048583020000601
Figure BDA0004048583020000611
Figure BDA0004048583020000621
实施例4：靶向增加SAB分子的激动活性
已经证明了靶向结构域对双特异性分子的激动活性的贡献。在CD137荧光素酶检测中，通过添加HT1080细胞的aPDL1-scCD137L-SAB的激动活性的极大增加是明显的(参见图29)。几乎每个HT1080细胞表达PD-L1，然而它们对于CD95L表达为阴性的。因此，这些细胞未能相应增加aCD95L-scCD137L-SAB(非靶向对照)的激动活性并不令人惊讶。然而，PD-L1靶向构建体aPDL1-scCD137L-SAB的激动的大量增加(比对于六价scCD137L-Fc所观察到的活性高约16倍)是令人惊讶的，并强调了这些分子作为靶向表达PD-L1的肿瘤的强效共刺激分子的潜力。aPDL1-scCD137L-SAB的这种增加的激动活性也已经通过添加其它表达PD-L1的癌症细胞系AsPC-1、LN-18和MDA-MB231(未示出)得到了证明。对于共刺激激动剂经常观察到钟形浓度依赖性。事实是，在这种情况下，钟形浓度依赖性是非常明显的，这可以用高度激动来解释：由表达PD-L1的细胞交联的激动分子的数量有一个最佳值，较高数量的配体三聚体导致配体/受体复合物中受体的减少。
类似地，在CD27荧光素酶检测中，通过添加MDA-MB231细胞的aPDL1-scCD27L-SAB的激动活性的极大增加是明显的(参见图30)。几乎每个MDA-MB231细胞表达PD-L1，然而它们对于CD95L表达为阴性的。因此，这些细胞未能增加aCD95L-scCD27L-SAB(非靶向对照)的激动活性并不令人惊讶。然而，PD-L1靶向构建体aPDL1-scCD27L-SAB的激动的大量增加(比对于六价scCD27L-Fc所观察到的活性高约3倍)是令人惊讶的，并强调了这些分子作为靶向表达PD-L1的肿瘤的强效共刺激分子的潜力。aPDL1-scCD27L-SAB的这种增加的激动活性也已经通过添加其它表达PD-L1的癌症细胞系LN-18和HT1080(未示出)得到了证明。
实施例5：双特异性CD137L分子激活T细胞增殖
使用T细胞激活检测，在图31中示出了scCD137L双特异性分子的生物活性。在存在抗CD3刺激的情况下，双特异性分子aPDL1-scCD137L-SAB和aCD95L-scCD137L-SAB导致相似的T细胞增殖。
实施例6：aPD-L1-scCD40L-SAB双特异性显示出对树突状细胞的优异刺激
使用未成熟树突状细胞激活检测，在图32和图33中示出了scCD40L双特异性分子和其它CD40激动剂的生物活性。在图32所示的实验中，将三价scCD40L和抗PD-L1抗体片段组合的双特异性分子aPDL1-scCD40L-SAB是树突状细胞的最有效激活剂，激活水平(CD86+/CD83+)为88.31％。因此，将两个部分组合在一个分子中远比将这些部分作为两个分别的分子添加更有效，即aPD-L1抗体+CD40L(三聚体)，其显示仅39.48％的激活水平。aCD40单克隆抗体和CD40L(三聚体)均显示中等激活水平(40.59％和46.10％)，而aPD-L1单克隆抗体具有与培养基对照相同的活性。
在图33中所示的实验中，将三价scCD40L和抗PD-L1抗体片段组合的双特异性分子是树突状细胞的非常有效的激活剂，激活水平(CD86+/CD83+)类似于scCD40L-Fc的激活水平：对于aPDL1-scCD40L-SAB为73.00％，对于aPDL1-scCD40L(三价)为87.18％和对于aPDL1-scCD40L-Fc为96.12％。相比之下，靶向CD95L而不是PD-L1的双特异性分子aCD95L-scCD40L-SAB仅具有中等激活水平，因为缺少表达PD-L1的单核细胞上的CD95L的表达。对于aCD40单克隆抗体和CD40L(三聚体)也看到低激活水平。
实施例7：GITR荧光素酶检测
使用GITR荧光素酶检测，在图34中示出了aPDL1-scGITRL双特异性分子的生物活性。通过与抗人Fc(x-连接)交联，三价GITR激动剂aPDL1-scGITRL的活性明显增强至对于六价GITR激动剂scGITRL-Fc所观察到的水平
本申请进一步特征为其权利要求和以下项目1-11。
项目1：一种多特异性TNF家族融合蛋白组装体，其至少包含
(a)单链TNF-SF受体结合结构域超家族配体，其融合至
(b)第一肽接头，其融合至
(c)第一异二聚化结构域，和
(d)抗原结合或相互作用蛋白质部分，其融合至
(e)第二肽接头，其融合至
(f)第二异二聚化结构域。
项目2：一种多特异性TNF家族融合蛋白组装体，其至少包含
(a)单链TNF-SF受体结合结构域，其融合至
(b)第一肽接头，其融合至
(c)第一异二聚化结构域，和
(d)和第二单链TNF-SF受体结合结构域，其融合至
(e)第二肽接头，其融合至
(f)第二异二聚化结构域。
项目3：一种多特异性TNF家族融合蛋白组装体，其至少包含
(a)抗体的功能性Fab结构域，其融合至
(b)单链TNF-SF受体结合结构域，
其中Fab片段(a)的恒定重链结构域的c-末端通过肽接头融合至单链TNF-SF受体结合。
项目4：一种多特异性TNF家族融合蛋白组装体，其包含
(a)由SEQ-ID33和SEQ-ID 46以及SEQ-ID:47组成的组装体，
(b)由SEQ-ID37和SEQ-ID46以及SEQ-ID:47组成的组装体，
(c)由SEQ-ID41和SEQ-ID 46以及SEQ-ID:47组成的组装体。
项目5：一种多特异性TNF家族融合蛋白组装体，其选自包括以下的列表：
(a)由SEQ-ID33和SEQ-ID 54以及SEQ-ID:55组成的组装体，
(b)由SEQ-ID37和SEQ-ID 54以及SEQ-ID:55组成的组装体，
(c)由SEQ-ID41和SEQ-ID 54以及SEQ-ID:55组成的组装体。
项目6：一种多特异性TNF家族融合蛋白组装体，其选自包括以下的列表：
(a)由SEQ-ID33和SEQ-ID50以及SEQ-ID:51组成的组装体，
(b)由SEQ-ID37和SEQ-ID50以及SEQ-ID:51组成的组装体，
(c)由SEQ-ID41和SEQ-ID50以及SEQ-ID:51组成的组装体。
项目7：一种核酸分子，其编码项目1部分a)-c)的蛋白质部分，以及一种核酸分子，其编码项目1部分d)-e)的蛋白质部分。
项目8.一种核酸分子，其编码项目2部分a)-c)的蛋白质部分，以及一种核酸分子，其编码项目2部分d)-e)的蛋白质部分。
项目9：一种核酸分子，其编码项目3中任一种蛋白质，或用于项目4-6中任一项的蛋白组装体的共表达的核酸分子。
项目10：一种宿主细胞，其包含项目7-9中任一项的核酸。
项目11：一种药物组合物，其至少包含权利要求1-6中任一项的多特异性蛋白组装体或7-9的核酸。
序列表
<110> O·黑尔(Hill, Oliver)
C·吉菲尔斯(Gieffers, Christian)
M·斯尔曼(Thiemann, Meinolf)
K·比丽安-弗雷(BILLIAN-FREY, Katharina)
阿珀吉尼科斯股份公司(Apogenix AG)
<120> 多特异性免疫调节剂
<130> BiTech
<160> 119
<170> BiSSAP 1.3.6
<210> 1
<211> 20
<212> PRT
<213> 人工序列
<220>
<223> 信号肽
<400> 1
Met Glu Thr Asp Thr Leu Leu Val Phe Val Leu Leu Val Trp Val Pro
1 5 10 15
Ala Gly Asn Gly
20
<210> 2
<211> 21
<212> PRT
<213> 人工序列
<220>
<223> 铰链1
<400> 2
Gly Ser Ser Ser Ser Ser Ser Ser Ser Gly Ser Cys Asp Lys Thr His
1 5 10 15
Thr Cys Pro Pro Cys
20
<210> 3
<211> 20
<212> PRT
<213> 人工序列
<220>
<223> 铰链2
<400> 3
Gly Ser Ser Ser Ser Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr
1 5 10 15
Cys Pro Pro Cys
20
<210> 4
<211> 19
<212> PRT
<213> 人工序列
<220>
<223> 铰链3
<400> 4
Gly Ser Ser Ser Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys
1 5 10 15
Pro Pro Cys
<210> 5
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> 铰链4
<400> 5
Gly Ser Ser Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro
1 5 10 15
Pro Cys
<210> 6
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> 铰链5
<400> 6
Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
1 5 10 15
<210> 7
<211> 20
<212> PRT
<213> 人工序列
<220>
<223> 铰链6
<400> 7
Gly Ser Ser Ser Ser Ser Ser Ser Ser Gly Ser Asp Lys Thr His Thr
1 5 10 15
Cys Pro Pro Cys
20
<210> 8
<211> 19
<212> PRT
<213> 人工序列
<220>
<223> 铰链7
<400> 8
Gly Ser Ser Ser Ser Ser Ser Ser Gly Ser Asp Lys Thr His Thr Cys
1 5 10 15
Pro Pro Cys
<210> 9
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> 铰链8
<400> 9
Gly Ser Ser Ser Ser Ser Ser Gly Ser Asp Lys Thr His Thr Cys Pro
1 5 10 15
Pro Cys
<210> 10
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> 铰链9
<400> 10
Gly Ser Ser Ser Ser Ser Gly Ser Asp Lys Thr His Thr Cys Pro Pro
1 5 10 15
Cys
<210> 11
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> 铰链10
<400> 11
Gly Ser Ser Ser Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys
1 5 10 15
<210> 12
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 铰链11
<400> 12
Gly Ser Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys
1 5 10
<210> 13
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 接头-1
<400> 13
Asp Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser
1 5 10
<210> 14
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> 接头-2
<400> 14
Asp Lys Thr His Gly Ser Gly Ser Ser Ser Ser
1 5 10
<210> 15
<211> 8
<212> PRT
<213> 人工序列
<220>
<223> 接头3
<400> 15
Asp Lys Thr His Gly Ser Gly Ser
1 5
<210> 16
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> 接头-4
<400> 16
Gly Ser Gly Ser Ser Ser Ser Ser Ser
1 5
<210> 17
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 接头-5
<400> 17
Gly Ser Gly Ser Ser Ser
1 5
<210> 18
<211> 4
<212> PRT
<213> 人工序列
<220>
<223> 接头-6
<400> 18
Gly Ser Gly Ser
1
<210> 19
<211> 8
<212> PRT
<213> 人工序列
<220>
<223> 接头-7
<400> 19
Gly Gly Gly Ser Gly Gly Gly Ser
1 5
<210> 20
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> Fc-N297S-杵
<400> 20
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
35 40 45
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
50 55 60
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
65 70 75 80
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
85 90 95
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
100 105 110
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu
115 120 125
Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr
130 135 140
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
145 150 155 160
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
165 170 175
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
180 185 190
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
195 200 205
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 21
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> Fc-N297S-臼
<400> 21
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
35 40 45
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
50 55 60
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
65 70 75 80
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
85 90 95
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
100 105 110
Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu
115 120 125
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr
130 135 140
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
145 150 155 160
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
165 170 175
Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
180 185 190
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
195 200 205
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 22
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> Fc-杵-US20160200833A1
<400> 22
Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
35 40 45
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
50 55 60
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
65 70 75 80
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
85 90 95
Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
100 105 110
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu
115 120 125
Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr
130 135 140
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
145 150 155 160
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
165 170 175
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
180 185 190
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
195 200 205
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 23
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> Fc-臼-US20160200833A1
<400> 23
Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
35 40 45
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
50 55 60
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
65 70 75 80
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
85 90 95
Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
100 105 110
Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu
115 120 125
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr
130 135 140
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
145 150 155 160
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
165 170 175
Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
180 185 190
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
195 200 205
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 24
<211> 217
<212> PRT
<213> 人工序列
<220>
<223> Cambridge-Fc-杵
<400> 24
Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
20 25 30
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
35 40 45
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
50 55 60
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
85 90 95
Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
100 105 110
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu
115 120 125
Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro
130 135 140
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
145 150 155 160
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
165 170 175
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
180 185 190
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 25
<211> 217
<212> PRT
<213> 人工序列
<220>
<223> Cambridge-Fc-臼
<400> 25
Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
1 5 10 15
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
20 25 30
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
35 40 45
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
50 55 60
Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
65 70 75 80
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
85 90 95
Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
100 105 110
Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu
115 120 125
Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro
130 135 140
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
145 150 155 160
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
165 170 175
Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
180 185 190
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
195 200 205
Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 26
<211> 106
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CL-kappa
<400> 26
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
1 5 10 15
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
20 25 30
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
35 40 45
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
50 55 60
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
65 70 75 80
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
85 90 95
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105
<210> 27
<211> 103
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1
<400> 27
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys
100
<210> 28
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1CH2sCH3-杵
<400> 28
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ser 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 Cys Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Trp 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> 29
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1CH2sCH3-RF-杵
<400> 29
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ser 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 Cys Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Trp 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 Arg Phe Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 30
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1CH2sCH3-臼
<400> 30
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ser 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 Cys Thr Leu Pro Pro Ser Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala 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 Val 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> 31
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1CH2sCH3-RF-臼
<400> 31
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ser 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 Cys Thr Leu Pro Pro Ser Arg Asp Glu
225 230 235 240
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala 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 Val 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 Arg Phe Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 32
<211> 675
<212> PRT
<213> 人工序列
<220>
<223> scCD40L-Fc-杵_a
<400> 32
Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser
1 5 10 15
Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu
20 25 30
Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu
35 40 45
Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser
50 55 60
Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg
65 70 75 80
Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys
85 90 95
Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln
100 105 110
Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser
115 120 125
His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly
130 135 140
Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser
145 150 155 160
Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr
165 170 175
Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val
180 185 190
Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser
195 200 205
Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu
210 215 220
Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr
225 230 235 240
His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly
245 250 255
Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp
260 265 270
Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu
275 280 285
Lys Leu Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val
290 295 300
Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu
305 310 315 320
Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly
325 330 335
Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln
340 345 350
Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile
355 360 365
Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu
370 375 380
Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser
385 390 395 400
Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe
405 410 415
Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr
420 425 430
Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Ser Ser Gly Ser Cys
435 440 445
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
450 455 460
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
465 470 475 480
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
485 490 495
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
500 505 510
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
515 520 525
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
530 535 540
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
545 550 555 560
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
565 570 575
Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
580 585 590
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
595 600 605
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
610 615 620
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
625 630 635 640
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
645 650 655
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
660 665 670
Pro Gly Lys
675
<210> 33
<211> 670
<212> PRT
<213> 人工序列
<220>
<223> scCD40L-Fc-杵_b
<400> 33
Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser
1 5 10 15
Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu
20 25 30
Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu
35 40 45
Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser
50 55 60
Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg
65 70 75 80
Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys
85 90 95
Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln
100 105 110
Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser
115 120 125
His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly
130 135 140
Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser
145 150 155 160
Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr
165 170 175
Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val
180 185 190
Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser
195 200 205
Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu
210 215 220
Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr
225 230 235 240
His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly
245 250 255
Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp
260 265 270
Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu
275 280 285
Lys Leu Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val
290 295 300
Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu
305 310 315 320
Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly
325 330 335
Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln
340 345 350
Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile
355 360 365
Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu
370 375 380
Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser
385 390 395 400
Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe
405 410 415
Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr
420 425 430
Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Asp Lys Thr His Thr
435 440 445
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
450 455 460
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
465 470 475 480
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
485 490 495
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
500 505 510
Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val
515 520 525
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
530 535 540
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
545 550 555 560
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
565 570 575
Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val
580 585 590
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
595 600 605
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
610 615 620
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
625 630 635 640
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
645 650 655
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
660 665 670
<210> 34
<211> 675
<212> PRT
<213> 人工序列
<220>
<223> scCD40L-Fc-臼_a
<400> 34
Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser
1 5 10 15
Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu
20 25 30
Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu
35 40 45
Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser
50 55 60
Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg
65 70 75 80
Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys
85 90 95
Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln
100 105 110
Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser
115 120 125
His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly
130 135 140
Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser
145 150 155 160
Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr
165 170 175
Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val
180 185 190
Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser
195 200 205
Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu
210 215 220
Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr
225 230 235 240
His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly
245 250 255
Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp
260 265 270
Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu
275 280 285
Lys Leu Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val
290 295 300
Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu
305 310 315 320
Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly
325 330 335
Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln
340 345 350
Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile
355 360 365
Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu
370 375 380
Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser
385 390 395 400
Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe
405 410 415
Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr
420 425 430
Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Ser Ser Gly Ser Cys
435 440 445
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
450 455 460
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
465 470 475 480
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
485 490 495
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
500 505 510
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
515 520 525
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
530 535 540
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
545 550 555 560
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
565 570 575
Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
580 585 590
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
595 600 605
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
610 615 620
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
625 630 635 640
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
645 650 655
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
660 665 670
Pro Gly Lys
675
<210> 35
<211> 670
<212> PRT
<213> 人工序列
<220>
<223> scCD40L-Fc-臼_b
<400> 35
Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser
1 5 10 15
Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu
20 25 30
Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu
35 40 45
Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser
50 55 60
Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg
65 70 75 80
Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys
85 90 95
Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln
100 105 110
Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser
115 120 125
His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly
130 135 140
Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser
145 150 155 160
Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr
165 170 175
Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val
180 185 190
Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser
195 200 205
Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu
210 215 220
Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr
225 230 235 240
His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly
245 250 255
Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp
260 265 270
Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu
275 280 285
Lys Leu Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val
290 295 300
Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu
305 310 315 320
Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly
325 330 335
Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln
340 345 350
Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile
355 360 365
Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu
370 375 380
Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser
385 390 395 400
Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe
405 410 415
Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr
420 425 430
Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Asp Lys Thr His Thr
435 440 445
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
450 455 460
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
465 470 475 480
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
485 490 495
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
500 505 510
Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val
515 520 525
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
530 535 540
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
545 550 555 560
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro
565 570 575
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val
580 585 590
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
595 600 605
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
610 615 620
Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
625 630 635 640
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
645 650 655
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
660 665 670
<210> 36
<211> 681
<212> PRT
<213> 人工序列
<220>
<223> scCD27L-Fc-杵_a
<400> 36
Gln Ser Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly
1 5 10 15
Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly
20 25 30
Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile
35 40 45
His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile
50 55 60
Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val
65 70 75 80
Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser
85 90 95
Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala
100 105 110
Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser
115 120 125
Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly
130 135 140
Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu
145 150 155 160
Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp
165 170 175
Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu
180 185 190
Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His
195 200 205
Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His
210 215 220
His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser
225 230 235 240
Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser
245 250 255
Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu
260 265 270
Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly
275 280 285
Val Gln Trp Val Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser
290 295 300
Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln
305 310 315 320
Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser
325 330 335
Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg
340 345 350
Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser
355 360 365
Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile
370 375 380
Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His
385 390 395 400
Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly
405 410 415
Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn
420 425 430
Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly Ser Gly
435 440 445
Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
450 455 460
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
465 470 475 480
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
485 490 495
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
500 505 510
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
515 520 525
Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
530 535 540
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
545 550 555 560
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
565 570 575
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu
580 585 590
Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro
595 600 605
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
610 615 620
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
625 630 635 640
Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
645 650 655
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
660 665 670
Lys Ser Leu Ser Leu Ser Pro Gly Lys
675 680
<210> 37
<211> 676
<212> PRT
<213> 人工序列
<220>
<223> scCD27L-Fc-杵_b
<400> 37
Gln Ser Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly
1 5 10 15
Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly
20 25 30
Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile
35 40 45
His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile
50 55 60
Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val
65 70 75 80
Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser
85 90 95
Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala
100 105 110
Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser
115 120 125
Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly
130 135 140
Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu
145 150 155 160
Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp
165 170 175
Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu
180 185 190
Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His
195 200 205
Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His
210 215 220
His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser
225 230 235 240
Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser
245 250 255
Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu
260 265 270
Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly
275 280 285
Val Gln Trp Val Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser
290 295 300
Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln
305 310 315 320
Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser
325 330 335
Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg
340 345 350
Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser
355 360 365
Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile
370 375 380
Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His
385 390 395 400
Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly
405 410 415
Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn
420 425 430
Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly Ser Gly
435 440 445
Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
450 455 460
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
465 470 475 480
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
485 490 495
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
500 505 510
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
515 520 525
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
530 535 540
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
545 550 555 560
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
565 570 575
Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val
580 585 590
Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
595 600 605
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
610 615 620
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
625 630 635 640
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
645 650 655
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
660 665 670
Ser Pro Gly Lys
675
<210> 38
<211> 681
<212> PRT
<213> 人工序列
<220>
<223> scCD27L-Fc-臼_a
<400> 38
Gln Ser Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly
1 5 10 15
Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly
20 25 30
Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile
35 40 45
His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile
50 55 60
Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val
65 70 75 80
Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser
85 90 95
Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala
100 105 110
Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser
115 120 125
Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly
130 135 140
Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu
145 150 155 160
Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp
165 170 175
Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu
180 185 190
Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His
195 200 205
Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His
210 215 220
His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser
225 230 235 240
Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser
245 250 255
Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu
260 265 270
Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly
275 280 285
Val Gln Trp Val Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser
290 295 300
Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln
305 310 315 320
Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser
325 330 335
Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg
340 345 350
Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser
355 360 365
Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile
370 375 380
Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His
385 390 395 400
Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly
405 410 415
Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn
420 425 430
Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly Ser Gly
435 440 445
Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro
450 455 460
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys
465 470 475 480
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val
485 490 495
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
500 505 510
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
515 520 525
Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
530 535 540
Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
545 550 555 560
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
565 570 575
Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu
580 585 590
Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro
595 600 605
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
610 615 620
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
625 630 635 640
Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
645 650 655
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
660 665 670
Lys Ser Leu Ser Leu Ser Pro Gly Lys
675 680
<210> 39
<211> 676
<212> PRT
<213> 人工序列
<220>
<223> scCD27L-Fc-臼_b
<400> 39
Gln Ser Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly
1 5 10 15
Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly
20 25 30
Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile
35 40 45
His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile
50 55 60
Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val
65 70 75 80
Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser
85 90 95
Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala
100 105 110
Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser
115 120 125
Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly
130 135 140
Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu
145 150 155 160
Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp
165 170 175
Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu
180 185 190
Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His
195 200 205
Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His
210 215 220
His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser
225 230 235 240
Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser
245 250 255
Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu
260 265 270
Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly
275 280 285
Val Gln Trp Val Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser
290 295 300
Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln
305 310 315 320
Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser
325 330 335
Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg
340 345 350
Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser
355 360 365
Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile
370 375 380
Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His
385 390 395 400
Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly
405 410 415
Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn
420 425 430
Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly Ser Gly
435 440 445
Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
450 455 460
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
465 470 475 480
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
485 490 495
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
500 505 510
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
515 520 525
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
530 535 540
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
545 550 555 560
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
565 570 575
Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
580 585 590
Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
595 600 605
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
610 615 620
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr
625 630 635 640
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
645 650 655
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
660 665 670
Ser Pro Gly Lys
675
<210> 40
<211> 618
<212> PRT
<213> 人工序列
<220>
<223> scGITRL-Fc-杵_a
<400> 40
Gln Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
1 5 10 15
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
20 25 30
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
35 40 45
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
50 55 60
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
65 70 75 80
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
85 90 95
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
100 105 110
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn
115 120 125
Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp
130 135 140
Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp
145 150 155 160
Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val
165 170 175
Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu
180 185 190
Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile
195 200 205
Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp
210 215 220
Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp
225 230 235 240
Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser
245 250 255
Gly Asn Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser
260 265 270
Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser
275 280 285
Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly
290 295 300
Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val
305 310 315 320
Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser
325 330 335
Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr
340 345 350
Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr
355 360 365
Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser
370 375 380
Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
385 390 395 400
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
405 410 415
