CN115151279A - 基于细胞因子的可生物活化药物及其使用方法 - Google Patents

基于细胞因子的可生物活化药物及其使用方法 Download PDF

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CN115151279A
CN115151279A CN202080097196.4A CN202080097196A CN115151279A CN 115151279 A CN115151279 A CN 115151279A CN 202080097196 A CN202080097196 A CN 202080097196A CN 115151279 A CN115151279 A CN 115151279A
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construct
seq
amino acid
various embodiments
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李跃升
芮凌云
徐静
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Keyou Gene Co
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Keyou Gene Co
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Abstract

本公开内容提供了一种基于细胞因子的可生物活化的药物构建体(“VitoKine”)平台，该平台旨在降低基于全身性机制的毒性，并使蛋白和细胞因子诸如IL‑15和IL‑2在治疗癌症、自身免疫性疾病、炎性疾病、病毒感染、移植和各种其他紊乱中具有更广泛的治疗用途。本发明的新的VitoKine构建体包含：1)组织或疾病部位靶向部分D1结构域(“D1”)、2)可生物活化部分D2结构域(“D2”)和遮蔽部分D3结构域(“D3”)。重要的是，因为VitoKine构建体的“活性部分”将保持惰性，直到被病变组织中上调的蛋白酶在局部活化，这将限制活性部分与非病变细胞和组织的外周中或细胞表面上的受体或靶的结合，以防止途径的过度活化并减少不期望的“组织外”“中靶”毒性。此外，在蛋白酶活化之前，VitoKine活性部分的惰性将显著降低潜在的抗原吸收或靶吸收，并从而延长体内半衰期且导致改进的生物分布、生物可利用度和治疗功效。

Description

基于细胞因子的可生物活化药物及其使用方法
背景
许多细胞因子已被评价为用于治疗疾病的潜在治疗剂。然而，它们对身体免疫系统的全身性过度刺激或过度抑制严重阻碍了它们的开发和临床用途。
白介素-2(IL-2)和白介素-15(IL-15)共享共有受体组分(γc和IL-2Rβ)和信号传导途径，并具有若干相似的功能。两种细胞因子都刺激T细胞的增殖；诱导细胞毒性T淋巴细胞(CTL)的产生；促进B细胞的增殖和免疫球蛋白的合成；以及诱导自然杀伤(NK)细胞的产生和持续。基于大量临床前研究以及多项临床评估，这两种细胞因子被认为是癌症、自身免疫性紊乱、炎性紊乱、移植和各种其他紊乱的潜在有价值的治疗剂。重组IL-2已被批准用于患有转移性肾细胞癌和恶性黑素瘤的患者。对于IL-15，有几项正在进行的肿瘤学临床试验，但尚未批准使用。此外，IL-2和IL-15两者分别具有第三个独特的非信号传导受体α-亚基：IL-2Rα(也称为CD25)或IL-15Rα，这可能有助于它们不同的受体特异性和生物功能。
重组人类IL-2是一种正被用于转移性黑素瘤和肾癌的有效的免疫疗法，约10％的患者有持久的响应。然而，短半衰期和严重毒性限制了IL-2的最佳给药。此外，IL-2还以较高的亲和力与其异源三聚体受体IL-2Rαβγ结合，优先扩增表达高组成型IL-2Rα水平的免疫抑制性调节性T细胞(Treg)。Treg的扩增可能代表了IL-2对癌症免疫疗法的不期望的影响。然而，IL-2即使在低剂量也能刺激Treg细胞的能力可以用于治疗自身免疫性紊乱和慢性炎性紊乱。最近发现，IL-2可以被修饰以选择性地刺激细胞毒性效应T细胞或Treg细胞。各种方法已经导致产生具有改进的和选择性的免疫调节活性的IL-2变体。
IL-2和IL-15两者都是有效的免疫效应细胞激动剂，并且关键的是，细胞毒性免疫细胞仅当在疾病部位(例如癌症部位)处或非常接近疾病部位时才被完全活化，以便仅特异性地破坏肿瘤细胞；或者细胞毒性免疫细胞仅当在炎性问题部位处或非常接近炎性问题部位时才被完全活化，以便局部地发挥抗自身免疫性紊乱和抗慢性炎性紊乱的作用。对于所有细胞因子、趋化因子和生长因子，改进对靶的特异性和选择性并保持健康细胞和组织完整且无损是非常重要的。
发明的公开内容
在一个方面，本发明提供了一种基于细胞因子的可生物活化的药物(“VitoKine”)平台，该平台旨在降低基于全身性机制的毒性，并使诸如IL-15和IL-2的细胞因子、趋化因子、激素和生长因子在治疗癌症、自身免疫性紊乱、炎性紊乱和各种其他紊乱中具有更广泛的治疗用途。VitoKine平台由如图1中描绘的构建体和如图2中描绘的提出的活化方法定义。参考图1，本发明的新的VitoKine构建体包含3个结构域：1)D1结构域(“D1”)，其选自由以下组成的组：组织靶向结构域、半衰期延长结构域或双重功能部分结构域，2)D2结构域(“D2”)，其为“活性部分结构域”，和3)D3结构域(“D3”)，其为“遮蔽部分结构域”。重要的是，VitoKine构建体的D2结构域保持几乎惰性或最低活性，直到被病变组织中上调的蛋白酶或疾病部位的水解局部活化，这将限制活性部分与非病变细胞或正常组织的外周中或细胞表面上的受体的结合，以防止途径的过度活化，并减少不期望的“组织外”“中靶”(“on-target”“off tissue”)毒性和不期望的靶吸收(target sink)。
在各种实施方案中，本发明的VitoKine构建体包含D1，该D1为靶向部分，诸如与肿瘤相关抗原(TAA)、或免疫检查点调节剂、或组织特异性抗原、细胞表面分子或细胞外基质蛋白或蛋白酶或任何翻译后修饰残基结合的抗体或抗体片段。在各种实施方案中，本发明的VitoKine构建体包含D1，该D1为靶向部分，诸如对病变细胞或组织表现出结合亲和力的蛋白或肽。设想在本发明的VitoKine构建体中用作D1的示例性抗体包括各种PD-1拮抗剂抗体、PD-L1阻断性抗体Tecentriq、抗CTLA4抗体伊匹单抗、激动性CD40抗体RO7009789、肿瘤抗原靶向抗体(包括针对纤连蛋白胞外域的L19、针对CD20的利妥昔单抗、针对Her-2的赫赛汀(Herceptin)、针对EGFR的西妥昔单抗)、用于肿瘤靶向和保留的抗FAP抗体、以及抗炎抗体(针对整联蛋白α4β7的维多珠单抗(Vedolizumab)和针对TNFα的修美乐(Humira))。
在各种实施方案中，D1是为拮抗性成纤维细胞活化蛋白(FAP)抗体的抗体或抗体片段。在各种实施方案中，抗体是包含SEQ ID NO:193和194中列出的氨基酸序列的人源化抗FAP抗体。在各种实施方案中，D1是针对免疫检查点调节剂的抗体或抗体片段。在各种实施方案中，抗体是拮抗性PD-1抗体或抗体片段。在各种实施方案中，抗体是包含SEQ ID NO:195和196中列出的氨基酸序列的拮抗性人源化PD-1抗体。在各种实施方案中，抗体是包含SEQ ID NO:197和198中列出的氨基酸序列的拮抗性人类PD-1抗体。在各种实施方案中，抗体是包含SEQ ID NO:275和276中列出的氨基酸序列的拮抗性人源化PD-1抗体。在各种实施方案中，抗体是包含SEQ ID NO:277和278中列出的氨基酸序列的拮抗性人类PD-1抗体。在各种实施方案中，抗体是拮抗性PD-L1抗体或抗体片段。在各种实施方案中，抗体是包含SEQID NO:279和280中列出的氨基酸序列的拮抗性人类PD-L1抗体。在各种实施方案中，VitoKine构建体包含SEQ ID NO:128-142、180-181、281-286、296-297和303-306中列出的氨基酸序列。
在各种实施方案中，本发明的VitoKine构建体包含D1，该D1是修饰的蛋白或肽，诸如聚糖修饰的，其对在病变细胞或组织上表达的特定受体诸如c型凝集素受体表现出结合亲和力。在各种实施方案中，本发明的VitoKine构建体包含D1，该D1作用为将细胞因子保持在组织部位处。在各种实施方案中，本发明的VitoKine构建体包含D1，该D1是双重功能的，例如组织靶向和保持。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是聚合物。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是半衰期延长部分。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是Fc结构域或其功能片段。
“Fc结构域”是指两个Fc结构域单体的二聚体，通常包含铰链区的全部或部分。在各种实施方案中，Fc结构域选自由以下组成的组：人类IgG1Fc结构域、人类IgG2 Fc结构域、人类IgG3 Fc结构域、人类IgG4 Fc结构域、IgAFc结构域、IgD Fc结构域、IgE Fc结构域、IgGFc结构域和IgM Fc结构域或它们的任何组合。在各种实施方案中，Fc结构域包含导致Fc结构域具有改变的补体结合特性或Fc受体结合特性的氨基酸变化。已知产生具有改变的补体结合特性或Fc受体结合特性的Fc结构域的氨基酸变化是本领域已知的。在各种实施方案中，用于制备VitoKine构建体的Fc结构域序列是SEQ ID NO:13中列出的人类IgG1-Fc结构域序列。在各种实施方案中，用于制备VitoKine构建体的Fc结构域序列是SEQ ID NO:14中列出的序列，SEQ ID NO:14包含消除(ablate)FcγR和C1q结合的氨基酸取代。在各种实施方案中，Fc结构域包括导致体内半衰期进一步延长的氨基酸变化。已知产生具有进一步延长的半衰期的Fc结构域的氨基酸变化是本领域已知的。在各种实施方案中，用于制备VitoKine构建体的Fc结构域序列是SEQ ID NO:156或166中列出的序列，两者都含有消除FcγR和C1q结合并延长体内半衰期的氨基酸取代。在各种实施方案中，用于制备VitoKine构建体的异源二聚体Fc结构域序列源自SEQ ID NO:15中列出的Knob-Fc结构域序列。在各种实施方案中，用于制备VitoKine构建体的异源二聚体Fc结构域序列源自SEQ ID NO:16中列出的Hole-Fc结构域序列。在各种实施方案中，用于制备VitoKine构建体的异源二聚体Fc结构域序列源自SEQ ID NO:167中列出的具有延长的体内半衰期序列的Knob-Fc结构域。在各种实施方案中，用于制备VitoKine构建体的异源二聚体Fc结构域序列源自SEQ ID NO:168中列出的、具有延长体内半衰期的序列的Hole-Fc结构域。
在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是蛋白。在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是细胞因子(选自包括但不限于以下的组：IL-1、IL-2、IL-4、IL-5、IL-6、IL-7、IL-9、IL-10、IL-12、IL-15、IL-23)和转化生长因子β(TGFβ)超家族例如TGFβ的配体(SEQ ID NO:24)。在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是IL-15。在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是IL-15变体(或突变体)，包含对IL-15多肽的一个或更多个氨基酸取代、缺失或插入。在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是IL-2。在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是IL-2变体(或突变体)，包含对IL-2多肽的一个或更多个氨基酸取代、缺失或插入。
在各种实施方案中，VitoKine构建体的D2结构域是IL-15结构域，该IL-15结构域包含如SEQ ID NO:2中列出的成熟的人类IL-15多肽(在本文中也称为huIL-15或IL-15野生型(wt))的序列。在各种实施方案中，IL-15结构域是IL-15变体(或突变体)，包含源自如SEQID NO:2中列出的成熟的人类IL-15多肽序列的序列。在各种实施方案中，IL-15结构域是IL-15变体(或突变体)，包含与SEQ ID NO:2具有至少80％、至少85％、至少90％或至少95％序列同源性的序列。本文使用天然氨基酸、天然氨基酸在成熟序列中的位置和变异氨基酸来指代IL-15的变体(或突变体)。例如，huIL-15“S58D”是指包含在SEQ ID NO:2的位置58处的S至D取代的人类IL-15。在各种实施方案中，IL-15变体作为IL-15激动剂发挥作用，如通过例如，与天然IL-15多肽相比，对IL-15Rβγc受体的结合活性增加展示的。在各种实施方案中，IL-15变体作为IL-15拮抗剂发挥作用，如通过以下展示的：例如，与天然IL-15多肽相比，对IL-15Rβγc受体的结合活性降低，或对IL-15Rβγc受体的结合活性相似或增加，但信号传导活性降低或消除。在各种实施方案中，与天然IL-15多肽相比，IL-15变体对IL-15Rβγc受体的结合亲和力增加或结合活性降低。在各种实施方案中，与天然IL-15序列相比，IL-15变体的序列具有至少一个(即，1个、2个、3个、4个、5个、6个、7个、8个、9个、10个或更多个)氨基酸变化。氨基酸变化可以包括IL-15多肽中的一个或更多个氨基酸取代、缺失或插入，诸如在与IL-15Rβ和/或IL-15Rβγc相互作用的IL-15的结构域中。在各种实施方案中，氨基酸变化是SEQ ID NO:2的位置30、31、32、58、62、63、67、68或108处的一个或更多个氨基酸取代。在各种实施方案中，氨基酸变化是成熟的人类IL-15序列的位置30处的D至T取代、位置31处的V至Y取代、位置32处的H至E取代、位置58处的S至D或G或H或R或Q或I或P取代、位置61处的T至D取代、位置63处的V至F或A或K或R取代、位置67处的I至V取代、位置68处的I至F或H或D或K或Q或G取代、位置108处的Q至A或M或S或E或K取代，或这些取代的任何组合。在各种实施方案中，氨基酸变化是在SEQ ID NO:2的N末端处的1个、或2个、或3个、或4个、5个或6个氨基酸缺失。在各种实施方案中，氨基酸变化是在SEQ ID NO:2的C末端处的1个、或2个、或3个、或4个、或5个、或6个、或7个、或8个、或9个或10个氨基酸缺失。在各种实施方案中，氨基酸变化是在SEQ ID NO:2的位置N95后的‘GS’、‘GSGS’(SEQ ID NO:116)、或‘GGSGG’(SEQ ID NO:119)或‘GSSGGSGGS’(SEQ ID NO:110)氨基酸插入。在各种实施方案中，IL-15结构域具有氨基酸取代、缺失和插入的任何组合。在各种实施方案中，VitoKine构建体将利用具有最佳的减弱效力的IL-15变体，因此导致对应的VitoKine构建体的固有基础活性降低。在各种实施方案中，IL-15变体包含SEQ ID NO:3、182-192和199-215中列出的氨基酸序列。
在各种实施方案中，本发明的VitoKine构建体的D2结构域包含IL-2多肽。在各种实施方案中，本发明的VitoKine构建体包含D2结构域，该D2结构域是包含一个或更多个氨基酸取代、缺失或插入的IL-2变体(或突变体)。在各种实施方案中，VitoKine构建体包含D2结构域，其中IL-2结构域包含如SEQ ID NO:8中列出的成熟的人类IL-2多肽(在本文中也称为huIL-2或IL-2野生型(wt))的序列。在各种实施方案中，IL-2结构域是IL-2变体(或突变体)，包含源自如SEQ ID NO:8中列出的成熟的人类IL-2多肽序列的序列。在各种实施方案中，IL-2结构域将是IL-2变体(或突变体)，包含与SEQ ID NO:8具有至少80％、至少85％、至少90％或至少95％序列同源性的序列。在各种实施方案中，IL-2变体起IL-2激动剂的作用。在各种实施方案中，IL-2变体起IL-2拮抗剂的作用。在各种实施方案中，氨基酸变化是SEQID NO:8的位置19、20、38、41、42、44、62、65、68、88、107、125或126处的一个或更多个氨基酸取代。在各种实施方案中，氨基酸变化是在成熟的人类IL-2序列的位置19处的L至D或H或N或P或Q或R或S或Y取代，位置20处的D至E或I或N或Q或S或T或Y取代，位置38处的R至E或A取代，位置41处的T至A或G或V取代，位置42处的F至A取代，位置44处的F至G或V取代，位置62处的E至A或F或H或L取代，位置65处的P至G或E或H或R或A或K或N或Q取代，位置68处的E至A或F或H或L或P取代，位置88处的N至D、E或G或I或M或Q或T或R取代，位置107处的Y至G或H或L或V取代，位置125处的S至E或H或K或I或L或V或W取代，位置126处的Q至D或E或K或L或M或N取代，或这些取代的任何组合。在各种实施方案中，VitoKine构建体将包含设计为具有减少/消除的与IL-2Rα结合的IL-2部分。在各种实施方案中，与天然IL-2多肽相比，IL-2变体具有降低的对IL-2Rβγc受体的结合活性。在各种实施方案中，与天然IL-2多肽相比，IL-2变体具有减少/消除的与IL-2Rα结合和改变的对IL-2Rβγc受体的结合活性。在各种实施方案中，具有减少/消除的与IL-2Rα结合的IL-2变体包含SEQ ID NO:232-247中列出的氨基酸序列。在各种实施方案中，设计了用于选择性扩增Teff细胞的含IL-2的VitoKine构建体，包括那些包含SEQ ID NO:59-61、271-274和286-291中列出的氨基酸序列的含IL-2的VitoKine构建体。在各种实施方案中，VitoKine构建体中的IL-2变体可以调节IL-2 VitoKine的固有基础活性，以在期望的抗肿瘤功效和不希望的全身性毒性之间实现最佳平衡。
在各种实施方案中，本发明的VitoKine构建体包含“遮蔽部分结构域”(D3)，该D3结构域是针对D2蛋白或细胞因子鉴定的同源受体/结合配偶体或任何结合配偶体。在各种实施方案中，D3结构域是D2结构域的同源受体/结合配偶体的变体。在各种实施方案中，与野生型同源受体/结合配偶体相比，D3结构域具有增强的与D2结构域的结合。在各种实施方案中，与野生型同源受体/结合配偶体相比，D3结构域具有减少或消除的与D2结构域的结合。在各种实施方案中，D3结构域是能够遮蔽D2的活性的蛋白、或肽、或抗体、或抗体片段。在各种实施方案中，D3结构域是DNA、RNA片段或聚合物，诸如PEG。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-15Rα细胞外结构域或其功能片段。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-15RαSushi结构域。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2Rα细胞外结构域或其功能片段。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2RαSushi结构域。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2RαSushi结构域的变体(突变体)。在各种实施方案中，IL-2RαSushi结构域变体(或突变体)包含对源自如SEQ ID NO:10中列出的序列的序列的氨基酸变化。在各种实施方案中，氨基酸变化是SEQ ID NO:10的位置36、38、42或43处的一个或更多个氨基酸取代。在各种实施方案中，氨基酸变化是在位置36处的R至A取代，在位置38处的K至E取代，在位置42处的L至G取代，在位置43处的Y至A取代。在各种实施方案中，D3结构域能够遮蔽D2的功能活性，直到D2在意图的治疗部位处被活化。在各种实施方案中，D3结构域被设计成在蛋白水解裂解和活化后有利于解离和扩散。
在各种实施方案中，VitoKine构建体的D1结构域、D2结构域和D3结构域通过蛋白酶可裂解的多肽接头序列连接。在各种实施方案中，VitoKine构建体的D1结构域、D2结构域和D3结构域通过不可裂解的多肽接头序列连接。在各种实施方案中，本发明的VitoKine构建体的L1和L2都是蛋白酶可裂解的肽接头。在各种实施方案中，本发明的VitoKine构建体的L1是蛋白酶可裂解的肽接头，并且L2是不可裂解的肽接头。在各种实施方案中，本发明的VitoKine构建体的L1是不可裂解的肽接头，并且L2是蛋白酶可裂解的肽接头。在各种实施方案中，本发明的VitoKine构建体的L1和L2都是不可裂解的接头。在各种实施方案中，不可裂解的接头富含G/S含量(例如接头中至少约60％、70％、80％、90％或更多的氨基酸是G或S)。每种肽接头序列可以独立地选择。在各种实施方案中，蛋白酶可裂解的接头选自SEQ IDNO:71-96和157-161中列出的序列的组。在各种实施方案中，蛋白酶可裂解的接头可以在可裂解接头的N末端或在可裂解接头的C末端或可裂解接头的两个末端具有可变长度的另外的肽间隔物。在各种实施方案中，不可裂解的接头选自SEQ ID NO:107-127中列出的序列的组。在各种实施方案中，接头是柔性的或刚性的，并且具有各种长度。
在各种实施方案中，VitoKine构建体的D2结构域和D3结构域被置于D1结构域的N末端，如图1A中描绘的。在各种实施方案中，VitoKine构建体的D2结构域和D3结构域被置于D1结构域的C末端，如图1B中描绘的。
在各种实施方案中，VitoKine构建体的D1结构域、D2结构域和D3结构域可以是单体或二聚体或二聚体和单体的组合，诸如D1是二聚体，并且D2和D3是单体。
在另一方面，本公开内容提供了用于治疗受试者的癌症或癌症转移的方法，该方法包括向有相应需要的受试者施用治疗有效量的本发明的药物组合物。在一种实施方案中，受试者是人类受试者。在各种实施方案中，癌症选自胰腺癌、胃癌、肝癌、乳腺癌、卵巢癌、结肠直肠癌、黑素瘤、白血病、骨髓增生异常综合征、肺癌、前列腺癌、脑癌、膀胱癌、头颈癌或横纹肌肉瘤或任何癌症。
在另一方面，本公开内容提供了一种用于治疗受试者的癌症或癌症转移的方法，该方法包括施用与第二疗法组合的治疗有效量的本发明的药物组合物，所述第二疗法选自由以下组成的组：细胞毒性化学疗法、免疫疗法、小分子激酶抑制剂靶向疗法、手术、放射疗法、干细胞移植、细胞疗法包括CAR-T、CAR-NK、iPS诱导的CAR-T或iPS诱导的CAR-NK、以及疫苗诸如卡介苗(Bacille Calmette-Guerine，BCG)。在各种实施方案中，组合疗法可以包括向受试者施用治疗有效量的免疫疗法，所述免疫疗法包括但不限于，使用针对特定肿瘤抗原的消耗性抗体的治疗；使用抗体-药物缀合物的治疗；使用针对共刺激性或共抑制性分子(免疫检查点)(诸如CTLA-4、PD-1、PD-L1、CD40、OX-40、CD137、GITR、LAG3、TIM-3、Siglec-7、Siglec-8、Siglec-9、Siglec-15和VISTA)的激动性抗体、拮抗性抗体或阻断性抗体的治疗；使用双特异性T细胞结合抗体
Figure BDA0003806684520000091
诸如博纳吐单抗(blinatumomab)的治疗；涉及施用生物响应调节剂(诸如IL-12、IL-21、GM-CSF、IFN-α、IFN-β和IFN-γ)的治疗；使用治疗性疫苗诸如sipuleucel-T的治疗；使用树突细胞疫苗或肿瘤抗原肽疫苗的治疗；使用嵌合抗原受体(CAR)-T细胞的治疗；使用CAR-NK细胞的治疗；使用肿瘤浸润淋巴细胞(TIL)的治疗；使用过继转移的抗肿瘤T细胞(离体扩增的T细胞和/或TCR转基因的T细胞)的治疗；使用TALL-104细胞的治疗；和使用免疫刺激剂诸如Toll样受体(TLR)激动剂CpG和咪喹莫特的治疗；以及使用疫苗诸如BCG的治疗；其中组合疗法提供了增加的对肿瘤细胞的效应细胞杀伤，即，当被共施用时，VitoKine构建体和免疫疗法之间存在协同作用。
在另一方面，本公开内容提供了用于治疗受试者的病毒感染的方法，该方法包括向有相应需要的受试者施用治疗有效量的本发明的药物组合物。在一种实施方案中，受试者是人类受试者。在各种实施方案中，病毒是HIV。
在另一方面，本公开内容提供了用于治疗受试者的病毒感染的方法，该方法包括施用与第二疗法组合的治疗有效量的本发明的药物组合物，所述第二疗法包括但不限于阿昔洛韦、伊柯鲁沙(Epclusa)、Mavyret、齐多夫定和恩福韦肽。
在另一方面，本公开内容提供了用于治疗受试者的自身免疫性疾病的方法，该方法包括向有相应需要的受试者施用治疗有效量的本发明的药物组合物。在一种实施方案中，受试者是人类受试者。在各种实施方案中，自身免疫性疾病选自由以下组成的组：系统性红斑狼疮(SLE)、寻常天疱疮、重症肌无力、溶血性贫血、血小板减少性紫癜、格雷夫斯病、干燥综合征、皮肌炎、桥本氏病、多发性肌炎、炎性肠病、多发性硬化(MS)、糖尿病、类风湿性关节炎和硬皮病。
在另一方面，本公开内容提供了用于治疗受试者的炎性疾病的方法，该方法包括向有相应需要的受试者施用治疗有效量的本发明的药物组合物。在一种实施方案中，受试者是人类受试者。在各种实施方案中，炎性疾病选自由以下组成的组：克罗恩病、溃疡性结肠炎、胶原性结肠炎、淋巴细胞性结肠炎、缺血性结肠炎、转移性结肠炎、白塞氏综合征(Behcet’s syndrome)和未定型结肠炎(indeterminate colitis)。
在各种实施方案中，炎性疾病选自由诸如以下的其他自身免疫性疾病和炎性疾病组成的组：贲门失弛缓症、成人斯蒂尔病、无丙种球蛋白血症、淀粉样变性、抗GBM/抗TBM肾炎、抗磷脂综合征、自身免疫性血管性水肿、自身免疫性自主神经失调、自身免疫性脑脊髓炎、自身免疫性内耳病、自身免疫性卵巢炎、自身免疫性睾丸炎、自身免疫性胰腺炎、自身免疫性视网膜病、自身免疫性荨麻疹、轴突和神经元神经病、巴洛病、白塞氏病、良性粘膜类天疱疮、Castleman病、查加斯病、慢性炎性脱髓鞘性多神经病、慢性复发性多病灶性骨髓炎、变应性肉芽肿血管炎、瘢痕性类天疱疮、科干综合征、柯萨奇病毒性心肌炎、CREST综合征、疱疹样皮炎、Devic病/视神经脊髓炎、盘状狼疮、德雷斯勒综合征、嗜酸细胞性食管炎、嗜酸细胞性筋膜炎、结节性红斑、混合性冷凝球蛋白血症、纤维化肺泡炎、巨细胞动脉炎、巨细胞心肌炎、过敏性紫癜、妊娠疱疹或妊娠性类天疱疮、IgA肾病、IgG4相关硬化性疾病、免疫相关不良事件、包涵体肌炎、间质性膀胱炎、幼年关节炎、幼年肌炎、兰伯特-伊顿综合征、白细胞破碎性血管炎、扁平苔藓、苔藓硬化、木质结膜炎、线性IgA病、慢性莱姆病、梅尼埃病、显微镜下多血管炎、混合结缔组织病、蚕蚀性角膜溃疡(Mooren’s Ulcer)、Mucha-Habermann病、多病灶性运动神经病、视神经炎、复发性风湿病、PANDAS、副肿瘤性小脑变性、ParryRomberg综合征、睫状体扁平部炎、Parsonage-Turner综合征、静脉周脑脊髓炎、POEMS综合征、结节性多动脉炎、多腺体综合征、风湿性多肌痛、心肌梗死后综合征、心包切开术后综合征、原发性硬化性胆管炎、孕酮皮炎、银屑病关节炎、纯红细胞再生障碍、坏疽性脓皮病、雷诺现象、反射性交感神经营养不良、复发性多软骨炎、腹膜后纤维化、巩膜炎、精子和睾丸自身免疫、僵人综合征、亚急性细菌性心内膜炎、Susac综合征、交感性眼炎、大动脉炎、血小板减少性紫癜、痛性眼肌麻痹综合征、横贯性脊髓炎、未分化结缔组织病、Vogt-Koyonagi-Harada病。
在另一方面，本公开内容提供了VitoKine构建体用于制备用于治疗癌症的药物的用途。
在另一方面，本公开内容提供了VitoKine构建体用于制备用于治疗病毒感染的药物的用途。
在另一方面，本公开内容提供了VitoKine构建体用于制备用于治疗自身免疫性疾病的药物的用途。
在另一方面，本公开内容提供了VitoKine构建体用于制备用于治疗炎症的药物的用途。
在另一方面，本公开内容提供了本发明的VitoKine构建体与能够治疗癌症、病毒感染或自身免疫性疾病或炎症的第二治疗剂或细胞疗法组合的用途。
在另一方面，本公开内容提供了分离的核酸分子，该分离的核酸分子包含编码本公开内容的VitoKine构建体的多核苷酸。在另一方面，本公开内容提供了包含本文描述的核酸的载体。在各种实施方案中，载体是表达载体。在另一方面，本公开内容提供了包含本公开内容的核酸的分离的细胞。在各种实施方案中，细胞是包含本公开内容的表达载体的宿主细胞。在另一方面，制备VitoKine构建体的方法通过在促进蛋白或多肽的表达的条件下培养宿主细胞来提供。
在另一方面，本公开内容提供了药物组合物，所述药物组合物包含与药学上可接受的载体混合的分离的VitoKine构建体。
附图简述
图1描绘了本发明的代表性VitoKine构建体形式。图1A描绘了D2结构域和D3结构域被置于D1结构域的C末端的VitoKine构建体。图1B描绘了D2结构域和D3结构域被置于D1结构域的N末端的VitoKine构建体。图1C-图1E进一步描绘了D2结构域和D3结构域被置于D1结构域的C末端的VitoKine构建体；其中D1结构域为抗体；(图1C)D2结构域和D3结构域被置于抗体异源二聚体重链的C末端的VitoKine构建体；(图1D)D2结构域和D3结构域被置于抗体同源二聚体重链的C末端的VitoKine构建体；(图1E)D2结构域和D3结构域被置于抗体轻链的C末端的VitoKine构建体。类似地，D2结构域和D3结构域可以被置于抗体重链(异源二聚体和同源二聚体两者)和轻链的N末端。
图2描绘了提出的VitoKine构建体的活化机制。示例性VitoKine包含两个蛋白酶可裂解的接头；由L1接头裂解导致的蛋白酶1活化产生活性形式1；由L2接头裂解导致的蛋白酶2活化产生活性形式2；由L1接头和L2接头裂解导致的两种蛋白酶活化产生活性形式3。图2A描绘了在蛋白水解后D3结构域保持与D2非共价复合的实例；图2B描绘了在蛋白酶裂解后D3从D2释放并扩散离开的实例。