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
420 425 430
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
435 440 445
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
450 455 460
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
465 470 475 480
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
485 490 495
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
500 505 510
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu
515 520 525
Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr
530 535 540
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
545 550 555 560
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
565 570 575
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
580 585 590
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
595 600 605
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
610 615
<210> 41
<211> 613
<212> PRT
<213> 人工序列
<220>
<223> scGITRL-Fc-杵_b
<400> 41
Gln Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
1 5 10 15
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
20 25 30
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
35 40 45
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
50 55 60
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
65 70 75 80
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
85 90 95
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
100 105 110
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn
115 120 125
Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp
130 135 140
Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp
145 150 155 160
Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val
165 170 175
Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu
180 185 190
Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile
195 200 205
Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp
210 215 220
Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp
225 230 235 240
Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser
245 250 255
Gly Asn Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser
260 265 270
Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser
275 280 285
Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly
290 295 300
Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val
305 310 315 320
Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser
325 330 335
Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr
340 345 350
Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr
355 360 365
Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser
370 375 380
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
385 390 395 400
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
405 410 415
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
420 425 430
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
435 440 445
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser
450 455 460
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
465 470 475 480
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
485 490 495
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
500 505 510
Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln
515 520 525
Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
530 535 540
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
545 550 555 560
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
565 570 575
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
580 585 590
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
595 600 605
Leu Ser Pro Gly Lys
610
<210> 42
<211> 618
<212> PRT
<213> 人工序列
<220>
<223> scGITRL-Fc-臼_a
<400> 42
Gln Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
1 5 10 15
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
20 25 30
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
35 40 45
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
50 55 60
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
65 70 75 80
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
85 90 95
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
100 105 110
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn
115 120 125
Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp
130 135 140
Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp
145 150 155 160
Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val
165 170 175
Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu
180 185 190
Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile
195 200 205
Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp
210 215 220
Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp
225 230 235 240
Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser
245 250 255
Gly Asn Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser
260 265 270
Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser
275 280 285
Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly
290 295 300
Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val
305 310 315 320
Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser
325 330 335
Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr
340 345 350
Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr
355 360 365
Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser
370 375 380
Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
385 390 395 400
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
405 410 415
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
420 425 430
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
435 440 445
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
450 455 460
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
465 470 475 480
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
485 490 495
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
500 505 510
Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu
515 520 525
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr
530 535 540
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
545 550 555 560
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
565 570 575
Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
580 585 590
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
595 600 605
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
610 615
<210> 43
<211> 613
<212> PRT
<213> 人工序列
<220>
<223> scGITRL-Fc-臼_b
<400> 43
Gln Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
1 5 10 15
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
20 25 30
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
35 40 45
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
50 55 60
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
65 70 75 80
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
85 90 95
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
100 105 110
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn
115 120 125
Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp
130 135 140
Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp
145 150 155 160
Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val
165 170 175
Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu
180 185 190
Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile
195 200 205
Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp
210 215 220
Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp
225 230 235 240
Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser
245 250 255
Gly Asn Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser
260 265 270
Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser
275 280 285
Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly
290 295 300
Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val
305 310 315 320
Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser
325 330 335
Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr
340 345 350
Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr
355 360 365
Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser
370 375 380
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
385 390 395 400
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
405 410 415
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
420 425 430
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
435 440 445
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser
450 455 460
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
465 470 475 480
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
485 490 495
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
500 505 510
Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
515 520 525
Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
530 535 540
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
545 550 555 560
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu
565 570 575
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
580 585 590
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
595 600 605
Leu Ser Pro Gly Lys
610
<210> 44
<211> 452
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-HC-杵
<400> 44
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
225 230 235 240
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
245 250 255
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
260 265 270
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
275 280 285
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
290 295 300
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
305 310 315 320
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
325 330 335
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
340 345 350
Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val
355 360 365
Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
370 375 380
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
385 390 395 400
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
405 410 415
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
420 425 430
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
435 440 445
Ser Pro Gly Lys
450
<210> 45
<211> 452
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-HC-臼
<400> 45
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
225 230 235 240
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
245 250 255
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
260 265 270
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
275 280 285
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
290 295 300
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
305 310 315 320
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
325 330 335
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
340 345 350
Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
355 360 365
Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
370 375 380
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
385 390 395 400
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr
405 410 415
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
420 425 430
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
435 440 445
Ser Pro Gly Lys
450
<210> 46
<211> 452
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-HC-RF-臼
<400> 46
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
225 230 235 240
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
245 250 255
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
260 265 270
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
275 280 285
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
290 295 300
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
305 310 315 320
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
325 330 335
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
340 345 350
Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
355 360 365
Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
370 375 380
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
385 390 395 400
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr
405 410 415
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
420 425 430
Met His Glu Ala Leu His Asn Arg Phe Thr Gln Lys Ser Leu Ser Leu
435 440 445
Ser Pro Gly Lys
450
<210> 47
<211> 219
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-LC
<400> 47
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 Ser Ile Arg Thr Ser
20 25 30
Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Lys Ala Ser Asp Leu Pro Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Ser Tyr Asp Phe Arg Asp Thr
85 90 95
Ile Asn Asn Gly His Ser Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
130 135 140
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
145 150 155 160
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
165 170 175
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
180 185 190
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
195 200 205
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 48
<211> 451
<212> PRT
<213> 人工序列
<220>
<223> aCEA-HC-杵
<400> 48
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 Thr Ser Gly Phe Thr Phe Thr Asp Tyr
20 25 30
Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu
35 40 45
Gly Phe Ile Gly Asn Lys Ala Asn Gly Tyr Thr Thr Glu Tyr Ser Ala
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Thr Leu Gln Ala Glu Asp Ser Ala Ile Tyr
85 90 95
Tyr Cys Thr Arg Asp Arg Gly Leu Arg Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 49
<211> 451
<212> PRT
<213> 人工序列
<220>
<223> aCEA-HC-臼
<400> 49
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 Thr Ser Gly Phe Thr Phe Thr Asp Tyr
20 25 30
Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu
35 40 45
Gly Phe Ile Gly Asn Lys Ala Asn Gly Tyr Thr Thr Glu Tyr Ser Ala
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Thr Leu Gln Ala Glu Asp Ser Ala Ile Tyr
85 90 95
Tyr Cys Thr Arg Asp Arg Gly Leu Arg Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 50
<211> 451
<212> PRT
<213> 人工序列
<220>
<223> aCEA-HC-RF-臼
<400> 50
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 Thr Ser Gly Phe Thr Phe Thr Asp Tyr
20 25 30
Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu
35 40 45
Gly Phe Ile Gly Asn Lys Ala Asn Gly Tyr Thr Thr Glu Tyr Ser Ala
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Thr Leu Gln Ala Glu Asp Ser Ala Ile Tyr
85 90 95
Tyr Cys Thr Arg Asp Arg Gly Leu Arg Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
225 230 235 240
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
275 280 285
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
305 310 315 320
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
340 345 350
Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn Arg Phe Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 51
<211> 213
<212> PRT
<213> 人工序列
<220>
<223> aCEA-LC
<400> 51
Gln Thr Val Leu Thr Gln Ser Pro Ser Ser Leu Ser Val Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Ser Val Thr Tyr Ile
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Leu Ala Pro Lys Ser Leu Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu
65 70 75 80
Asp Ile Ala Thr Tyr Tyr Cys Gln His Trp Ser Ser Lys Pro Pro Thr
85 90 95
Phe Gly Gln Gly Thr Lys Val Glu Val Lys Arg Thr Val Ala Ala Pro
100 105 110
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
115 120 125
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
130 135 140
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
145 150 155 160
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
165 170 175
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
180 185 190
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
195 200 205
Asn Arg Gly Glu Cys
210
<210> 52
<211> 450
<212> PRT
<213> 人工序列
<220>
<223> aPD-L1-HC-杵
<400> 52
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
340 345 350
Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
355 360 365
Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 53
<211> 450
<212> PRT
<213> 人工序列
<220>
<223> aPD-L1-HC-臼
<400> 53
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys
340 345 350
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
355 360 365
Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 54
<211> 450
<212> PRT
<213> 人工序列
<220>
<223> aPD-L1-HC-RF-臼
<400> 54
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
225 230 235 240
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
325 330 335
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys
340 345 350
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
355 360 365
Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430
Glu Ala Leu His Asn Arg Phe Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445
Gly Lys
450
<210> 55
<211> 217
<212> PRT
<213> 人工序列
<220>
<223> aPD-L1-LC
<400> 55
Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr
20 25 30
Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser
85 90 95
Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Glu Ile Lys Arg Thr
100 105 110
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
115 120 125
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
130 135 140
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
145 150 155 160
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
165 170 175
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
180 185 190
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
195 200 205
Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 56
<211> 676
<212> PRT
<213> 人工序列
<220>
<223> aCEA-hc-scCD40L-RBD
<400> 56
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 Thr Ser Gly Phe Thr Phe Thr Asp Tyr
20 25 30
Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu
35 40 45
Gly Phe Ile Gly Asn Lys Ala Asn Gly Tyr Thr Thr Glu Tyr Ser Ala
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Thr Leu Gln Ala Glu Asp Ser Ala Ile Tyr
85 90 95
Tyr Cys Thr Arg Asp Arg Gly Leu Arg Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125
Val Phe Pro Phe Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala
130 135 140
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
145 150 155 160
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
180 185 190
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
195 200 205
Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
210 215 220
Asp Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Gln Ile Ala
225 230 235 240
Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln
245 250 255
Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu
260 265 270
Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile
275 280 285
Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala
290 295 300
Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg
305 310 315 320
Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly
325 330 335
Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala
340 345 350
Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr
355 360 365
Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Gly Asn
370 375 380
Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr
385 390 395 400
Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn
405 410 415
Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln
420 425 430
Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu
435 440 445
Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro
450 455 460
Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser
465 470 475 480
Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu
485 490 495
Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln
500 505 510
Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly
515 520 525
Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu
530 535 540
Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr
545 550 555 560
Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu
565 570 575
Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe
580 585 590
Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu
595 600 605
Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala
610 615 620
Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu
625 630 635 640
Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val
645 650 655
Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly
660 665 670
Leu Leu Lys Leu
675
<210> 57
<211> 677
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-hc-scCD40L-RBD
<400> 57
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Gln Ile
225 230 235 240
Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu
245 250 255
Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr
260 265 270
Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr
275 280 285
Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln
290 295 300
Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu
305 310 315 320
Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys
325 330 335
Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly
340 345 350
Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly
355 360 365
Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Gly
370 375 380
Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys
385 390 395 400
Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser
405 410 415
Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg
420 425 430
Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg
435 440 445
Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser
450 455 460
Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser
465 470 475 480
Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe
485 490 495
Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser
500 505 510
Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu
515 520 525
Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser
530 535 540
Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly
545 550 555 560
Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln
565 570 575
Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr
580 585 590
Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser
595 600 605
Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala
610 615 620
Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His
625 630 635 640
Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn
645 650 655
Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe
660 665 670
Gly Leu Leu Lys Leu
675
<210> 58
<211> 675
<212> PRT
<213> 人工序列
<220>
<223> aPDL1-hc-scCD40L-RBD
<400> 58
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Gln Ile Ala Ala
225 230 235 240
His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp
245 250 255
Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu
260 265 270
Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr
275 280 285
Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro
290 295 300
Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile
305 310 315 320
Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln
325 330 335
Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser
340 345 350
Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly
355 360 365
Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Gly Asn Gly
370 375 380
Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr
385 390 395 400
Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn
405 410 415
Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly
420 425 430
Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala
435 440 445
Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly
450 455 460
Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala
465 470 475 480
Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu
485 490 495
Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val
500 505 510
Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser
515 520 525
Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala
530 535 540
Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr
545 550 555 560
Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr
565 570 575
Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys
580 585 590
Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser
595 600 605
Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn
610 615 620
Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly
625 630 635 640
Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr
645 650 655
Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu
660 665 670
Leu Lys Leu
675
<210> 59
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> aCD137-VH
<400> 59
Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr
20 25 30
Tyr Trp Ser Trp Ile Arg Gln Ser Pro Glu Lys Gly Leu Glu Trp Ile
35 40 45
Gly Glu Ile Asn His Gly Gly Tyr Val Thr Tyr Asn Pro Ser Leu Glu
50 55 60
Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu
65 70 75 80
Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Asp Tyr Gly Pro Gly Asn Tyr Asp Trp Tyr Phe Asp Leu Trp Gly
100 105 110
Arg Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 60
<211> 110
<212> PRT
<213> 人工序列
<220>
<223> aCD137-VL
<400> 60
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro
85 90 95
Ala Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
<210> 61
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> aMeso-VH
<400> 61
Glu Val Gln Leu Val 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 Tyr Thr Phe Thr Thr Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Gly Tyr Ile Arg Pro Ser Thr Gly Tyr Thr Glu Tyr Asn Gln Lys Phe
50 55 60
Lys Asp Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala 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 Ser Arg Trp Leu Leu Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 62
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> aMeso-VL
<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 Lys Ser Ser Gln Ser Val Leu Tyr Ser
20 25 30
Ser Asn Gln Lys Asn Tyr Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys
35 40 45
Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
50 55 60
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys His Gln
85 90 95
Tyr Leu Ser Ser Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
Arg
<210> 63
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> aCD25-VH
<400> 63
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 Ser Tyr
20 25 30
Arg Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Tyr Ile Asn Pro Ser Thr Gly Tyr Thr Glu Tyr Asn Gln Lys Phe
50 55 60
Lys Asp Lys Ala Thr Ile Thr Ala Asp Glu Ser Thr Asn 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 Gly Gly Gly Val Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu
100 105 110
Thr Val Ser Ser
115
<210> 64
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> aCD25-VL
<400> 64
Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Ile Ser Tyr Met
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr
35 40 45
Thr Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp
65 70 75 80
Asp Phe Ala Thr Tyr Tyr Cys His Gln Arg Ser Thr Tyr Pro Leu Thr
85 90 95
Phe Gly Ser Gly Thr Lys Val Glu Val Lys Arg
100 105
<210> 65
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> aPD1-a-VH
<400> 65
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser
100 105 110
Ser
<210> 66
<211> 108
<212> PRT
<213> 人工序列
<220>
<223> aPD1-a-VL
<400> 66
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105
<210> 67
<211> 120
<212> PRT
<213> 人工序列
<220>
<223> aPD1-b-VH
<400> 67
Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
20 25 30
Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe
50 55 60
Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr
65 70 75 80
Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 68
<211> 112
<212> PRT
<213> 人工序列
<220>
<223> aPD1-b-VL
<400> 68
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser
20 25 30
Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
35 40 45
Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala
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 Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg
85 90 95
Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
<210> 69
<211> 439
<212> PRT
<213> 人工序列
<220>
<223> scCD40L-RBD
<400> 69
Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser
1 5 10 15
Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu
20 25 30
Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu
35 40 45
Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser
50 55 60
Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg
65 70 75 80
Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys
85 90 95
Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln
100 105 110
Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser
115 120 125
His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly
130 135 140
Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser
145 150 155 160
Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr
165 170 175
Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val
180 185 190
Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser
195 200 205
Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu
210 215 220
Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr
225 230 235 240
His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly
245 250 255
Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp
260 265 270
Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu
275 280 285
Lys Leu Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val
290 295 300
Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu
305 310 315 320
Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly
325 330 335
Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln
340 345 350
Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile
355 360 365
Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu
370 375 380
Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser
385 390 395 400
Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe
405 410 415
Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr
420 425 430
Ser Phe Gly Leu Leu Lys Leu
435
<210> 70
<211> 445
<212> PRT
<213> 人工序列
<220>
<223> scCD27L-RBD
<400> 70
Gln Ser Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly
1 5 10 15
Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly
20 25 30
Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile
35 40 45
His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile
50 55 60
Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val
65 70 75 80
Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser
85 90 95
Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala
100 105 110
Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser
115 120 125
Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly
130 135 140
Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu
145 150 155 160
Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp
165 170 175
Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu
180 185 190
Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His
195 200 205
Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His
210 215 220
His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser
225 230 235 240
Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser
245 250 255
Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu
260 265 270
Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly
275 280 285
Val Gln Trp Val Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser
290 295 300
Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln
305 310 315 320
Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser
325 330 335
Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg
340 345 350
Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser
355 360 365
Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile
370 375 380
Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His
385 390 395 400
Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly
405 410 415
Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn
420 425 430
Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro
435 440 445
<210> 71
<211> 382
<212> PRT
<213> 人工序列
<220>
<223> scGITRL-RBD
<400> 71
Gln Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
1 5 10 15
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
20 25 30
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
35 40 45
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
50 55 60
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
65 70 75 80
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
85 90 95
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
100 105 110
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn
115 120 125
Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp
130 135 140
Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp
145 150 155 160
Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val
165 170 175
Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu
180 185 190
Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile
195 200 205
Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp
210 215 220
Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp
225 230 235 240
Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser
245 250 255
Gly Asn Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser
260 265 270
Lys Trp Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser
275 280 285
Asp Trp Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly
290 295 300
Gln Val Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val
305 310 315 320
Arg Leu Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser
325 330 335
Lys Ile Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr
340 345 350
Ile Asp Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr
355 360 365
Tyr Trp Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
370 375 380
<210> 72
<211> 472
<212> PRT
<213> 人工序列
<220>
<223> scCD137L- RBD
<400> 72
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
145 150 155 160
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
165 170 175
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
180 185 190
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
195 200 205
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
210 215 220
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
225 230 235 240
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
245 250 255
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
260 265 270
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
275 280 285
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
290 295 300
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
305 310 315 320
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
325 330 335
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
340 345 350
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
355 360 365
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
370 375 380
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
385 390 395 400
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
405 410 415
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
420 425 430
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
435 440 445
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
450 455 460
Thr Val Leu Gly Leu Phe Arg Val
465 470
<210> 73
<211> 462
<212> PRT
<213> 人工序列
<220>
<223> scLIGHT-RBD
<400> 73
Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr
1 5 10 15
Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe
20 25 30
Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala
35 40 45
Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys
50 55 60
Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr
65 70 75 80
Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro
85 90 95
Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe
100 105 110
Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg
115 120 125
Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr
130 135 140
Phe Gly Ala Phe Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro
145 150 155 160
Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly
165 170 175
Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu
180 185 190
Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr
195 200 205
Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu
210 215 220
Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro
225 230 235 240
Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala
245 250 255
Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val
260 265 270
Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu
275 280 285
Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe
290 295 300
Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro Ala Ala His Leu
305 310 315 320
Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu Trp
325 330 335
Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr His Asp
340 345 350
Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys
355 360 365
Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr Ile
370 375 380
Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu Glu
385 390 395 400
Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Ser Ser
405 410 415
Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu Glu
420 425 430
Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val Arg
435 440 445
Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val
450 455 460
<210> 74
<211> 120
<212> PRT
<213> 人工序列
<220>
<223> aPD-L1-VH
<400> 74
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 75
<211> 111
<212> PRT
<213> 人工序列
<220>
<223> aPD-L1-VL
<400> 75
Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr
20 25 30
Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser
85 90 95
Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
<210> 76
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> Streptag-II 元件-1
<400> 76
Ser Ser Ser Ser Ser Ser Ala Trp Ser His Pro Gln Phe Glu Lys
1 5 10 15
<210> 77
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> Streptag-II 元件-2
<400> 77
Gly Ser Ser Ser Ser Ser Ser Ala Trp Ser His Pro Gln Phe Glu Lys
1 5 10 15
<210> 78
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> 铰链17
<400> 78
Gly Ser Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro
1 5 10 15
Pro Cys
<210> 79
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> 铰链12
<400> 79
Gly Ser Gly Ser Gly Gly Gly Ser Asp Lys Thr His Thr Cys Pro Pro
1 5 10 15
Cys
<210> 80
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> 铰链13
<400> 80
Gly Ser Gly Ser Gly Gly Gly Ser Thr His Thr Cys Pro Pro Cys
1 5 10 15
<210> 81
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> 铰链14
<400> 81
Gly Ser Gly Ser Thr His Thr Cys Pro Pro Cys
1 5 10
<210> 82
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> 铰链15
<400> 82
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys
1 5 10
<210> 83
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> 铰链16
<400> 83
Gly Ser Gly Ser Ser Ser Gly Ser Asp Lys Thr His Thr Cys Pro Pro
1 5 10 15
Cys
<210> 84
<211> 674
<212> PRT
<213> 人工序列
<220>
<223> scCD40L-Fc-杵_c
<400> 84
Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser
1 5 10 15
Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu
20 25 30
Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val Lys Arg Gln Gly Leu