图3描绘了蛋白A纯化后示例性IL-15 Fc VitoKine P-0315的蛋白谱，A)在还原剂不存在和存在下的SDS-PAGE、以及B)尺寸排阻色谱图。
图4描绘了与高活性IL-15融合蛋白P-0198相比IL-15 Fc VitoKine P-0172的结合和功能活性。(A)通过ELISA测定测量的与IL-2Rβ的结合活性；(B-C)通过FACS分析新鲜的人类PBMC的人类CD8+T细胞(B)和NK细胞(C)上的CD69表达的诱导。
图5描绘了与高活性IL-15融合蛋白P-0166相比单体IL-15 Fc VitoKine P-0170的功能活性。新鲜的人类PBMC的人类CD8+T细胞上CD69表达的诱导通过FACS来测量和分析。
图6描绘了与高活性IL-15/IL-15RαFc融合蛋白P-0165相比，具有不同接头长度的说明性IL-15 Fc VitoKine构建体(P-0204、P-0205和P-0206)对人类PBMC的A)CD8+T细胞和B)NK(CD56+)细胞上的CD69表达的诱导。
图7描绘了与完全活性的IL-15/IL-15RαFc融合蛋白P-0207和P-0217相比，具有不同的L1接头和L2接头的示例性IL-15 Fc VitoKine构建体(P-0202、P-0203和P-0204)引起的人类PBMC中的NK(CD56+)细胞的增殖。
图8描绘了与IL-15/IL-15RαFc融合蛋白P-0156相比，通过FACS测量的具有不同的L2接头序列组成的说明性IL-15 Fc VitoKine构建体(P-0351、P-0488和P-0489)引起的人类PBMC中的A)CD8+T细胞和B)NK(CD56+)细胞的增殖。
图9描绘了使用不同量的MMP-2进行IL-15 Fc VitoKine P-0315体外蛋白水解的SDS-PAGE分析。
图10描绘了在不同条件下使用uPA进行IL-15 Fc VitoKine P-0203体外蛋白水解的SDS-PAGE分析，以确定完全裂解的合适反应条件。
图11描绘了A)被uPA体外蛋白水解之前和之后的IL-15 Fc VitoKine P-0203的SDS-PAGE分析。B)经uPA消化和蛋白A纯化去除含Fc的片段后，活化的VitoKine P-0203的蛋白谱。
图12描绘了A)被MMP-2体外蛋白水解之前和之后的IL-15 Fc VitoKine P-0315的SDS-PAGE分析。凝胶还显示了经MMP-2消化和蛋白A纯化的P-0315的谱；B)由MMP-2消化、随后蛋白A纯化产生的VitoKine P-0315的活性形式2的蛋白谱；C)由MMP-2和uPA双重蛋白水解、随后以流通模式通过蛋白A纯化产生的VitoKine P-0315的活性形式3的蛋白谱。
图13描绘了通过分析A)CD56+NK细胞和B)CD8+T细胞上活化标志物CD69的诱导，对蛋白酶(uPA)活化的IL-15 Fc VitoKine P-0203的活性评估。高活性的IL-15融合蛋白P-0165被包括在内作为阳性对照。
图14描绘了通过分析A)CD56+NK细胞和B)CD8+T细胞上活化标志物CD69的诱导，对蛋白酶活化的IL-15 Fc VitoKine P-0315的两种形式的活性评估。P-0315活性形式2由MMP-2消化产生，并且P-0315活性形式3由MMP-2和uPA两者双重蛋白水解产生。与P-0315的活性形式2结构相似的高活性IL-15融合蛋白P-0313被包括在内作为阳性对照。
图15描绘了通过分析A)CD56+NK细胞和B)CD8+T细胞上增殖标志物Ki67的诱导，对MMP-2活化的IL-15 Fc VitoKine P-0315(活性形式2)的活性评估。包含不可裂解的L1和L2接头且与P-0315共有相同的L2接头长度的P-0351被包括在内用于比较。
图16描绘了在Balb/C小鼠中单次注射后，可裂解的IL-15 Fc VitoKine P-0315、不可裂解的IL-15 Fc VitoKine P-0351对外周血中的A)CD8+T、B)NK细胞和C)白细胞的扩增的剂量依赖效应和时间依赖效应。完全活性的IL-15 Fc融合蛋白P-0313被包括在内用于比较。在第-1天、第3天、第5天和第7天收集血液以便通过FACS分析进行淋巴细胞表型分析。数据表示为平均值±SEM。通过双因素ANOVA检验、随后Tukey事后检验进行统计分析。****p<0.0001，***p<0.001，*p<0.05，在各自的时间点与PBS组相比。
图17描绘了在P-0315、P-0351、P-0313或PBS对照的4×Q5D剂量后一天小鼠CT26肺转移模型中肺转移结节的抑制。第一次给药在注射CT26细胞后一天开始。除非另外说明，否则所有比较均与PBS组相比；****p<0.0001；**p<0.01；*p<0.05。
图18描绘了CT26转移小鼠的总血液淋巴细胞中的A)CD8+T细胞％和B)NK细胞％。在P-0315、P-0351、P-0313或PBS对照的三次Q5D腹膜内注射后4天，通过流式细胞术确定细胞数目。所有比较均与PBS组相比；****p<0.0001；**p<0.01；*p<0.05。
图19描绘了在已建立的CT26鼠结肠直肠癌肿瘤模型中，与完全活性的IL-15 Fc融合体P-0313相比，IL-15 Fc VitoKine P-0315的抗肿瘤功效。示出了在两次Q5D处理后，以下组的个体小鼠中皮下CT26肿瘤的生长曲线：A)媒介物PBS组、B)0.1mg/kg P-0315组、C)0.1mg/kg P-0313组和(D)每个处理组随时间推移的平均肿瘤体积±SEM。所有比较均与媒介物处理相比；n＝11只/组；****P<0.0001。
图20描绘了在CT26鼠结肠直肠癌肿瘤模型中，IL-15 Fc VitoKine P-0315或高活性IL-15 Fc融合体P-0313处理后小鼠外周血的免疫药效学谱分析。在肿瘤植入后11天开始的两次Q5D处理后，在第19天通过流式细胞术确定A)NK细胞和B)CD8+T细胞中增殖标志物Ki67的增加百分比。****P<0.0001，与PBS相比。
图21描绘了在CT26鼠结肠直肠癌肿瘤模型中，P-0315或P-0313处理后小鼠外周血的免疫药效学谱分析。在肿瘤植入后11天开始的两次Q5D处理后，在第19天通过流式细胞术确定每μl全血的循环总白细胞(A)、NK细胞(B)和CD8+T细胞(C)的数目的增加。****P<0.0001，与PBS相比。图22描绘了在CT26鼠结肠直肠癌肿瘤模型中，P-0315或P-0313处理后脾的免疫药效学谱分析。在肿瘤植入后11天开始的两次Q5D处理后，在第19天通过流式细胞术确定脾的总白细胞(A)、NK细胞(B)和CD8+T细胞(C)的数目的增加。****P<0.0001，*P<0.05，与PBS相比。
图23描绘了通过分析A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导，对在位置V63、I68和Q108处包含一个或两个氨基酸取代的各种IL-15/IL-15RαFc融合蛋白的活性评估。高效的IL-15/IL-15RαFc融合蛋白P-0313被包括在内用于比较。
图24描绘了通过分析A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导，对IL-15 Q108S/IL-15RαFc融合蛋白P-0764及其对应的Fc VitoKine(被称为P-0682)的活性评估。高效的IL-15/IL-15RαFc融合蛋白P-0313被包括在内用于比较。
图25描绘了通过分析A)CD56+NK细胞和B)CD8+T细胞上增殖标志物Ki67的诱导，对不可裂解的IL-15 Fc VitoKine P-0351和基准的活性比较。
图26描绘了蛋白A纯化后示例性IL-2 VitoKine P-0320的蛋白谱，A)在还原剂不存在和存在下的SDS-PAGE、以及B)尺寸排阻色谱图。
图27描绘了通过分析新鲜的人类PBMC中的A)CD4+Foxp3+/CD25高Treg和B)CD4+Foxp3-/CD25低CD4常规T细胞亚群中的pStat5水平，对两种IL-2 Fc VitoKine P-0320(IL-2融合在Fc的C末端)和P-0329(IL-2融合在Fc的N末端)的活性评估。具有高活性的IL-2 Fc融合蛋白P-0250被包括在内作为阳性对照。
图28描绘了A)对IL-2 VitoKine P-0382及其通过MMP-2消化活化随后通过Ni-Excel纯化的SDS-PAGE分析，以及B)通过蛋白A以结合及洗脱模式纯化的经MMP-2活化的P-0382的蛋白谱。
图29描绘了通过分析新鲜的人类PBMC中的A)CD4+Foxp3+/CD25高Treg和B)CD4+Foxp3-/CD25低CD4常规T(Tconv)细胞亚群中的pStat5水平，对蛋白酶活化的IL-2 FcVitoKine P-0382的活性评估。两种活化的样品通过用Ni-Excel树脂纯化以去除蛋白酶(P-0382活性形式1)或通过蛋白A纯化以去除蛋白酶和由蛋白水解产生的IL-2RαSushi结构域(P-0382活性形式2)。具有高活性的IL-2 Fc融合蛋白P-0250被包括在内作为阳性对照。
图30描绘了通过分析新鲜的人类PBMC中的A)CD4+Foxp3+/CD25高Treg和B)CD4+Foxp3-/CD25低CD4 Tconv细胞亚群中的pStat5水平，对MMP-2蛋白水解之前和之后的IL-2Fc VitoKine P-0398的活性评估。P-0382与P-0398的区别仅在于L2接头长度，并且具有高活性的IL-2 Fc融合蛋白P-0250被包括在内用于比较。
图31描绘了在位置P65处包含不同氨基酸取代的IL-2变体Fc融合蛋白P-0704、P-0707、P-0708和P-0709与IL-2Rα的ELISA结合。野生型IL-2对应物P-0689被包括在内用于比较。
图32描绘了通过分析新鲜的人类PBMC中CD8+T细胞上增殖标志物Ki67的诱导，对P-0704和P-0689的活性评估。P-0704是失去与IL-2Rα的结合活性的IL-2 P65R Fc融合蛋白，并且P-0689是野生型IL-2对应物。
图33描绘了各种IL-2RαSushi变体P-0751、P-0752和P-0753与单价野生型IL-2 Fc融合蛋白P-0689的ELISA结合。P-0757包含野生型IL-2RαSushi并被包括在内用于比较。
图34描绘了对具有野生型IL-2RαSushi作为D3结构域(P-0701)或IL-2RαSushi变体之一作为D3结构域(P-0754、P-0755和P-0756)的IL-2 Fc VitoKine的活性评估。失去与IL-2Rα的结合活性但完全保留IL-2Rβγ活性的IL-2 P65R Fc融合蛋白P-0704被包括在内作为对照。通过分析用流式细胞术确定的A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导来评估活性。
图35描绘了对IL-2 P65R Fc融合蛋白P-0704及其对应的具有野生型IL-2RαSushi作为D3结构域(P-0745)或IL-2RαSushi变体之一作为D3结构域(P-0807、P-0808和P-0809)的Fc VitoKine的活性评估。通过分析用流式细胞术确定的A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导来评估活性。
图36描绘了对具有IL-2RαSushi L42G变体作为D3结构域的IL-2 Fc VitoKine P-0755在体外蛋白酶活化之前和之后的活动评估。完全保留IL-2Rβγ活性的IL-2 P65R Fc融合蛋白P-0704被包括在内作为对照。通过分析新鲜的人类PBMC中A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导来评估活性。
图37描绘了通过用流式细胞术分析新鲜的人类PBMC中A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导，对IL-15 Fc VitoKine P-0315与IL-15抗体VitoKine P-0485的活性评估。
图38描绘了对由各种IL-15 VitoKine与它们各自的非Vitokine融合对应物相比诱导的CD8+T细胞上Ki67表达的流式细胞术分析。A)在人类PBMC中IL-15 Fc VitoKine P-0315对比对应物Fc融合体；B)在人类PBMC中IL-15抗体VitoKine P-0875对比对应物抗体融合体P-0870和Fc融合体P-0773；C)在食蟹猴PBMC中P-0875对比P-0773。
图39描绘了通过分析用流式细胞术确定的A)CD8+T细胞和B)CD56+NK细胞上增殖标志物Ki67的诱导，对各种IL-2抗体VitoKine P-0800、P-0830、P-0831和P-0802与非Vitokine IL-2抗体融合体P-0782的活性评估。这四种IL-2抗体VitoKine的区别仅在于IL-2部分与IL-2Rα具有不同水平的结合强度。
图40用以下分析描绘了IL-2抗体VitoKine P-0872的蛋白酶裂解和活化：A)还原性SDS-PAGE凝胶和B)流式细胞术分析人类PBMC中CD8+T细胞上Ki67表达的剂量依赖性诱导。P-0872包含单价IL-2部分和连接D2结构域和D3结构域的单个蛋白酶可裂解的接头。
图41用以下分析描绘了IL-2抗体VitoKine P-0929的蛋白酶裂解和活化：A)还原性SDS-PAGE凝胶和B)流式细胞术分析人类PBMC中CD8+T细胞上Ki67表达的剂量依赖性诱导。P-0929包含二价IL-2部分和两个蛋白酶可裂解的接头。
图42以ELISA形式描绘了基于IL-2Rβ的封闭肽(L01、L02、L03、L04和L05)与IL-15的结合。
图43描绘了IL-15融合蛋白(P-0153、P-0159、P-0160和P-0161)与固定在板上的IL-2Rβ的结合。P-0159、P-0160和P-0161包含不同的基于IL-2Rβ的封闭肽。
图44描绘了四种IL-2 VitoKine(P-0320、P-0382、P-0362和P-0379)(图40B-图40E)与它们的对应物Fc融合蛋白P-0250(图44A)的尺寸排阻色谱图。与P-0250差异在于IL-2中具有单个氨基酸取代S125I的P-0531被包括在内用于比较(图44F)。
图45描绘了包含不同D3结构域的IL-15 Fc VitoKine P-0389(A)和P-0315(B)的SDS-PAGE凝胶。
图46描绘了在已建立的MC38鼠结肠癌模型中，IL-2 PD-1拮抗剂抗体VitoKine P-0922A、P-0928A和P-0929A与它们的不可裂解的对应物P-0877相比的抗肿瘤功效。每组的个体小鼠在单次处理后的第7天的肿瘤尺寸被图示出。
用于进行本公开内容的方式
本公开内容提供了新的“VitoKine”构建体作为平台技术，以降低全身性中靶毒性并提高意图用于治疗癌症、病毒感染、自身免疫性疾病或炎性疾病的细胞因子的治疗指数。VitoKine平台由如图1中描绘的构建体和如图2中描绘的提出的活化方法定义。参考图1，本发明的新的VitoKine构建体包含3个结构域：1)D1结构域(“D1”)，其选自由以下组成的组：组织靶向结构域、半衰期延长结构域、免疫检查点调节剂靶向结构域或双重功能部分结构域，2)D2结构域(“D2”)，其为“活性部分结构域”，和3)D3结构域(“D3”)，其为“遮蔽部分结构域”。重要的是，D3结构域能够遮蔽或减弱D2的功能活性，直到D2在意图的治疗部位处被活化。
这三个结构域使用接头来连接，所述接头具有可变的长度和刚性、偶联有蛋白酶可裂解序列，所述蛋白酶可裂解序列是特定蛋白酶亚型的肽底物，所述特定蛋白酶亚型在疾病部位中具有升高或失调的表达，因此允许在疾病部位处露出或释放功能性D2结构域。接头长度和组成经过优化，以驱动D2结构域对其受体的可及性的最佳遮蔽，以减少其全身性参与，同时保持VitoKine在血液循环中的稳定性，并允许在意图的疾病部位遇到特定蛋白酶后进行有效裂解。“VitoKine”的设计也基于细胞因子与其同源受体的分子相互作用的知识进行合理操纵。细胞因子受体通常作为由两个至四个受体亚基组成的寡聚体复合物发挥作用。不同的亚基执行专门的功能，诸如配体结合或信号转导。细胞因子受体的α亚基是赋予配体特异性的结合受体，增强配体与信号传导受体的相互作用，并将信号传导受体从低亲和力转化为高亲和力。因此，VitoKine的D3结构域优选地是D2结构域的同源结合受体。裂解后，D3结构域可以与D2结构域解离或再缔合，并在局部完全恢复D2结构域的结合和信号传导活性。因此，D3结构域可以在调节D2结构域的功能中具有双重作用。当VitoKine未活化时，D3结构域使D2结构域保持惰性，并且当VitoKine被裂解和活化时，D3结构域可以参与D2功能。然而，D3结构域可以是能够遮蔽D2活性的任何蛋白、肽、抗体、抗体片段或聚合物或核苷酸。
在另一方面，D3结构域的添加还可以导致VitoKine构建体的可开发性谱显著改进，表达产量增加，以及聚集倾向降低。
D1结构域可以是半衰期延长结构域，以延长VitoKine的循环半衰期，此外还可以作为额外的结构域来遮蔽D2结构域的功能活性。D1结构域也可以是疾病或组织靶向基序，其将VitoKine特异性地引导至感兴趣的部位，并将VitoKine的活化限制在局部，以进一步提高治疗指数。因此，“VitoKine”平台允许细胞因子在意图的部位选择性地活化，并且具有降低全身性毒性的益处，同时增加在疾病部位处的治疗效果，从而提高其治疗指数。
VitoKine构建体的D2结构域是活性部分，但保持惰性，直到被病变组织中上调的蛋白酶在局部活化；这将限制活性部分与非病变细胞或组织的外周中或细胞表面上的受体的结合，以防止途径的过度活化并减少不期望的“组织外”“中靶”毒性。此外，在蛋白酶活化之前，VitoKine活性部分的惰性将显著降低潜在的抗原吸收(antigen sink)，并且从而延长体内半衰期，以及导致在意图的治疗部位处的生物分布、生物可利用度和功效的改进。此外，基于本发明，VitoKine平台可以增强蛋白的可开发性谱，包括但不限于，诸如当使用同源受体α作为D3结构域时，提高表达水平和降低聚集倾向。
尽管对于大多数VitoKine，可裂解的接头是优选的，以限制全身性活化并在施用后在意图的部位处释放活性结构域，但不可裂解的接头也可能是期望的，以提供低效力但有药理学活性的VitoKine的持续全身性暴露并提高治疗功效。
在示例性实施方案中，VitoKine构建体包含基于IL-15、基于IL-15变体、基于IL-2或基于IL-2变体的活性部分(D2)。对于这些基于IL-15和/或IL-2的VitoKine构建体，每种细胞因子受体的独特且非信号传导的α-亚基经由蛋白酶可裂解的接头被用作遮蔽部分结构域(D3)之一，以可逆地遮蔽细胞因子活性。取决于每种受体复合物的不同特性和意图用VitoKine分子治疗的不同疾病适应症的不同需求，在接头被蛋白酶裂解后，遮蔽性α-亚基可以优选地通过非共价缔合与活化的细胞因子复合(例如，对于IL-15)或者优选地解离掉(例如，对于IL-2)。因此，对α-受体进行氨基酸修饰以调节与其同源细胞因子的结合亲和力可能是需要的且是有益的。
在示例性实施方案中，VitoKine构建体包含基于IL-15、基于IL-15变体、基于IL-2或基于IL-2变体的活性部分(D2)。对于这些基于IL-15和/或IL-2的VitoKine构建体，共有的受体β亚基或基于受体β的封闭肽被用作经由蛋白酶可裂解的接头的遮蔽部分结构域(D3)，以可逆地遮蔽细胞因子活性。
这种经由可活化的接头将同源受体、蛋白、抗体、抗体片段、结合肽与细胞因子偶联以遮蔽其功能活性直到在意图的治疗部位处被活化的概念可以针对各种细胞因子进行定制，包括但不限于IL-4、IL-7、IL-9、IL-10、IL-12、IL-22、IL-23和TGFβ、趋化因子诸如CXCR3、或各种生长因子诸如TNF家族、TGFα和TGFβ以及激素。同样的概念也可以应用于其他蛋白来产生前蛋白，以实现对疾病部位的增强的靶向性并扩大治疗效用。
定义
术语“多肽”、“肽”和“蛋白”在本文中可互换使用，以指氨基酸残基的聚合物。在各种实施方案中，“肽”、“多肽”和“蛋白”是其α碳通过肽键连接的氨基酸链。因此链的一个末尾(氨基末端)处的末端氨基酸具有游离氨基基团，而链的另一个末尾(羧基末端)处的末端氨基酸具有游离羧基基团。如本文使用的，术语“氨基末端”(缩写为N末端)是指在肽的氨基末端处的氨基酸上的游离α-氨基基团，或指在肽中的任何其他位置处的氨基酸的α-氨基基团(参与肽键时的氨基基团)。类似地，术语“羧基末端”(缩写为C末端)是指肽的羧基末端上的游离羧基基团，或肽中任何其他位置处的氨基酸的羧基基团。肽还包括基本上任何多氨基酸，包括但不限于肽模拟物(peptide mimetic)，诸如通过醚键而非酰胺键连接的氨基酸。
本公开内容的多肽包括已经以任何方式和出于任何原因被修饰的多肽，例如，以：(1)降低对蛋白水解的敏感性，(2)降低对氧化的敏感性，(3)改变形成蛋白复合物的结合亲和力，(4)改变结合亲和力，和(5)赋予或改变其他物理化学特性或功能特性。
如本文使用的氨基酸“取代”是指用不同的氨基酸替换多肽中亲本多肽序列的特定位置处的一个氨基酸。氨基酸取代可以使用本领域熟知的遗传方法或化学方法来产生。例如，单氨基酸取代或多氨基酸取代(例如，保守氨基酸取代)可以在天然存在的序列中进行(例如，在形成分子间接触的结构域以外的多肽部分中)。“保守氨基酸取代”是指多肽中的氨基酸被功能上相似的氨基酸取代。以下六组各自包含互相为保守取代的氨基酸：
1)丙氨酸(A)、丝氨酸(S)和苏氨酸(T)
2)天冬氨酸(D)和谷氨酸(E)
3)天冬酰胺(N)和谷氨酰胺(Q)
4)精氨酸(R)和赖氨酸(K)
5)异亮氨酸(I)、亮氨酸(L)、甲硫氨酸(M)和缬氨酸(V)
6)苯丙氨酸(F)、酪氨酸(Y)和色氨酸(W)
“非保守氨基酸取代”是指这些类别之一的成员取代为来自另一类别的成员。在进行这样的改变时，根据各种实施方案，可以考虑氨基酸的亲水指数(hydropathic index)。基于氨基酸的疏水性和电荷特征，每种氨基酸已经被指定了亲水指数。它们是：异亮氨酸(+4.5)；缬氨酸(+4.2)；亮氨酸(+3.8)；苯丙氨酸(+2.8)；半胱氨酸/胱氨酸(+2.5)；甲硫氨酸(+1.9)；丙氨酸(+1.8)；甘氨酸(-0.4)；苏氨酸(-0.7)；丝氨酸(-0.8)；色氨酸(-0.9)；酪氨酸(-1.3)；脯氨酸(-1.6)；组氨酸(-3.2)；谷氨酸(-3.5)；谷氨酰胺(-3.5)；天冬氨酸(-3.5)；天冬酰胺(-3.5)；赖氨酸(-3.9)和精氨酸(-4.5)。
本领域理解亲水氨基酸指数在赋予蛋白相互作用的生物功能中的重要性(参见例如，Kyte等人,1982,J.Mol.Biol.157:105-131)。已知，某些氨基酸可以被具有相似亲水指数或评分的其他氨基酸取代，并且仍保持相似的生物活性。在基于亲水指数进行改变时，在各种实施方案中，包括了其亲水指数在±2内的氨基酸的取代。在各种实施方案中，包括在±1内的那些，并且在各种实施方案中，包括在±0.5内的那些。
本领域还理解，可以基于亲水性有效地进行相似氨基酸的取代，特别是当从而产生的生物功能蛋白或肽被意图用于在免疫学实施方案中使用时，如本文所公开的。在各种实施方案中，蛋白的最大局部平均亲水性(如由其相邻氨基酸的亲水性决定的)与其免疫原性和抗原性，即，与蛋白的生物学特性相关。
以下亲水性值(hydrophilicity values)被指定至这些氨基酸残基：精氨酸(+3.0)；赖氨酸(+3.0)；天冬氨酸(+3.0.+-.1)；谷氨酸(+3.0.+-.1)；丝氨酸(+0.3)；天冬酰胺(+0.2)；谷氨酰胺(+0.2)；甘氨酸(0)；苏氨酸(-0.4)；脯氨酸(-0.5.+-.1)；丙氨酸(-0.5)；组氨酸(-0.5)；半胱氨酸(-1.0)；甲硫氨酸(-1.3)；缬氨酸(-1.5)；亮氨酸(-1.8)；异亮氨酸(-1.8)；酪氨酸(-2.3)；苯丙氨酸(-2.5)和色氨酸(-3.4)。在基于相似亲水性值进行改变时，在各种实施方案中，包括其亲水性值在±2内的氨基酸的取代，在各种实施方案中，包括在±1内的那些，并且在各种实施方案中，包括在±0.5内的那些。
示例性的氨基酸取代在表1中列出。
表1
Figure BDA0003806684520000241
技术人员将能够使用公知技术确定如本文列出的合适的多肽变体。在各种实施方案中，本领域技术人员可以通过靶向被认为对于活性不重要的区域来鉴定分子中可以被改变而不破坏活性的适合区域。在其他实施方案中，技术人员可以鉴定在相似多肽间保守的分子的残基和部分。在另外的实施方案中，即使可能对生物活性或对结构重要的区域也可以经历保守氨基酸取代，而不破坏生物活性或不会不利地影响多肽结构。
此外，本领域技术人员可以回顾鉴定相似多肽中对活性或结构重要的残基的结构-功能研究。鉴于这样的比较，技术人员可以预测对应于相似多肽中对活性或结构重要的氨基酸残基的多肽中的氨基酸残基的重要性。本领域技术人员可以选择对这样预测的重要氨基酸残基进行化学上相似的氨基酸取代。
本领域技术人员还可以分析相似多肽中与该结构相关的三维结构和氨基酸序列。鉴于这样的信息，本领域技术人员可以预测多肽的氨基酸残基在其三维结构方面的排列。在各种实施方案中，本领域技术人员可以选择不对预测在多肽表面上的氨基酸残基进行根本改变(radical changes)，因为此类残基可能参与与其他分子的重要相互作用。此外，本领域技术人员可以产生在每个期望的氨基酸残基处包含单个氨基酸取代的测试变体。然后可以使用本领域技术人员已知的活性测定来筛选变体。这些变体可以用于收集关于合适变体的信息。例如，如果人们发现，特定氨基酸残基的改变导致破坏的、不期望地减少的或不合适的活性，则可以避免具有这样的改变的变体。换句话说，基于从这样的常规实验收集的信息，本领域技术人员可以容易地确定应避免在该处单独的或与其他突变组合的进一步取代的氨基酸。
如本文使用的术语“多肽片段”和“截短的多肽”是指与对应的全长蛋白相比具有氨基末端缺失和/或羧基末端缺失的多肽。在各种实施方案中，片段的长度可以是例如，至少5个、至少10个、至少25个、至少50个、至少100个、至少150个、至少200个、至少250个、至少300个、至少350个、至少400个、至少450个、至少500个、至少600个、至少700个、至少800个、至少900个或至少1000个氨基酸。在各种实施方案中，片段的长度还可以是例如，最多1000个、最多900个、最多800个、最多700个、最多600个、最多500个、最多450个、最多400个、最多350个、最多300个、最多250个、最多200个、最多150个、最多100个、最多50个、最多25个、最多10个或最多5个氨基酸。片段还可以在其任一个或两个末端处包含一个或更多个另外的氨基酸，例如，来自不同的天然存在的蛋白的氨基酸的序列(例如，Fc或亮氨酸拉链结构域)或人工氨基酸序列(例如，人工接头序列)。
本文使用的术语“多肽变体”、“杂合多肽(hybrid polypeptide)”和“多肽突变体”是指包含这样的氨基酸序列的多肽，其中相对于另一个多肽序列，一个或更多个氨基酸残基被插入到该氨基酸序列中、从该氨基酸序列中缺失和/或被取代到该氨基酸序列中。在各种实施方案中，待被插入、缺失或取代的氨基酸残基的数目可以是例如，至少1个、至少2个、至少3个、至少4个、至少5个、至少10个、至少25个、至少50个、至少75个、至少100个、至少125个、至少150个、至少175个、至少200个、至少225个、至少250个、至少275个、至少300个、至少350个、至少400个、至少450个或至少500个氨基酸的长度。本公开内容的杂合体包括融合蛋白。
多肽的“衍生物”是已经被化学修饰的多肽，所述化学修饰例如，缀合至另一个化学部分(诸如例如，聚乙二醇、白蛋白(例如，人类血清白蛋白))、磷酸化和糖基化。
术语“％序列同一性”与术语“％同一性”在本文中可互换使用并且指当使用序列比对程序比对时，在两个或更多个肽序列之间的氨基酸序列同一性的水平或在两个或更多个核苷酸序列之间的核苷酸序列同一性的水平。例如，如本文使用的，通过定义的算法确定的80％同一性与80％序列同一性意指相同的意思，并且意指给定序列与另一长度的另一序列至少80％相同。在各种实施方案中，％同一性选自例如，与给定序列至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％、至少95％或至少99％或更大的序列同一性。在各种实施方案中，％同一性在例如约60％至约70％、约70％至约80％、约80％至约85％、约85％至约90％、约90％至约95％或约95％至约99％的范围内。
术语“％序列同源性”与术语“％同源性”在本文中可互换使用并且指当使用序列比对程序比对时，在两个或更多个肽序列之间的氨基酸序列同源性的水平或在两个或更多个核苷酸序列之间的核苷酸序列同源性的水平。例如，如本文使用的，通过定义的算法确定的80％同源性与80％序列同源性意指相同的意思，并且因此给定序列的同源物具有相对于给定序列的长度大于80％的序列同源性。在各种实施方案中，％同源性选自例如，与给定序列至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％、至少95％或至少99％或更大的序列同源性。在各种实施方案中，％同源性在例如约60％至约70％、约70％至约80％、约80％至约85％、约85％至约90％、约90％至约95％或约95％至约99％的范围内。
可以用于确定两个序列之间的同一性的示例性计算机程序包括但不限于在互联网在NCBI的网站上公开地可得的一套BLAST程序，例如BLASTN、BLASTX和TBLASTX、BLASTP和TBLASTN。另参见Altschul等人,J.Mol.Biol.215:403-10,1990(特别参考公布的默认设置，即参数w＝4、t＝17)和Altschul等人,Nucleic Acids Res.,25:3389-3402,1997。当相对于GenBank蛋白序列和其他公开数据库中的氨基酸序列评价给定氨基酸序列时，通常使用BLASTP程序进行序列检索。BLASTX程序优选用于针对在GenBank蛋白序列和其他公开数据库中的氨基酸序列检索在所有阅读框中已经被翻译的核酸序列。使用开放空位(gap)罚分为11.0和延伸空位罚分为1.0的默认参数并且使用BLOSUM-62矩阵运行BLASTP和BLASTX两者。
除了计算序列同一性百分比以外，BLAST算法还进行两个序列之间的相似性的统计分析(参见例如，Karlin&Altschul,Proc.Natl.Acad.Sci.USA,90:5873-5787,1993)。由BLAST算法提供的相似性的一个量度是最小总概率(P(N))，所述最小总概率提供在两个核苷酸序列或氨基酸序列之间的匹配偶然发生的概率的指示。例如，如果在测试核酸与参考核酸的比较中，最小总概率为例如小于约0.1、小于约0.01或小于约0.001，则该核酸被认为与参考序列相似。
如本文使用的术语“修饰”是指对肽骨架(例如氨基酸序列)的任何操作或对多肽的翻译后修饰(例如糖基化)。
如本文使用的术语“knob-into-hole修饰”是指两个免疫球蛋白重链的CH3结构域之间的界面内的修饰。在一种实施方案中，“knob-into-hole修饰”包含一条抗体重链中的氨基酸取代T366W和任选的氨基酸取代S354C，以及另一条抗体重链中的氨基酸取代T366S、L368A、Y407V和任选的Y349C。knob-into-hole技术在例如美国专利第5,731,168号；美国专利第7,695,936号；Ridgway等人,Prot Eng 9,617-621(1996)和Carter,J Immunol Meth248,7-15(2001)中描述。
如本文使用的术语“可生物活化药物”或“VitoKine”意指这样的化合物，该化合物是一种药物前体，在被施用至受试者后，经由一些化学加工或生理加工在体内释放药物，使得可生物活化药物转化为对靶组织有活性的产物。可生物活化药物是经历生物活化然后表现出其药理学作用的任何化合物。因此，可生物活化药物可被视为包含专门的无毒保护性基团的药物，所述专门的无毒保护性基团以暂时的方式用于改变或消除母体分子的不期望的特性。
如本文使用的术语“融合蛋白”是指包含两个或更多个最初编码不同蛋白的基因的融合多肽分子，其中融合蛋白的组分通过肽键直接彼此连接或通过肽接头彼此连接。如本文使用的术语“融合”是指通过肽键直接连接的组分或经由一个或更多个肽接头连接的组分。
“接头”是指共价地或通过离子键、范德华力或氢键连接两个其他分子的分子，例如在5'末端处与一个互补序列杂交并且在3'末端处与另一个互补序列杂交从而连接两个非互补序列的核酸分子。“可裂解的接头”是指可以被降解、消化或以其他方式切断以使通过该可裂解的接头连接的两个组分分开的接头。可裂解的接头通常被酶，通常是肽酶、蛋白酶、核酸酶、脂肪酶等裂解。可裂解的接头还可以通过环境因素，诸如例如温度、pH、盐浓度等的改变来裂解。
如本文使用的术语“肽接头”是指包含一个或更多个氨基酸，通常为约1-30个氨基酸的肽。肽接头是本领域已知的或在本文中被描述。合适的非免疫原性接头肽包括，例如，(G4S)n、(SG4)n或G4(SG4)n肽接头。“n”通常是1和10之间，通常2和4之间的数字。
“药物组合物”是指适于在动物中药物使用的组合物。药物组合物包含药理学有效量的活性剂和药学上可接受的载体。“药理学有效量”是指有效产生预期药理学结果的剂的量。“药学上可接受的载体”是指任何标准的药物载体、媒介物、缓冲剂和赋形剂，诸如磷酸盐缓冲盐水溶液、5％右旋糖的水性溶液，和乳液，诸如油/水乳液或水/油乳液，和各种类型的润湿剂和/或佐剂。合适的药学载体和制剂在Remington's Pharmaceutical Sciences,第21版2005,Mack Publishing Co,Easton中描述。“药学上可接受的盐”是可以被配制成用于药学用途的化合物的盐，包括例如金属盐(钠、钾、镁、钙等)和氨的盐或有机胺的盐。
如本文使用的，“治疗(treatment)”(及其语法变化形式诸如“治疗(treat)”或“治疗(treating)”)是指试图改变被治疗的个体的疾病的自然过程的临床干预，并且临床干预可以用于预防来进行或者在临床病理过程期间进行。期望的治疗效果包括但不限于，防止疾病的发生或复发、减轻症状、减轻疾病的任何直接或间接病理后果、防止转移、降低疾病进展速度、改善或缓和疾病状态以及缓解或改善预后。如本文使用的，“减轻”疾病、紊乱或状况意指降低疾病、紊乱或状况的症状的严重性和/或发生频率。另外，本文中述及“治疗”包括述及治愈性、缓解性和预防性治疗。
如本文使用的术语“有效量”或“治疗有效量”是指足以治疗特定紊乱、状况或疾病，诸如改善、缓和、减轻和/或延迟其一种或更多种症状的化合物或组合物的量。述及癌症或其他不需要的细胞增殖，有效量包括足以实现以下的量：(i)降低癌细胞的数目；(ii)降低肿瘤尺寸；(iii)在一定程度抑制、延缓、减缓并且优选地停止癌细胞浸润到外周器官中；(iv)抑制(即，在一定程度减缓并且优选地停止)肿瘤转移；(v)抑制肿瘤生长；(vi)预防或延迟肿瘤的发生和/或复发；和/或(vii)在一定程度减轻一种或更多种与癌症相关的症状。有效量可以以一次或更多次施用来施用。
措辞“施用”或“引起施用(cause to be administered)”是指由医学专业人员(例如医师)或控制患者的医疗护理的人员采取的控制和/或允许向患者施用所讨论的剂/化合物的行动。引起施用可以包括诊断和/或确定合适的治疗方案，和/或为患者开特定的剂/化合物的处方。这样的开处方可以包括例如起草处方形式、注释医疗记录等。当本文中描述施用时也设想了“引起施用”。
术语“患者”、“个体”和“受试者”可以互换地使用，并且指哺乳动物，优选人类或非人类灵长类动物，但也指家养哺乳动物(例如犬科动物或猫科动物)、实验室哺乳动物(例如小鼠、大鼠、兔、仓鼠、豚鼠)和农业哺乳动物(例如马科动物、牛科动物、猪、绵羊(ovine))。在各种实施方案中，患者可以是在医院、精神病护理机构如门诊或其他临床环境中的医师或其他卫生工作者的护理下的人类(例如，成年男性、成年女性、青少年男性、青少年女性、男性儿童、女性儿童)。在各种实施方案中，患者可以是免疫功能低下的患者或免疫系统减弱的患者，包括但不限于患有原发性免疫缺陷、AIDS的患者；服用某些免疫抑制药物的癌症患者和移植患者；以及患有影响免疫系统的遗传性疾病(例如，先天性无丙种球蛋白血症、先天性IgA缺乏症)的患者。在各种实施方案中，患者患有免疫原性癌症，包括但不限于膀胱癌、肺癌、黑素瘤和报道具有高突变率的其他癌症(Lawrence等人,Nature,499(7457):214-218,2013)。
术语“免疫疗法”是指包括但不限于以下的癌症治疗：使用针对特定肿瘤抗原的耗尽性抗体的治疗；使用抗体-药物缀合物的治疗；使用针对共刺激分子或共抑制分子(免疫检查点)(诸如CTLA-4、PD-1、PDL-1、CD40、OX-40、CD137、GITR、LAG3、TIM-3、SIRPα、CD47、GITR、ICOS、CD27、Siglec 7、Siglec 8、Siglec 9、Siglec 15和VISTA、CD276、CD272、TIM-3、B7-H4)的激动性抗体、拮抗性抗体或阻断性抗体的治疗；使用双特异性T细胞结合抗体
Figure BDA0003806684520000301
诸如博纳吐单抗的治疗；涉及施用生物响应调节剂(诸如IL-2、IL-4、IL-7、IL-10、IL-12、IL-15、IL-21、IL-22、GM-CSF、IFN-αIFN-β和IFN-γ、TGF-β拮抗剂或TGF-β陷阱)的治疗；使用治疗性疫苗诸如sipuleucel-T的治疗；使用治疗性病毒(包括但不限于溶瘤病毒诸如T-vec)的治疗；使用树突细胞疫苗或肿瘤抗原肽疫苗或新抗原疫苗的治疗；使用NK细胞的治疗；使用嵌合抗原受体(CAR)-T细胞的治疗；使用CAR-NK细胞的治疗；使用DC或T细胞的治疗；使用iPS诱导的NK细胞的治疗；使用iPS诱导的T细胞的治疗和使用疫苗诸如卡介苗(BCG)的治疗；使用肿瘤浸润淋巴细胞(TIL)的治疗；使用过继转移的抗肿瘤T细胞(离体扩增的T细胞和/或TCR-T细胞)的治疗；使用TALL-104细胞的治疗；以及使用免疫刺激剂诸如Toll样受体(TLR)激动剂CpG、TLR7、TLR8、TLR9和咪喹莫特的治疗。
“耐受性或难治性癌症”是指不响应于先前的抗癌疗法的肿瘤细胞或癌症，所述先前的抗癌疗法包括例如化学疗法、手术、放射疗法、干细胞移植和免疫疗法。肿瘤细胞可以在治疗的开始时是耐受性或难治性的，或者它们可以在治疗期间变成耐受性或难治性的。难治性肿瘤细胞包括在治疗的开始时不响应的肿瘤，或最初持续短的时间段响应但最终不能响应治疗的肿瘤。难治性肿瘤细胞还包括响应于使用抗癌疗法的治疗但不能响应于随后几轮疗法的肿瘤。为了本发明的目的，难治性肿瘤细胞还包括通过使用抗癌疗法的治疗表现出被抑制但在治疗停止后长达5年，有时长达10年或更长时间而复发的肿瘤。抗癌疗法可以使用单独的化学治疗剂、单独的放射、单独的靶向疗法、单独的手术或其组合。为了便于描述而非限制，应理解，难治性肿瘤细胞与耐受性肿瘤是可互换的。
术语“肿瘤相关抗原”(TAA)是指，例如，相对于大多数正常细胞，由癌细胞选择性表达或在癌细胞中过表达的细胞表面抗原。如本文使用的术语“TAA变体”和“TAA突变体”是指包含这样的氨基酸序列的TAA，其中相对于另一个TAA序列，一个或更多个氨基酸残基被插入到该氨基酸序列中、从该氨基酸序列中缺失和/或被取代到该氨基酸序列中。在各种实施方案中，待被插入、缺失或取代的氨基酸残基的数目可以是例如，至少1个、至少2个、至少3个、至少4个、至少5个、至少10个、至少25个、至少50个、至少75个、至少100个、至少125个、至少150个、至少175个、至少200个、至少225个、至少250个、至少275个、至少300个、至少350个、至少400个、至少450个或至少500个氨基酸的长度。
术语“新抗原”是指例如免疫系统先前未暴露于其的细胞表面抗原，尤其是通过放射、化学疗法、病毒感染、致瘤性转化/突变、药物代谢等改变宿主抗原而产生的细胞表面抗原，相对于大多数正常细胞，所述细胞表面抗原由癌细胞选择性表达或在癌细胞中过表达。
如本文使用的术语“抗体”以最宽泛的意义使用，并涵盖各种抗体结构(IgG1、2、3或4、IgM、IgA、IgE)，包括但不限于单克隆抗体、多克隆抗体、多特异性抗体(例如双特异性抗体或双功能抗体)和抗体片段，条件为它们表现出期望的抗原结合活性。
如本文使用的，术语“抗体片段”是指除完整抗体之外的分子，其包含完整抗体的一部分并且与完整抗体所结合的抗原结合。抗体片段的实例包括但不限于Fv、Fab、Fab'、Fab'-SH、F(ab')2、双抗体、线性抗体、单链抗体分子(例如scFv)以及单结构域抗体。
如本文使用的术语“Fab片段”是指包含轻链的VL结构域和恒定结构域(CL)以及重链的VH结构域和第一恒定结构域(CH1)的免疫球蛋白片段。
如本文使用的术语“可变区”或“可变结构域”是指通常参与免疫球蛋白或抗体与抗原结合的免疫球蛋白或抗体重链或轻链的结构域。免疫球蛋白或抗体的重链和轻链的可变结构域(分别为VH和VL)通常具有相似的结构，其中每个结构域包含四个保守的框架区(FR)和三个互补决定区(CDR)。
如本文使用的“人类免疫球蛋白”是具有这样的氨基酸序列的免疫球蛋白，该氨基酸序列对应于由人类或人类细胞产生的或利用人类免疫球蛋白组库或其他人类免疫球蛋白编码序列衍生自非人类来源的免疫球蛋白的氨基酸序列。人类免疫球蛋白的这一定义明确排除了包含非人类抗原结合残基的人源化免疫球蛋白。
如本文使用的术语“Fc结构域”或“Fc区”用于定义免疫球蛋白重链的C末端区域，其包含至少一部分恒定区。该术语包括天然序列Fc区和变体Fc区。IgG Fc区包含IgG CH2和IgG CH3结构域。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(protuberance)”(“knob”)并且在其另一条链中具有相应的引入的“空腔(cavity)”(“hole”)的CH3结构域；参见美国专利第5,821,333号，通过引用明确并入本文)。这样的变体CH3结构域可以用于促进如本文描述的两个不同的免疫球蛋白重链的异源二聚体化。除非本文中另外规定，否则Fc区或恒定区中氨基酸残基的编号根据EU编号系统。
如本文使用的术语“效应子功能”是指可归因于免疫球蛋白的Fc区的那些生物活性，所述Fc区随免疫球蛋白亚型而不同。免疫球蛋白效应子功能的实例包括：C1q结合和补体依赖性细胞毒性(CDC)、Fc受体结合、抗体依赖性细胞介导的细胞毒性(ADCC)、抗体依赖性细胞吞噬作用(ADCP)、细胞因子分泌、免疫复合物介导的抗原呈递细胞的抗原摄取、细胞表面受体(例如B细胞受体)的下调和B细胞活化。
如本文使用的术语“调节性T细胞”或“Treg细胞”意指可以抑制其他T细胞(效应T细胞)的响应的专门类型的CD4+T细胞。Treg细胞的特征在于CD4、IL-2受体(CD25)的α亚基和转录因子叉头盒P3(FOXP3)的表达(Sakaguchi,Annu Rev Immunol 22,531-62(2004))，并在诱导和维持对抗原(包括由肿瘤表达的抗原)的外周自身耐受性中起关键作用。
如本文使用的术语“常规CD4+T细胞”意指调节性T细胞之外的CD4+T细胞。
如本文使用的术语“Treg细胞的选择性活化”意指基本上不伴随其他T细胞亚群(诸如CD4+辅助T细胞、CD8+细胞毒性T细胞、NK T细胞)或自然杀伤(NK)细胞的活化的Treg细胞的活化。
如本文使用的，“特异性结合”意指对抗原的结合是选择性的，并且可以与不需要的或非特异性的相互作用区分开。免疫球蛋白结合特定抗原的能力可以通过酶联免疫吸附测定(ELISA)或本领域技术人员熟悉的其他技术例如表面等离子体共振(SPR)技术来测量。
如本文使用的术语“亲和力”或“结合亲和力”是指分子(例如抗体)的单个结合位点与其结合配偶体(例如抗原)之间的非共价相互作用的总和的强度。分子X对其配偶体Y的亲和力一般可以通过解离常数(KD)来呈现，解离常数是解离速率常数和结合速率常数(分别为koff和kon)的比率。用于测量亲和力的一种特殊方法是表面等离子体共振(SPR)。
如本文使用的，术语“降低的结合”是指各自相互作用的亲和力的降低，如例如通过SPR测量的。相反，“增加的结合”是指各自相互作用的结合亲和力的增加。
如本文使用的术语“聚合物”通常包括但不限于均聚物；共聚物，诸如例如嵌段、接枝、无规和交替共聚物；和三元共聚物；以及它们的混合物和变化形式(modifications)。此外，除非另外明确限定，否则术语“聚合物”应包括材料的所有可能的几何构型。这些构型包括但不限于全同立构、间同立构和无规对称。
“多核苷酸”是指包含核苷酸单元的聚合物。多核苷酸包括天然存在的核酸，诸如脱氧核糖核酸(“DNA”)和核糖核酸(“RNA”)以及核酸类似物。核酸类似物包括包含以下的那些：非天然存在的碱基，以天然存在的磷酸二酯键以外的连接与其他核苷酸衔接(engage)的核苷酸，或包含通过除了磷酸二酯键以外的连接来附接的碱基的核苷酸。因此，核苷酸类似物包括例如并且不限于，硫代磷酸酯、二硫代磷酸酯、磷酸三酯(phosphorotriester)、氨基磷酸酯(phosphoramidate)、硼烷磷酸酯(boranophosphate)、甲基膦酸酯、手性甲基膦酸酯、2-O-甲基核糖核苷酸、肽核酸(PNA)等。这样的多核苷酸可以例如使用自动化的DNA合成仪来合成。术语“核酸”通常是指大的多核苷酸。术语“寡核苷酸”通常是指短的多核苷酸，通常不大于约50个核苷酸。将理解的是，当核苷酸序列由DNA序列(即，A、T、G、C)表示时，这还包括其中“U”代替“T”的RNA序列(即，A、U、G、C)。
本文使用常规符号描述多核苷酸序列：单链多核苷酸序列的左手末端为5'-末端；双链多核苷酸序列的左手方向被称作5'-方向。向新生RNA转录物5'至3'添加核苷酸的方向被称作转录方向。具有与mRNA相同的序列的DNA链被称作“编码链”；在具有与从该DNA转录的mRNA相同的序列的DNA链上并且位于RNA转录物的5'-末端的5'的序列被称作“上游序列”；在具有与RNA相同的序列的DNA链上并且在编码RNA转录物的3'-末端的3'的序列被称作“下游序列”。
“互补的”是指两个多核苷酸的相互作用表面的拓扑相容性或匹配在一起。因此，这两个分子可以被描述为互补的，并且更进一步地，接触表面特征是彼此互补的。如果第一多核苷酸的核苷酸序列与第二多核苷酸的多核苷酸结合配偶体的核苷酸序列基本上相同，或者如果第一多核苷酸可以与第二多核苷酸在严格杂交条件下杂交，则第一多核苷酸与第二多核苷酸是互补的。
“与...特异性杂交”或“特异性杂交”或“与...选择性杂交”是指当特定核苷酸序列存在于复杂混合物(例如，总细胞)DNA或RNA中时，在严格条件下核酸分子优先与该序列的结合、双链体化或杂交。术语“严格条件”是指这样的条件，在该条件下探针将优先与其靶子序列杂交，并且在较小程度上与其它序列杂交或根本不与其他序列杂交。在核酸杂交实验诸如DNA杂交和RNA杂交的背景下，“严格杂交”和“严格杂交洗涤条件”为序列依赖性的，并且在不同的环境参数下是不同的。对核酸杂交的广泛指导可见于：Tijssen,1993,Laboratory Techniques in Biochemistry and Molecular Biology--Hybridizationwith Nucleic Acid Probes,第I部分，第2章，“Overview of principles ofhybridization and the strategy of nucleic acid probe assays”,Elsevier,N.Y.；Sambrook等人,2001,Molecular Cloning:A Laboratory Manual,Cold Spring HarborLaboratory，第3增补版,NY；以及Ausubel等人，编著，现行版，Current Protocols inMolecular Biology,Greene Publishing Associates and Wiley Interscience,NY。
通常，高度严格的杂交和洗涤条件被选择为比在定义的离子强度和pH的特定序列的热解链点(Tm)低约5℃。Tm为50％的靶序列与完全匹配的探针杂交的温度(在定义的离子强度和pH条件下)。非常严格的条件被选择为等于特定探针的Tm。用于在DNA印迹或RNA印迹中的滤器上进行具有多于约100个互补残基的互补核酸的杂交的严格杂交条件的实例为在42℃，50％福尔马林与1mg肝素，过夜进行杂交。高度严格的洗涤条件的实例为在72℃，0.15M NaCl，持续约15分钟。严格洗涤条件的实例为在65℃，0.2 x SSC洗涤，持续15分钟。参见Sambrook等人对于SSC缓冲液的描述。可以进行低严格洗涤以去除背景探针信号，然后进行高严格洗涤。对于例如多于约100个核苷酸的双链体的示例性中度严格洗涤为在45℃，1 x SSC，持续15分钟。对于例如多于约100个核苷酸的双链体的示例性低严格洗涤为在40℃，4-6 x SSC，持续15分钟。通常，在特定杂交测定中，与对不相关探针观察到的信噪比相比2x(或更高)的信噪比指示检测到特异性杂交。
“引物”是指能够与指定的多核苷酸模板特异性杂交并提供用于合成互补多核苷酸的起始点的多核苷酸。当多核苷酸引物被置于诱导合成的条件下(即，在核苷酸、互补多核苷酸模板和用于聚合的剂诸如DNA聚合酶的存在下)时，这样的合成发生。引物通常为单链的，但可以为双链的。引物通常为脱氧核糖核酸，但许多合成的和天然存在的引物可用于许多应用。引物与模板互补，引物被设计成与模板杂交以用作合成起始的位点，但不需要反映模板的精确序列。在这样的情况下，引物与模板的特异性杂交取决于杂交条件的严格度。引物可以用例如显色性、放射性或萦光性的部分标记并用作可检测的部分。
当用于述及多核苷酸使用时，“探针”是指能够与另一多核苷酸的指定序列特异性杂交的多核苷酸。探针与靶互补多核苷酸特异性杂交，但不需要反映模板的精确互补序列。在这样的情况下，探针与靶的特异性杂交取决于杂交条件的严格度。探针可以用例如显色性、放射性或萦光性的部分标记并用作可检测的部分。在其中探针提供用于合成互补多核苷酸的起始点的情况下，探针也可以是引物。
“载体”是可以被用于将与其连接的另一核酸引入到细胞中的多核苷酸。一种类型的载体是“质粒”，其是指可以将另外的核酸区段连接到其中的线性或环状双链DNA分子。另一种类型的载体是病毒载体(例如，复制缺陷型逆转录病毒、腺病毒和腺相关病毒)，其中另外的DNA区段可以被引入到病毒基因组中。某些载体能够在引入了它们的宿主细胞中自主复制(例如，包含细菌复制起点的细菌载体和附加型哺乳动物载体(episomal mammalianvector))。其他载体(例如，非附加型哺乳动物载体)在引入到宿主细胞中后被整合到宿主细胞的基因组中，并且从而与宿主基因组一起复制。“表达载体”是可以指导选择的多核苷酸的表达的一种类型的载体。
“调控序列”是影响与其可操作地连接的核酸的表达(例如，表达的水平、时机或位置)的核酸。调控序列可以例如直接对被调控的核酸发挥其作用，或通过一种或更多种其他分子(例如，与调控序列和/或核酸结合的多肽)的作用发挥其作用。调控序列的实例包括启动子、增强子和其他表达控制元件(例如多腺苷酸化信号)。调控序列的另外的实例描述于例如Goeddel,1990,Gene Expression Technology:Methods in Enzymology 185,Academic Press,San Diego；Calif.和Baron等人,1995,Nucleic Acids Res.23:3605-06中。如果调控序列影响核苷酸序列的表达(例如，表达的水平、时机或位置)，则核苷酸序列与调控序列“可操作地连接”。
“宿主细胞”是可以被用于表达本公开内容的多核苷酸的细胞。宿主细胞可以是原核生物，例如大肠杆菌(E.coli)，或者宿主细胞可以是真核生物，例如单细胞真核生物(例如，酵母或其他真菌)、植物细胞(例如，烟草或番茄植物细胞)、动物细胞(例如，人类细胞、猴细胞、仓鼠细胞、大鼠细胞、小鼠细胞或昆虫细胞)或杂交瘤。通常，宿主细胞是可以用编码多肽的核酸转化或转染的培养的细胞，所述核酸然后可以在宿主细胞中被表达。措辞“重组宿主细胞”可以被用于表示已经用待表达的核酸转化或转染的宿主细胞。宿主细胞还可以是包含核酸但不以期望的水平表达该核酸的细胞，除非调控序列被引入到宿主细胞中，使得调控序列变成与该核酸可操作地连接。应理解，术语宿主细胞不仅指特定的受试者细胞，而且还指这样的细胞的后代或潜在后代。因为某些修饰可以由于例如突变或环境影响而在随后世代中发生，这样的后代实际上可能与亲本细胞不相同，但仍然被包括在如本文使用的该术语的范围内。
术语“分离的分子”(其中分子是例如多肽或多核苷酸)是这样的分子：该分子凭借其起源或衍生的来源(1)不与在其天然状态中伴随其的、天然地缔合的组分缔合，(2)基本上不含来自相同物种的其他分子，(3)由来自不同物种的细胞表达，或(4)不存在于自然界中。因此，化学合成的、或在与其天然起源的细胞不同的细胞系统中表达的分子将是与其天然缔合的组分“分离”的。也可以通过使用本领域熟知的纯化技术分离使分子基本上不含天然缔合的组分。分子纯度或均一性可以通过本领域熟知的许多手段来测定。例如，多肽样品的纯度可以使用本领域熟知的技术使用聚丙烯酰胺凝胶电泳和对凝胶染色以使多肽可视化来测定。为了某些目的，可以通过使用HPLC或本领域熟知的用于纯化的其他手段来提供更高的分辨率。
当样品的至少约60％至75％表现为单一种类的多肽时，蛋白或多肽是“基本上纯的”、“基本上均一的”或“基本上纯化的”。多肽或蛋白可以是单体的或多聚体的。基本上纯的多肽或蛋白将通常包含约50％、60％、70％、80％或90％W/W的蛋白样品，更通常约95％并且优选地将超过99％纯。蛋白纯度或均一性可以通过本领域熟知的许多手段来指示，所述手段诸如蛋白样品的聚丙烯酰胺凝胶电泳，随后用本领域熟知的染色剂对凝胶进行染色后使单个多肽条带可视化。为了某些目的，可以通过使用HPLC或本领域熟知的用于纯化的其他手段来提供更高的分辨率。
如本文使用的术语“标记”或“标记的”是指在抗体中掺入另一种分子。在一种实施方案中，标记物是可检测的标志物，例如掺入放射性标记的氨基酸或附接至可以通过标记的亲和素检测的生物素基部分的多肽(例如，包含萦光标志物的链霉亲和素或可以通过光学方法或量热法检测的酶活性)。在另一种实施方案中，标记物或标志物可以是治疗性的，例如药物缀合物或毒素。标记多肽和糖蛋白的各种方法是本领域中已知的并且可以被使用。用于多肽的标记物的实例包括但不限于以下：放射性同位素或放射性核素(例如3H、14C、15N、35S、90Y、99Tc、111In、125I、131I)；萦光标记物(例如FITC、罗丹明、镧系元素萦光团)；酶标记物(例如辣根过氧化物酶、β-半乳糖苷酶、萤光素酶、碱性磷酸酶)；化学发光标志物；生物素基基团；由第二报告物识别的预定的多肽表位(例如亮氨酸拉链配对序列、第二抗体的结合位点、金属结合结构域、表位标签)；磁性剂(magnetic agent)，诸如钆螯合物；毒素诸如百日咳毒素、紫杉醇、细胞松弛素B、短杆菌肽D、溴化乙锭、吐根碱、丝裂霉素、依托泊苷、替尼泊苷、长春新碱、长春花碱、秋水仙碱、多柔比星、柔红霉素、二羟基蒽二酮(dihydroxyanthracin dione)、米托蒽醌、光神霉素、放线菌素D、1-去氢睾酮、糖皮质激素、普鲁卡因、丁卡因、利多卡因、普萘洛尔(propranolol)和嘌呤霉素及其类似物或同系物。在各种实施方案中，标记物通过各种长度的间隔臂(spacer arm)附接，以降低潜在的空间位阻。
如本文使用的术语“异源”是指非天然的或在自然界不存在的组合或状态，所述组合或状态例如可以通过用源自另一来源的组分或状态替换现有的天然组分或状态来实现。类似地，在除了其中蛋白天然地表达的生物体之外的生物体中的蛋白表达构成异源表达系统和异源蛋白。
应理解，本文描述的本公开内容的方面和实施方案包括“由这些方面和实施方案组成”和/或“主要由这些方面和实施方案组成”。
本文中述及“约”值或参数包括(并且描述)针对该值或参数本身的变化。例如，述及“约X”的描述包括对“X”的描述。
除非上下文另外明确规定，否则如本文和所附的权利要求中使用的单数形式“一”、“或”和“该”包括复数指代物。应理解，本文描述的本公开内容的方面和变化形式包括“由这些方面和变化形式组成”和/或“主要由这些方面和变化形式组成”。
VitoKine平台的描述
本发明提供了一种基于细胞因子的可生物活化药物(“VitoKine”)平台，该平台旨在降低基于全身性机制的毒性，并导致蛋白(例如细胞因子)的更广泛的治疗效用。参考图1，本发明的新的VitoKine构建体包含D1结构域、“活性部分结构域”(D2)和“遮蔽部分结构域”(D3)，所述D1结构域为靶向结构域、免疫检查点调节剂靶向结构域、半衰期延长结构域或双重或多功能部分结构域。所提出的活化VitoKine D2结构域的方法在图2中描绘。重要的是，因为VitoKine构建体的D2将保持惰性或减弱的活性，直到被病变组织中上调的蛋白酶在局部活化，这将限制活性部分与非病变细胞的外周中或细胞表面上的受体的结合，以防止途径的过度活化并减少不期望的“组织外”“中靶”毒性。此外，在蛋白酶活化之前，VitoKine活性部分的惰性将显著降低潜在的抗原吸收或靶吸收(antigen or targetsink)，并从而延长体内半衰期且导致改进的在意图治疗部位处的生物分布和生物可利用度。
D1结构域(“靶向结构域、半衰期延长结构域或双重或多功能部分结构域”)
在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是针对肿瘤相关抗原的抗体或抗体片段或蛋白或肽的形式的靶向部分。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是针对免疫检查点调节剂的抗体、抗体片段、蛋白或肽。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是作为自身免疫调节剂的抗体或抗体片段或蛋白或肽。在各种实施方案中，本发明的VitoKine构建体包含D1，该D1结构域作用为将D2结构域保持在组织部位，诸如肿瘤微环境(TME)或炎性组织部位。在各种实施方案中，本发明的VitoKine构建体包含D1，该D1是双重功能的，例如组织靶向和保留。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是聚合物。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是半衰期延长部分。在各种实施方案中，本发明的VitoKine构建体包含D1结构域，该D1结构域是Fc结构域。
Fc结构域
IgG类的免疫球蛋白是人类血液中最丰富的蛋白之一。它们的循环半衰期可长达21天。已经报道了将IgG的Fc区与另一种蛋白(诸如各种细胞因子和受体)的结构域组合的融合蛋白(参见例如，Capon等人,Nature,337:525-531,1989；Chamow等人,TrendsBiotechnol.,14:52-60,1996)；美国专利第5,116,964号和第5,541,087号)。原型融合蛋白是通过IgG Fc的铰链区中的半胱氨酸残基连接的同源二聚体蛋白，产生类似于IgG分子、没有重链可变区和CH1结构域以及轻链的分子。包含Fc结构域的融合蛋白的二聚体性质在提供与其他分子的更高级相互作用(即二价或双特异性结合)方面可能是有利的。由于结构同源性，Fc融合蛋白表现出与具有相似同种型的人类IgG相当的体内药代动力学谱。
术语“Fc”是指包含完整抗体的非抗原结合片段的序列的分子或序列，无论是单体形式还是多聚体形式。天然Fc的原始免疫球蛋白来源优选地为人类来源，并且可以是任何免疫球蛋白，尽管IgG1和IgG2是优选的。天然Fc由单体多肽组成，单体多肽可以通过共价(即二硫键)和非共价缔合连接成二聚体或多聚体形式。天然Fc分子的单体亚基之间的分子间二硫键的数目取决于类别(例如IgG、IgM、IgA、IgE)或亚类(例如IgG1、IgG2、IgG3、IgG4、IgA1、IgGA2)而从1至4变化。天然Fc的一种实例是由木瓜蛋白酶消化IgG产生的二硫键合的二聚体(参见Ellison等人(1982),Nucleic Acids Res.10:4071-9)。如本文使用的术语“天然Fc”是单体、二聚体和多聚体形式的总称。Fc结构域包含蛋白A、蛋白G、各种Fc受体和补体蛋白的结合位点。