35 40 45
Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser Asn Arg Glu Ala Ser
50 55 60
Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg
65 70 75 80
Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr His Ser Ser Ala Lys
85 90 95
Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly Val Phe Glu Leu Gln
100 105 110
Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp Pro Ser Gln Val Ser
115 120 125
His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly
130 135 140
Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val Ile Ser Glu Ala Ser
145 150 155 160
Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly Tyr Tyr Thr
165 170 175
Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln Leu Thr Val
180 185 190
Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr Phe Cys Ser
195 200 205
Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser Leu Ser Leu
210 215 220
Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala Ala Asn Thr
225 230 235 240
His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His Leu Gly Gly
245 250 255
Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn Val Thr Asp
260 265 270
Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe Gly Leu Leu
275 280 285
Lys Leu Gly Ser Gly Ser Gly Asn Gly Ser Gln Ile Ala Ala His Val
290 295 300
Ile Ser Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu
305 310 315 320
Lys Gly Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly
325 330 335
Lys Gln Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln
340 345 350
Val Thr Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile
355 360 365
Ala Ser Leu Ser Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu
370 375 380
Arg Ala Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser
385 390 395 400
Ile His Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe
405 410 415
Val Asn Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr
420 425 430
Ser Phe Gly Leu Leu Lys Leu Gly Ser Gly Ser Ser Ser Gly Ser Asp
435 440 445
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
450 455 460
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
465 470 475 480
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
485 490 495
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
500 505 510
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg
515 520 525
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
530 535 540
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
545 550 555 560
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
565 570 575
Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
580 585 590
Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
595 600 605
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
610 615 620
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
625 630 635 640
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
645 650 655
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
660 665 670
Gly Lys
<210> 85
<211> 708
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V1-Fc-杵_a
<400> 85
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
145 150 155 160
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
165 170 175
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
180 185 190
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
195 200 205
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
210 215 220
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
225 230 235 240
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
245 250 255
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
260 265 270
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
275 280 285
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
290 295 300
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
305 310 315 320
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
325 330 335
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
340 345 350
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
355 360 365
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
370 375 380
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
385 390 395 400
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
405 410 415
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
420 425 430
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
435 440 445
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
450 455 460
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Ser Ser Gly Ser
465 470 475 480
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
485 490 495
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
500 505 510
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
515 520 525
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
530 535 540
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
545 550 555 560
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
565 570 575
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
580 585 590
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
595 600 605
Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val
610 615 620
Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
625 630 635 640
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
645 650 655
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
660 665 670
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
675 680 685
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
690 695 700
Ser Pro Gly Lys
705
<210> 86
<211> 703
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V1-Fc-杵_b
<400> 86
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
145 150 155 160
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
165 170 175
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
180 185 190
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
195 200 205
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
210 215 220
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
225 230 235 240
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
245 250 255
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
260 265 270
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
275 280 285
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
290 295 300
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
305 310 315 320
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
325 330 335
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
340 345 350
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
355 360 365
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
370 375 380
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
385 390 395 400
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
405 410 415
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
420 425 430
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
435 440 445
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
450 455 460
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Asp Lys Thr His
465 470 475 480
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
485 490 495
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
500 505 510
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
515 520 525
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
530 535 540
Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser
545 550 555 560
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
565 570 575
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
580 585 590
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
595 600 605
Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
610 615 620
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
625 630 635 640
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
645 650 655
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
660 665 670
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
675 680 685
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
690 695 700
<210> 87
<211> 708
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V1-Fc-臼_a
<400> 87
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
145 150 155 160
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
165 170 175
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
180 185 190
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
195 200 205
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
210 215 220
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
225 230 235 240
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
245 250 255
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
260 265 270
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
275 280 285
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
290 295 300
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn Gly Ser
305 310 315 320
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
325 330 335
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
340 345 350
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
355 360 365
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
370 375 380
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
385 390 395 400
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
405 410 415
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
420 425 430
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
435 440 445
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
450 455 460
Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Ser Ser Gly Ser
465 470 475 480
Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
485 490 495
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
500 505 510
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
515 520 525
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
530 535 540
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr
545 550 555 560
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
565 570 575
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
580 585 590
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
595 600 605
Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
610 615 620
Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
625 630 635 640
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
645 650 655
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr
660 665 670
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
675 680 685
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
690 695 700
Ser Pro Gly Lys
705
<210> 88
<211> 641
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V1-Fc-臼_b
<400> 88
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
1 5 10 15
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
20 25 30
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
35 40 45
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
50 55 60
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
65 70 75 80
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn
85 90 95
Gly Ser Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
100 105 110
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
115 120 125
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
130 135 140
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
145 150 155 160
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
165 170 175
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
180 185 190
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
195 200 205
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
210 215 220
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
225 230 235 240
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Gly Asn
245 250 255
Gly Ser Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
260 265 270
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
275 280 285
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
290 295 300
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
305 310 315 320
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
325 330 335
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
340 345 350
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
355 360 365
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
370 375 380
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
385 390 395 400
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Gly Ser Gly Ser Asp Lys
405 410 415
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
420 425 430
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
435 440 445
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
450 455 460
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
465 470 475 480
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val
485 490 495
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
500 505 510
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
515 520 525
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr
530 535 540
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser
545 550 555 560
Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
565 570 575
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
580 585 590
Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys
595 600 605
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
610 615 620
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
625 630 635 640
Lys
<210> 89
<211> 715
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V2-Fc-杵_a
<400> 89
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Asn Gly
145 150 155 160
Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
165 170 175
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
180 185 190
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
195 200 205
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
210 215 220
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
225 230 235 240
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
245 250 255
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
260 265 270
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
275 280 285
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
290 295 300
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly
305 310 315 320
Asn Gly Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val
325 330 335
Leu Leu Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala
340 345 350
Gly Val Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu
355 360 365
Leu Val Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu
370 375 380
Leu Arg Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala
385 390 395 400
Leu His Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala
405 410 415
Leu Thr Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala
420 425 430
Phe Gly Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu
435 440 445
Gly Val His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu
450 455 460
Thr Gln Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly
465 470 475 480
Ser Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro
485 490 495
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
500 505 510
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
515 520 525
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
530 535 540
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
545 550 555 560
Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
565 570 575
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
580 585 590
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
595 600 605
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp
610 615 620
Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe
625 630 635 640
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
645 650 655
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
660 665 670
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
675 680 685
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
690 695 700
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
705 710 715
<210> 90
<211> 710
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V2-Fc-杵_b
<400> 90
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Asn Gly
145 150 155 160
Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
165 170 175
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
180 185 190
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
195 200 205
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
210 215 220
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
225 230 235 240
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
245 250 255
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
260 265 270
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
275 280 285
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
290 295 300
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly
305 310 315 320
Asn Gly Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val
325 330 335
Leu Leu Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala
340 345 350
Gly Val Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu
355 360 365
Leu Val Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu
370 375 380
Leu Arg Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala
385 390 395 400
Leu His Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala
405 410 415
Leu Thr Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala
420 425 430
Phe Gly Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu
435 440 445
Gly Val His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu
450 455 460
Thr Gln Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly
465 470 475 480
Ser Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
485 490 495
Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
500 505 510
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
515 520 525
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
530 535 540
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser
545 550 555 560
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
565 570 575
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
580 585 590
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
595 600 605
Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn
610 615 620
Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
625 630 635 640
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
645 650 655
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
660 665 670
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
675 680 685
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
690 695 700
Ser Leu Ser Pro Gly Lys
705 710
<210> 91
<211> 715
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V2-Fc-臼_a
<400> 91
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Asn Gly
145 150 155 160
Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
165 170 175
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
180 185 190
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
195 200 205
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
210 215 220
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
225 230 235 240
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
245 250 255
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
260 265 270
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
275 280 285
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
290 295 300
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly
305 310 315 320
Asn Gly Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val
325 330 335
Leu Leu Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala
340 345 350
Gly Val Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu
355 360 365