在各种实施方案中，术语“Fc变体”是指从天然Fc修饰、但仍然包含补救受体FcRn的结合位点的分子或序列。国际申请WO 97/34631(1997年9月25日公布)和WO 96/32478描述了示例性的Fc变体以及与补救受体的相互作用，并特此通过引用并入。此外，天然Fc包含可被去除的位点，因为它们提供本发明的融合分子不需要的结构特征或生物活性。因此，在各种实施方案中，术语“Fc变体”包括缺乏一个或更多个天然Fc位点或残基的分子或序列，所述Fc位点或残基影响或参与(1)二硫键的形成，(2)与所选宿主细胞的不相容性，(3)在所选宿主细胞中表达时的N末端或C末端异质性，(4)糖基化，(5)与补体的相互作用，诸如CDC，(6)与补救受体以外的Fc受体的结合，或(7)抗体依赖性细胞毒性(ADCC)。
术语“Fc结构域”包括如上文定义的天然Fc和Fc变体分子和序列。如同Fc变体和天然Fc，术语“Fc结构域”包括单体或多聚体形式的分子，无论是从完整抗体消化的还是通过重组基因表达或通过其他方式产生的。在各种实施方案中，“Fc结构域”是指两个Fc结构域单体(SEQ ID NO:13)的二聚体，其通常包括全部或部分的铰链区。在各种实施方案中，Fc结构域可以被突变以缺乏效应子功能。在各种实施方案中，Fc结构域中的每个Fc结构域单体包含CH2抗体恒定结构域中的氨基酸取代，以减少Fc结构域和Fcγ受体之间的相互作用或结合。在各种实施方案中，Fc结构域的每个亚基包含两个氨基酸取代，这两个氨基酸取代减少与活化性Fc受体的结合和/或效应子功能，其中所述氨基酸取代是L234A和L235A。在各种实施方案中，Fc结构域的每个亚基包含三个氨基酸取代，这三个氨基酸取代减少与活化性Fc受体的结合和/或效应子功能，其中所述氨基酸取代是L234A、L235A和G237A(SEQ ID NO:14)。在各种实施方案中，Fc结构域的每个亚基包含三个氨基酸取代，这三个氨基酸取代减少与活化性Fc受体的结合和/或效应子功能，其中所述氨基酸取代是L234A、L235A和P329A。
在各种实施方案中，Fc结构域可以被突变以进一步延长体内半衰期。在各种实施方案中，Fc结构域的每个亚基包含增强与人类FcRn的结合的三个氨基酸取代，其中所述氨基酸取代是M252Y、S254T和T256E(SEQ ID NO:156)，公开于美国专利公布第7,658,921号中。在各种实施方案中，Fc结构域的每个亚基包含增强与人类FcRn的结合的一个氨基酸取代，其中所述氨基酸取代是N434A(SEQ ID NO:166)，公开于美国专利公布第7,371,826号中。在各种实施方案中，Fc结构域的每个亚基包含增强与人类FcRn的结合的一个氨基酸取代，其中所述氨基酸取代是M428L和N434S，公开于美国专利公布第8,546,543号中。在各种实施方案中，半衰期延长突变可以与减少与活化性Fc受体的结合和/或效应子功能的氨基酸取代组合。
在各种实施方案中，Fc结构域中的两个Fc结构域单体中的每一个包含促进两个单体异源二聚体化的氨基酸取代。在各种其他实施方案中，Fc结构域单体的异源二聚体化可以通过在两个Fc结构域单体中引入不同但相容的取代诸如“knob-into-hole”残基对来促进。“knob-into-hole”技术也公开于美国专利公布第8,216,805号中。在又另一种实施方案中，一个Fc结构域单体包含knob突变T366W，并且另一个Fc结构域单体包含hole突变T366S、L358A和Y407V。在各种实施方案中，引入形成稳定化的二硫桥的两个Cys残基(S354C在“knob”侧，并且Y349C在“hole”侧)(SEQ ID NO:15和16)。异源二聚体Fc的使用可以产生单价VitoKine构建体。
疾病相关靶或肿瘤相关抗原抗体和蛋白/肽结合物
在各种实施方案中，D1可以是呈以下形式的靶向部分：针对肿瘤相关抗原(TAA)的抗体或对病变细胞或病变组织表现出结合亲和力的另一种蛋白或肽。TAA可以是期望针对其的免疫应答的任何分子、大分子、分子的组合等。TAA可以是包含多于一个多肽亚基的蛋白。例如，蛋白可以为二聚体、三聚体或更高级多聚体。在各种实施方案中，蛋白的两个或更多个亚基可以以共价键诸如例如二硫键连接。在各种实施方案中，蛋白的亚基可以以非共价相互作用保持在一起。因此，TAA可以是技术人员希望针对其诱导免疫应答的任何肽、多肽、蛋白、核酸、脂质、碳水化合物或有机小分子或其任何组合。在各种实施方案中，TAA为包含以下的肽：约5个、约6个、约7个、约8个、约9个、约10个、约11个、约12个、约13个、约14个、约15个、约16个、约17个、约18个、约19个、约20个、约25个、约30个、约35个、约40个、约45个、约50个、约55个、约60个、约65个、约70个、约75个、约80个、约85个、约90个、约95个、约100个、约150个、约200个、约250个、约300个、约400个、约500个、约600个、约700个、约800个、约900或约1000个氨基酸。在各种实施方案中，肽、多肽或蛋白是通常通过注射向受试者施用的分子。在各种实施方案中，在施用后，肿瘤特异性抗体或结合蛋白用作靶向部分，以将VitoKine导向病变部位，诸如癌症部位，在该部位，活性结构域可以被释放并与其在病变细胞或病变组织上的同源受体相互作用。
任何前述标志物可以用作本发明的VitoKine构建体的疾病相关靶或TAA靶。在各种实施方案中，设想用于本公开内容的VitoKine构建体和方法的一种或更多种疾病相关靶或其变体或TAA、TAA变体或TAA突变体选自或源自表2中提供的列表。
表2
Figure BDA0003806684520000431
Figure BDA0003806684520000441
Figure BDA0003806684520000451
肿瘤相关抗原的另外的实例包括TRP-1、TRP-2、MAG-1、MAGE-3、BAGE、GAGE-1、GAGE-2、p15(58)、CEA、RAGE、NY-BSO(LAGE)、SCP-1、Hom/Mel-40、H-Ras、BCR-ABL、E2A-PRL、H4-RET、IGH-IGK、MYL-RAR、人类乳头瘤病毒(HPV)抗原E6和E7、TSP-180、MAGE-4、MAGE-5、MAGE-6、p185erbB2、p180erbB-3、c-met nm-23H1、PSA、TAG-72-4、CA 19-9、CA 72-4、CAM17.1、Numa、K-ras、β-联蛋白、CDK4、Muni-1、p16、TAGE、PSCA、CT7、端粒酶、43-9F、5T4、791Tgp72、β-HCG、BCA225、BTAA、CA 15-3(CA 27.29\BCAA)、CA 195、CA 242、CA-50、CAM43、CD68\KF1、CO-029、FGF-5、G250、Ga733(EpCAM)、HTgp-175、M344、MA-50、MG7-Ag、MOV18、NB/70K、NY-CO-1、RCAS1、SDCCAG16、TA-90(Mac-2结合蛋白\亲环蛋白C相关蛋白)、TAAL6、TAG72、TLP和TPS。
免疫检查点调节剂
已经报道了在各种免疫细胞上表达的许多免疫检查点蛋白抗原，包括例如，CD152(由活化的CD8+T细胞、CD4+T细胞和调节性T细胞表达)、CD279(在肿瘤浸润性淋巴细胞上表达，由活化的T细胞(CD4和CD8两者)、调节性T细胞、活化的B细胞、活化的NK细胞、无变应性T细胞、单核细胞、树突细胞表达)、CD274(在T细胞、B细胞、树突细胞、巨噬细胞、血管内皮细胞、胰岛细胞上表达)和CD223(由活化的T细胞、调节性T细胞、无变应性T细胞、NK细胞、NKT细胞和浆细胞样树突细胞表达)(参见，例如，Pardoll,D.,Nature Reviews Cancer,12:252-264,2012)。结合被确定为免疫检查点蛋白的抗原的抗体是本领域技术人员已知的。例如，本领域已经描述了各种抗CD276抗体(参见，例如，美国专利公布第20120294796号(Johnson等人)及其中引用的参考文献)；本领域已经描述了各种抗CD272抗体(参见，例如，美国专利公布第20140017255号(Mataraza等人)及其中引用的参考文献)；本领域已经描述了各种抗CD152/CTLA-4抗体(参见，例如，美国专利公布第20130136749号(Korman等人)及其中引用的参考文献)；本领域已经描述了各种抗LAG-3/CD223抗体(参见，例如，美国专利公布第20110150892号(Thudium等人)及其中引用的参考文献)；本领域已经描述了各种抗CD279/PD-1抗体(参见例如，美国专利第7,488,802号(Collins等人)及其中引用的参考文献)；本领域已经描述了各种抗PD-L1抗体(参见例如，美国专利公布第20130122014号(Korman等人)及其中引用的参考文献)；本领域已经描述了各种抗TIM-3抗体(参见，例如，美国专利公布第20140044728号(Takayanagi等人)及其中引用的参考文献)；并且本领域已经描述了各种抗B7-H4抗体(参见，例如，美国专利公布第20110085970号(Terrett等人)及其中引用的参考文献)。这些参考文献中的每一篇关于其中教导的特定抗体和序列通过引用以其整体特此并入。
在各种实施方案中，D1可包含表现出与存在于免疫细胞表面上的免疫检查点蛋白抗原结合的抗体、抗体片段或蛋白或肽。在各种实施方案中，免疫检查点蛋白抗原选自由以下组成但不限于以下的组：CD276、CD272、CD152、CD223、CD279、CD274、CD40、SIRPα、CD47、OX-40、GITR、ICOS、CD27、4-1BB、TIM-3、B7-H4、Siglec-7、Siglec-8、Siglec-9、Siglec-15和VISTA。
在各种实施方案中，D1可以包含针对存在于肿瘤细胞表面上的免疫检查点蛋白抗原的抗体，所述免疫检查点蛋白抗原选自由以下组成但不限于以下的组：PD-L1、B7-H3和B7-H4。
在各种实施方案中，D1是为拮抗性成纤维细胞活化蛋白(FAP)抗体的抗体或抗体片段。在各种实施方案中，抗体是包含SEQ ID NO:193和194中列出的氨基酸序列的人源化抗FAP抗体。在各种实施方案中，D1是针对免疫检查点调节剂的抗体或抗体片段。在各种实施方案中，抗体是拮抗性PD-1抗体或抗体片段。在各种实施方案中，抗体是包含SEQ ID NO:195和196中列出的氨基酸序列的拮抗性人源化PD-1抗体。在各种实施方案中，抗体是包含SEQ ID NO:197和198中列出的氨基酸序列的拮抗性人类PD-1抗体。在各种实施方案中，抗体是包含SEQ ID NO:275和276中列出的氨基酸序列的拮抗性人源化PD-1抗体。在各种实施方案中，抗体是包含SEQ ID NO:277和278中列出的氨基酸序列的拮抗性人类PD-1抗体。在各种实施方案中，抗体是拮抗性PD-L1抗体或抗体片段。在各种实施方案中，抗体是包含SEQID NO:279和280中列出的氨基酸序列的拮抗性人类PD-L1抗体。在各种实施方案中，抗体VitoKine构建体包含SEQ ID NO:128-142、180-181、281-286、296-297和303-306中列出的氨基酸序列。
自身免疫性紊乱和炎性紊乱的调节剂
在各种炎性组织或免疫细胞上高度表达的任何前述蛋白可用作本发明的VitoKine构建体的自身免疫/炎性疾病靶。在各种实施方案中，设想用于本公开内容的VitoKine构建体和方法的一种或更多种自身免疫/炎性疾病靶、其变体或其突变体/同种型选自或源自表3中提供的列表。这些靶也可以作为癌症靶向应用。
表3
自身免疫性紊乱和炎性紊乱或癌症的靶
Figure BDA0003806684520000481
Figure BDA0003806684520000491
在各种实施方案中，D1靶向部分可以是炎性组织特异性抗体、抗体片段、表现出与病变细胞或疾病微环境结合的另一种蛋白或肽，诸如TNF、TNFR、整联蛋白A4β7、IL-6Rα、BLYS、TSLP。在各种实施方案中，抗体VitoKine构建体包含SEQ ID NO:143-146中列出的氨基酸序列。
聚合物
在各种实施方案中，D1可以是聚合物，例如聚乙二醇(PEG)。在各种实施方案中，使用常规的化学方法，例如化学缀合，可以将聚合物例如PEG共价附接在N末端或C末端或内部位置处。在各种实施方案中，聚合物，例如PEG，可以经由位点特异性缀合或细胞因子的其他氨基酸或工程化的特定氨基酸取代共价附接在D2结构域的N末端处。
半衰期延长部分
在各种实施方案中，其他半衰期延长部分可以在本发明中用作D1结构域以增加VitoKine的血清半衰期。半衰期延长部分包括但不限于Fc结构域、Fc变体、抗体、抗体片段(Fab、ScFv)和EXTEN(Schellenberger等人,Nat.Biotechnol.27:1 186-1 192,2009)和人类血清白蛋白蛋白。
D2结构域(“活性部分结构域”)
D2是VitoKine构建体的活性部分，D2的活性被可逆地遮蔽在构建体中，并可在疾病部位被蛋白酶裂解后恢复。该活性部分可以是任何蛋白，包括但不限于任何天然或变体白介素或细胞因子多肽。重要的是，因为VitoKine构建体的“活性部分”将保持惰性或减弱的活性，直到被病变组织中上调的蛋白酶在局部活化，这将限制活性部分与非病变细胞的外周中或细胞表面上的受体的结合，以防止途径的过度活化并减少不期望的“组织外”“中靶”毒性。此外，在蛋白酶活化之前，VitoKine活性部分的惰性将显著降低潜在的抗原吸收或靶吸收，并从而延长体内半衰期且导致改进的在意图治疗部位处的生物分布和暴露。
IL-15
白介素-15(IL-15)是一种细胞因子，由两个独立的研究组根据其在中和性抗IL-2抗体的存在下刺激IL-2依赖性CTLL-2 T细胞系增殖的能力鉴定出(Steel等人,Trends inPharmacological Sciences,33(1):35-41,2012)。IL-15和IL-2在体外具有相似的生物学特性，与它们的共有受体(R)信号传导组分(IL-2/15Rβγc)一致。然而，IL-15与IL-2的特异性是由独特的私有的α链受体提供的，这些α链受体使IL-15Rαβγ和IL-2Rαβγ异源三聚体高亲和力受体复合物完整，并且从而允许取决于所表达的配体和高亲和力受体的不同的响应性。令人感兴趣地，IL-15转录物和IL-15Rα转录物两者具有比IL-2/IL-2Rα广泛得多的组织分布。此外，各种复杂的转录后调控机制严格控制IL-15表达。因此，基于复杂的调控以及IL-15和IL-15Rα表达的不同模式，这种受体/配体对的关键的体内功能可能不同于IL-2和IL-2Rα的体内功能。迄今为止，检查IL-15的生物学的研究已经鉴定出若干关键的非冗余作用，诸如IL-15在自然杀伤(NK)细胞、NK-T细胞和肠上皮内淋巴细胞的发育过程中的重要性和功能。IL-15在自身免疫过程(诸如类风湿性关节炎)和恶性肿瘤(诸如成人T细胞白血病)期间的作用，表明IL-15的失调可能导致对宿主的有害影响(Fehniger等人,Blood,97:14-32,2001)。
如本文使用的，术语“天然IL-15”和“天然白介素-15”在蛋白或多肽的上下文中是指任何天然存在的哺乳动物白介素-15氨基酸序列，包括未成熟形式或前体形式和成熟形式。各个物种的天然哺乳动物白介素-15的氨基酸序列的GenBank登录号的非限制性实例包括NP_032383(小家鼠(Mus musculus)，未成熟形式)、AAB60398(猕猴(macaca mulatta)，未成熟形式)、NP_000576(人类，未成熟形式)、CAA62616(人类，未成熟形式)、AAI00964(人类，未成熟形式)和AAH18149(人类)。在本发明的各种实施方案中，天然IL-15是天然存在的哺乳动物IL-15的未成熟或前体形式。在其他实施方案中，天然IL-15是天然存在的哺乳动物IL-15的成熟形式。在各种实施方案中，天然IL-15是天然存在的人类IL-15的前体形式。在各种实施方案中，天然IL-15是天然存在的人类IL-15的成熟形式。在各种实施方案中，天然IL-15蛋白/多肽是分离或纯化的。在各种实施方案中，基于IL-15的结构域D2源自SEQ IDNO:1中列出的人类IL-15前体序列的氨基酸序列：
Figure BDA0003806684520000511
在各种实施方案中，基于IL-15的结构域D2包含SEQ ID NO:2中列出的人类IL-15成熟形式序列的氨基酸序列：
Figure BDA0003806684520000512
在各种实施方案中，基于IL-15的结构域D2是IL-15变体(或突变体)，该IL-15变体(或突变体)包含源自如SEQ ID NO:2中列出的成熟的人类IL-15多肽序列的序列。本文使用天然氨基酸、天然氨基酸在成熟序列中的位置和变异氨基酸来指代IL-15的变体(或突变体)。例如，“huIL-15 S58D”是指包含在SEQ ID NO:2的位置58处的S至D取代的人类IL-15。在各种实施方案中，本发明的D2结构域包含为IL-15变体(本文中也称为IL-15突变体结构域)的IL-15结构域。在各种实施方案中，IL-15变体包含与天然(或野生型)IL-15蛋白不同的氨基酸序列。在各种实施方案中，IL-15变体结合IL-15Rα多肽，并且起IL-15激动剂或拮抗剂的作用。在各种实施方案中，具有激动剂活性的IL-15变体具有超级激动剂活性。在各种实施方案中，IL-15变体可以起IL-15激动剂或拮抗剂的作用，与其与IL-15Rα的缔合无关。IL-15激动剂通过与野生型IL-15相比生物活性相当或增加来例示。IL-15拮抗剂通过与野生型IL-15相比生物活性降低或通过抑制IL-15介导的响应的能力来例示。在各种实施方案中，IL-15变体以增加或减少的活性与IL-15Rβγc受体结合。在各种实施方案中，与天然IL-15序列相比，IL-15变体的序列具有至少一个氨基酸变化，例如取代或缺失，这样的变化导致IL-15激动剂或拮抗剂活性。在各种实施方案中，氨基酸取代/缺失位于与IL-15Rβ和/或γC相互作用的IL-15的结构域中。在各种实施方案中，氨基酸取代/缺失不影响与IL-15Rα多肽的结合或产生IL-15变体的能力。基于已知的IL-15结构、IL-15与同源分子诸如具有已知结构的IL-2的比较，通过本文提供的合理或随机诱变和功能测定或其他经验方法，可以鉴定产生IL-15变体的合适的氨基酸取代/缺失。此外，合适的氨基酸取代可以是保守或非保守的改变和另外氨基酸的插入。在各种实施方案中，本发明的IL-15变体在SEQ ID NO:2中列出的成熟的人类IL-15序列的位置30、31、32、58、62、63、67、68或108处包含一个或多于一个氨基酸缺失或者一个或更多个氨基酸取代。在各种实施方案中，D30T(“D30”是指天然的成熟的人类IL-15序列中的氨基酸和残基位置，并且“T”是指IL-15变体中该位置处取代的氨基酸残基)、V31Y、H32E、S58H、S58I、S58P、S58R、S58Q、T62D、V63A、V63F、V63K、V63R、I67V、I68H、I68F、I68Q、I68G、I68K、I68D、Q108A、Q108S、Q108E、Q108K或Q108M取代导致具有拮抗剂活性的IL-15变体，并且S58D取代导致具有激动剂活性的IL-15变体。在各种实施方案中，IL-15变体在SEQ ID NO:2的N末端处包含1个、或2个、或3个、或4个、5个或6个氨基酸缺失。在各种实施方案中，IL-15变体在SEQ ID NO:2的C末端处包含1个、或2个、或3个、或4个、或5个、或6个、或7个、或8个、或9个、或10个氨基酸缺失。在各种实施方案中，IL-2变体包含在SEQ ID NO:2的位置N95后的‘GS’(SEQ ID NO:12)、或‘GGSGG’(SEQ ID NO:153)或‘GSSGGSGGS’(SEQ ID NO:154)氨基酸插入。在各种实施方案中，IL-15变体包含SEQ ID NO：3、182-192和199-215中列出的氨基酸序列。
在表4中提供了示例性的IL-15 Fc VitoKine构建体：
表4
蛋白ID SEQ ID NO：
P-0351 25
P-0170 26+15
P-0172 27
P-0202 28
P-0203 29
P-0204 30
P-0205 31
P-0206 32
P-0315 33
P-0316 34
P-0350 35
P-0354 36
P-0355 37
P-0385 38
P-0386 39
P-0387 40
P-0388 41
P-0389 42
P-0397 43
P-0660 162
P-0488 163
P-0489 164
P-0661 165
P-0650 169
P-0651 170
P-0662 171+15
P-0663 172+167
P-0664 173+167
P-0665 174+167
P-0682 230
P-0806 231
在各种实施方案中，IL-15抗体VitoKine或IL-15 Fc融合分子包含两个或更多个异源二聚体链，如表5中列出的：
表5
Figure BDA0003806684520000541
在各种实施方案中，基于IL-15的D2结构域包含IL-15构建体，该IL-15构建体包含SEQ ID NO:12中列出的IL-2Rβ或基于IL-2Rβ的封闭肽，该封闭肽选自具有SEQ ID NO:66-70中列出的氨基酸序列的构建体。
在各种实施方案中，基于IL-15的D2结构域包含IL-15构建体，该IL-15构建体包含基于IL-2Rβ的封闭肽，并具有两个或更多个异源二聚体链，如表6中列出的：
表6
Figure BDA0003806684520000551
IL-2
白介素-2(IL-2)是一种经典的Th1细胞因子，由T细胞通过T细胞抗原受体和共刺激分子CD28活化后产生。IL-2的调节通过作用于IL-2启动子以产生新的基因转录的信号传导途径和转录因子的活化而发生，但也涉及对IL-2mRNA的稳定性的调节。IL-2结合多链受体，包括通过Jak-STAT途径介导信号传导的高度调节的α链及β和γ链。IL-2向T细胞、B细胞和NK细胞递送活化、生长和分化信号。IL-2在介导活化诱导的T细胞的细胞死亡中也很重要，这一功能为终止免疫应答提供了关键机制。一种商业上可获得的未糖基化的人类重组IL-2产品阿地白介素(可从Prometheus Laboratories Inc.,San Diego Calif.以商标为
Figure BDA0003806684520000552
的des-丙氨酰基-1,丝氨酸-125人类白介素-2获得)，已被批准用于向罹患转移性肾细胞癌和转移性黑素瘤的患者施用。IL-2还被建议用于在罹患或感染丙型肝炎病毒(HCV)、人类免疫缺陷病毒(HIV)、急性髓性白血病、非霍奇金淋巴瘤、皮肤T细胞淋巴瘤、幼年类风湿性关节炎、特应性皮炎、乳腺癌和膀胱癌的患者中施用。不幸的是，短半衰期和严重毒性限制了IL-2的最佳给药。
如本文使用的，术语“天然IL-2”和“天然白介素-2”在蛋白或多肽的上下文中是指任何天然存在的哺乳动物白介素-2氨基酸序列，包括未成熟形式或前体形式和成熟形式。各个物种的天然哺乳动物白介素-2的氨基酸序列的GenBank登录号的非限制性实例包括NP_032392.1(小家鼠，未成熟形式)、NP_001040595.1(猕猴，未成熟形式)、NP_000577.2(人类，前体形式)、CAA01199,1(人类，未成熟形式)、AAD48509.1(人类，未成熟形式)和AAB20900.1(人类)。在本发明的各种实施方案中，天然IL-2是天然存在的哺乳动物IL-2的未成熟或前体形式。在其他实施方案中，天然IL-2是天然存在的哺乳动物IL-2的成熟形式。在各种实施方案中，天然IL-2是天然存在的人类IL-2的前体形式。在各种实施方案中，天然IL-2是天然存在的人类IL-2的成熟形式。在各种实施方案中，基于IL-2的结构域D2源自SEQID NO:6中列出的人类IL-2前体序列的氨基酸序列：
Figure BDA0003806684520000561
在各种实施方案中，基于IL-2的结构域D2包含SEQ ID NO:8中列出的在位置125处包含半胱氨酸至丝氨酸的取代的人类IL-2成熟形式野生型序列的氨基酸序列，但是与天然存在的IL-2相比，不改变IL-2受体结合：
Figure BDA0003806684520000562
在各种实施方案中，基于IL-2的结构域D2是IL-2变体(或突变体)，该IL-2变体(或突变体)包含源自如SEQ ID NO:8中列出的成熟的人类IL-2多肽序列的序列。在各种实施方案中，IL-2变体包含与天然(或野生型)IL-2蛋白不同的氨基酸序列。在各种实施方案中，IL-2变体结合IL-2Rα多肽，并且起IL-2激动剂或拮抗剂的作用。在各种实施方案中，具有激动剂活性的IL-2变体具有超级激动剂活性。在各种实施方案中，IL-2变体可以起IL-2激动剂或拮抗剂的作用，与其与IL-2Rα的缔合无关。IL-2激动剂通过与野生型IL-2相比生物活性相当或增加来例示。IL-2拮抗剂通过与野生型IL-2相比生物活性降低或通过抑制IL-2介导的响应的能力来例示。在各种实施方案中，与天然IL-2序列相比，IL-2变体的序列具有至少一个氨基酸变化，例如取代或缺失，这样的变化导致IL-2激动剂或拮抗剂活性。在各种实施方案中，IL-2变体具有源自SEQ ID NO:8的氨基酸序列，具有减少/消除的与IL-2Rα的结合，以便选择性地使用于治疗癌症的效应T细胞(Teff)活化和增殖；示例性氨基酸取代在表7中列出。在各种实施方案中，具有减少/消除的与IL-2Rα结合的IL-2变体包含SEQ ID NO:232-247中列出的氨基酸序列。在各种实施方案中，IL-2变体具有源自SEQ ID NO:8的氨基酸序列，具有减少的与IL-2Rβ和/或γc的结合，并且在使用于治疗自身免疫性疾病的调节性T细胞(Treg)活化和增殖方面具有增强的选择性；示例性氨基酸取代在表7中列出。如本领域技术人员将理解的，所有的突变可以任选地和独立地以任何方式组合，以实现最佳的亲和力和活性调节。
表7
Figure BDA0003806684520000571
在表8中提供了示例性的基于IL-2的VitoKine构建体：
表8
Figure BDA0003806684520000572
Figure BDA0003806684520000581
在各种实施方案中，活性部分选自由以下组成但不限于以下的序列的组的氨基酸序列：白介素-4(IL-4)(SEQ ID NO:17)、白介素-7(IL-7)(SEQ ID NO:18)、白介素-9(IL-9)(SEQ ID NO:19)、白介素-10(IL-10)(SEQ ID NO:20)、白介素-12α(IL-12α)(SEQ ID NO:21)、白介素-12β(IL-12β)(SEQ ID NO:22)、白介素-23α(IL-23α)(SEQ ID NO:23)和TGFβ(SEQ ID NO:24)。在各种实施方案中，活性部分是异源二聚体人类IL-12细胞因子，其包含作为链1的SEQ ID NO:21和作为链2的SEQ ID NO:22。在各种实施方案中，活性部分是异源二聚体人类IL-23细胞因子，其包含作为链1的SEQ ID NO:23和作为链2的SEQ ID NO:22。
D3结构域(“遮蔽部分结构域”)
D3结构域是“遮蔽部分结构域”，并主要用于在特定VitoKine构建体中可逆地遮蔽D2结构域的活性。D3结构域能够遮蔽D2的功能活性，直到在意图的治疗部位处被活化。在各种实施方案中，本发明的VitoKine构建体包含“遮蔽部分结构域”(D3)，该D3是D2蛋白或细胞因子的同源受体/结合配偶体。在各种实施方案中，D3结构域是D2结构域的同源受体/结合配偶体的变体或特异性结合物，诸如肽或抗体片段。在各种实施方案中，与野生型同源受体/结合配偶体相比，D3结构域具有增强的与D2结构域的结合。在各种实施方案中，与野生型同源受体/结合配偶体相比，D3结构域具有减少或消除的与D2结构域的结合。在各种实施方案中，D3结构域是能够遮蔽D2的活性的蛋白、或肽、或抗体、或抗体片段。在各种实施方案中，D3结构域是通过可裂解接头连接的DNA、RNA片段或诸如PEG的聚合物。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-15Rα细胞外结构域或其功能片段或变体。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-15RαSushi结构域(SEQ ID NO:5的氨基酸1-65)。在各种优选的实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是在Sushi结构域的C末端含有1-30个另外的IL-15Rα残基的IL-15RαSushi+结构域(例如，SEQ ID NO:5)。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2Rα细胞外结构域或其功能片段。在各种优选的实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2RαSushi结构域。在各种优选的实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2RαSushi结构域的变体。在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是IL-2Rβ细胞外结构域或IL-2Rβ-衍生的封闭肽。在各种实施方案中，D3结构域能够遮蔽D2的功能活性，直到D2在意图的治疗部位处被活化。
IL-15受体α
IL-15受体是一种I型细胞因子受体，由与IL-2受体共有的beta(β)和gamma(γ)亚基以及以高亲和力结合IL-15的alpha(α)亚基组成。全长人类IL-15Rα是一种1型跨膜蛋白，具有32个AA的信号肽、173个AA的细胞外结构域、21个AA的跨膜结构域、37个AA的细胞质尾和多个N-连接或O-连接的糖基化位点(Anderson等人,J.Biol Chem,270:29862-29869,1995)。先前已经展示了，对应于IL-15Rα完整细胞外结构域的IL-15Rα链的天然可溶性形式起高亲和力IL-15拮抗剂的作用。然而，与该发现形成鲜明对比的是，已展示，具有对IL-15的大部分结合亲和力的IL-15Rα的重组可溶性sushi结构域，通过IL-15Rβ/γ异源二聚体增强其结合和生物效应(增殖和防止凋亡)，起有效的IL-15激动剂的作用，而sushi结构域不影响IL-15结合和IL-15Rα/β/γ三部分膜受体的功能。这些结果表明，如果是天然产生的，这样的可溶性sushi结构域可能参与IL-15的反式呈递机制(Mortier等人,J.Biol Chem,281(3):1612-1619,2006)。