Leu Val Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu
370 375 380
Leu Arg Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala
385 390 395 400
Leu His Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala
405 410 415
Leu Thr Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala
420 425 430
Phe Gly Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu
435 440 445
Gly Val His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu
450 455 460
Thr Gln Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly
465 470 475 480
Ser Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro
485 490 495
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
500 505 510
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
515 520 525
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
530 535 540
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
545 550 555 560
Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
565 570 575
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
580 585 590
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
595 600 605
Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp
610 615 620
Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe
625 630 635 640
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
645 650 655
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
660 665 670
Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
675 680 685
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
690 695 700
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
705 710 715
<210> 92
<211> 710
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V2-Fc-臼_b
<400> 92
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Asn Gly
145 150 155 160
Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
165 170 175
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
180 185 190
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
195 200 205
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
210 215 220
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
225 230 235 240
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
245 250 255
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
260 265 270
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
275 280 285
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
290 295 300
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly
305 310 315 320
Asn Gly Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val
325 330 335
Leu Leu Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala
340 345 350
Gly Val Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu
355 360 365
Leu Val Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu
370 375 380
Leu Arg Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala
385 390 395 400
Leu His Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala
405 410 415
Leu Thr Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala
420 425 430
Phe Gly Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu
435 440 445
Gly Val His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu
450 455 460
Thr Gln Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly
465 470 475 480
Ser Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
485 490 495
Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
500 505 510
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
515 520 525
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
530 535 540
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser
545 550 555 560
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
565 570 575
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
580 585 590
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
595 600 605
Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn
610 615 620
Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile
625 630 635 640
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
645 650 655
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys
660 665 670
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
675 680 685
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
690 695 700
Ser Leu Ser Pro Gly Lys
705 710
<210> 93
<211> 707
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V3-Fc-杵_a
<400> 93
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly
145 150 155 160
Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro
165 170 175
Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly
180 185 190
Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala
195 200 205
Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala
210 215 220
Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu
225 230 235 240
Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro
245 250 255
Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg
260 265 270
Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr
275 280 285
Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val
290 295 300
Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly Met Phe
305 310 315 320
Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu Ser
325 330 335
Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly Leu
340 345 350
Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly Val
355 360 365
Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly Glu
370 375 380
Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg Ser
385 390 395 400
Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro Ala
405 410 415
Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu Leu
420 425 430
His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu Ala
435 440 445
Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu Gly
450 455 460
Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Ser Ser Gly Ser Cys
465 470 475 480
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
485 490 495
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
500 505 510
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
515 520 525
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
530 535 540
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
545 550 555 560
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
565 570 575
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
580 585 590
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
595 600 605
Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
610 615 620
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
625 630 635 640
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
645 650 655
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
660 665 670
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
675 680 685
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
690 695 700
Pro Gly Lys
705
<210> 94
<211> 702
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V3-Fc-杵_b
<400> 94
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly
145 150 155 160
Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro
165 170 175
Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly
180 185 190
Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala
195 200 205
Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala
210 215 220
Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu
225 230 235 240
Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro
245 250 255
Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg
260 265 270
Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr
275 280 285
Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val
290 295 300
Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly Met Phe
305 310 315 320
Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu Ser
325 330 335
Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly Leu
340 345 350
Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly Val
355 360 365
Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly Glu
370 375 380
Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg Ser
385 390 395 400
Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro Ala
405 410 415
Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu Leu
420 425 430
His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu Ala
435 440 445
Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu Gly
450 455 460
Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Asp Lys Thr His Thr
465 470 475 480
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
485 490 495
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
500 505 510
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
515 520 525
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
530 535 540
Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val
545 550 555 560
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
565 570 575
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
580 585 590
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
595 600 605
Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val
610 615 620
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
625 630 635 640
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
645 650 655
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
660 665 670
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
675 680 685
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
690 695 700
<210> 95
<211> 707
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V3-Fc-臼_a
<400> 95
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly
145 150 155 160
Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro
165 170 175
Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly
180 185 190
Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala
195 200 205
Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala
210 215 220
Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu
225 230 235 240
Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro
245 250 255
Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg
260 265 270
Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr
275 280 285
Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val
290 295 300
Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly Met Phe
305 310 315 320
Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu Ser
325 330 335
Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly Leu
340 345 350
Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly Val
355 360 365
Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly Glu
370 375 380
Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg Ser
385 390 395 400
Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro Ala
405 410 415
Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu Leu
420 425 430
His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu Ala
435 440 445
Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu Gly
450 455 460
Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Ser Ser Gly Ser Cys
465 470 475 480
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
485 490 495
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
500 505 510
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
515 520 525
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
530 535 540
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr
545 550 555 560
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
565 570 575
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
580 585 590
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
595 600 605
Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
610 615 620
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
625 630 635 640
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
645 650 655
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
660 665 670
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
675 680 685
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
690 695 700
Pro Gly Lys
705
<210> 96
<211> 702
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V3-Fc-臼_b
<400> 96
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly
145 150 155 160
Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro
165 170 175
Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly
180 185 190
Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala
195 200 205
Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala
210 215 220
Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu
225 230 235 240
Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro
245 250 255
Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg
260 265 270
Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr
275 280 285
Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val
290 295 300
Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly Met Phe
305 310 315 320
Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu Ser
325 330 335
Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly Leu
340 345 350
Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly Val
355 360 365
Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly Glu
370 375 380
Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg Ser
385 390 395 400
Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro Ala
405 410 415
Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu Leu
420 425 430
His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu Ala
435 440 445
Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu Gly
450 455 460
Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Asp Lys Thr His Thr
465 470 475 480
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
485 490 495
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
500 505 510
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
515 520 525
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
530 535 540
Lys Pro Arg Glu Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val
545 550 555 560
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
565 570 575
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
580 585 590
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro
595 600 605
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val
610 615 620
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
625 630 635 640
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
645 650 655
Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
660 665 670
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
675 680 685
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
690 695 700
<210> 97
<211> 698
<212> PRT
<213> 人工序列
<220>
<223> scLIGHT-Fc-杵_a
<400> 97
Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr
1 5 10 15
Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe
20 25 30
Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala
35 40 45
Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys
50 55 60
Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr
65 70 75 80
Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro
85 90 95
Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe
100 105 110
Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg
115 120 125
Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr
130 135 140
Phe Gly Ala Phe Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro
145 150 155 160
Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly
165 170 175
Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu
180 185 190
Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr
195 200 205
Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu
210 215 220
Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro
225 230 235 240
Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala
245 250 255
Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val
260 265 270
Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu
275 280 285
Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe
290 295 300
Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro Ala Ala His Leu
305 310 315 320
Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu Trp
325 330 335
Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr His Asp
340 345 350
Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys
355 360 365
Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr Ile
370 375 380
Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu Glu
385 390 395 400
Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Ser Ser
405 410 415
Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu Glu
420 425 430
Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val Arg
435 440 445
Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val Gly Ser
450 455 460
Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
465 470 475 480
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
485 490 495
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
500 505 510
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
515 520 525
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
530 535 540
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
545 550 555 560
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
565 570 575
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
580 585 590
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu
595 600 605
Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr
610 615 620
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
625 630 635 640
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
645 650 655
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
660 665 670
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
675 680 685
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
690 695
<210> 98
<211> 693
<212> PRT
<213> 人工序列
<220>
<223> scLIGHT-Fc-杵_b
<400> 98
Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr
1 5 10 15
Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe
20 25 30
Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala
35 40 45
Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys
50 55 60
Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr
65 70 75 80
Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro
85 90 95
Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe
100 105 110
Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg
115 120 125
Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr
130 135 140
Phe Gly Ala Phe Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro
145 150 155 160
Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly
165 170 175
Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu
180 185 190
Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr
195 200 205
Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu
210 215 220
Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro
225 230 235 240
Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala
245 250 255
Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val
260 265 270
Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu
275 280 285
Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe
290 295 300
Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro Ala Ala His Leu
305 310 315 320
Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu Trp
325 330 335
Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr His Asp
340 345 350
Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys
355 360 365
Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr Ile
370 375 380
Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu Glu
385 390 395 400
Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Ser Ser
405 410 415
Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu Glu
420 425 430
Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val Arg