如本文使用的，术语“天然IL-15Rα”和“天然白介素-15受体α”在蛋白或多肽的上下文中是指任何天然存在的哺乳动物白介素-15受体α("IL-15Rα")氨基酸序列，包括未成熟形式或前体形式和成熟形式以及天然存在的同种型。各种天然哺乳动物IL-15Rα的氨基酸序列的GenBank登录号的非限制性实例包括NP_002180(人类)、ABK41438(猕猴)、NP_032384(小家鼠)、Q60819(小家鼠)、CA141082(人类)。在各种实施方案中，天然IL-15Rα是天然存在的哺乳动物IL-15Rα多肽的未成熟形式。在各种实施方案中，天然IL-15Rα是天然存在的哺乳动物IL-15Rα多肽的成熟形式。在各种实施方案中，天然IL-15Rα是天然存在的哺乳动物IL-15Rα多肽的形式。在各种实施方案中，天然IL-15Rα是天然存在的哺乳动物IL-15Rα多肽的全长形式。在各种实施方案中，天然IL-15Rα是天然存在的人类IL-15Rα多肽的未成熟形式。在各种实施方案中，天然IL-15Rα是天然存在的人类IL-15Rα多肽的成熟形式。在各种实施方案中，天然IL-15Rα是天然存在的人类IL-15Rα多肽的全长形式。在各种实施方案中，天然IL-15Rα蛋白或多肽是分离或纯化的。在各种实施方案中，IL-15Rα结构域源自SEQ ID NO:4中列出的人类IL-15Rα序列的氨基酸序列：
Figure BDA0003806684520000601
在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是包含成熟的人类IL-15Rα多肽的氨基酸序列的IL-15RαSushi+结构域，如SEQ ID NO:5中列出的：
Figure BDA0003806684520000611
在各种实施方案中，重现了全长IL-15Rα(SEQ ID NO:4)的大部分结合亲和力的IL-15的截短的同源共受体IL-15RαSushi+(SEQ ID NO:5)被用作D3结构域，以通过调节连接IL-15和IL-15RαSushi+的可裂解或不可裂解的接头来遮蔽IL-15的活性，以制备IL-15VitoKine。如技术人员可以理解的，D3结构域的长度可以不同于SEQ ID NO:5中列出的序列，条件为该D3结构域能重现全长IL-15α(SEQ ID NO:4)的大部分结合活性，即该D3结构域为功能片段。IL-15 VitoKine设计的独特之处在于充分利用了IL-15途径的独特特征，包括IL-15和IL-15α之间异常高的亲和力(30pM)，以及IL-15α的复合增强了IL-15在体内的活性。在通过疾病部位处上调的蛋白酶裂解连接IL-15和IL-15αSushi+的接头后，预计IL-15αShushi+或源自IL-15αECD的任何功能片段保持与IL-15的非共价缔合并增强IL-15活性。
IL-2受体
IL-2受体(IL-2R)是一种在某些免疫细胞诸如淋巴细胞的表面上表达的异源三聚体蛋白，结合并响应于一种称为IL-2的细胞因子。IL-2R具有三个亚基：α(CD25)、β(CD122)和γc(CD132，这是与其他五种细胞因子受体IL-4R、IL-7R、IL-9R、IL-15R和IL-21R共有的链)。人类受体的α链(别名：Tac抗原或p55)由染色体10p14-15上的基因IL-2RA编码。人类受体的β链(IL-2RB，CD122)的基因位于染色体22q11.2-12上，而共有的IL-2RγC链(IL-2RG)的基因位于染色体Xq13上。受体的所有三个亚基的组装对于信号转导进入B细胞和T细胞是重要的。IL-2R存在(暂时或永久地)于几乎所有造血细胞(包括淋巴谱系T、B和NK细胞，以及髓性细胞，如巨噬细胞、单核细胞和中性粒细胞)的细胞表面上。信号通过Janus激酶—Jak1和Jak3传递到细胞中。受体的β链的胞质内部分的磷酸化使得STAT-3和STAT-5因子能够形成同源二聚体。STAT-3和STAT-5的同源二聚体显示出增加的对细胞核的亲和力，在细胞核中它们与特定的DNA元件结合，增强IL-2依赖性基因的转录。
如本文使用的，术语“天然IL-2Rα”和“天然白介素-2受体α”在蛋白或多肽的上下文中是指任何天然存在的哺乳动物白介素-2受体α("IL-2Rα")氨基酸序列，包括未成熟形式或前体形式和成熟形式以及天然存在的同种型。各种天然哺乳动物IL-2Rα的氨基酸序列的GenBank登录号的非限制性实例包括NP_032393.3(小家鼠)、CAK26553.1(人类)和NP_000408.1(人类)。在各种实施方案中，天然IL-2Rα是天然存在的哺乳动物IL-2Rα多肽的未成熟形式。在各种实施方案中，天然IL-2Rα是天然存在的哺乳动物IL-2Rα多肽的成熟形式。在各种实施方案中，天然IL-2Rα是天然存在的哺乳动物IL-2Rα多肽的形式。在各种实施方案中，天然IL-2Rα是天然存在的哺乳动物IL-2Rα多肽的全长形式。在各种实施方案中，天然IL-2Rα是天然存在的人类IL-2Rα多肽的未成熟形式。在各种实施方案中，天然IL-2Rα是天然存在的人类IL-2Rα多肽的成熟形式。在各种实施方案中，天然IL-2Rα是天然存在的人类IL-2Rα多肽的全长形式。在各种实施方案中，天然IL-2Rα蛋白或多肽是分离或纯化的。在各种实施方案中，IL-2Rα结构域源自SEQ ID NO:9中列出的人类IL-2Rα序列的氨基酸序列：
Figure BDA0003806684520000621
在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是包含成熟的人类IL-2Rα多肽的氨基酸序列的IL-2RαSushi结构域，如SEQ ID NO:10中列出的：
Figure BDA0003806684520000622
在各种实施方案中，IL-2RαSushi(SEQ ID NO:10)用于遮蔽IL-2活性，以制备IL-2VitoKine。与包含单个sushi结构域的IL-15Rα不同，IL-2Rα包含由接头隔开的两个sushi结构域。在各种实施方案中，IL-2 VitoKine包含IL-2RαSushi变体，该IL-2RαSushi变体包含氨基酸取代以破坏IL-2Rα和IL-2之间的特异性非共价相互作用，从而降低IL-2Rα与IL-2的结合亲和力。虽然天然IL-2Rα以30nM的中等亲和力与IL-2结合，但在裂解接头后，IL-2Rα仍有可能不解离。IL-2Rα与IL-2的缔合可以降低IL-2的活性和/或T细胞亚群的平衡向不期望的方向的倾斜。随着亲和力降低突变(例如R36A、K38E或L42G或Y43A或这些取代的任何组合)被引入IL-2RαSushi中，IL-2RαSushi结构域可能在蛋白酶裂解接头后从IL-2解离离开。
在各种实施方案中，本发明的VitoKine构建体包含D3结构域，该D3结构域是包含如SEQ ID NO:267-270中列出的氨基酸序列的IL-2RαSushi结构域变体之一。
L1接头和L2接头
可裂解的接头
可裂解的接头或对疾病相关酶敏感的接头可以包含部分，例如蛋白底物，其能够被与非疾病组织相比在疾病部位以升高的水平存在的蛋白酶特异性地裂解。文献中有报道，在各种类型的癌症例如实体瘤中，具有已知底物的酶的水平增加。参见例如，La Rocca等人,Brit.J.Cancer 90:1414-1421和Ducry等人,Bioconjug.Chem.21:5-13,2010，其中每篇文献通过引用以其整体并入本文。在各种实施方案中，能够裂解蛋白酶可裂解接头的蛋白酶选自由以下组成的组：金属蛋白酶例如基质金属蛋白酶(MMP)1-28，和丝氨酸蛋白酶例如尿激酶型纤溶酶原激活物(uPA)和蛋白裂解酶(Matriptase)、半胱氨酸蛋白酶例如豆荚蛋白、天冬氨酸蛋白酶以及组织蛋白酶蛋白酶。在表9中提供了示例性蛋白酶底物肽序列：
表9
Figure BDA0003806684520000641
在表10中提供了示例性蛋白酶底物肽序列，其可用作蛋白酶可裂解接头，在可裂解接头的C末端或N末端或两个末端上具有或不具有不同长度的肽间隔物：
表10
蛋白酶 底物肽 SEQ ID NO：
MMP-2，7，9，14 SPLGLAGS 71
MMP-2、7、9、14，蛋白裂解酶 EPLELRAG 72
蛋白裂解酶、uPA、豆荚蛋白 LSGRSDNH 73
MMP-2 GPLGIAGQ 74
MMP-2，14 GTAHLMGG 75
MMP-14 RIGSLRTA 76
MMP-14 SGRSENIRTA 157
MMP-2，9 GPLGMLSQ 77
MMP-9，uPA RPSASRSA 78
MMP PLGLAG 79
uPA LGGSGRSANAILE 80
uPA GGSGRSANAI 81
uPA SGRSA 82
豆荚蛋白 AANL 83
豆荚蛋白 GPTNKVR 158
组织蛋白酶C GFFY 84
组织蛋白酶D GPICFRLG 85
组织蛋白酶E RQAGFSL 86
蛋白裂解酶 RQARAVGG 159
前列腺特异性抗原 HSSKLQ 87
在各种实施方案中，蛋白酶是MMP-9或MMP-2。在另外的特定实施方案中，蛋白酶是uPA。在另外的特定实施方案中，蛋白酶是MMP-14。在另外的特定实施方案中，蛋白酶是豆荚蛋白。在各种实施方案中，一个VitoKine分子包含两种不同的蛋白酶可裂解接头。在各种实施方案中，蛋白酶可裂解的接头包含蛋白酶识别序列‘GPLGMLSQ’(SEQ ID NO：77)。在各种实施方案中，蛋白酶可裂解的接头包含蛋白酶识别序列‘LGGSGRSANAILE’(SEQ ID NO：80)。在各种实施方案中，蛋白酶可裂解的接头包含蛋白酶识别序列‘SGRSENIRTA’(SEQ ID NO：157)。在各种实施方案中，蛋白酶可裂解的接头包含蛋白酶识别序列‘GPTNKVR’(SEQ IDNO：158)。在各种实施方案中，接头(例如，可裂解的接头)可以被肿瘤相关蛋白酶裂解。在各种实施方案中，在除癌症之外的疾病诸如炎性疾病中，可裂解接头可以被其他疾病特异性蛋白酶裂解。
在各种实施方案中，肽间隔物可以被掺入蛋白酶可裂解序列的任一侧或蛋白酶可裂解序列的两侧，或者作为不具有蛋白酶底物位点的不可裂解接头。肽间隔物用于将可裂解接头定位成对负责裂解的酶更可及。间隔物的长度可以被改变或优化，以平衡酶促裂解的可及性和可逆地遮蔽D2结构域以免发挥其生物活性所需的空间限制。间隔物可以包含1-100个氨基酸。合适的肽间隔物是本领域已知的，并且包括但不限于含有柔性氨基酸残基诸如甘氨酸和丝氨酸的肽接头。在各种实施方案中，间隔物可以包含1个至12个氨基酸，包括基序G、S、GS、GSGS(SEQ ID NO：116)、GGS(SEQ ID NO：117)、GSGS(SEQ ID NO：121)、GSGSGS(SEQ ID NO：122)、GSGSGSGS(SEQ ID NO：123)、GSGSGSGSGS(SEQ ID NO：124)或GSGSGSGSGSGS(SEQ ID NO：125)。在其他实施方案中，间隔物可以包含基序(GGGGS)(SEQ IDNO：118)n，其中n是1至10的整数。在其他实施方案中，间隔物还可以包含除甘氨酸和丝氨酸之外的氨基酸。
在表11中提供了示例性蛋白酶可裂解接头，其具有位于蛋白酶底物肽(加下划线)的侧翼的间隔肽：
表11
蛋白酶可裂解的接头 SEQ ID NO：
GGGSGGGGSGGGGS<u>LSGRSDNH</u>GGSGGGGS 88
GSSSGRSENIRTAGT 89
GGGGSGGGGSGGGS<u>LGGSGRSANAILE</u>GGSGGGGS 90
GGGGSGGGGS<u>LGGSGRSANAILE</u>GGGGS 91
GGGGS<u>LGGSGRSANAILE</u>GGS 92
GGGSGPTNKVRGGS 93
GGS<u>GPLGMLSQ</u>GGGS 94
G<u>GPLGMLSQ</u>S 95
GG<u>GPLGMLSQ</u>GGS 96
G<u>GPTNKVR</u>GS 160
G<u>RQARAVGG</u>S 161
GGG<u>SGRSENIRTA</u>GG 298
在各种实施方案中，可裂解接头可以通过除蛋白水解以外的机制活化，所述蛋白水解包括但不限于水解，诸如可以在不同的pH条件下经由受控的释放机制脱落的可释放的PEG化聚合物。
不可裂解的接头
不可裂解的接头在蛋白结构域之间提供共价连接和另外的结构和/或空间柔性。如本领域已知的，含有柔性氨基酸残基诸如甘氨酸和丝氨酸的肽接头可用作不可裂解的接头。在各种实施方案中，不可裂解的接头可以包含1-100个氨基酸。在各种实施方案中，间隔物可以包含基序GS、GSGS(SEQ ID NO：116)、GGS(SEQ ID NO：117)、GGGGS(SEQ ID NO：118)、GGSG(SEQ ID NO：119)或SGGG(SEQ ID NO：120)。在其他实施方案中，接头可以包含基序(GGGGS)(SEQ ID NO：118)n，其中n是1至10的整数。在其他实施方案中，接头还可以包含除甘氨酸和丝氨酸之外的氨基酸。在另一种实施方案中，不可裂解的接头可以是简单的化学键，例如酰胺键(例如通过PEG的化学缀合)。不可裂解的接头在生理条件下以及在病变部位诸如癌症部位或炎性疾病部位是稳定的。
在表12中提供了示例性的不可裂解的接头：
表12
接头序列 SEQ ID NO：
EPKSSDKTHTSPPS 107
GGGSGGGSGGGS 108
GGGS 109
GSSGGSGGS 110
GSSGT 111
GGGGSGGGGSGGGS 112
AEAAAKEAAAKEAAAKA 113
GGGGSGGGGSGGGGSGGGGS 114
GGGSGGGS 115
GSGS 116
GGS 117
GGGGS 118
GGSGG 119
SGGG 120
GSGS 121
GSGSGS 122
GSGSGSGS 123
GSGSGSGSGS 124
GSGSGSGSGSGS 125
GGGGSGGGGS 126
GGGGSGGGGSGGGGS 127
可裂解的接头和不可裂解的接头的组合
在各种实施方案中，L1接头和L2接头可以都是可裂解接头或都是不可裂解的接头或者是可裂解接头和不可裂解的接头的组合，以产生不同形式的D2结构域的活性部分，从而实现不同的治疗意图或平衡风险/收益比或顺从细胞因子的不同特性。通过接头裂解而释放的示例性活性形式在图2中描绘。分别源自L1的裂解和L1及L2两者的裂解的活性形式1和活性形式3是短效细胞因子，取决于D3构象而具有不同程度的功能活性。从半衰期延长或疾病组织靶向部分D1的裂解和释放会增加活化的D2结构域的局部浓度。在局部作用后，短效活性形式可以从体循环中迅速消除，以减少毒性。相比之下，源自L2裂解的活性形式2是一种功能完全恢复的、长效的和组织靶向的保存的细胞因子(conserved cytokine)，其在疾病部位持续保留更长时间和增强的功效。
多核苷酸
在另一方面，本公开内容提供了分离的核酸分子，该分离的核酸分子包含编码以下的多核苷酸：本公开内容的IL-15、IL-15变体、IL-15Rα、IL-15Rα变体、IL-2、IL-2变体、IL-2Rα、IL-2Rα变体、Fc、Fc变体、靶向TAA或免疫检查点调节剂的抗体、抗体片段或VitoKine构建体。主题核酸可以是单链的或双链的。这样的核酸可以是DNA或RNA分子。DNA包括例如cDNA、基因组DNA、合成的DNA、通过PCR扩增的DNA及其组合。编码VitoKine构建体的基因组DNA获取自基因组文库，该基因组文库对许多物种是可得的。合成的DNA从化学合成重叠的寡核苷酸片段然后组装片段以重构编码区和侧翼序列的部分或全部而可获得。RNA可以从指导高水平合成mRNA的原核表达载体(诸如使用T7启动子和RNA聚合酶的载体)获得。本公开内容的DNA分子包括全长基因以及多核苷酸及其片段。全长基因还可以包含编码N末端信号序列的序列。此类核酸可以例如在用于制备新的VitoKine构建体的方法中使用。
在各种实施方案中，分离的核酸分子包含本文描述的多核苷酸，并且还包含编码本文描述的至少一种异源蛋白的多核苷酸。在各种实施方案中，核酸分子还包含编码本文描述的接头或铰链接头的多核苷酸。
在各种实施方案中，本公开内容的重组核酸可以可操作地连接至表达构建体中的一种或更多种调控核苷酸序列。调控序列是本领域所认知的并且被选择以指导VitoKine构建体的表达。相应地，术语调控序列包括启动子、增强子和其他表达控制元件。在Goeddel；Gene Expression Technology:Methods in Enzymology,Academic Press,San Diego,Calif.(1990)中描述了示例性的调控序列。通常，所述一种或更多种调控核苷酸序列可以包括但不限于启动子序列、前导或信号序列、核糖体结合位点、转录起始和终止序列、翻译起始和终止序列、以及增强子或激活序列。本公开内容设想了本领域已知的组成型或诱导型启动子。启动子可以是天然存在的启动子或者组合多于一个启动子的元件的杂合启动子。表达构建体可以存在于细胞中的附加体诸如质粒上，或者表达构建体可以插入染色体中。在各种实施方案中，表达载体包含选择性标志物基因以允许选择转化的宿主细胞。选择性标志物基因是本领域熟知的并且将随着使用的宿主细胞变化。
在本公开内容的另一方面，主题核酸在表达载体中提供，所述表达载体包含编码VitoKine构建体并可操作地连接到至少一个调控序列的核苷酸序列。术语“表达载体”是指用于从多核苷酸序列表达多肽的质粒、噬菌体、病毒或载体。适于在宿主细胞中表达的载体是容易地可得的并利用标准重组DNA技术将核酸分子插入到载体中。此类载体可以包括各种各样的表达控制序列，当被可操作地连接至DNA序列时，所述表达控制序列控制该DNA序列的表达，并可以在这些载体中使用以表达编码VitoKine构建体的DNA序列。此类有用的表达控制序列包括，例如，SV40的早期启动子和晚期启动子、tet启动子、腺病毒或巨细胞病毒介导的早期启动子、RSV启动子、lac系统、trp系统、TAC或TRC系统、其表达由T7 RNA聚合酶指导的T7启动子、λ噬菌体的主要操纵子和启动子区域、fd外壳蛋白的控制区域、3-磷酸甘油酸激酶或其他糖酵解酶的启动子、酸性磷酸酶例如PhoS的启动子、酵母a-交配因子的启动子、杆状病毒系统的多角体启动子和已知控制原核细胞或真核细胞或它们的病毒的基因表达的其他序列及其各种组合。应理解，表达载体的设计可能取决于诸如待转化的宿主细胞的选择和/或期望表达的蛋白的类型的因素。此外，还应考虑载体的拷贝数、控制该拷贝数的能力、以及载体编码的任何其他蛋白诸如抗生素标志物的表达。适用于VitoKine的表达的示例性表达载体是包含VitoKine多核苷酸的pDSRa及其衍生物，以及本领域已知的或下文描述的任何另外的合适的载体。
本公开内容的重组核酸可以通过将克隆的基因或其一部分连接到适于在原核细胞、真核细胞(酵母、鸟类、昆虫或哺乳动物)或两者中表达的载体中来产生。用于产生重组VitoKine构建体的表达媒介物包括质粒和其他载体。例如，合适的载体包括以下类型的质粒：用于在原核细胞诸如大肠杆菌中表达的pBR322衍生的质粒、pEMBL衍生的质粒、pEX衍生的质粒、pBTac衍生的质粒和pUC衍生的质粒。
一些哺乳动物表达载体包含利于载体在细菌中增殖的原核序列以及在真核细胞中表达的一种或更多种真核转录单元两者。pcDNAI/amp、pcDNAI/neo、pRc/CMV、pSV2gpt、pSV2neo、pSV2-dhfr、pTk2、pRSVneo、pMSG、pSVT7、pko-neo和pHyg衍生的载体为适于转染真核细胞的哺乳动物表达载体的实例。这些载体中的一些被来自细菌质粒诸如pBR322的序列修饰，以利于在原核细胞和真核细胞两者中复制和药物抗性选择。可选地，病毒诸如牛乳头瘤病毒的衍生物(BPV-1)或Epstein-Barr病毒的衍生物(pHEBo、pREP衍生的和p205)可以用于蛋白在真核细胞中的瞬时表达。其他病毒(包括逆转录病毒)表达系统的实例可见于下文基因疗法递送系统的描述中。在质粒的制备方面和宿主生物体的转化方面所采用的各种方法是本领域熟知的。对于适于原核细胞和真核细胞两者的其他表达系统以及一般的重组程序，参见Sambrook、Fritsch和Maniatis的Molecular Cloning A Laboratory Manual,第2版(Cold Spring Harbor Laboratory Press,1989)第16和17章。在一些情形下，可能期望通过使用杆状病毒表达系统来表达重组多肽。此类杆状病毒表达系统的实例包括pVL衍生的载体(诸如pVL1392、pVL1393和pVL941)、pAcUW衍生的载体(诸如pAcUW1)以及pBlueBac衍生的载体(诸如含有B-gal的pBlueBac III)。
在各种实施方案中，载体将被设计成用于在CHO细胞中产生主题VitoKine构建体，诸如Pcmv-Script载体(Stratagene,La Jolla,Calif.)、pcDNA4载体(Invitrogen,Carlsbad,Calif.)和pCI-neo载体(Promega,Madison,Wis.)。将明显的是，主题基因构建体可以被用于引起主题VitoKine构建体在增殖于培养基中的细胞中表达，例如以产生用于纯化的蛋白，包括融合蛋白或变体蛋白。
本公开内容还涉及用重组基因转染的宿主细胞，所述重组基因包含编码一种或更多种主题VitoKine构建体的氨基酸序列的核苷酸序列。宿主细胞可以是原核细胞或真核细胞。例如，本公开内容的VitoKine构建体可以在细菌细胞诸如大肠杆菌、昆虫细胞(例如，使用杆状病毒表达系统)、酵母或哺乳动物细胞中表达。其他合适的宿主细胞是本领域技术人员已知的，诸如中国仓鼠卵巢(CHO)细胞或人胚肾293(HEK293)细胞。
相应地，本公开内容还涉及产生主题VitoKine构建体的方法。例如，可以在允许发生VitoKine构建体的表达的适当条件下培养用编码VitoKine构建体的表达载体转染的宿主细胞。VitoKine构建体可以从含有VitoKine构建体的细胞分泌和从含有VitoKine构建体的细胞和培养基的混合物分离。可选地，VitoKine构建体可以保留在细胞质中或保留在膜级分中，并收获、裂解细胞和分离蛋白。细胞培养物包括宿主细胞、培养基和其他副产物。用于细胞培养的合适培养基是本领域熟知的。
本公开内容的多肽和蛋白可以根据本领域技术人员熟知的蛋白纯化技术来纯化。这些技术在一个层面上涉及蛋白质级分和非蛋白质级分的粗分级。将肽或多肽与其他蛋白分离后，可以利用色谱技术和电泳技术进一步纯化感兴趣的肽或多肽，以实现部分或完全纯化(或纯化至均一)。如本文使用的术语“分离的多肽”或“纯化的多肽”意图指可与其他组分分离的组合物，其中多肽被纯化至相对于其天然可获得状态的任何程度。因此纯化的多肽还指脱离其可能天然存在的环境的多肽。通常，“纯化的”将指已经经历分级以去除各种其他组分的多肽组合物，并且多肽该组合物基本上保留了其表达的生物活性。当使用术语“基本上纯化的”时，该指定将指这样的肽或多肽组合物，其中多肽或肽形成组合物的大部分组分，诸如构成组合物中的蛋白的约50％、约60％、约70％、约80％、约85％或约90％或更多。
适用于纯化的各种技术将是本领域技术人员熟知的。这些技术包括，例如，用硫酸铵、PEG、抗体(免疫沉淀)等沉淀或通过热变性沉淀，然后离心；色谱，诸如亲和色谱(蛋白A柱)、离子交换色谱、凝胶过滤色谱、反相色谱、羟基磷灰石色谱、疏水相互作用色谱；等电聚焦；凝胶电泳；以及这些技术的组合。如本领域一般已知的，认为进行各种纯化步骤的顺序可以改变，或者某些步骤可以省略，并且仍得到用于制备基本上纯化的多肽的合适方法。
药物组合物
在另一方面，本公开内容提供了药物组合物，所述药物组合物包含与药学上可接受的载体混合的VitoKine构建体。此类药学上可接受的载体是本领域普通技术人员所熟知并理解的，并且已被广泛描述(参见例如，Remington's Pharmaceutical Sciences,第18版,A.R.Gennaro编著,Mack Publishing Company,1990)。可以包括药学上可接受的载体以用于改变、保持或维持，例如，pH、摩尔渗透压浓度、黏度、澄清度、颜色、等渗性、气味、无菌性、稳定性、溶解或释放速率、组合物的吸收或渗透的目的。这样的药物组合物可以影响多肽的物理状态、稳定性、体内释放速率以及体内清除速率。合适的药学上可接受的载体包括但不限于，氨基酸(诸如甘氨酸、谷氨酰胺、天冬酰胺、精氨酸或赖氨酸)；抗菌剂；抗氧化剂(诸如抗坏血酸、亚硫酸钠或亚硫酸氢钠)；缓冲剂(诸如硼酸盐、碳酸氢盐、Tris-HCl、柠檬酸盐、磷酸盐、其他有机酸)；填充剂(bulking agents)(诸如甘露醇或甘氨酸)；螯合剂(诸如乙二胺四乙酸(EDTA))；复合剂(complexing agent)(诸如咖啡因、聚乙烯吡咯烷酮、β-环糊精或羟丙基-β-环糊精)；填料；单糖；二糖和其他碳水化合物(诸如葡萄糖、甘露糖或糊精)；蛋白(诸如血清白蛋白、明胶或免疫球蛋白)；着色剂；调味剂和稀释剂；乳化剂；亲水性聚合物(诸如聚乙烯吡咯烷酮)；低分子量多肽；盐形成抗衡离子(诸如钠)；防腐剂(诸如苯扎氯铵、苯甲酸、水杨酸、硫柳汞、苯乙醇、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、氯己定、山梨酸或过氧化氢)；溶剂(诸如甘油、丙二醇或聚乙二醇)；糖醇(诸如甘露醇或山梨醇)；悬浮剂；表面活性剂或润湿剂(诸如普朗尼克、PEG、山梨醇酐酯(sorbitan esters)、聚山梨醇酯诸如聚山梨醇酯20、聚山梨醇酯80、曲拉通(triton)、氨丁三醇、卵磷脂、胆固醇、tyloxapal)；稳定性增强剂(蔗糖或山梨醇)；张度增强剂(诸如碱金属卤化物(优选氯化钠或氯化钾)、甘露醇和山梨醇)；递送媒介物；稀释剂；赋形剂和/或药物佐剂(pharmaceutical adjuvants)。
药物组合物中的主要媒介物或载体实质上可以是水性的或非水性的。例如，合适的媒介物或载体可以是可能补充有用于肠胃外施用的组合物中常见的其他材料的注射用水、生理盐水溶液或人工脑脊液。中性缓冲盐水或与血清白蛋白混合的盐水是另外的示例性媒介物。其他的示例性药物组合物包含约pH 7.0-8.5的Tris缓冲液或约pH 4.0-5.5的乙酸盐缓冲液，其还可以包含山梨醇或山梨醇的合适替代物。在本公开内容的一种实施方案中，可以通过将选择的具有期望程度的纯度的组合物与任选的制剂用剂(formulationagent)(Remington's Pharmaceutical Sciences,同上)混合来制备组合物以便以冻干的块状物或水性溶液形式储存。另外，可以使用合适的赋形剂诸如蔗糖将治疗组合物配制为冻干物。最佳的药物组合物将取决于例如意图的施用途径、递送形式和期望的剂量由本领域普通技术人员确定。
当设想肠胃外施用时，治疗性药物组合物可以呈在药学上可接受的媒介物中包含期望的VitoKine构建体的无致热原、肠胃外可接受的水性溶液的形式。用于肠胃外注射的特别合适的媒介物是无菌蒸馏水，其中多肽被配制为合适储存的无菌、等渗溶液。在各种实施方案中，适于可注射施用的药物制剂可以在水性溶液，优选地在生理上相容的缓冲液诸如Hanks溶液、Ringer溶液或生理缓冲盐水中配制。水性注射悬浮液可以包含增加悬浮液黏度的物质，诸如羧甲基纤维素钠、山梨醇或右旋糖酐。此外，活性化合物的悬浮液可以被制备为合适的油性注射悬浮液。任选地，悬浮液还可以包含合适的稳定剂或增加化合物的溶解度并允许制备高度浓缩的溶液的剂。
在各种实施方案中，可以使用胶体分散系统将治疗性药物组合物配制为用于靶向递送。胶体分散系统包括大分子复合体、纳米胶囊、微球、珠、和基于脂质的系统，基于脂质的系统包括水包油乳液、胶束、混合的胶束和脂质体。在脂质体产生中有用的脂质的实例包括磷脂酰化合物，诸如磷脂酰甘油、磷脂酰胆碱、磷酯酰丝氨酸、磷脂酰乙醇胺、鞘脂、脑苷脂和神经节苷脂。示例性磷脂包括卵磷脂、二棕榈酰磷脂酰胆碱和二硬脂酰磷脂酰胆碱。脂质体的靶向还可以基于例如器官特异性、细胞特异性和细胞器特异性并且是本领域已知的。
在各种实施方案中，设想了药物组合物的口服施用。以该形式施用的药物组合物可以使用或不使用固体剂型诸如片剂和胶囊的复合中通常使用的那些载体配制。在用于口服施用的固体剂型(胶囊、片剂、丸剂、糖衣丸、粉末、颗粒等)中，本公开内容的一种或更多种治疗化合物可以与一种或更多种药学上可接受的载体诸如柠檬酸钠或磷酸氢钙(dicalcium phosphate)和/或以下中的任一种混合：(1)填料或增量剂(extender)，诸如淀粉、乳糖、蔗糖、葡萄糖、甘露醇和/或硅酸；(2)粘合剂，诸如例如羧甲基纤维素、藻酸盐、明胶、聚乙烯吡咯烷酮、蔗糖和/或阿拉伯树胶；(3)保湿剂，诸如甘油；(4)崩解剂，诸如琼脂-琼脂、碳酸钙、马铃薯或木薯淀粉、藻酸、某些硅酸盐和碳酸钠；(5)溶液阻滞剂，诸如石蜡；(6)吸收加速剂，诸如季铵化合物；(7)润湿剂，诸如例如鲸蜡醇和单硬脂酸甘油酯；(8)吸收剂，诸如高岭土和膨润土；(9)润滑剂，诸如滑石、硬脂酸钙、硬脂酸镁、固体聚乙二醇、月桂基硫酸钠及其混合物；以及(10)着色剂。在胶囊、片剂和丸剂的情况下，药物组合物还可以包含缓冲剂。相似类型的固体组合物还可以用作使用诸如乳糖(lactose)或乳糖(milksugar)以及高分子量聚乙二醇等的赋形剂软填充和硬填充的明胶胶囊中的填料。用于口服施用的液体剂型包括药学上可接受的乳液、微乳液、溶液、悬浮液、糖浆剂和酏剂。除了活性成分之外，液体剂型可以包含本领域中常用的惰性稀释剂，诸如水或其他的溶剂、增溶剂和乳化剂诸如乙醇、异丙醇、碳酸乙酯、乙酸乙酯、苄醇、苯甲酸苄酯、丙二醇、1,3-丁二醇、油(特别地，棉籽油、花生油、玉米油、胚芽油、橄榄油、蓖麻油和芝麻油)、甘油、四氢糠醇、聚乙二醇和脱水山梨醇的脂肪酸酯及其混合物。除惰性稀释剂之外，口服组合物还可以包含辅料(adjuvants)，诸如润湿剂、乳化剂和悬浮剂、增甜剂、调味剂、着色剂、增香剂和防腐剂。
在各种实施方案中，设想了将药物组合物表面施用(topical administration)至皮肤或施用至黏膜。表面制剂还可以包含已知作为皮肤或角质层渗透增强剂有效的各种各样的剂中的一种或更多种。这些剂的实例是2-吡咯烷酮、N-甲基-2-吡咯烷酮、二甲基乙酰胺、二甲基甲酰胺、丙二醇、甲醇或异丙醇、二甲基亚砜和氮酮。还可以包含另外的剂以使制剂在化妆品上可接受。这些剂的实例是脂肪、蜡、油、染料、芳香剂、防腐剂、稳定剂和表面活性剂。还可以包含角质软化剂(keratolytic agent)，诸如本领域已知的那些角质软化剂。实例是水杨酸和硫磺。用于表面或经皮施用的剂型包括粉末、喷雾剂、软膏剂(ointments)、糊剂、霜剂(creams)、洗剂、凝胶、溶液、贴剂和吸入剂。可以在无菌条件下将活性化合物与药学上可接受的载体以及与可能需要的任何防腐剂、缓冲剂或喷射剂混合。除了本公开内容的主题化合物(例如VitoKine构建体)之外，软膏剂、糊剂、霜剂和凝胶可以包含赋形剂，诸如动物和植物脂肪、油、蜡、石蜡、淀粉、黄芪胶、纤维素衍生物、聚乙二醇、硅酮、膨润土、硅酸、滑石和氧化锌或其混合物。
本文中设想的用于使用的另外的药物组合物包括在持续递送或控制递送的制剂中包含多肽的制剂。在各种实施方案中，药物组合物可以被配制成纳米颗粒、作为缓释水凝胶或掺入溶瘤病毒中。这样的纳米颗粒方法包括，例如，包封在包含作为药物载体的具有疏水主链和亲水支链的聚合物的纳米颗粒中，包封在微米颗粒中，以乳剂插入到脂质体中，以及与其他分子缀合。纳米颗粒的实例包括用壳聚糖和卡波姆包衣的粘膜粘附性(mucoadhesive)纳米颗粒(Takeuchi等人,Adv.Drug Deliv.Rev.47(1):39-54,2001)以及包含带电荷的组合聚酯、聚(2-磺丁基-乙烯醇)和聚(D,L-乳酸-共-羟基乙酸)的纳米颗粒(Jung等人,Eur.J.Pharm.Biopharm.50(1):147-160,2000)。基于白蛋白的纳米颗粒组合物已被开发为用于递送疏水性药物诸如紫杉烷的药物递送系统。参见，例如，美国专利第5,916,596号、第6,506,405号、第6,749,868号、第6,537,579号、第7,820,788号和第7,923,536号。
Figure BDA0003806684520000751
(白蛋白稳定的紫杉醇纳米颗粒制剂)于2005年在美国获得批准，并随后在其他多个国家获得批准，用于治疗转移性乳腺癌。
用于配制各种其他持续递送或控制递送工具诸如脂质体运载体、生物可侵蚀的微粒或多孔珠和贮库型注射剂的技术也是本领域技术人员已知的。
待被治疗上使用的药物组合物的有效量将取决于例如治疗背景和治疗目标。本领域技术人员将理解，用于治疗的适当剂量水平将因此部分地取决于所递送的分子、多肽所用于的适应症、施用途径、以及患者的尺寸(体重、体表或器官尺寸)和状况(年龄和总体健康)而变化。相应地，临床医师可以调整剂量并改变施用途径以获得最佳治疗效果。典型的剂量可以取决于上文提及的因素而在从约0.0001mg/kg至最多约100mg/kg或更多的范围内。多肽组合物可以优选地注射或静脉内施用。长效药物组合物可以取决于特定制剂的半衰期和清除速率每三天至每四天一次、每周一次或每两周一次施用。给药频率将取决于使用的制剂中的多肽的药代动力学参数。通常，施用组合物直至达到实现期望的效果的剂量。因此组合物可以作为单剂量或作为随时间的多剂量(以相同或不同的浓度/剂量)或作为持续的输注施用。常规地进行适当剂量的进一步改进。适当的剂量可以通过使用适当的剂量-响应数据来确定。
药物组合物的施用途径根据已知方法，例如口服；通过静脉内、腹膜内、瘤内、脑内(实质内)、脑室内、肌内、眼内、动脉内、门静脉内、病灶内途径，髓内、鞘内、心室内、囊泡内、经皮、皮下或腹膜内注射；以及鼻内、肠内、表面、舌下、尿道、阴道或直肠方式；通过持续释放系统或通过植入装置。当期望时，组合物可以通过团注(bolus)注射施用，或通过输注持续施用，或通过植入装置施用。可选地或另外地，组合物可以经由期望的分子已经吸附或包封至其上的膜、海绵状物或另一种适当的材料的植入局部施用。当使用植入装置时，可以将该装置植入到任何合适的组织或器官中，并且期望的分子的递送可以经由扩散施用、缓释团注或持续施用。
治疗用途
本公开内容提供了治疗受试者的癌细胞的方法，包括向所述受试者施用药学上可接受的载体中的治疗有效量(作为单一疗法或在组合疗法方案中)的本公开内容的VitoKine构建体，其中此类施用抑制癌细胞的生长和/或增殖。特别地，本公开内容的VitoKine构建体在治疗以癌症为特征的紊乱中是有用的。此类紊乱包括但不限于实体瘤，诸如乳腺癌、呼吸道癌、脑癌、生殖器官的癌症、消化道的癌症、尿道的癌症、眼癌、肝癌、皮肤癌、头颈癌、甲状腺癌、甲状旁腺癌以及它们的远端转移，淋巴瘤，肉瘤，多发性骨髓瘤和白血病。乳腺癌的实例包括但不限于侵袭性导管癌、侵袭性小叶癌、原位导管癌和原位小叶癌。呼吸道的癌症的实例包括但不限于小细胞肺癌和非小细胞肺癌，以及支气管腺瘤和胸膜肺母细胞瘤(pleuropulmonary blastoma)。脑癌的实例包括但不限于脑干和下丘脑胶质瘤、小脑和大脑星形细胞瘤、神经母细胞瘤、髓母细胞瘤、室管膜瘤以及神经外胚层和松果体肿瘤。男性/雄性生殖器官的肿瘤包括但不限于前列腺癌和睾丸癌。女性/雌性生殖器官的肿瘤包括但不限于子宫内膜癌、宫颈癌、卵巢癌、阴道癌和外阴癌以及子宫的肉瘤。消化道的肿瘤包括但不限于肛门癌、结肠癌、结肠直肠癌、食管癌、胆囊癌、胃癌、肝癌、乳腺癌、胰腺癌、直肠癌、小肠癌和唾液腺癌。尿道的肿瘤包括但不限于膀胱癌、阴茎癌、肾癌、肾盂癌、输尿管癌和尿道癌。眼癌包括但不限于眼内黑素瘤和视网膜母细胞瘤。肝癌的实例包括但不限于肝细胞癌(具有或不具有纤维板层变体的肝细胞癌)、胆管癌(肝内胆管癌)和混合性肝细胞胆管癌。皮肤癌包括但不限于鳞状细胞癌、卡波济氏肉瘤(Kaposi's sarcoma)、恶性黑素瘤、梅克尔细胞癌(Merkel cell skin cancer)和非黑素瘤皮肤癌。头颈癌包括但不限于鼻咽癌以及唇和口腔癌。淋巴瘤包括但不限于AIDS相关淋巴瘤、非霍奇金淋巴瘤、皮肤T细胞淋巴瘤、霍奇金病和中枢神经系统淋巴瘤。肉瘤包括但不限于软组织肉瘤、骨肉瘤、恶性纤维组织细胞瘤、淋巴肉瘤和横纹肌肉瘤。白血病包括但不限于急性髓性白血病、急性淋巴母细胞白血病、慢性淋巴细胞白血病、慢性髓细胞性白血病和毛细胞白血病。在各种实施方案中，癌症将是具有TGF-β家族成员(诸如激活蛋白A、肌生成抑制蛋白、TGF-β和GDF15)的高表达的癌症，例如胰腺癌、胃癌、肝癌、乳腺癌、卵巢癌、结肠直肠癌、黑素瘤白血病、肺癌、前列腺癌、脑癌、膀胱癌和头颈癌。
在各种实施方案中，VitoKine构建体可用作治疗所有种类癌症的单一剂，所述癌症包括但不限于非小细胞肺癌、小细胞肺癌、黑素瘤、肾细胞癌、尿路上皮癌、肝癌、乳腺癌、胰腺癌、结肠直肠癌、胃癌、前列腺癌和肉瘤。
在另一方面，本公开内容提供了用于治疗受试者的自身免疫性疾病的方法，该方法包括向所述受试者施用在药学上可接受的载体中的治疗有效量(作为单一疗法或在组合疗法方案中)的本公开内容的VitoKine构建体。“自身免疫性疾病”是指源于并针对个体自身组织的非恶性疾病或紊乱。自身免疫性疾病或紊乱的实例包括但不限于，炎性反应，诸如炎性皮肤疾病，包括银屑病和皮炎(例如特应性皮炎)；与炎性肠病(诸如克罗恩病和溃疡性结肠炎)相关的反应；皮炎；过敏性状况，诸如湿疹和哮喘；类风湿性关节炎；系统性红斑狼疮(SLE)(包括但不限于狼疮性肾炎、皮肤狼疮)；糖尿病(例如1型糖尿病或胰岛素依赖型糖尿病)；多发性硬化和幼年发作糖尿病。
在另一方面，本公开内容提供了用于治疗受试者的炎性疾病的方法，该方法包括向所述受试者施用在药学上可接受的载体中的治疗有效量(作为单一疗法或在组合疗法方案中)的本公开内容的VitoKine构建体。“炎性疾病”包括与急性或慢性炎症相关的所有疾病。急性炎症是身体对有害刺激的初始反应，并且由血浆和白细胞(诸如例如粒细胞)从血液到损伤组织中的增加的移动引起。许多生物化学事件传播炎性反应并使炎性反应成熟，所述炎性反应涉及损伤组织内的局部血管系统、免疫系统和各种细胞。长期的炎症被称为慢性炎症，其导致存在于炎症部位处的细胞类型的进行性变化，并且其特征在于来自炎症过程的组织的同时破坏和愈合。炎性疾病的实例是本领域熟知的。在各种实施方案中，炎性疾病选自由以下组成的组：炎性肠病、银屑病和细菌性脓毒症。如本文使用的，术语“炎性肠病”是指结肠和小肠的一组炎性状况，包括，例如，克罗恩病、溃疡性结肠炎、胶原性结肠炎、淋巴细胞性结肠炎、缺血性结肠炎、改道性结肠炎(diversion colitis)、白塞氏综合征和未定型结肠炎。
在另一方面，本公开内容提供了用于治疗受试者的病毒感染的方法，该方法包括向所述受试者施用在药学上可接受的载体中的治疗有效量(作为单一疗法或在组合疗法方案中)的本公开内容的VitoKine构建体。在各种实施方案中，待治疗的病毒感染可以由感染原引起，所述感染原包括但不限于细菌、真菌、原生动物和病毒。可以根据本文描述的方法预防、治疗和/或控制的病毒性疾病包括但不限于由以下引起的那些：甲型肝炎病毒、乙型肝炎病毒、丙型肝炎病毒、流感病毒、水痘病毒、腺病毒、单纯疱疹I型(HSY-I)病毒、单纯疱疹II型(HSY-II)病毒、牛瘟病毒、鼻病毒、埃可病毒、轮状病毒、呼吸道合胞病毒、乳头瘤病毒、乳多空病毒、巨细胞病毒、echinovirus、虫媒病毒、汉坦病毒、柯萨奇病毒、腮腺炎病毒、麻疹病毒、风疹病毒、脊髓灰质炎病毒、天花病毒、Epstein Barr病毒、人类免疫缺陷病毒I型(HIV-I)、人类免疫缺陷病毒II型(HIV-II)和病毒性疾病诸如病毒性脑膜炎、脑炎、登革热或天花的病原体。
可以根据本文描述的方法预防、治疗和/或控制的由细菌(例如大肠杆菌(Escherichia coli)、肺炎克雷伯菌(Klebsiella pneumoniae)、金黄色葡萄球菌(Staphylococcus aureus)、粪肠球菌(Enterococcus faecalis)、白色念珠菌(Candidaalbicans)、普通变形杆菌(Proteus vulgaris)、草绿色链球菌(Staphylococcusviridans)和铜绿假单胞菌(Pseudomonas aeruginosa))引起的细菌性疾病包括但不限于分枝杆菌、立克次氏体、支原体、奈瑟氏球菌、肺炎链球菌(S.pneumonia)、伯氏疏螺旋体(Borrelia burgdorferi)(莱姆病)、炭疽杆菌(Bacillus anthracis)(炭疽)、破伤风、链球菌、葡萄球菌、分枝杆菌、百日咳、霍乱、鼠疫、白喉、衣原体、金黄色葡萄球菌和军团菌(legionella)。
可以根据本文描述的方法预防、治疗和/或控制的由原生动物引起的原生动物疾病包括但不限于利什曼虫、kokzidioa、锥虫(trypanosoma)或疟疾。
可以根据本文描述的方法预防、治疗和/或控制的由寄生物引起的寄生物疾病包括但不限于衣原体和立克次氏体。
“治疗有效量”或“治疗有效剂量”是指将使被治疗的紊乱的一种或更多种症状缓解至一定程度的被施用的治疗剂的量。
治疗有效剂量可以通过确定IC50从细胞培养物测定初始地评估。然后，可以在动物模型中配制剂量以实现包括如在细胞培养物中确定的IC50的循环血浆浓度范围。此类信息可以被用于更精确地确定在人类中有用的剂量。血浆中的水平可以例如通过HPLC测量。确切的组合物、施用途径和剂量可以由个体医师鉴于受试者的状况来选择。
可以调整剂量方案以提供最佳的期望的响应(例如，治疗响应或预防响应)。例如，可以施用单次团注，可以随时间施用若干分次剂量(多个或重复或维持)并且剂量可以根据治疗情境的迫切需要所指示的按比例减少或增加。为了便于施用和剂量的一致性，配制呈剂量单位形式的肠胃外组合物是特别有益的。如本文使用的剂量单位形式是指适合作为用于待治疗的哺乳动物受试者的单一剂量的物理分散的单位；每一个单位包含与需要的药物载体一起的被计算产生期望的治疗作用的预定的量的活性化合物。本公开内容的剂量单位形式的规格将主要由抗体的独特特征和待实现的特定治疗作用或预防作用决定。
因此，技术人员将理解，基于本文提供的公开内容，根据治疗领域熟知的方法调整剂量和给药方案。即，最大可耐受剂量可以被容易地确定，并且向受试者提供可检测的治疗益处的有效量也可以被确定，向受试者提供可检测的治疗益处而施用每种剂的时间要求同样可以被确定。因此，尽管在本文中例举了某些剂量和施用方案，这些实例绝非限制在实践本公开内容时可以向受试者提供的剂量和施用方案。
应注意，剂量值可以随着待减轻的状况的类型和严重性而变化，并且可以包括单个剂量或多于一个剂量。还应理解，对于任何特定受试者，应根据个体需要和施用组合物或监督组合物的施用的人士的专业判断随时间调整具体剂量方案，并且本文列出的剂量范围仅是示例性的并且不意图限制所要求保护的组合物的范围或实践。另外，本公开内容的组合物的剂量方案可以基于多个因素，包括疾病的类型、受试者的年龄、体重、性别、医学状况、状况的严重性、施用途径和使用的特定抗体。因此，剂量方案可以广泛变化，但可以使用标准方法常规地确定。例如，剂量可以基于药代动力学或药效学参数调整，所述参数可以包括临床作用诸如毒性作用和/或实验值。因此，本公开内容包括如技术人员确定的受试者内的剂量递增(intra-subject dose-escalation)。确定适当的剂量和方案是相关技术领域熟知的并且将被理解为一旦提供本文所公开的教导，则由技术人员掌握。
对于本公开内容的VitoKine或VitoKine变体的治疗有效量或预防有效量的示例性的、非限制性的每天给药范围可以为0.0001至100mg/kg体重、0.0001至90mg/kg体重、0.0001至80mg/kg体重、0.0001至70mg/kg体重、0.0001至60mg/kg体重、0.0001至50mg/kg体重、0.0001至40mg/kg体重、0.0001至30mg/kg体重、0.0001至20mg/kg体重、0.0001至10mg/kg体重、0.0001至5mg/kg体重、0.0001至4mg/kg体重、0.0001至3mg/kg体重、0.0001至2mg/kg体重、0.0001至1mg/kg体重、0.001至50mg/kg体重、0.001至40mg/kg体重、0.001至30mg/kg体重、0.001至20mg/kg体重、0.001至10mg/kg体重、0.001至5mg/kg体重、0.001至4mg/kg体重、0.001至3mg/kg体重、0.001至2mg/kg体重、0.001至1mg/kg体重、0.010至50mg/kg体重、0.010至40mg/kg体重、0.010至30mg/kg体重、0.010至20mg/kg体重、0.010至10mg/kg体重、0.010至5mg/kg体重、0.010至4mg/kg体重、0.010至3mg/kg体重、0.010至2mg/kg体重、0.010至1mg/kg体重、0.1至50mg/kg体重、0.1至40mg/kg体重、0.1至30mg/kg体重、0.1至20mg/kg体重、0.1至10mg/kg体重、0.1至5mg/kg体重、0.1至4mg/kg体重、0.1至3mg/kg体重、0.1至2mg/kg体重、0.1至1mg/kg体重、1至50mg/kg体重、1至40mg/kg体重、1至30mg/kg体重、1至20mg/kg体重、1至10mg/kg体重、1至5mg/kg体重、1至4mg/kg体重、1至3mg/kg体重、1至2mg/kg体重或1至1mg/kg体重。应注意，剂量值可以随着待被缓解的状况的类型和严重性变化。还应理解，对于任何特定受试者，应根据个体需要和施用组合物或监督组合物的施用的人士的专业判断随时间调整具体剂量方案，并且本文列出的剂量范围仅是示例性的并且不意图限制所要求保护的组合物的范围或实践。
本公开内容的药物组合物的毒性和治疗指数可以在细胞培养物或实验动物中通过标准制药学程序确定，例如，确定LD50(对群体的50％致死的剂量)和ED50(对群体的50％治疗有效的剂量)。毒性剂量和治疗有效剂量之间的剂量比是治疗指数，并且治疗指数可以表示为比率LD50/ED50。表现出大治疗指数的组合物通常是优选的。
VitoKine构建体药物组合物的施用的给药频率取决于疗法的性质和被治疗的特定疾病。受试者可以以规律间隔治疗，诸如每周一次或每月一次，直到实现期望的治疗结果。示例性给药频率包括但不限于：无间断地每周一次；每隔一周地每周一次；每2周一次；每3周一次；无间断地每周一次，持续2周，然后每月一次；无间断地每周一次，持续3周，然后每月一次；每月一次；每两个月一次；每3个月一次；每4个月一次；每5个月一次；或每6个月一次，或每年一次。
组合疗法
如本文使用的，提及本公开内容的VitoKine构建体和一种或更多种其他治疗剂时，术语“共施用(co-administration)”、“共施用(co-administered)”和“与...组合(incombination with)”意图意指，并且的确是指并且包括以下：本公开内容的VitoKine构建体和一种或更多种治疗剂的此类组合同时施用至需要治疗的受试者，其中这样的组分被一起配制成单一剂型，所述单一剂型在基本上相同的时间将所述组分释放至所述受试者；本公开内容的VitoKine构建体和一种或更多种治疗剂的此类组合基本上同时施用至需要治疗的受试者，其中这样的组分彼此分开配制成单独的剂型，所述单独的剂型在基本上相同的时间被所述受试者服用，届时所述组分基本上在相同的时间释放至所述受试者；本公开内容的VitoKine构建体和一种或更多种治疗剂的此类组合顺序施用至需要治疗的受试者，其中这样的组分彼此分开配制成单独的剂型，所述单独的剂型被所述受试者在连续的时间服用，每次施用之间具有显著的时间间隔，届时所述组分在基本上不同的时间释放至所述受试者；以及，本公开内容的VitoKine构建体和一种或更多种治疗剂的此类组合顺序施用至需要治疗的受试者，其中这样的组分一起配制成单一剂型，所述单一剂型以受控方式释放所述组分，届时所述组分在相同和/或不同的时间同时、连续和/或重叠地释放至所述受试者，其中每个部分可以通过相同或不同的途径施用。
在另一方面，本公开内容提供了一种用于治疗受试者的癌症或癌症转移的方法，该方法包括施用与第二疗法组合的治疗有效量的本发明的药物组合物，所述第二疗法包括但不限于免疫疗法、细胞毒性化学疗法、小分子激酶抑制剂靶向疗法、手术、放射疗法和干细胞移植。例如，这样的方法可以被用于预防性癌症预防、预防手术后癌症复发和转移，以及作为其他常规癌症疗法的辅助手段。本公开内容认识到，常规癌症疗法(例如，化学疗法、放射疗法、光疗法、免疫疗法和手术)的有效性可以通过本文描述的组合方法的使用来增强。
大量的常规化合物已经被证明具有抗赘生物活性。这些化合物已经被用作化学疗法中的药剂以缩小实体瘤、预防转移和进一步生长、或减少白血病或骨髓恶性肿瘤中的恶性T细胞的数目。尽管化学疗法在治疗各种类型的恶性肿瘤中是有效的，但许多抗赘生物化合物诱导不期望的副作用。已经显示出，当将两种或更多种不同的治疗组合时，治疗可以协同地工作，并且允许降低每种治疗的剂量，从而降低每种化合物在较高的剂量产生的有害副作用。在其他情况下，对于治疗是难治性的恶性肿瘤可以响应于两种或更多种不同治疗的组合疗法。
在各种实施方案中，第二抗癌剂，诸如化学治疗剂将被施用至患者。示例性化学治疗剂的列表包括但不限于，柔红霉素、更生霉素(dactinomycin)、多柔比星、博莱霉素、丝裂霉素、氮芥、苯丁酸氮芥、美法仑、环磷酰胺、6-巯基嘌呤、6-硫鸟嘌呤、苯达莫司汀、阿糖胞苷(CA)、5-氟尿嘧啶(5-FU)、氟尿苷(5-FUdR)、氨甲蝶呤(MTX)、秋水仙碱、长春新碱、长春花碱、依托泊苷、替尼泊苷、顺铂、卡铂、奥沙利铂、喷司他丁、克拉屈滨、阿糖胞苷、吉西他滨、普拉曲沙、米托蒽醌、己烯雌酚(DES)、氟达拉滨(fluradabine)、异环磷酰胺、羟基脲紫杉烷(hydroxyureataxanes)(诸如紫杉醇和多西他赛(doxetaxel))和/或蒽环类抗生素，以及剂的组合，诸如但不限于DA-EPOCH、CHOP、CVP或FOLFOX。在各种实施方案中，这样的化疗剂的剂量包括但不限于约10mg/m2、20mg/m2、30mg/m2、40mg/m2、50mg/m2、60mg/m2、75mg/m2、80mg/m2、90mg/m2、100mg/m2、120mg/m2、150mg/m2、175mg/m2、200mg/m2、210mg/m2、220mg/m2、230mg/m2、240mg/m2、250mg/m2、260mg/m2和300mg/m2中的任一种。
在各种实施方案中，本公开内容的组合治疗方法还可以包括向受试者施用治疗有效量的免疫疗法，所述免疫疗法包括但不限于，使用针对特定肿瘤抗原的消耗性抗体的治疗；使用抗体-药物缀合物的治疗；使用针对共刺激性或共抑制性分子(免疫检查点)的激动性抗体、拮抗性抗体或阻断性抗体的治疗，诸如包括但不限于针对CTLA-4、PD-1、PDL-1、CD40、OX-40、CD137、GITR、LAG3、TIM-3、SIRPα、CD47、GITR、ICOS、CD27、Siglec 7、Siglec 8、Siglec 9、Siglec 15和VISTA、CD276、CD272、TIM-3、B7-H4的抗体；使用双特异性T细胞结合抗体
Figure BDA0003806684520000841
诸如博纳吐单抗的治疗；涉及施用生物响应调节剂(诸如IL-2、IL-7、IL-10、IL-12、IL-15、IL-21、IL-22、GM-CSF、IFN-α、IFN-β、IFN-γ、TGF-β拮抗剂或TGF-β陷阱)的治疗；使用治疗性疫苗(包括但不限于溶瘤病毒，诸如T-vec)或使用治疗性疫苗诸如sipuleucel-T的治疗；使用树突细胞疫苗或肿瘤抗原肽或新抗原疫苗的治疗；使用嵌合抗原受体(CAR)-T细胞的治疗；使用CAR-NK细胞的治疗；使用NK细胞的治疗；使用iPS诱导的NK细胞的治疗；使用iPS诱导的T细胞的治疗；使用iPS诱导的CAR-T细胞或iPS诱导的CAR-NK细胞的治疗；使用肿瘤浸润淋巴细胞(TIL)的治疗；使用过继转移的抗肿瘤T细胞(离体扩增的T细胞和/或TCR-T细胞)的治疗；使用TALL-104细胞的治疗；和使用免疫刺激剂诸如Toll样受体(TLR)激动剂CpG、TLR7、TLR8、TLR9和疫苗诸如卡介苗(BCG)和咪喹莫特的治疗；其中组合疗法提供了增加的对肿瘤细胞的效应细胞杀伤，即，当被共施用时，VitoKine构建体和免疫疗法之间存在协同作用。
在各种实施方案中，本公开内容的组合治疗方法还可以包括向受试者施用治疗有效量的用于自身免疫性疾病、炎性疾病和其他免疫紊乱的抗炎剂，包括但不限于使用针对特定免疫细胞的消耗性抗体的治疗；使用调节性抗体(激动性、拮抗性或阻断性)作为针对靶(配体或其受体)的免疫应答靶调节剂的治疗，所述靶包括但不限于IL-1α、IL-1β或IL-1R、IL-4或IL-4R、IL-5或IL-5R、IL-6或IL-6R、IL-8或IL-8R、IL-7或IL-7R、IL-10或IL-10R、IL-11或IL-11R、IL-12或IL-12R、IL-17或IL-17R、IL-18或IL-18R、IL-21或IL-21R、IL-22或IL-22R、IL-23或IL-23R、MCSF或MCSF-R、GM-CSF或GM-CSFR、IFN-α、IFN-β、IFN-γ、TGF-α、TGF-β或TGF-β、TNF家族或其相关受体、整联蛋白家族(例如α4β7)、TSLP、补体5(C5)或C5a、IgE、APRIL、TACI、BCMA、CD20、CD22、CD40/CD40L、B7H1、B7H2、ICOS、BAFF、BCR、BLys、B7RP1、TLR7、TLR8、TLR8、TLR9；使用调节性小分子(激动性或拮抗性)作为针对靶的免疫应答靶调节剂的治疗，所述靶包括但不限于NFkB、Jak1、Jak2、Jak3、Tyk2、Syk、BTK、PIK3、环加氧酶2和NMDA受体；其中组合治疗提供了增加的调节免疫应答的功效，即，当共施用时，VitoKine构建体和抗炎疗法之间存在协同作用。
在各种实施方案中，组合疗法包括同时施用在同一药物组合物中或在单独的药物组合物中的VitoKine构建体和第二剂组合物。在各种实施方案中，顺序施用VitoKine构建体组合物和第二剂组合物，即，在施用第二剂组合物之前或之后施用VitoKine构建体组合物。在各种实施方案中，VitoKine构建体组合物和第二剂组合物的施用是同时的，即，VitoKine构建体组合物和第二剂组合物的施用时间段彼此重叠。在各种实施方案中，VitoKine构建体组合物和第二剂组合物的施用是非同时的。例如，在各种实施方案中，终止VitoKine构建体组合物的施用，然后施用第二剂组合物。在各种实施方案中，终止施用第二剂组合物，然后施用VitoKine构建体组合物。
提供以下实施例以更充分地说明本公开内容，但不应解释为限制本公开内容的范围。
实施例1
IL-15 Fc VitoKine构建体的构建和产生
目标是设计IL-15 VitoKine构建体，该构建体将保持惰性，直到被癌症或病变组织中上调的蛋白酶在局部活化。本文描述了具有被可逆地遮蔽在Fc结构域和IL-15RαSushi+(SEQ ID NO:5)之间的作为活性部分的野生型IL-15(SEQ ID NO:2)或IL-15突变蛋白(mutein)(例如SEQ ID NO:3)的VitoKine。这些构建体包括一个或两个被肿瘤特异性蛋白酶识别的可裂解接头。在表达蛋白酶的肿瘤细胞的存在下，连接Fc和IL-15突变蛋白的接头和/或连接IL-15和IL-15αSushi+的接头会被裂解，并从而恢复IL-15活性。值得注意的是，由于IL-15和IL-15α之间异常高的亲和力(KD＝30pM)，蛋白水解后释放的IL-15αSushi+预计保持与IL-15的非共价缔合。产生并在图1中示意性地描绘了具有各种接头和肽间隔物的组合的IL-15 Fc VitoKine构建体，它们各自的序列列出为SEQ ID NO:25-43、162-165和169-174。
对所有基因进行密码子优化以便在哺乳动物细胞中表达，合成这些基因并亚克隆到接受者哺乳动物表达载体(GenScript)中。蛋白表达由CMV启动子驱动，并且合成的SV40聚腺苷酸(polyA)信号序列存在于CDS的3'端。在构建体的N末端处工程化有前导序列，以确保合适的信号传导和加工以便分泌。
构建体通过使用聚乙烯亚胺(PEI，25,000MW线性，Polysciences)用哺乳动物表达载体共转染悬浮生长的HEK293-F细胞来产生。如果有两种或更多种表达载体，则载体以1:1的比转染。对于转染，将HEK293细胞在无血清FreeStyleTM293表达培养基(ThermoFisher)中培养。对于在1000ml摇瓶(工作体积330mL)中的产生，将HEK293细胞以0.8×106个细胞/ml的密度接种，24小时后进行转染。将总量为330μg DNA的表达载体与16.7ml Opti-mem培养基(ThermoFisher)混合。在添加稀释于16.7ml Opti-mem培养基中的0.33mg PEI后，将混合物涡旋15sec，并随后在室温孵育10min。然后将DNA/PEI溶液添加至细胞，并在37℃在具有8％的CO2气氛的培养箱中孵育。在第4天，将丁酸钠(Millipore Sigma)以2mg/L的终浓度添加至细胞培养物，以帮助维持蛋白表达。培养6天后，收集上清液用于通过以2200rpm离心20min进行纯化。将溶液进行无菌过滤(0.22μm过滤器，Corning)。使用蛋白A亲和色谱从细胞培养物上清液纯化所分泌的蛋白。
对于亲和色谱，将上清液加载到用25ml磷酸盐缓冲盐水pH 7.2(ThermoFisher)平衡的HiTrap MabSelectSure蛋白A FF柱(CV＝5mL,GE Healthcare)上。通过用5倍柱体积的PBS pH 7.2洗涤来去除未结合的蛋白，并用25mM柠檬酸钠、25mM氯化钠，pH 3.2洗脱目标蛋白。蛋白溶液通过添加3％的1M Tris pH 10.2来中和。靶蛋白用
Figure BDA0003806684520000861
Ultra-15浓缩器10KDa NMWC(Merck Millipore Ltd.)浓缩。
在具有还原剂和不存在还原剂的情况下，通过SDS-PAGE和用考马斯亮蓝(Coomassie)(ImperialR stain)染色来分析所纯化的构建体的纯度和分子量。根据制造商的说明使用
Figure BDA0003806684520000862
预制凝胶系统(4-12％或8-16％Bis-Tris,ThermoFisher)。纯化的蛋白样品的蛋白浓度通过测量280nm处的UV吸光度(Nanodrop分光光度计，ThermoFisher)除以基于氨基酸序列计算的摩尔消光系数来确定。构建体的聚集体含量在Agilent 1200高效液相色谱(HPLC)系统上进行分析。在25℃，使用150mM磷酸钠pH 7.0作为流动相，将样品注射到AdvanceBio尺寸排阻柱(
Figure BDA0003806684520000863
4.6x150mm,2.7μm,LC柱,Agilent)上。
P-0315是一种二聚体C末端IL-15 Fc VitoKine，在L1接头和L2接头中分别含有uPA和MMP裂解序列。IL-15是S58D变体蛋白。作为展示IL-15 Fc VitoKine的蛋白谱的实例，对P-0315(SEQ ID NO:33)的SDS-PAGE分析在图3A中示出。尺寸排阻色谱图在图3B中。
实施例2
IL-15的体外活性在VitoKine形式中被有效地遮蔽
IL-15 VitoKine P-0172(SEQ ID NO:27)包含通过短GS肽接头连接的IL-15/IL-15RαSushi+融合多肽，在同源二聚体融合形式中，所述肽接头经由uPA可裂解接头连接到同源二聚体Fc结构域的C末端。P-0198是一种二聚C末端Fc-IL-15融合蛋白，具有非共价复合的IL-15αSushi。这两种分子在Fc和IL-15融合体之间具有相似的构型，在IL-15αSushi掺入方面具有重大差异。一种通过短的GS接头融合(P-0172)，而另一种通过非共价性为游离的(P-0198)。通过酶联免疫吸附测定(ELISA)确定P-0172对IL-2Rβ的结合活性，并与P-0198(包含SEQ ID NO:45、44和5)比较，P-0198是一种高活性的IL-15/IL-15Rα-Fc融合蛋白。