435 440 445
Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val Gly Ser
450 455 460
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
465 470 475 480
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
485 490 495
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
500 505 510
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
515 520 525
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser
530 535 540
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
545 550 555 560
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
565 570 575
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
580 585 590
Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln
595 600 605
Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
610 615 620
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
625 630 635 640
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
645 650 655
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
660 665 670
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
675 680 685
Leu Ser Pro Gly Lys
690
<210> 99
<211> 698
<212> PRT
<213> 人工序列
<220>
<223> scLIGHT-Fc-臼_a
<400> 99
Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr
1 5 10 15
Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe
20 25 30
Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala
35 40 45
Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys
50 55 60
Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr
65 70 75 80
Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro
85 90 95
Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe
100 105 110
Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg
115 120 125
Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr
130 135 140
Phe Gly Ala Phe Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro
145 150 155 160
Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly
165 170 175
Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu
180 185 190
Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr
195 200 205
Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu
210 215 220
Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro
225 230 235 240
Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala
245 250 255
Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val
260 265 270
Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu
275 280 285
Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe
290 295 300
Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro Ala Ala His Leu
305 310 315 320
Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu Trp
325 330 335
Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr His Asp
340 345 350
Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys
355 360 365
Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr Ile
370 375 380
Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu Glu
385 390 395 400
Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Ser Ser
405 410 415
Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu Glu
420 425 430
Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val Arg
435 440 445
Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val Gly Ser
450 455 460
Gly Ser Ser Ser Gly Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
465 470 475 480
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
485 490 495
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
500 505 510
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
515 520 525
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
530 535 540
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
545 550 555 560
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
565 570 575
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
580 585 590
Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu
595 600 605
Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr
610 615 620
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
625 630 635 640
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
645 650 655
Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
660 665 670
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
675 680 685
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
690 695
<210> 100
<211> 693
<212> PRT
<213> 人工序列
<220>
<223> scLIGHT-Fc-臼_b
<400> 100
Glu Val Asn Pro Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr
1 5 10 15
Gly Ser Gly Gly Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe
20 25 30
Leu Arg Gly Leu Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala
35 40 45
Gly Tyr Tyr Tyr Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys
50 55 60
Pro Leu Gly Leu Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr
65 70 75 80
Pro Arg Tyr Pro Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro
85 90 95
Cys Gly Arg Ala Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe
100 105 110
Leu Gly Gly Val Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg
115 120 125
Val Leu Asp Glu Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr
130 135 140
Phe Gly Ala Phe Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro
145 150 155 160
Ala Ala His Leu Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly
165 170 175
Pro Leu Leu Trp Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu
180 185 190
Ser Tyr His Asp Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr
195 200 205
Ile Tyr Ser Lys Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu
210 215 220
Ala Ser Thr Ile Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro
225 230 235 240
Glu Glu Leu Glu Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala
245 250 255
Thr Ser Ser Ser Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val
260 265 270
Val His Leu Glu Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu
275 280 285
Arg Leu Val Arg Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe
290 295 300
Met Val Gly Ser Gly Ser Gly Asn Gly Ser Asn Pro Ala Ala His Leu
305 310 315 320
Thr Gly Ala Asn Ser Ser Leu Thr Gly Ser Gly Gly Pro Leu Leu Trp
325 330 335
Glu Thr Gln Leu Gly Leu Ala Phe Leu Arg Gly Leu Ser Tyr His Asp
340 345 350
Gly Ala Leu Val Val Thr Lys Ala Gly Tyr Tyr Tyr Ile Tyr Ser Lys
355 360 365
Val Gln Leu Gly Gly Val Gly Cys Pro Leu Gly Leu Ala Ser Thr Ile
370 375 380
Thr His Gly Leu Tyr Lys Arg Thr Pro Arg Tyr Pro Glu Glu Leu Glu
385 390 395 400
Leu Leu Val Ser Gln Gln Ser Pro Cys Gly Arg Ala Thr Ser Ser Ser
405 410 415
Arg Val Trp Trp Asp Ser Ser Phe Leu Gly Gly Val Val His Leu Glu
420 425 430
Ala Gly Glu Glu Val Val Val Arg Val Leu Asp Glu Arg Leu Val Arg
435 440 445
Leu Arg Asp Gly Thr Arg Ser Tyr Phe Gly Ala Phe Met Val Gly Ser
450 455 460
Gly Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
465 470 475 480
Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
485 490 495
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
500 505 510
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
515 520 525
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ser Ser
530 535 540
Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
545 550 555 560
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
565 570 575
Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
580 585 590
Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln
595 600 605
Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
610 615 620
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
625 630 635 640
Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu
645 650 655
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
660 665 670
Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser
675 680 685
Leu Ser Pro Gly Lys
690
<210> 101
<211> 683
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-hc-scCD27L-RBD
<400> 101
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Glu Ser
225 230 235 240
Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln
245 250 255
Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser
260 265 270
Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg
275 280 285
Asp Gly Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser
290 295 300
Ser Thr Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile
305 310 315 320
Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His
325 330 335
Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly
340 345 350
Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn
355 360 365
Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly Ser Gly
370 375 380
Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu Leu Gln
385 390 395 400
Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly
405 410 415
Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys
420 425 430
Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His Ile Gln
435 440 445
Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His His Pro
450 455 460
Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser
465 470 475 480
Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg
485 490 495
Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly
500 505 510
Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln
515 520 525
Trp Val Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly
530 535 540
Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp
545 550 555 560
Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu
565 570 575
His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly
580 585 590
Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr
595 600 605
Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser
610 615 620
Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly
625 630 635 640
Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr
645 650 655
Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp
660 665 670
Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro
675 680
<210> 102
<211> 681
<212> PRT
<213> 人工序列
<220>
<223> aPDL1-hc-scCD27L-RBD
<400> 102
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Glu Ser Leu Gly
225 230 235 240
Trp Asp Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp
245 250 255
Pro Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu
260 265 270
His Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly
275 280 285
Ile Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr
290 295 300
Thr Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser
305 310 315 320
Pro Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly
325 330 335
Cys Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr
340 345 350
Leu Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp
355 360 365
Glu Thr Phe Phe Gly Val Gln Trp Val Arg Pro Gly Ser Gly Ser Gly
370 375 380
Asn Gly Ser Glu Ser Leu Gly Trp Asp Val Ala Glu Leu Gln Leu Asn
385 390 395 400
His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro
405 410 415
Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln
420 425 430
Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr
435 440 445
Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr
450 455 460
Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu
465 470 475 480
Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr
485 490 495
Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu
500 505 510
Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val
515 520 525
Arg Pro Gly Ser Gly Ser Gly Asn Gly Ser Glu Ser Leu Gly Trp Asp
530 535 540
Val Ala Glu Leu Gln Leu Asn His Thr Gly Pro Gln Gln Asp Pro Arg
545 550 555 560
Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly
565 570 575
Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr
580 585 590
Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala
595 600 605
Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala
610 615 620
Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr
625 630 635 640
Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys
645 650 655
Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr
660 665 670
Phe Phe Gly Val Gln Trp Val Arg Pro
675 680
<210> 103
<211> 620
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-hc-scGITRL-RBD
<400> 103
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
115 120 125
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
130 135 140
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
145 150 155 160
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
165 170 175
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
180 185 190
Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
195 200 205
His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser
210 215 220
Cys Asp Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Gln Pro
225 230 235 240
Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser
245 250 255
Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile
260 265 270
Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala
275 280 285
Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys
290 295 300
Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly
305 310 315 320
Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn
325 330 335
Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu
340 345 350
Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn Gly Ser
355 360 365
Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
370 375 380
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
385 390 395 400
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
405 410 415
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
420 425 430
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
435 440 445
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
450 455 460
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
465 470 475 480
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn
485 490 495
Gly Ser Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp
500 505 510
Gln Met Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp
515 520 525
Lys Leu Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val
530 535 540
Ala Pro Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu
545 550 555 560
Tyr Lys Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile
565 570 575
Gln Asn Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp
580 585 590
Leu Ile Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp
595 600 605
Gly Ile Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
610 615 620
<210> 104
<211> 618
<212> PRT
<213> 人工序列
<220>
<223> aPDL1-hc-scGITRL-RBD
<400> 104
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
145 150 155 160
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
180 185 190
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
210 215 220
Lys Thr His Gly Ser Gly Ser Ser Ser Ser Ser Ser Gln Pro Cys Met
225 230 235 240
Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser Ser Glu
245 250 255
Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile Leu Gln
260 265 270
Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala Asn Tyr
275 280 285
Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys Asp Met
290 295 300
Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly Gly Thr
305 310 315 320
Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn Ser Glu
325 330 335
His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu Leu Ala
340 345 350
Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn Gly Ser Glu Pro
355 360 365
Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met Ala Ser
370 375 380
Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu Glu Ile
385 390 395 400
Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro Asn Ala
405 410 415
Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys Asn Lys
420 425 430
Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn Val Gly
435 440 445
Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile Phe Asn
450 455 460
Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile Ile Leu
465 470 475 480
Leu Ala Asn Pro Gln Phe Ile Ser Gly Ser Gly Ser Gly Asn Gly Ser
485 490 495
Glu Pro Cys Met Ala Lys Phe Gly Pro Leu Pro Ser Lys Trp Gln Met
500 505 510
Ala Ser Ser Glu Pro Pro Cys Val Asn Lys Val Ser Asp Trp Lys Leu
515 520 525
Glu Ile Leu Gln Asn Gly Leu Tyr Leu Ile Tyr Gly Gln Val Ala Pro
530 535 540
Asn Ala Asn Tyr Asn Asp Val Ala Pro Phe Glu Val Arg Leu Tyr Lys
545 550 555 560
Asn Lys Asp Met Ile Gln Thr Leu Thr Asn Lys Ser Lys Ile Gln Asn
565 570 575
Val Gly Gly Thr Tyr Glu Leu His Val Gly Asp Thr Ile Asp Leu Ile
580 585 590
Phe Asn Ser Glu His Gln Val Leu Lys Asn Asn Thr Tyr Trp Gly Ile
595 600 605
Ile Leu Leu Ala Asn Pro Gln Phe Ile Ser
610 615
<210> 105
<211> 479
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V2-RBD
<400> 105
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Asn Gly
145 150 155 160
Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
165 170 175
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
180 185 190
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
195 200 205
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
210 215 220
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
225 230 235 240
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
245 250 255
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
260 265 270
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
275 280 285
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
290 295 300
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly
305 310 315 320
Asn Gly Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val
325 330 335
Leu Leu Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala
340 345 350
Gly Val Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu
355 360 365
Leu Val Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu
370 375 380
Leu Arg Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala
385 390 395 400
Leu His Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala
405 410 415
Leu Thr Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala
420 425 430
Phe Gly Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu
435 440 445
Gly Val His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu
450 455 460
Thr Gln Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu
465 470 475
<210> 106
<211> 471
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V3-RBD
<400> 106
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly
145 150 155 160
Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro
165 170 175
Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly
180 185 190
Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala
195 200 205
Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala
210 215 220
Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu
225 230 235 240
Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro
245 250 255
Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg
260 265 270
Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr
275 280 285
Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val
290 295 300
Leu Gly Leu Phe Arg Val Thr Pro Glu Gly Ser Gly Ser Gly Met Phe
305 310 315 320
Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu Ser
325 330 335
Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly Leu
340 345 350
Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly Val
355 360 365
Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly Glu
370 375 380
Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg Ser
385 390 395 400
Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro Ala
405 410 415
Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu Leu
420 425 430
His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu Ala
435 440 445
Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu Gly
450 455 460
Leu Phe Arg Val Thr Pro Glu
465 470
<210> 107
<211> 476
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V4-RBD
<400> 107
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Gly Ser Gly Asn Gly Ser
145 150 155 160
Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile
165 170 175
Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser
180 185 190
Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val
195 200 205
Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg
210 215 220
Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu
225 230 235 240
Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val
245 250 255
Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe
260 265 270
Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His
275 280 285
Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly
290 295 300
Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro Gly Ser Gly Asn Gly
305 310 315 320
Ser Gly Ser Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu
325 330 335
Ile Asp Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val
340 345 350
Ser Leu Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val
355 360 365
Val Ala Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg
370 375 380
Arg Val Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His
385 390 395 400
Leu Gln Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr
405 410 415
Val Asp Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly
420 425 430
Phe Gln Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val
435 440 445
His Leu His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln
450 455 460
Gly Ala Thr Val Leu Gly Leu Phe Arg Val Thr Pro
465 470 475
<210> 108
<211> 468
<212> PRT
<213> 人工序列
<220>
<223> scCD137L-V5-RBD
<400> 108
Gln Gly Met Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp
1 5 10 15
Gly Pro Leu Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu
20 25 30
Thr Gly Gly Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala
35 40 45
Lys Ala Gly Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val
50 55 60
Val Ala Gly Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln
65 70 75 80
Pro Leu Arg Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp
85 90 95
Leu Pro Pro Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln
100 105 110
Gly Arg Leu Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu
115 120 125
His Thr Glu Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala
130 135 140
Thr Val Leu Gly Leu Phe Arg Val Thr Pro Gly Ser Gly Ser Gly Met
145 150 155 160
Phe Ala Gln Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu
165 170 175
Ser Trp Tyr Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly
180 185 190
Leu Ser Tyr Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly
195 200 205
Val Tyr Tyr Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly
210 215 220
Glu Gly Ser Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg
225 230 235 240
Ser Ala Ala Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro
245 250 255
Ala Ser Ser Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu
260 265 270
Leu His Leu Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu
275 280 285
Ala Arg Ala Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu
290 295 300
Gly Leu Phe Arg Val Thr Pro Gly Ser Gly Ser Gly Met Phe Ala Gln
305 310 315 320
Leu Val Ala Gln Asn Val Leu Leu Ile Asp Gly Pro Leu Ser Trp Tyr
325 330 335
Ser Asp Pro Gly Leu Ala Gly Val Ser Leu Thr Gly Gly Leu Ser Tyr
340 345 350
Lys Glu Asp Thr Lys Glu Leu Val Val Ala Lys Ala Gly Val Tyr Tyr
355 360 365
Val Phe Phe Gln Leu Glu Leu Arg Arg Val Val Ala Gly Glu Gly Ser
370 375 380
Gly Ser Val Ser Leu Ala Leu His Leu Gln Pro Leu Arg Ser Ala Ala
385 390 395 400
Gly Ala Ala Ala Leu Ala Leu Thr Val Asp Leu Pro Pro Ala Ser Ser
405 410 415
Glu Ala Arg Asn Ser Ala Phe Gly Phe Gln Gly Arg Leu Leu His Leu
420 425 430
Ser Ala Gly Gln Arg Leu Gly Val His Leu His Thr Glu Ala Arg Ala
435 440 445
Arg His Ala Trp Gln Leu Thr Gln Gly Ala Thr Val Leu Gly Leu Phe
450 455 460
Arg Val Thr Pro
465
<210> 109
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> Fc-N297S
<400> 109
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
35 40 45
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
50 55 60
Glu Gln Tyr Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
65 70 75 80
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
85 90 95
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
100 105 110
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
115 120 125
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
130 135 140
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
145 150 155 160
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
165 170 175
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
180 185 190
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
195 200 205
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 110
<211> 218
<212> PRT
<213> 人工序列
<220>
<223> FC-WT
<400> 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
1 5 10 15
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
20 25 30
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
35 40 45
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
50 55 60
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
65 70 75 80
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
85 90 95
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
100 105 110
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu
115 120 125
Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
130 135 140
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
145 150 155 160
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
165 170 175
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
180 185 190
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
195 200 205
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
210 215
<210> 111
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1CH2CH3-RF
<400> 111
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
180 185 190
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
195 200 205
Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
210 215 220
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 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 Arg Phe Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 112
<211> 330
<212> PRT
<213> 人工序列
<220>
<223> IGG1-CH1CH2sCH3-RF
<400> 112
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
65 70 75 80
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95
Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys
100 105 110
Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
115 120 125
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
145 150 155 160
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
165 170 175
Glu Gln Tyr Ser 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 Arg Phe Thr
305 310 315 320
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 113
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> aCEA-a-VH
<400> 113
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 Thr Ser Gly Phe Thr Phe Thr Asp Tyr
20 25 30
Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu
35 40 45
Gly Phe Ile Gly Asn Lys Ala Asn Gly Tyr Thr Thr Glu Tyr Ser Ala
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Ser Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Thr Leu Gln Ala Glu Asp Ser Ala Ile Tyr
85 90 95
Tyr Cys Thr Arg Asp Arg Gly Leu Arg Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 114
<211> 107
<212> PRT
<213> 人工序列
<220>
<223> aCEA-a-VL
<400> 114
Gln Thr Val Leu Thr Gln Ser Pro Ser Ser Leu Ser Val Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Ser Val Thr Tyr Ile
20 25 30
His Trp Tyr Gln Gln Lys Pro Gly Leu Ala Pro Lys Ser Leu Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu
65 70 75 80
Asp Ile Ala Thr Tyr Tyr Cys Gln His Trp Ser Ser Lys Pro Pro Thr
85 90 95
Phe Gly Gln Gly Thr Lys Val Glu Val Lys Arg
100 105
<210> 115
<211> 121
<212> PRT
<213> 人工序列
<220>
<223> aCEA-b-VH
<400> 115
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Gly Phe Ile Arg Asn Lys Ala Asn Gly Gly Thr Thr Glu Tyr Ala Ala
50 55 60
Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr
65 70 75 80
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
85 90 95
Tyr Cys Ala Arg Asp Arg Gly Leu Arg Phe Tyr Phe Asp Tyr Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 116
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> aCEA-b-VL
<400> 116
Gln Ala Val Leu Thr Gln Pro Ala Ser Leu Ser Ala Ser Pro Gly Ala
1 5 10 15
Ser Ala Ser Leu Thr Cys Thr Leu Arg Arg Gly Ile Asn Val Gly Ala
20 25 30
Tyr Ser Ile Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Pro Pro Gln Tyr
35 40 45
Leu Leu Arg Tyr Lys Ser Asp Ser Asp Lys Gln Gln Gly Ser Gly Val
50 55 60
Ser Ser Arg Phe Ser Ala Ser Lys Asp Ala Ser Ala Asn Ala Gly Ile
65 70 75 80
Leu Leu Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys
85 90 95
Met Ile Trp His Ser Gly Ala Ser Ala Val Phe Gly Gly Gly Thr Lys
100 105 110
Leu Thr Val Leu
115
<210> 117
<211> 122
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-VH
<400> 117
Glu Val Gln Leu Val 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 Ser Phe Ser Asp His
20 25 30
Tyr Trp Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
35 40 45
Val Ala Cys Ile Tyr Thr Ala Asp Ser Asp Ser Tyr Tyr Ala Asp Ser
50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Ser Ser Lys Asn Thr Leu
65 70 75 80
Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr
85 90 95
Cys Ala Arg Asn Gly Ala Tyr Ala Gly Gly Pro Tyr Gly Asp Leu Trp
100 105 110
Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120
<210> 118
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> aCD95L-VL
<400> 118
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 Ser Ile Arg Thr Ser
20 25 30
Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Lys Ala Ser Asp Leu Pro Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Ser Tyr Asp Phe Arg Asp Thr
85 90 95
Ile Asn Asn Gly His Ser Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 110
Arg
<210> 119
<211> 416
<212> PRT
<213> 人工序列
<220>
<223> scCD27L-V2-RBD
<400> 119
Asp Val Ala Glu Leu Gln Leu Asp His Thr Gly Pro Gln Gln Asp Pro
1 5 10 15
Arg Leu Tyr Trp Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His
20 25 30
Gly Pro Glu Leu Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile
35 40 45
Tyr Met Val His Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr
50 55 60
Ala Ser Arg His His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro
65 70 75 80
Ala Ser Arg Ser Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys
85 90 95
Thr Ile Ala Ser Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu
100 105 110
Cys Thr Asn Leu Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu
115 120 125
Thr Phe Phe Gly Val Gln Trp Val Gly Ser Gly Ser Asp Val Ala Glu
130 135 140
Leu Gln Leu Asp His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp
145 150 155 160
Gln Gly Gly Pro Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu
165 170 175
Asp Lys Gly Gln Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His
180 185 190
Ile Gln Val Thr Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His
195 200 205
His Pro Thr Thr Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser
210 215 220
Ile Ser Leu Leu Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser
225 230 235 240
Gln Arg Leu Thr Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu
245 250 255
Thr Gly Thr Leu Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly
260 265 270
Val Gln Trp Val Gly Ser Gly Ser Asp Val Ala Glu Leu Gln Leu Asp
275 280 285
His Thr Gly Pro Gln Gln Asp Pro Arg Leu Tyr Trp Gln Gly Gly Pro
290 295 300
Ala Leu Gly Arg Ser Phe Leu His Gly Pro Glu Leu Asp Lys Gly Gln
305 310 315 320
Leu Arg Ile His Arg Asp Gly Ile Tyr Met Val His Ile Gln Val Thr
325 330 335
Leu Ala Ile Cys Ser Ser Thr Thr Ala Ser Arg His His Pro Thr Thr
340 345 350
Leu Ala Val Gly Ile Cys Ser Pro Ala Ser Arg Ser Ile Ser Leu Leu
355 360 365
Arg Leu Ser Phe His Gln Gly Cys Thr Ile Ala Ser Gln Arg Leu Thr
370 375 380
Pro Leu Ala Arg Gly Asp Thr Leu Cys Thr Asn Leu Thr Gly Thr Leu
385 390 395 400
Leu Pro Ser Arg Asn Thr Asp Glu Thr Phe Phe Gly Val Gln Trp Val
405 410 415

Claims (42)

1.一种双特异性TNF超家族融合蛋白组装体，其至少包含
(g)单链TNF-SF受体结合结构域，其融合至
(h)第一肽接头，其融合至
(i)第一(异)二聚化结构域，和
(j)抗原结合或相互作用蛋白质部分，其融合至
(k)第二肽接头，其融合至
(l)第二(异)二聚化结构域。
2.根据权利要求1所述的双特异性TNF超家族融合蛋白组装体，其中
二聚化结构域c)和f)的异二聚化由CH3结构域执行，所述CH3结构域独立地选自IgGl或IgG2或IgG3或IgG4或IgA或IgD衍生的CH3结构域，并由铰链链间半胱氨酸额外稳定化。
3.根据权利要求1至2中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中所述单链TNF-SF受体结合结构域a)选自由单链CD40L、单链GITRL、单链OX40L、单链LIGHT、单链TL1A、单链CD137L，单链CD27L或单链TRAIL组成的组。
4.根据权利要求3所述的双特异性TNF超家族融合蛋白组装体，
其中所述单链TNF-SF受体结合结构域a)选自由SEQ ID NO:69-73和105-108或其变体组成的组。
5.根据权利要求1-4中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中所述抗原结合或相互作用蛋白质部分d)是IgG抗体衍生的重链和轻链、或抗体片段，所述抗体片段选自由Fv、Fab、Fab'、Fab'-SH、F(ab')2；双抗体、三抗体、四抗体、交叉Fab片段；或单链抗体(例如scFv)或单结构域抗体组成的组。
6.根据权利要求1-5中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中所述抗原结合或相互作用蛋白质部分d)是IgG抗体衍生的重链和轻链，具有选自由抗PD-L1、抗CD137、抗间皮素、抗CD25、抗PD-1、抗CEA或抗CD95L组成的组的特异性。
7.根据权利要求1-6中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中所述抗原结合或相互作用蛋白质部分d)是IgG抗体衍生的重链和轻链，其具有选自由以下组成的组的特异性：抗PD-L1(SEQ-ID:74aPD-L1-VH，SEQ-ID:75aPD-L1-VL)、或抗CD137(SEQ-ID:59aCD137-VH，SEQ-ID:60aCD137-VL)、或抗间皮素(SEQ-ID:61aMeso-VH，SEQ-ID:62aMeso-VL)、或抗CD25(SEQ-ID:63aCD25-VH，SEQ-ID:64aCD25-VL)、或抗PD-1(SEQ-ID:65aPD1-a-VH，SEQ-ID:66aPD1-a-VL，SEQ-ID:67aPD1-b-VH，SEQ-ID:68aPD1-b-VL)、或抗CEA(SEQ-ID:113aCEA-a-VH，SEQ-ID:114aCEA-a-VL，SEQ-ID:115aCEA-b-VH，SEQ-ID:116aCEA-b-VL；SEQ-ID:56aCEA-hc-scCD40L-RBD/SEQ-ID:51aCEA-LC)。
8.根据前述权利要求中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:33(scCD40L-Fc-杵_b)或SEQ-ID:84(scCD40L-Fc-杵_c)的scCD40L-RBD，以及其中d)的IgG衍生的抗体包含SEQ-ID:55(aPD-L1-LC)和/或SEQ-ID:54(aPD-L1-HC-RF-臼)。
9.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:39(scCD27L-Fc-杵_b)的scCD27L-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:55(aPD-L1-LC)和/或SEQ-ID:54(aPD-L1-HC-RF-臼)。
10.根据前述权利要求1-7所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:41(scGITRL-Fc-杵_b)的scGITRL-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:55(aPD-L1-LC)和/或SEQ-ID:54(aPD-L1-HC-RF-臼)。
11.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:86(scCD137L-V1-Fc-杵_b)或SEQ-ID:90(scCD137L-V2-Fc-杵_b)或SEQ-ID:94(scCD137L-V3-Fc-杵_b)的scCD137L-Fc，或者其中scCD137L-Fc包含SEQ-ID:107(scCD137L-V4-RBD)或SEQ-ID:108(scCD137L-V5-RBD)的TNFSF模块，以及
其中d)的IgG衍生的抗体包含SEQ-ID:55(aPD-L1-LC)和/或SEQ-ID:54(aPD-L1-HC-RF-臼)。
12.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:98(scLIGHT-Fc-杵_b)的scLIGHT-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:55(aPD-L1-LC)和/或SEQ-ID:54(aPD-L1-HC-RF-臼)。
13.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:33(scCD40L-Fc-杵_b)或SEQ-ID:84(scCD40L-Fc-杵_c)的scCD40L RBD，以及其中d)的IgG衍生的抗体包含SEQ-ID:47(aCD95L-LC)和/或SEQ-ID:46(aCD95L-HC-RF-臼)。
14.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:39(scCD27L-Fc-杵_b)的scCD27L-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:47(aCD95L-LC)和/或SEQ-ID:46(aCD95L-HC-RF-臼)。
15.根据前述权利要求1-7所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:41(scGITRL-Fc-杵_b)的scGITRL-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:47(aCD95L-LC)和/或SEQ-ID:46(aCD95L-HC-RF-臼)。
16.根据前述权利要求1-7所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:86(scCD137L-V1-Fc-杵_b)或SEQ-ID:90(scCD137L-V2-Fc-杵_b)或SEQ-ID:94(scCD137L-V3-Fc-杵_b)的scCD137L-Fc，或者其中scCD137L-Fc包含SEQ-ID:107(scCD137L-V4-RBD)或SEQ-ID:108(scCD137L-V5-RBD)的TNFSF模块，以及
其中d)的IgG衍生的抗体包含SEQ-ID:47(aCD95L-LC)和/或SEQ-ID:46(aCD95L-HC-RF-臼)。
17.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:98(scLIGHT-Fc-杵_b)的scLIGHT-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:47(aCD95L-LC)和/或SEQ-ID:46(aCD95L-HC-RF-臼)。
18.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:33(scCD40L-Fc-杵_b)或SEQ-ID:84(scCD40L-Fc-杵_c)的scCD40L RBD，以及其中d)的IgG衍生的抗体包含SEQ-ID:51(aCEA-LC)和/或SEQ-ID:50(aCEA-HC-RF-臼)。
19.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:39(scCD27L-Fc-杵_b)的scCD27L-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:51(aCEA-LC)和/或SEQ-ID:50(aCEA-HC-RF-臼)。
20.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:41(scGITRL-Fc-杵_b)的scGITRL-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:51(aCEA-LC)和/或SEQ-ID:50(aCEA-HC-RF-臼)。
21.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:86(scCD137L-V1-Fc-杵_b)或SEQ-ID:90(scCD137L-V2-Fc-杵_b)或SEQ-ID:94(scCD137L-V3-Fc-杵_b)的scCD137L-Fc，或者其中scCD137L-Fc包含SEQ-ID:107(scCD137L-V4-RBD)或SEQ-ID:108(scCD137L-V5-RBD)的TNFSF模块，以及
其中d)的IgG衍生的抗体包含SEQ-ID:51(aCEA-LC)和/或SEQ-ID:50(aCEA-HC-RF-臼)。
22.根据前述权利要求1-7中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中三价单链TNFSF-RBD-Fc是SEQ-ID:98(scLIGHT-Fc-杵_b)的scLIGHT-Fc，以及其中d)的IgG衍生的抗体包含SEQ-ID:51(aCEA-LC)和/或SEQ-ID:50(aCEA-HC-RF-臼)。
23.根据前述权利要求中任一项所述的双特异性TNF超家族融合蛋白组装体，
其中二聚化CH3结构域选自表示为SEQ-ID:44、SEQ-ID:45、SEQ-ID:48、SEQ-ID:49、SEQ-ID:52、SEQ-ID:53、SEQ-ID:28、SEQ-ID:29、SEQ-ID:30、SEQ-ID:31、SEQ-ID:111、SEQ-ID:112、SEQ-ID:20、SEQ-ID:21、SEQ-ID:22、SEQ-ID:23、SEQ-ID:24、SEQ-ID:25、SEQ-ID:109或SEQ-ID:110的部分的CH3结构域。
24.一种多特异性TNF家族融合蛋白组装体，其至少包含
(a)单链TNF-SF受体结合结构域，其融合至
(b)第一肽接头，其融合至
(c)第一异二聚化结构域，和
(d)和第二单链TNF-SF受体结合结构域，其融合至
(e)第二肽接头，其融合至
(f)第二异二聚化结构域。
25.一种多特异性TNF家族融合蛋白组装体，其至少包含
(a)抗体的功能性Fab结构域，其融合至
(b)单链TNF-SF受体结合结构域，
(c)其中Fab片段(a)的恒定重链结构域的c-末端通过肽接头融合至单链TNF-SF受体结合。
26.根据权利要求25所述的多特异性TNF家族融合蛋白组装体，其中c)的肽接头选自由SEQ-ID NO:13-19组成的组。
27.根据权利要求25或26所述的多特异性免疫调节剂，其将抗PD-L1(aPDL1)靶向与CD40激动相结合，由此成熟蛋白组装体包含SEQ-ID:58(aPDL1-hc-scCD40L-RBD)和/或SEQ-ID:55(aPD-L1-LC)。
28.根据权利要求25或26所述的多特异性免疫调节剂，其将抗PD-L1(aPDL1)靶向与CD27激动相结合，由此成熟蛋白组装体包含SEQ-ID:102(aPDL1-hc-scCD27L-RBD)和/或SEQ-ID:55(aPD-L1-LC)或其变体，特别地变体包含SEQ-ID:119作为scCD27L-RBD的变体。
29.根据权利要求25或26所述的多特异性免疫调节剂，其将抗PD-L1(aPDL1)靶向与GITR激动相结合，由此成熟蛋白组装体包含SEQ-ID:104(aPDL1-hc-scGITRL-RBD)和/或SEQ-ID:55(aPD-L1-LC)。
30.根据权利要求25或26所述的多特异性免疫调节剂，其将抗CD95L(aCD95L)靶向与CD40激动相结合，由此成熟蛋白组装体包含SEQ-ID:57(aCD95L-hc-scCD40L-RBD)和/或SEQ-ID:47(aCD95L-LC)。
31.根据权利要求25或26所述的多特异性免疫调节剂，其将抗CD95L(aCD95L)靶向与CD27激动相结合，由此成熟蛋白组装体包含SEQ-ID:101(aCD95L-hc-scCD27L-RBD)和/或SEQ-ID:47(aCD95L-LC)或其变体，特别地变体包含SEQ-ID:119作为scCD27L-RBD的变体。
32.根据权利要求25或26所述的多特异性免疫调节剂，其将抗CD95L(aCD95L)靶向与GITR激动相结合，由此成熟蛋白组装体包含SEQ-ID:103(aCD95L-hc-scGITRL-RBD)和/或SEQ-ID:47(aCD95L-LC)。
33.根据权利要求25或26的多特异性免疫调节剂，其将抗CD95L(aCD95L)靶向与CD137激动或与HVEM/LTbR-激动相结合，由此成熟蛋白组装体包含SEQ-ID:72(scCD137L-RBD)、SEQ-ID:105(scCD137L-V2-RBD)、SEQ-ID:106(scCD137L-V3-RBD)、SEQ-ID:107(scCD137L-V4-RBD)、SEQ-ID:108(scCD137L-V5-RBD)或SEQ-ID:73(scLIGHT-RBD)、SEQ-ID:103(aCD95L-hc-scGITRL-RBD)和SEQ-ID:47(aCD95L-LC)。
34.根据权利要求25-33中任一项所述的多特异性免疫调节剂，其中Fab靶向结构域选自抗CD137靶向(SEQ-ID:59aCD137-VH，SEQ-ID:60aCD137-VL)、或抗间皮素靶向(SEQ-ID:61aMeso-VH、SEQ-ID:62aMeso-VL)、或抗CD25靶向(SEQ-ID:63aCD25-VH、SEQ-ID:64aCD25-VL)、或抗PD-1靶向(SEQ-ID:65aPD1-a-VH，SEQ-ID:66aPD1-a-VL，SEQ-ID:67aPD1-b-VH，SEQ-ID:68aPD1-b-VL)、或抗CEA靶向(SEQ-ID:113aCEA-a-VH，SEQ-ID:114aCEA-a-VL，SEQ-ID:115aCEA-b-VH，SEQ-ID:116aCEA-b-VL；SEQ-ID:56aCEA-hc-scCD40L-RBD/SEQ-ID:51aCEA-LC)。
35.根据权利要求25-34中任一项所述的多特异性免疫调节剂，其中scTNFSF RBD选自根据SEQ ID NO:69-73和105-108的scCD40L、scGITRL、scOX40L、scLIGHT、scTL1A、scCD137L、scCD27L或scTRAIL或其变体作为激动结构域。
36.一种核酸分子，其编码权利要求1-23中任一项的部分a)-c)蛋白质部分，和/或一种核酸分子，其编码权利要求1-23中任一项的部分d)-e)蛋白质部分。
37.一种核酸，其编码权利要求24部分a)-c)的蛋白质部分，和/或一种核酸，其编码权利要求24部分d)-e)的蛋白质部分。
38.一种核酸，其编码至少一种根据权利要求25-35中任一项所述的蛋白质。
39.根据权利要求36-38中任一项所述的核酸分子，其用于蛋白质或蛋白组装体的表达或共表达。
40.一种载体，其包含至少一种根据权利要求36-39中任一项所述的核酸分子。
41.一种宿主细胞，其包含根据权利要求36-40中任一项所述的核酸。
42.一种药物组合物，其至少包含根据权利要求1-35中任一项所述的多特异性蛋白组装体或多特异性蛋白质。
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