简而言之，将IL-2Rβ-ECD(SEQ ID NO:12)以1μg/孔包被到Nunc Maxisorp 96孔微孔板的孔上。在4℃孵育过夜并用superblock(ThermoFisher)封闭后，以100μl/孔向每个孔添加从100nM开始的3倍系列稀释的IL-15化合物。在室温孵育1小时后，向每个孔添加100μl/孔的山羊抗人类IgG Fc-HRP(在稀释剂中1:5000稀释)，并在室温孵育1小时。每一步骤后，将孔彻底抽吸，并用PBS/0.05％吐温-20洗涤三次。最后，向每个孔添加100μl TMB底物(ThermoFisher)，将板在室温在黑暗中显影10分钟，并添加100μl/孔的停止溶液(2N硫酸，Ricca Chemical)。确定450nm处的吸光度，并使用Prism软件(GraphPad)拟合曲线。如图4A中说明的，与P-0198相比，VitoKine P-0172以显著降低的效力与IL-2Rβ结合(12.2nM对比0.21nM)，这可能是由于IL-15和IL-15αSushi之间的短共价连接导致的空间限制，表明VitoKine平台中的IL-15αSushi有效地遮蔽了IL-15结构域与其受体结合。
通过FACS分析检查来自新鲜的人类外周血单核细胞(PBMC)的人类NK细胞和CD8+T细胞上IL-15介导的CD69表达的诱导，进一步评估了与P-0198相比IL-15 VitoKine P-0172的功能活性。CD69是一种细胞表面糖蛋白，在包括NK细胞和T细胞的淋巴活化期间中被早期诱导。
简而言之，通过Ficoll-Hypaque离心法从购自Oklahoma Blood Institute的血沉棕黄层分离人类PBMC。将纯化的人类PBMC用每种IL-15测试化合物的系列稀释液处理，并在37℃孵育48小时。通过以300xg离心收集细胞，并重悬于FACS缓冲液中。通过添加人类TruStain FcX(1:50稀释)封闭Fc受体后，用抗人类CD56-FITC抗体、抗人类CD69-PE抗体和抗人类CD8-APC抗体(1:50稀释)对细胞进行染色。在室温与抗体一起孵育30分钟后，收集细胞并洗涤，重悬于FACS缓冲液中，并通过流式细胞术分析。CD69表达通过对CD56+NK和CD8+T细胞进行门控来确定，并且数据表示为门控的群体中CD69阳性细胞的％。
如图4B和图4C中展示的，VitoKine P-0172对CD8+T细胞和NK细胞的CD69活化显著降低，并且仅在所测试的最高浓度可测量到，效力比P-0198的效力低至少2-3个对数。这表明在VitoKine形式中IL-15活性被有效遮蔽。遮蔽效应在PBMC CD69活化测定中比在IL-2RβELISA结合测定中更明显，表明在生理条件下IL-15活性的严重损害比在ELISA的体外重建条件下更明显。由于空间限制的存在，VitoKine严重破坏了IL-15与淋巴细胞上表达的IL-2Rβ和γC复合物的结合，并因此导致低效的途径活化和严重受损的活性。
还检查了单体IL-15 Fc VitoKine的生物活性。P-0170(SEQ ID NO:26和15)是P-0172的单体对应物，具有相同的接头和融合构型。与高活性的IL-15 Fc融合蛋白P-0166相比，P-0170显示出显著降低的活化CD8+T细胞上的CD69的能力(图5)，表明单体VitoKine平台还有效地遮蔽了D2结构域中的IL-15的生物活性。
实施例3
IL-15和IL-15αSushi+之间以及Fc和IL15之间不同接头长度和组成的IL-15 FcVitoKine遮蔽效率的比较
如图1中描绘的，经由肽接头L1和L2将人类IL-15融合在两个不同的结构域诸如半衰期延长Fc结构域和IL-15的同源高亲和力共受体α结构域之间来构建IL-15 VitoKine。检查连接Fc和IL-15对比连接IL-15和IL-15αSushi结构域的两个接头以及接头的长度和组成对IL-15生物活性的差异影响，以获得期望的活性损伤。
对新鲜的人类PBMC的免疫细胞亚群的活化标志物CD69进行FACS分析，以评估IL-15和IL-15αSushi+之间不同的不可裂解接头(L2)长度的IL-15 VitoKine。遵循与实施例2中相同的方案。
P-0204(SEQ ID NO:30)、P-0205(SEQ ID NO:31)和P-0206(SEQ ID NO:32)是在Fc和IL-15之间共有相同的uPA可裂解接头序列(L1)的IL-15Fc VitoKine，但是在三种Vitokine中连接IL-15和IL-15αSushi+结构域的接头(L2)长度不同，并且分别是(GGGGS)3(SEQ ID NO:127)、(GGGGS)2(SEQ ID NO:126)和GGGGS(SEQ ID NO:118)。
如图6中展示的，IL-15和IL-15αSushi+之间从5个至15个氨基酸的不同接头长度的IL-15 Fc VitoKine都导致活化CD8+T细胞(图6A)或NK细胞(图6B)的效力显著降低。比较P-0206、P-0205和P-0204的效力，明显的是，连接IL-15和IL-15αSushi+结构域的接头长度越短，VitoKine变得越无活性；表明活性降低的程度可以通过L2接头长度进一步调节；总之，我们通过调节IL-15和IL-15α之间的接头(L2)长度以产生适当水平的空间限制，将IL-15遮蔽在Fc结构域和IL-15的同源高亲和力共受体α之间，从而使IL-15活性几乎完全丧失。
在图7中说明了还检查了连接Fc和IL-15的接头(L1)对VitoKine生物活性的影响。P-0204和P-0203(SEQ ID NO:29)在IL-15和IL-15Rα之间共有相同的15-氨基酸柔性(G4S)3接头(SEQ ID NO:112)(L2)，但L1接头的长度不同；P-0203在连接Fc和IL-15的uPA底物肽的侧翼包含比P-0204长7个富含GS的残基的肽间隔物。尽管连接Fc和IL-15的L1接头长度不同，但P-0204和P-0203的生物活性相似(图7)，表明当跨13个至35个氨基酸残基时，连接Fc和IL-15的L1接头对IL-15活性的损害影响最小。然而，L1接头长度少于13个氨基酸或多于35个氨基酸或在不同的细胞因子的背景下可能影响对D2结构域的遮蔽活性。在同一项研究中，包括了P-0202，P-0202与P-0203共有相同的连接Fc和IL-15的L1接头，但P-0202的连接IL-15和IL-15αSushi结构域的L2接头与P-0203相比短13个氨基酸。P-0202显示出比P-0203更低的生物活性，证实了在VitoKine的遮蔽活性方面，接头L2比接头L1更重要。
通过测量IL-15 VitoKine处理后NK细胞和CD8 T细胞的细胞核中的Ki67表达，研究了接头组成或接头肽序列对VitoKine活性的影响。Ki67是细胞增殖的标志物，并建立了离体人类PBMC测定。简而言之，将纯化的人类PBMC用IL-15 VitoKine化合物的系列稀释液处理，并在37℃孵育5天。在第5天，用FACS缓冲液(1％FBS/PBS)洗涤细胞一次，并首先用Fc封闭剂和表面标志物抗体(包括抗人类CD56-FITC和抗人类CD8-APC(1:50稀释))进行染色。在30分钟的孵育和洗涤后，用200μl/孔的1×Foxp3固定及透化工作溶液将细胞沉淀充分重悬，并在室温在黑暗中孵育30分钟。离心后，向每个孔添加200μl的1×透化缓冲液进行另一次洗涤。用抗人类Ki67-PE(1:25稀释)将细胞沉淀重悬于透化缓冲液中。在室温孵育30分钟后，收集细胞并洗涤，重悬于FACS缓冲液中，并通过流式细胞术进行分析。数据表示为门控的群体中Ki67阳性细胞的％。
由于与L1接头相比，L2接头对IL-15 VitoKine活性施加更显著的影响，因此我们检查了L2接头的不同序列组成对IL-15 VitoKine生物活性的影响。P-0351(SEQ ID NO:25)、P-0488(SEQ ID NO:163)和P-0489(SEQ ID NO:164)都共有相同的连接Fc和IL-15的(G4S)3接头(L1)。连接IL-15和IL-15Rα的接头长度都是10个氨基酸，但具有不同的序列。接头在P-0351中是(G4S)2，在P-0488中是MMP-14底物肽(SEQ ID NO:157)或在P-0489中是豆荚蛋白底物肽(SEQ ID NO:160)。
如图8中展示的，与高活性IL-15/IL-15RαFc融合蛋白P-0156(SEQ ID NO:175和176)相比，所有三种IL-15 VitoKine在增殖CD8+T细胞(图8A)或NK细胞(图8B)方面具有严重受损的效力。不同的肽接头序列对各VitoKine的生物活性有细微的影响(图8A和图8B)，这可能是由于每个接头肽的结构柔性引起的。L2接头肽越刚性，它对VitoKine分子施加的结构限制就越多，这可以导致更严重的活性损伤。然而，L2接头序列组成对VitoKine活性的影响是边缘性的，并且数据支持不同的可裂解接头可以作为L2接头被掺入，以有效遮蔽D2结构域的活性，从而扩大VitoKine的设计和应用范围。
总之，这些数据共同展示，连接IL-15(D2)和IL-15RαSushi+(D3)结构域的L2接头在遮蔽D2活性以产生惰性VitoKine中发挥重要作用。活性惰性的水平可以通过调整L2接头长度和改变接头序列/柔性来进一步调节。可裂解的L2接头长度和序列的选择应在特定蛋白酶在意图的疾病适应症部位的存在、底物肽对蛋白酶的可及性和期望的蛋白水解速率之间进行平衡。
实施例4
确定完全体外蛋白酶裂解的合适反应条件
最初的体外蛋白酶裂解实验使用IL-15 Fc VitoKine构建体P-0315和P-0203来进行，以分别确定MMP-2和uPA的蛋白酶可裂解性和最佳裂解条件。P-0315(SEQ ID NO:33)包含连接Fc和IL-15结构域的uPA可裂解接头(L1)以及连接IL-15和IL-15RαSushi+结构域的MMP-2/9可裂解接头(L2)。P-0203(SEQ ID NO:29)包含连接Fc和IL-15结构域的单个蛋白酶可裂解接头(uPA)(L1)。P-0203中IL-15和IL-15RαSushi+结构域之间的接头是柔性的(G4S)3接头。重组人类uPA和MMP-2购自BioLegend。MMP-2以潜伏形式(latent form)提供，并根据制造商的说明通过对氨基苯汞乙酸盐(APMA，Millipore Sigma)活化。
对于通过MMP-2进行蛋白水解裂解，在37℃将4μg P-0315与30ng、100ng或300ng的APMA活化的MMP-2一起在制造商推荐的测定缓冲液(100mM Tris、20mM CaCl2、300mM NaCl、0.1％(w/v)Brij 35，pH 7.5)中孵育3小时。为了停止反应，将SDS-PAGE加载染料添加到反应中，并将混合物在95℃加热5分钟。为了评估裂解，将消化的样品在4％-12％Tris-BisSDS-PAGE凝胶上分离。对未处理和处理的样品的比较显示，在用所有测试浓度的MMP-2处理后，IL-15 VitoKine被完全裂解。这是由尺寸变化和在SDS page凝胶中出现～9KDa的清晰条带所指示的(图9)，所述～9KDa的尖锐条带是从P-0315裂解下的IL-15RαSushi+结构域。
uPA的可裂解性通过使用P-0203来评估。首先，将不同的量的uPA添加到20μl PBSpH 7.2缓冲液中的2μg的P-0203，并将反应混合物在37℃孵育2小时。在图10A中说明了用0、25ng、50ng、100ng和300ng的uPA进行的裂解。图10A中的三个箭头代表非还原(NR)样品，并且表示Fc链随着uPA蛋白水解的变化。在“部分切割(Partial cut)”中，IL-15/IL-15RαSushi+融合多肽从两条Fc链中的仅一条上被裂解下，而在“完全切割(Full cut)”中，IL-15/IL-15RαSushi+融合多肽从两条Fc链上被裂解下。图10A中圈出的模糊条带是从Fc上裂解下的IL-15/IL-15RαSushi+融合多肽，并且该模糊外观很可能是由于糖基化引起的。在还原(R)样品中，上方条带是连接至IL-15/IL-15RαSushi+融合多肽的Fc链，并且下方清晰条带是IL-15/IL-15RαSushi+融合多肽已被裂解下来的Fc链。
SDS-PAGE凝胶清晰地显示，随着uPA量的增加，在非还原样品中完全切割的蛋白的量逐渐增加。同样，还原样品中裂解的Fc链的量增加，表明裂解水平增加。然而，没有条件导致完全裂解。为了实现完全消化，将类似的uPA消化反应孵育更长时间。图10B示出了在37℃用50ng、100ng和300ng的uPA裂解24小时的2μg P-0203。数据表明，100ng uPA和24小时的孵育导致几乎完全的裂解。
实施例5
蛋白酶裂解IL-15 Fc VitoKine P-0203以获得活化的IL-15产物
VitoKine P-0203(SEQ ID NO:29)含有连接Fc和IL-15的、在两端的侧翼带有间隔肽的uPA底物肽接头(SEQ ID NO:90)，并且第二个15-氨基酸柔性接头(GGGGS)3(SEQ IDNO:127)连接IL-15和IL-15RαSushi+结构域。通过在37℃将100μg的VitoKine P-0203与5μg重组人类uPA(BioLegend)在500μl PBS pH 7.2缓冲液中一起孵育24小时来实现体外蛋白酶裂解。为了停止反应，添加25μl的Ni-Excel树脂(在PBS中平衡的50％的浆液，GEHealthcare)以从溶液中去除加6-His标签的uPA。同时，还向反应物添加50μlMabSelectSure蛋白A树脂(在PBS中平衡的50％的浆液，GE Healthcare)，以去除裂解的Fc级分和未切割或未完全消化的P-0203。在室温与两种亲和树脂一起孵育15min后，通过离心去除树脂，并且回收含有蛋白酶活化的P-0203，即IL-15/IL-15RαSushi+融合多肽(在图2中示意性地例示为活性形式1)的流通物。如在图11A和图11B中可以观察到的，活化的P-0203片段与模糊条带一起迁移，模糊条带很可能是由于糖基化引起的。
实施例6
蛋白酶裂解IL-15 Fc VitoKine P-0315以获得活化的IL-15产物
VitoKine P-0315(SEQ ID NO:33)包含连接Fc和IL-15的uPA底物肽接头(SEQ IDNO:92)，和在IL-15和IL-15RαSushi+结构域之间的第二个10-氨基酸MMP-2/9可裂解接头(SEQ ID NO:95)。P-0315中的IL-15结构域含有S58D取代，以增强与受体β亚基的结合。通过蛋白酶消化产生P-0315的两种活化形式。
通过使用MMP-2的体外蛋白酶裂解获得了P-0315的一种活化形式(在图2中示意性地例示为活性形式2)。简而言之，根据制造商的说明将660ng的潜伏性MMP-2(BioLegend)以APMA(Millipore Sigma)活化，更换缓冲液，并添加到0.4ml制造商推荐的测定缓冲液(100mM Tris、20mM CaCl2、300mM NaCl、0.1％(w/v)Brij 35，pH 7.5)中的P-0315(80μg)中。在37℃孵育3小时后，向反应物中添加50μl MabSelectSure蛋白A树脂(在PBS中平衡的50％浆液，GE Healthcare)。用25mM柠檬酸钠、25mM氯化钠pH 3.2洗脱期望的活化形式1。蛋白通过添加3％的1M Tris pH 10.2来中和。为了评估裂解，将样品在4％-12％Tris-BisSDS-PAGE凝胶上分离(图12A)。在泳道1中显示了在还原剂的存在下MMP-2消化前的P-0315，并且泳道2和3是在MMP-2蛋白水解后但在蛋白A纯化前的非还原和还原的P-0315。IL-15Rα-sushi+结构域在凝胶上在9KDa处以尖锐条带出现，证实了MMP-2在MMP-2/9底物肽接头处的有效裂解。蛋白A纯化后，样品(泳道4和5)显示出相同的迁移模式。该数据表明，从共价连接释放的IL-15RαSushi+结构域保持与IL-15非共价缔合，IL-15与Fc融合，如图2中的以活性形式2描绘的；这样的缔合足够强，可以承受蛋白A洗脱过程中的低pH条件。图12B进一步说明了这种活化形式的两种非共价缔合的组分。
通过用uPA和MMP-2两者对P-0315进行蛋白酶裂解获得了P-0315的另一种活化形式(在图2中示意性地例示为活性形式3)。简而言之，将100μg P-0315与5μg uPA在400μlPBS pH 7.2缓冲液中孵育20小时。然后将含有200mM Tris、40mM CaCl2、450mM NaCl、0.2％(w/v)Brij 35，pH 7.5的等体积缓冲液添加到反应物中，以将缓冲液调整至接近制造商推荐的MMP-2测定缓冲液(100mM Tris、20mM CaCl2、300mM NaCl、0.1％(w/v)Brij 35，pH7.5)。通过APMA活化潜伏性MMP-2(660ng)，将缓冲液交换为测定缓冲液，添加到反应物中，并在37℃孵育3小时。添加Ni-Excel树脂(50μl在PBS中平衡的50％的浆液，GE Healthcare)以从溶液中去除加His标签的MMP-2和uPA。同时，向反应物添加100μl MabSelectSure蛋白A树脂(在PBS中平衡的50％的浆液，GE Healthcare)，以去除裂解的Fc级分和剩余的未切割或未完全消化的P-0315。在室温与两种亲和树脂一起孵育15min后，通过离心除去树脂，并且回收含有具有图2中的示意图的P-0315的活性形式3的流通物。如图12C中说明的，P-0315的活性形式3包含如预计的来自双重蛋白水解反应的IL-15/IL-15RαSushi+非共价复合物；IL-15以模糊条带形式迁移，而IL-15RαSushi+是～9KDa处的尖锐条带，如在活性形式2中观察到的(图12B)。
实施例7
通过人类PBMC测定对蛋白酶活化的IL-15 Fc VitoKine的活性评估
如在实施例2中详述的，对来自新鲜的人类PBMC的免疫细胞亚群的活化标志物CD69进行FACS分析，以评估蛋白酶活化的IL-15 VitoKine的活性。P-0203及其由uPA消化产生的相应活化形式(P-0203活化形式；在图2中示意性地例示为活性形式1)的比较，在图13中说明。蛋白酶活化前VitoKine的活性比高活性IL-15/IL-15RαFc融合蛋白P-0165低约3个对数，这与实施例3中描述的VitoKine活性一致。uPA消化显著恢复了活化CD56+NK细胞(图13A)和CD8+T细胞(图13B)两者的效力，但仍明显低于P-0165的效力，这可能是由于IL-15和IL-15α结构域的共价连接引起的。延长连接IL-15和IL-15α的柔性接头的长度被预计增强活化形式的效力。矛盾的是，接头长度的延长也可能会降低D3结构域的活性遮蔽效率，并因此导致更高基础活性的VitoKine构建体。
通过测量活化新鲜的人类PBMC的免疫细胞亚群中CD69的活化来评估另一种IL-15Fc VitoKine P-0315及其两种活化形式的生物活性。如在图14中观察到的，未裂解的P-0315的活性几乎无法在所测试的浓度测量到，证实了在VitoKine形式中活性部分的有效遮蔽。如图2中说明的，P-0315的活性形式2包含与由MMP-2裂解释放的IL-15RαSushi+结构域非共价复合的Fc-融合的IL-15；P-0315的活性形式2在结构上类似于阳性对照P-0313，P-0313是一种高效的IL-15 IL-15RαFc融合蛋白。如图2中描绘的，P-0315的活性形式3包含由uPA从Fc结构域裂解下的游离IL-15结构域，和由MMP-2裂解释放的IL-15RαSushi+结构域，两者形成非共价复合物。两种活化形式的P-0315在活化CD56+NK细胞(图14A)和CD8+T细胞(图14B)两者方面显示出完全或接近完全的效力恢复；活性形式3比活性形式2中度地更活跃。当期望肿瘤微环境中意图的途径的瞬时活化时，活性形式3中Fc结构域的缺乏可以是有益的。
MMP-2蛋白水解之前和之后的P-0315活性还通过测量处理后的NK细胞(图15A)和CD8+T细胞(图15B)的细胞核中的Ki67表达来研究。包含两个不可裂解的柔性接头的P-0351被包括在内用于比较。这些数据进一步展示了VitoKine的活性惰性以及在体外蛋白水解活化后在NK细胞和CD8+T细胞两者中约3个对数的效力恢复。P-0351和P-0315具有相同活性的观察结果表明，P-0315中的两个可裂解接头在产生、表达和储存期间中保持完整，并且对各自的蛋白酶具有特异性。
总之，IL-15 VitoKine P-0315被MMP-2/9和/或uPA裂解导致分子的活化，并且细胞因子活性恢复到与高活性IL-15化合物P-0313相似的水平，具有亚纳摩尔范围内的EC50。
实施例8
IL-15 Fc VitoKine在健康小鼠中具有最小的全身性细胞因子效应
VitoKine平台技术的目标是降低全身性中靶毒性和增强治疗窗口。VitoKine将活性细胞因子遮蔽为惰性状态，并阻止活性细胞因子与非病变细胞的外周中或细胞表面上的受体结合。因此，VitoKine平台限制了细胞因子途径的过度活化，并降低了不期望的“组织外”“中靶”毒性。意图VitoKine被病变组织中上调的蛋白酶在局部活化。为了评估这一假设，将蛋白酶可裂解和不可裂解的VitoKine施用到健康小鼠中，并与高活性IL-15Fc融合蛋白比较，来评价它们的全身性细胞因子效应。
P-0313(SEQ ID NO:47和5)是作为阳性对照的完全活性的IL-15/IL-15RαFc融合分子。P-0315(SEQ ID NO:33)是含有两个蛋白酶可裂解接头的IL-15 Fc VitoKine。P-0351(SEQ ID NO:25)是含有两个不可裂解接头的IL-15 Fc VitoKine。媒介物(PBS)被包括在内作为阴性对照。将化合物以0.1mg/kg和0.3mg/kg的剂量一次单一i.p.注射到健康的BALB/c小鼠(8-10周龄，n＝6只/组)中来提供。在给药前(第-1天)或给药后第3天、第5天和第7天采集血液样品以进行免疫表型分析。
通过BD pharm裂解缓冲液裂解红细胞后，通过台盼蓝死细胞排除法计数总的存活单核血细胞。用纯化的抗小鼠CD16/CD32(1:50稀释)封闭Fc受体后，用抗小鼠CD3-FITC、抗小鼠CD49b-APC和抗小鼠CD8-Percpcy5.5(1:50稀释)对细胞进行染色。孵育30分钟后，收集细胞并洗涤，重悬于FACS缓冲液中，并通过流式细胞术进行分析。
如图16中示出的，完全活性的IL-15 Fc融合蛋白P-0313以剂量依赖的方式在两个测试剂量显著扩增了外周血细胞毒性CD8+T细胞(图16A)、NK细胞(图16B)和总白细胞(图16C)。在第3天观察到细胞扩增，在第5天达到峰值，并且在第7天返回接近基线。相比之下，在整个7天的研究中，可裂解的(P-0315)和不可裂解的(P-0351)VitoKine都没有显示CD8 T细胞的增加。在用高剂量的可裂解的VitoKine P-0315处理的小鼠中观察到NK细胞扩增的轻微和延迟的增加。P-0351和低剂量的P-0315在测试的任何靶向细胞群体中没有显示出增加的迹象。总的来说，与活性分子P-0313相比，这两种测试的VitoKine显示出最小的全身性活化和靶向淋巴细胞群体的扩增，并展示出在外周中对IL-15的活性的成功的遮蔽和延迟。
实施例9
在小鼠中用IL-15 Fc VitoKine抑制结肠癌细胞肺转移
在小鼠CT26肺转移模型中研究了IL-15 Fc VitoKine分子的抗转移功效和免疫应答。简而言之，将1×105个小鼠结肠癌细胞CT26-WT(ATCC CRL-2638)静脉内注射到雌性Balb/C小鼠(9-11周龄)中。次日(第1天)开始经由腹膜内注射的四次Q5D处理。处理组(总共6个，n＝7只/组)包括0.3mg/kg P-0315、0.3mg/kg P-0351和0.1mg/kg P-0313。P-0315(SEQID NO:33)是含有两个蛋白酶可裂解接头的IL-15 Fc VitoKine。P-0351(SEQ ID NO:25)是不可裂解的IL-15 Fc VitoKine。P-0313(SEQ ID NO:47和5)是完全活性的IL-15/IL-15RαFc融合分子。媒介物(PBS)被包括在内作为阴性对照。在第17天，将所有小鼠处死进行组织收获。将肺用15％印度墨汁填充(inflate)，并在Fekete溶液(10％甲醛、5％冰乙酸和60％乙醇)中脱色。对肺部肿瘤结节进行计数，并且通过处理组和媒介物对照之间肿瘤结节的不同数目来表示抗转移作用。
如图17中说明的，P-0313在抑制肺转移的形成和生长方面具有显著效果。在0.1mg/kg，P-0313处理导致对肺转移的接近完全的抑制。可裂解的VitoKine P-0315展示出对肺结节发展的70％的抑制；所有三种剂量(0.3mg/kg、1mg/kg或3mg/kg)的抗转移功效相当。不可裂解的VitoKine P-0351在减少转移发展方面展示出相对较弱但仍显著的效果，表明在高剂量具有一定的固有基础活性。然而，P-0315展示出明显优于P-0351的抗转移功效(p<0.05；图17)，表明蛋白水解裂解P-0315中的一个或两个接头并且随后IL-15的活性形式的释放可能有助于P-0315的体内功效优于P-0351。肿瘤转移发展可以导致肿瘤微环境附近的蛋白水解活性的增加。
在第15天(第三次处理后4天)，通过对小鼠外周血的流式细胞术分析来研究IL-15化合物处理后的免疫应答。与对照相比，在用活性IL-15 Fc融合蛋白P-0313处理的小鼠中观察到CD8+T细胞的扩增，但在用可裂解的VitoKine P-0315或不可裂解的VitoKine P-0351处理的小鼠中均未观察到，表明在VitoKine不引起全身性CD8+T细胞升高的情况下观察到了VitoKine的抗结肠癌转移功效(图17和图18A)。然而，在重复给药后，外周血NK细胞在所有三种IL-15化合物处理的组中均升高，不可裂解的VitoKine组中增加最明显(图18B)。在VitoKine处理组中，NK细胞而不是CD8+T细胞的全身性扩增的增加表明，NK细胞比CD8+T细胞对IL-15处理更敏感，并且VitoKine的固有基础活性可能导致NK细胞扩增。因此，调整IL-15 VitoKine的给药浓度以减少残留的全身效应至关重要。在P-0351组中NK细胞的明显增加还表明，低效力的不可裂解的VitoKine可以微弱但持续地活化该途径，并导致长期的免疫应答。
实施例10
IL-15 Fc VitoKine P-0315抑制小鼠中已建立的CT26肿瘤生长，具有最少的全身细胞因子活化
在CT26鼠结肠直肠癌肿瘤模型中，研究了与完全活性的IL-15/IL-15Rα-Fc融合蛋白P-0313相比，IL-15 Fc VitoKine P-0315的抗肿瘤功效和免疫应答。简而言之，在雌性Balb/C小鼠(10-12周龄)的右腹肋皮下注射1×105个CT26细胞。在第11天，当平均肿瘤体积为～70mm3时，小鼠被随机分为三组(n＝11只/组)，并在随机分组的同一天接受腹膜内注射0.1mg/kg的媒介物(PBS)或P-0315或P-0313。在第16天进行各自测试剂的一次另外的腹膜内注射(2×Q5D)。使用卡尺每周测量肿瘤三次，并且肿瘤体积计算如下：体积＝0.5x(宽度)2x(长度)。为了研究免疫应答，在第19天在肝素处理过的管中收集非终末外周血。在第21天，将所有小鼠处死进行组织收获。
如图19A中示出的，PBS处理的小鼠快速地发展出大的皮下肿瘤，而用P-0315或P-0313处理小鼠在延迟肿瘤生长方面大致等效(图19B和图19C)。在肿瘤接种后的第21天，对照处理的小鼠中的平均肿瘤体积为～1000mm3，对比用P-0315或P-0313处理的小鼠中的平均肿瘤体积为～450mm3(****，P<0.0001；单因素ANOVA与Tukey事后检验)(图19D)。值得注意的是，最初P-0313比P-0315显示出更大的肿瘤负荷降低，但是随着处理进行，差异逐渐减小(tapered off)。P-0315的延迟的抗肿瘤作用可能是由于发展出适当的量的蛋白酶以接近和裂解底物肽接头并活化VitoKine所花费的时间。
接下来，通过流式细胞术研究了与P-0313和媒介物相比，P-0315对外周血中的CD8+T细胞和NK细胞的增殖的影响。对P-0315对外周和脾的总WBC的群体和淋巴细胞亚群(CD8+T细胞和NK细胞)的影响也进行了类似的评估。
对荷瘤小鼠注射完全活性的IL-15/IL-15RαFc融合蛋白P-0313，在外周血和脾中都诱导了明显的淋巴细胞增殖和扩增(图20-图22)。与PBS组相比，在P-0313处理后，外周NK细胞中的Ki67增殖增加到4倍(61％对比15％；图20A)，并且CD8+细胞中的Ki67增殖增加到5.3倍(46％对比8.6％；图20B)。同样，P-0313处理导致外周血(图21A-图21C)和脾(图22A-图22C)中总白细胞、NK细胞和CD8+T细胞的显著细胞扩增。例如，观察到外周总WCB细胞数目扩增到6倍，并且CD8+T细胞数目扩增到5倍；NK细胞数目显著增加到85倍。在脾中，也观察到NK细胞的最明显的细胞扩增(10倍)，其次是CD8+T细胞，扩增到2.9倍。总脾WBC中度地扩增到1.7倍。细胞毒性CD8+T细胞和NK细胞的强烈活化与IL-15的总体免疫调节特性一致，并且有效的免疫应答可能是体内P-0313抗肿瘤活性的主要贡献者。然而，血液中显著改变的淋巴细胞亚群可能引起毒性和降低治疗指数。
与P-0313形成鲜明对比，用IL-15 Fc VitoKine P-0315处理导致血液中淋巴细胞亚群的内稳态的最小改变。在图20中展示了外周NK细胞和CD8+T细胞的Ki67增殖的观察结果，在图21中展示了外周血中总白细胞、NK细胞和CD8+T细胞的细胞扩增的观察结果。在P-0315处理后，唯一值得注意的免疫药效学作用是脾中NK细胞数目增加到4倍(图22B)。由于在延迟已建立的CT26肿瘤生长方面，P-0315与P-0313大致等效(图19A-图19D)，因此P-0315的体内抗肿瘤活性可能是由肿瘤微环境附近VitoKine的可裂解接头蛋白水解和随后的活化引起的。由于活化的VitoKine仅存在于肿瘤附近，因此外周淋巴细胞对施用惰性VitoKine分子的响应远不如完全活性的P-0313明显。
总的来说，以P-0315示例的IL-15 Fc VitoKine能够有效延迟肿瘤生长，而不会显著改变血液和脾中淋巴细胞亚群的增殖和扩增。因此，通常与完全活性细胞因子相关的途径的过度活化、不期望的“组织外”“中靶”毒性以及不希望的靶吸收可以通过VitoKine形式来防止或减少，而不会损害抗肿瘤作用。
实施例11
下调IL-15部分的效力以使对应的VitoKine的固有基础活性最小化
尽管在完全活性的二价IL-15S58D/IL-15RαFc融合体P-0313和其对应的FcVitoKine P-0315之间有超过3个对数的活性减弱，但是P-0315仍然具有能够以50-100nMEC50离体刺激效应细胞的固有基础活性(在图15中例示)。在高体内剂量，VitoKine的固有基础活性可以潜在地导致具有持续的体内药效学作用的外周受体刺激，并且可能引起全身性毒性。因此，本发明人推断，下调IL-15部分的效力可以使对应的Vitokine的固有基础活性最小化。
一组具有破坏IL-15Rβγ的氨基酸取代的IL-15突变蛋白被表达为IL-15/IL-15Rα(非共价)Fc合蛋白，并且在人类PBMC测定中筛选效力减弱，如先前描述的。使用P-0313作为对照分子。具有示例性的单个或组合的残基V63、I68和Q108的IL-15氨基酸变化的突变蛋白导致CD8 T细胞增殖效力减弱，总结在表13中。与P-0313相比，这些IL-15变体/IL-15RαFc融合蛋白展示出广泛的效力减弱，从5倍跨至～6700倍。图23A和图23B进一步说明了用一些代表性IL-15变体/IL-15RαFc融合体(包括P-0736、P-0772、P-0737、P-0768、P-0793和P-0764)处理后，CD8 T细胞和NK细胞上Ki67表达的百分比。在图23中显示的融合蛋白中的IL-15氨基酸变化总结在表13中。
表13
示例性的单个或组合的IL-15氨基酸取代破坏了IL15Rβγ相互作用，并且产生在刺激CD8 T细胞上Ki67表达方面具有降低的效力的变体
Figure BDA0003806684520001011
此外，当包含以SEQ ID NO:182-192例示的氨基酸缺失或插入或者氨基酸取代和缺失/插入的组合的IL-15变体被表达为IL-15/IL-15RαSushi Fc融合体时，展示出不同水平的效力减弱(数据未示出)。这样的IL-15部分可以类似地用于IL-15 VitoKine形式中，以最佳地调节固有基础活性。如本领域技术人员将理解的，所有突变(氨基酸取代、缺失和插入)可以任选地和独立地以任何方式组合，以实现最佳的活性调节。
为了测试下调IL-15部分的效力是否确实使对应的VitoKine的固有基础活性最小化，显示出显著减弱的活性的IL-15 Q108S作为D2结构域被掺入VitoKine中，称为P-0682。除了含有不同的IL-15变体之外，P-0682与P-0315的差异还在于具有不可裂解的柔性L1接头。但除此之外，这两种VitoKine是相同的。如图24A和图24B所图示的，P-0682即使在1μM的最高测试浓度，也完全失去了诱导CD8 T细胞或NK细胞上Ki67表达的活性。P-0764是P-0682的Fc融合对应物，类似于P-0682的活化形式。本发明的汇总数据表明，在离体测定中，IL-15VitoKine平台在细胞因子效力方面通常产生1000倍至2000倍的减弱，并且因此估计基于IL-15 Q108S的VitoKine P-0682诱导CD8 T细胞和NK细胞的EC50值分别为100μM和20μM。因此，数据确实支持IL-15效力减弱导致对应的Vitokine的固有基础活性降低的推断。
由于IL-15 VitoKine的固有基础活性成比例地取决于细胞因子部分的活性，因此可以通过掺入具有不同效力水平的IL-15部分来调节，诸如表13中列出的那些。例如，包含IL-15 V63A/I68H作为D2结构域的IL-15 Fc VitoKine P-0806(SEQ ID NO:231)，预计具有在P-0315和P-0682之间的中等基础活性，估计在诱导CD8 T细胞方面EC50为2-3μM。重要的是，IL-15VitoKine固有基础活性的可调节性使得VitoKine活性在裂解之前的惰性和活化之后的效力之间能够达到最佳平衡，从而有利于获得期望的抗肿瘤功效，同时使不希望的全身性毒性最小化。
实施例12
不可裂解的VitoKine作为细胞因子的效力减弱的形式
本领域已知，在体外有效的细胞因子可能不会在体内引起最强的淋巴细胞响应。高效的细胞因子常常与较强的受体刺激、内化和脱敏、信号传导减弱、增殖和功能、以及细胞死亡增加或克隆消耗相关。因此，可能非常需要效力减弱的细胞因子来防止过度强烈的淋巴细胞活化，并实现持续和增强的体内药效学作用和抗肿瘤功效。
在体外与完全活性的IL-15化合物相比，不可裂解的IL-15 Fc VitoKine P-0351表现出显著的效力降低，但P-0351在小鼠CT26肺转移模型中显示出抗转移功效和显著的NK细胞响应(实施例8)。因此，不可裂解的VitoKine构建体可用作具有持续活性的效力减弱的细胞因子，以优化体内药效学。
在诱导NK细胞和CD8+T细胞两者(图25A和图25B)中的Ki67增殖方面，P-0351表现出与基准分子(SEQ ID NO:177和178)相同的效力，基准分子相当于专利申请WO2018071919A1中的XENP024306。XENP024306是一种IL-15/IL-15RαFc融合分子，在IL-15中含有氨基酸取代(D30N/E64Q/N65D)并且在Fc中含有半衰期延长突变。据报道，XENP024306的IL-15链中的三重突变导致体外效力降低200倍，但XENP024306被展示为在体内更具活性，这可能是由于优化的体内药效学。
同样，通过避免或减少通常与完全活性细胞因子相关的过度活化和不希望的靶吸收，预计P-0351的效力减弱会导致更持续的暴露以改进药效学(PD)。因此，P-0351的半衰期延长的对应物P-0651(SEQ ID NO:170)能促进更长的半衰期并进一步延长体内药效学作用。
实施例13
用于选择性扩增调节性T细胞的IL-2 Fc VitoKine(Treg IL-2 VitoKine)的构建和产生
目标是设计IL-2 VitoKine构建体，该构建体将保持惰性，直到被炎性部位处上调的蛋白酶在局部活化。据报道，与效应T细胞相比优先刺激Treg的低剂量野生型IL-2和与IL-2Rβ结合亲和力降低的IL-2突变蛋白扩大了选择性窗口。这些分子可被开发为用于预防自身免疫性疾病的治疗剂。干扰IL-2Rβ和/或γC结合且不影响与IL-2Rα相互作用的其他突变也可以扩大Treg活化高于Teff的选择性窗口。
IL-2 Fc VitoKine包含被用作活性部分的野生型IL-2或具有增加的刺激Treg高于效应T细胞的选择性的IL-2突变蛋白，所述活性部分被可逆地遮蔽在Fc结构域和IL-2RαSushi(SEQ ID NO:10)之间。IL-2Rα(SEQ ID NO:9)包含由天然肽接头区域隔开的两个sushi结构域。IL-2 VitoKine构建体包含一个或两个被蛋白酶识别的可裂解接头，所述蛋白酶被报道在炎性紊乱部位处上调。虽然连接Fc和IL-2/突变蛋白的接头既可以是可裂解的也可以是不可裂解的，但连接IL-2和IL-2RαSushi的接头优选地是能够被蛋白酶特异性地裂解的。
选择性刺激Treg的IL-2突变蛋白活性被预计在蛋白酶裂解后、IL-2Rα从IL-2释放和扩散离开后恢复。由于IL-2Rα和IL-2之间的nM结合亲和力，在接头裂解后IL-2RαSushi有可能保持与IL-2非共价缔合；因此，IL-2与Treg细胞上的IL-2Rα的相互作用保持被阻断。为了解决这个潜在的问题，设计了在与IL-2的界面处具有氨基酸取代的IL-2Rα突变蛋白，以削弱其与IL-2的结合。因此，在接头的蛋白酶裂解后，IL-2RαSushi突变体能解离，并且然后从IL-2扩散离开，这是一种与图2A中说明的活化机制略有不同的活化机制(在图2B中示意性地说明)。
在IL-2Rα结构域的位置38处(即K38E)和位置43处(即Y43A)进行了代表性的氨基酸取代。在与IL-2相互作用的残基上具有取代的其他IL-2Rα变体被预计破坏IL-2和IL-2Rα的相互作用，并且也可以被掺入。如本领域技术人员将理解的，所有的突变可以任选地和独立地以任何方式组合，以实现最佳的亲和力调节。产生了包含不同接头组成、野生型或变体IL-2和野生型或变体IL-2RαSushi的IL-2 VitoKine分子，并且它们各自的序列以SEQ IDNO:49-65列出。
基因合成、表达载体构建以及蛋白产生、纯化和表征遵循实施例1中详述的相同程序进行。作为展示IL-2 VitoKine的蛋白谱的实例，对P-0320的SDS-PAGE分析在图26A中示出。图26B中的尺寸排阻色谱图表明，在没有精制步骤(polishing step)的初始蛋白A捕获步骤之后，存在<5％的聚集。低聚集倾向表明IL-2 VitoKine具有有利的可开发性谱。
实施例14
Treg IL-2 Fc VitoKine体外活性评估
通过测量新鲜的人类PBMC中特定T细胞亚群中的磷酸化STAT5(pStat5)水平来确定IL-2 VitoKine对T细胞的生物活性。已知Stat5参与由IL-2结合跨膜IL-2RβγC复合物诱导的下游细胞内信号传导。使用针对pStat5肽的抗体通过流式细胞术测量固定且透化的细胞中的pStat5的水平。简而言之，通过Ficoll-Hypaque离心法从购自Oklahoma BloodInstitute的健康供体的血沉棕黄层分离人类PBMC。在37℃将2×105个PBMC用测试化合物的系列稀释液处理30分钟。然后根据制造商的说明，用Foxp3/转录因子染色缓冲液组(EBIO)处理细胞。然后用Cytofix缓冲液固定细胞并用Perm缓冲液III(BD Biosciences)透化并且然后洗涤。在通过添加人类TruStain FcX(1:50稀释)封闭Fc受体后，以制造商推荐的浓度用抗CD25-PE、抗FOXP3-APC、抗pSTAT5-FITC和抗CD4-PerCP-Cy5.5抗体的混合物将细胞在室温染色60分钟。然后收集细胞并洗涤，重悬于FACS缓冲液中，并通过流式细胞术进行分析。分别将Treg和CD4+常规T细胞亚群的流式细胞术数据门控为Foxp3+/CD25高和Foxp3-/CD25低组。数据表示为门控的群体中pStat5阳性细胞的百分比。
评估了与P-0250(SEQ ID NO:48)相比，IL-2 VitoKine P-0320(SEQ ID NO:49)和P-0329(SEQ ID NO:62)的pStat5活化。P-0320含有野生型IL-2结构域，该野生型IL-2结构域的N末端经由uPA可裂解的接头与Fc结构域融合，并且该野生型IL-2结构域的C末端通过柔性(GGGGS)3(SEQ ID NO:127)接头与IL-2RαSushi结构域连接。P-0329含有野生型IL-2结构域，该野生型IL-2结构域的C末端经由uPA可裂解的接头与Fc结构域融合，并且该野生型IL-2结构域的N末端通过柔性(GGGGS)3(SEQ ID NO:127)接头与IL-2RαSushi结构域连接。P-0250是一种高活性的IL-2 Fc融合蛋白。对于所测试的化合物，Treg和CD4+常规T细胞(Tconv)亚群中pStat5阳性细胞的百分比在图27中说明。清楚地观察到，与完全活性的IL-2融合蛋白相比，对于两种IL-2 VitoKine，Treg中的pStat5活化显著降低，并且CD4+Tconv细胞的pStat5活化几乎测量不到。这些数据清楚地展示了VitoKine形式中IL-2活性的有效遮蔽。
实施例15
IL-2 Fc VitoKine的蛋白酶活化及体外活性评估
IL-2 Fc VitoKine P-0382(SEQ ID NO:51)包含连接Fc和IL-2的柔性GGGSGGGS接头(SEQ ID NO:115)，以及在IL-2和IL-2RαSushi结构域之间的10-氨基酸MMP-2/9可裂解接头(SEQ ID NO:77)。P-0382中的IL-2RαSushi结构域含有设计为降低IL-2RαSushi结构域对IL-2的结合亲和力的氨基酸取代(K38E)，以确保IL-2RαSushi结构域在蛋白酶裂解接头后解离并随后从IL-2扩散离开。
通过使用MMP-2的体外蛋白酶裂解来活化P-0382。简而言之，首先根据制造商的说明将3.3μg的潜伏性MMP-2(BioLegend)以APMA(Millipore Sigma)活化，然后将其更换缓冲液，并添加到0.4ml制造商推荐的测定缓冲液(100mM Tris、20mM CaCl2、300mM NaCl、0.1％(w/v)Brij35，pH 7.5)中的120μg P-0382中。在37℃孵育20hrs后，将一半的反应物用MabSelectSure蛋白A树脂纯化，并且用25mM柠檬酸钠、25mM氯化钠pH 3.2洗脱活化的VitoKine。蛋白通过添加3％的1M Tris pH 10.2来中和。将另一半样品与Ni-Excel树脂一起孵育，以便通过去除加His标签的MMP-2蛋白来停止反应，并通过经由离心去除Ni树脂来收集活化的VitoKine。蛋白A纯化是为了证实在从多肽链裂解下后，IL-2RαSushi结构域不与IL-2非共价缔合，如图2B示意性地说明的。在图28中示出了在4％-12％Tris-Bis SDS-PAGE上对样品进行评估。尽管与结构上相似的IL-15VitoKine(例如P-0315)相比，蛋白酶的量增加且反应时间延长，但该反应并未导致完全的裂解。对经蛋白A纯化和未经蛋白A纯化的MMP-2处理的样品的比较(图29A和图29B)确实证实了IL-2RαSushi结构域从共价连接释放并且未与Fc-IL-2融合多肽在蛋白A中共纯化。
尽管裂解不完全，但是将两种MMP-2活化的样品(一种为Ni-Excel流通物(活性形式1)并且另一种为蛋白A洗脱物(活性形式2))在实施例14中描述的pStat5活化测定中进行评估，并且数据在图29中说明。P-0382在Treg中的活性非常低，并且对于CD4+Tconv细胞几乎测量不到，再次证实了IL-2 VitoKine形式中活性部分的有效遮蔽。两种活化的样品显示出活性几乎完全恢复。与P-0250相比效力稍低，可能是由于蛋白水解不完全。
活性形式1样品中MMP-2裂解的IL-2RαSushi结构域的存在表现为未改变活化的IL-2 VitoKine的活性，因为活性形式1和活性形式2在诱导Treg细胞和Tconv细胞两者的pStat5磷酸化方面具有相当的效力(图28A和图28B)。数据表明，由MMP-2裂解产生的IL-2RαSushi结构域不与IL-2缔合，并且应当不干扰IL-2与淋巴细胞上表达的受体复合物的结合。
P-0382的MMP-2蛋白水解未产生完全的裂解，并且推断可裂解接头的延长可以使得底物肽对于负责裂解的蛋白酶更可及。将P-0382中的10-氨基酸接头(SEQ ID NO:95)替换为含有额外的侧翼残基的15-氨基酸的MMP-2/9-可裂解接头(SEQ ID NO:94)，并且产生了新的VitoKine构建体P-0398(SEQ ID NO:52)。遵循上文详述的相同方案，通过使用MMP-2的体外蛋白酶裂解来活化P-0398。三倍低的量的MMP-2(对180μg P-0398为1.5μg MMP-2，而对120μg P-0382为3.3μg MMP-2)导致了P-0398的完全消化，证据为在SDS-PAGE凝胶上只存在“完全切割”条带(数据未示出)。
通过蛋白A纯化去除了IL-2RαSushi结构域的活化的P-0398的生物活性在pStat5测定中被确定(图29A和图29B)。活化的P-0398在序列和结构上类似于IL-2 Fc融合分子P-0250，并且它们在诱导Treg细胞和Tconv细胞两者中的Stat5磷酸化方面具有几乎相同的效力。虽然两种VitoKine(P-0382和P-0398)由于与IL-2RαSushi结构域的共价连接而具有显著受损的生物活性(4个对数)，但似乎存在一种趋势，即包含更长L2接头的P-0398活性稍高。与IL-15 Fc VitoKine的观察结果类似，IL-2 VitoKine的活性惰性水平可以通过调整L2接头长度来进一步调节。同样，可裂解的L2接头长度和序列的选择应在特定蛋白酶在意图的疾病适应症部位的存在、底物肽对蛋白酶的可及性和期望的蛋白水解速率之间进行平衡。
总之，与IL-15 VitoKine相比，IL-2 VitoKine需要更长的L2接头来获得最佳的酶可及性，以实现完全的蛋白水解。MMP-2对示例性IL-2VitoKine构建体P-0382和P-0398的裂解导致分子的完全活化。活化的IL-2VitoKine实现了与高活性IL-2 Fc融合化合物P-0250相似的生物活性。
实施例16
用于选择性扩增效应T细胞的IL-2 Fc VitoKine(Teff IL-2 VitoKine)的构建和产生
目标是设计IL-2 VitoKine构建体，该构建体将保持惰性，直到被仅在肿瘤部位处存在或上调的蛋白酶在局部活化。IL-2对Treg的优先扩增代表了IL-2对癌症免疫疗法的不期望的影响，因为Treg可以抑制效应T细胞响应。除了信号传导受体βγ亚基外，Treg上IL-2Rα的高且组成性的表达是IL-2优先扩增Treg的原因。为了克服这些限制，设计为不再与IL-2Rα结合的IL-2变体预计不会优先活化Treg，而只在CD8+T细胞和NK细胞也被活化时的浓度活化Treg。遵循这一概念，设计了一组在与IL-2Rα相互作用的残基处包含一个或若干个氨基酸取代的IL-2变体。残基R38、T41、F42、F44、E62、P65、E68和Y107位于与IL-2Rα的界面处并与多于一个IL-2Rα残基形成氢键/盐桥或疏水相互作用(Mathias Rickert,等人(2005)Science 308,1477-80)。这些位点处的氨基酸取代预计会破坏与IL-2Rα的相互作用，并产生具有减少或消除的与IL-2Rα结合的IL-2变体。
一组IL-2突变蛋白被表达为与Fc同源二聚体或Fc异源二聚体的C末端融合体，并以酶联免疫吸附测定(ELISA)筛选与IL-2Rα的结合。简而言之，将IL-2Rα-ECD以0.1μg/孔包被到孔上。在4℃孵育过夜并封闭后，以100μl/孔向每个孔添加系列稀释的IL-2 Fc融合蛋白。在室温孵育1小时后，向每个孔添加100μl/孔的山羊抗人类IgG Fc-HRP(在稀释液中1:5000稀释)，并在室温孵育1小时。在添加100μl TMB底物后，将板在室温在黑暗中显影10分钟，并添加100μl/孔的停止溶液。确定450nm处的吸光度，并使用Prism软件(GraphPad)拟合曲线。二价和单价IL-2部分的野生型IL-2 Fc融合蛋白的S125I等效物P-0531(SEQ ID NO:248)和P-0689(SEQ ID NO:249和168)分别被包括在内作为对照。
导致减少或消除的结合的示例性的单个或组合的IL-2氨基酸变化总结在表14中。在图31中图示出了与P-0689相比，几个代表性的IL-2变体单价Fc融合蛋白P-0704、P-0707、P-0708和P-0709与IL-2Rα的ELISA结合曲线。
表14
示例性的单个或组合的IL-2氨基酸取代破坏了IL-2Rα相互作用，并且产生具有与IL-2Rα减少/消除的结合的IL-2变体
Figure BDA0003806684520001081
除了包含破坏IL-2Rα的氨基酸取代之外，表14中的所有IL-2变体还包含S125I突变，以显著增强蛋白表达和减少聚集倾向。如本领域技术人员将理解的，调节它们对IL-2Rα的亲和力的任何另外的组合突变都符合本发明的精神和范围，无论是否改变它们对IL-2受体的特定组分的亲和力。
此外，虽然具有减少/消除的CD25结合，但这些IL-2变体保留了对二聚体IL-2Rβγ受体的完全结合和功能活性，并能够通过保留的IL-2Rβγ信号传导来活化效应细胞。这在人类PBMC测定中通过P-0704例示。消除了IL-2Rα结合的单价IL-2 P65R Fc融合蛋白P-0704与其野生型IL-2对应物P-0689在诱导CD8 T细胞上Ki67表达方面同等有效(图32)。在未来的实例中，P-0704和P-0689将可互换地用作具有完全IL-2 Teff效力的对照。
Teff IL-2 Fc VitoKine构建体包含作为活性部分的具有减少/消除的与IL-2Rα结合的IL-2变体，所述活性部分被可逆地遮蔽在Fc结构域和IL-2RαSushi(SEQ ID NO:10)之间。这些构建体包含一个或两个被蛋白酶识别的可裂解的接头，所述蛋白酶被报道在各种类型的癌症例如实体瘤中上调。虽然连接Fc和IL-2突变蛋白的接头既可以是可裂解的也可以是不可裂解的，但连接IL-2和IL-2RαSushi的接头优选地能够被蛋白酶特异性地裂解。由于VitoKine中的IL-2部分被设计成不再与IL-2Rα结合，因此VitoKine的遮蔽D3部分IL-2Rα可能在体内蛋白水解裂解后扩散离开(图28)，导致IL-2突变蛋白活性的恢复。在图1中示意性地描绘了掺入作为活性部分的不同IL-2突变蛋白的IL-2 VitoKine分子。构建并产生了用于选择性扩增Teff细胞的示例性IL-2 Fc VitoKine分子，并且它们各自的序列以SEQ ID NO:59-61和271-274列出。
基因合成、表达载体构建以及蛋白产生、纯化和表征遵循如实施例1中详述的相同程序进行。
实施例17
包含IL-2Rα变体作为遮蔽D3部分的IL-2 VitoKine
IL-2Rα变体被设计为通过在与IL-2相互作用的残基处掺入突变来减少与IL-2的结合，使得D3部分在体内蛋白水解后将容易地扩散离开。三种示例性IL-2RαSushi变体(SEQID NO:267-269)被表达为单价Fc融合蛋白，分别对应于P-0751(Y43A)、P-0752(L42G)和P-0753(R36A)。在ELISA中评估了这三种IL-2RαSushi变体Fc融合体的个体突变对与IL-2结合的影响，与它们的野生型IL-2RαSushi对应物融合体(被称为P-0757)进行了比较。
简而言之，将IL-2RαSushi变体单价Fc融合蛋白以1μg/孔包被到Nunc Maxisorp96孔微孔板的孔上。在4℃孵育过夜并用1％BSA封闭后，以100μl/孔向每个孔添加系列稀释的IL-2S125I单价Fc融合蛋白P-0689。在室温孵育1小时后，将100μl/孔的生物素抗IL-2抗体克隆B33-2(BD biosciences)以1μg/ml添加到每个孔中，并在室温孵育1小时。随后，向每个孔添加100μl/孔的以1:500稀释的亲和素-HRP(BioLegend)并孵育30min。最后，向每个孔添加100μl TMB底物；将板在室温显影，并添加100μl/孔的停止溶液。确定450nm处的吸光度，并使用Prism软件(GraphPad)拟合曲线。
如表15中总结的和图33图示的，IL-2Rα氨基酸取代Y43A、L42G和R36A都导致破坏与IL-2的相互作用。Y43A导致与IL-2的结合略微减少(8.1倍)，R36A突变在结合EC50方面产生更显著的346倍减少，并且L42G突变导致与IL-2结合的中等或35倍的减少。
表15
IL-2Rα氨基酸变化对与IL-2结合的影响
Figure BDA0003806684520001101
以上3种IL-2Rα变体及其野生型对应物被用于构建4种Teff IL-2 Fc VitoKine分子，这些分子均包含作为D2结构域的单体IL-2S125I(野生型等效物)和15个氨基酸的MMP2/9可裂解的L2接头。随后通过评价它们与P-0704相比在人类PBMC测定中诱导CD8 T细胞和NK细胞上Ki67表达的效力来评估IL-2Rα变体的遮蔽效率，P-0704是与P-0689具有等同效力的Fc融合体，P-0689是这些组Vitokine的Fc融合对应物。数据总结在表16中并且在图34A和图34B中图示。
表16
各种IL-2 VitoKine构建体的活性比较
Figure BDA0003806684520001111
如图34展示的，当与完全活性的Fc融合对应物P-0704相比时，VitoKine P-0701中作为遮蔽D3结构域的野生型IL-2RαSushi在诱导CD8+T细胞和NK细胞增殖方面产生显著的3个对数的活性降低。预测在IL-2RαSushi中掺入破坏IL-2-结合的突变将减弱D3结构域的遮蔽效应，并从而减少VitoKine的活性惰性。还预期，遮蔽效应减少的程度将与IL-2和IL-2RαSushi变体之间的结合强度降低的水平有关。
如图34A和表16所示，与P-0701相比，IL-2RαSushi变体Y43A和R36A作为它们各自的Fc VitoKine(P-0754和P-0756)中的D3结构域在诱导CD8 T细胞增殖方面确实显示出减弱的遮蔽效应，并从而减少了VitoKine的活性惰性。在NK细胞中观察到相同的活性趋势(图34B)。然而，IL-2RαSushi变体Y43A仅略微减少了与IL-2的结合(8.1倍)，而R36A取代导致IL-2结合的明显得多的346倍减少，但它们各自的VitoKine也表现出类似减弱的遮蔽效应，这与遮蔽效应减少的程度应与IL-2和IL-2RαSushi变体之间结合强度降低的水平相关的预测相矛盾。更引人注目的是，尽管与IL-2的结合减少了35倍，但IL-2RαSushi L42G变体与其野生型对应物相比没有损害其遮蔽效应，并从而几乎完全保留了其对应的VitoKine P-0755的活性惰性(图34A和图34B)。在IL-2Rα中掺入破坏IL-2的突变显示出对遮蔽效应的不同水平的影响并且与结合强度变化的程度不相关，这样的实验观察结果是预期不到的，但在多个实验中得到证实。这可能涉及IL-2和IL-2Rα结构域之间在空间水平上的最佳相互作用。IL-2RαSushi L42G变体被选为IL-2 VitoKine的优选D3结构域，因为IL-2RαSushi L42G变体保留的维持对应VitoKine的活性惰性的遮蔽效应，同时由于IL-2RαSushi L42G变体与IL-2的结合减弱，可以在体内蛋白水解后容易地扩散离开以获得完全活性。同时，当期望调节IL-2 VitoKine的固有基础活性以实现期望的抗肿瘤功效和不希望的全身性毒性之间的最佳平衡时，IL-2Rα变体，例如R36A，可以用作D3结构域。
此外，构建了包含作为D2结构域的IL-2 P65R和作为D3结构域的野生型IL-2RαSushi(P-0745)、IL-2RαSushi Y43A(P-0807)、IL-2RαSushi L42G(P-0808)或IL-2RαSushiR36A(P-0809)的IL-2 Fc VitoKine，并评估其诱导CD8 T细胞和NK细胞增殖的活性。如图35A和图35B中描绘的，当与这些组VitoKine的对应Fc融合体P-0704相比时，所有VitoKine都显示出相当的10倍至20倍的活性降低。由于P65R突变消除了IL-2与IL-2Rα的结合，因此所述较低的遮蔽效应可能是由于IL-2与IL-2RαSushi之间的空间互补，并且IL-2Rα中任何其他破坏IL-2的突变不会导致对遮蔽效应的任何进一步的影响。
此外，包含作为D2结构域的IL-2和作为D3结构域的IL-2RαSushi L42G的IL-2 FcVitoKine P-0755按照实施例15中详述的方法经由体外MMP-2裂解被活化，并在人类PBMC测定中进行评估。在D3结构域裂解和扩散离开后，活化的P-0755与其Fc融合对应物P-0689相似，有少量额外的残基用作蛋白酶底物的一部分并保留在IL-2部分的C末端。如图36A和图36B展示的，P-0755作为VitoKine实现了接近3个对数的活性降低，并能够被活化以恢复刺激效应细胞(包括CD8 T细胞和NK细胞)增殖的全部效力。
总之，构建并评价了包含作为遮蔽D3结构域的IL-2RαSushi变体的示例性TeffIL-2 Fc VitoKine。IL-2RαSushi L42G变体被选择为优选D3结构域，因为IL-2RαSushiL42G变体在VitoKine中保留的野生型遮蔽效应，并且由于IL-2RαSushi L42G变体与IL-2的结合减弱，在蛋白水解后更容易扩散离开以实现完全活化。当期望较高的IL-2 VitoKine固有基础活性时，可以采用其他具有降低的遮蔽能力的IL-2RαSushi突变，以实现期望的抗肿瘤功效和不希望的全身性毒性之间的最佳平衡。
实施例18
抗体VitoKine的构建、表达和纯化
重组抗体-细胞因子融合蛋白(免疫细胞因子)的使用有望通过将细胞因子靶向疾病部位来提高细胞因子的治疗指数。然而，将完全活性的细胞因子与抗体融合可以导致外周活化和缺乏肿瘤靶向性。VitoKine在意图治疗部位处活化之前的活性惰性使抗体VitoKine成为一种新的创新形式的免疫细胞因子。除了肿瘤靶向抗体之外，绕过肿瘤微环境中的免疫抑制效应的免疫检查点阻断抗体或增强现有应答的免疫刺激抗体也可用于构建抗体VitoKine，这可导致免疫系统抗肿瘤活性的进一步增强。此外，靶向炎性问题部位的抗体VitoKine可用于治疗抗自身免疫和慢性炎性紊乱。
遵循这一概念，构建了包含作为D2结构域的野生型或变体IL-15或者野生型或变体IL-2的抗体VitoKine蛋白。示例性抗体包括各种PD-1拮抗剂抗体(包括各种人类/人源化PD-1拮抗剂抗体(SEQ ID NO:195-198和275-278))、PD-L1阻断抗体阿特珠单抗(SEQ IDNO:279-280)、抗CTLA4抗体伊匹单抗、激动性CD40抗体RO7009789、肿瘤抗原靶向抗体(包括针对纤连蛋白胞外域的L19、针对CD20的利妥昔单抗、针对Her-2的赫赛汀、针对EGFR的西妥昔单抗)、用于肿瘤靶向和保留的抗FAP抗体(SEQ ID NO:193-194)以及抗炎抗体(针对整联蛋白α4β7的维多珠单抗和针对TNFα的修美乐)。示例性抗体VitoKine的序列以SEQ ID NO:128-146、180-181、281-286、296-297和303-306列出。
基因合成、表达载体构建以及蛋白产生、纯化和表征遵循实施例1中详述的相同程序进行。示例性的IL-15和IL-2抗体VitoKine显示出与它们的对应物Fc VitoKine相似的表达谱，诸如产率和聚集倾向。
实施例19
IL-15抗体VitoKine的离体功能生物活性评估
示例性抗PDL1抗体IL-15 VitoKine P-0485(SEQ ID NO:180和181)的生物活性通过在用IL-15 VitoKine化合物处理人类PBMC后测量CD8+T细胞(图37A)和NK细胞(图37B)中的Ki67表达来测试。P-0485与其Fc VitoKine对应物P-0315共有相同的L1接头和L2接头以及D2结构域和D3结构域。P-0485表现为具有稍高的效力，这可能是由PD-L1阻断引起的淋巴细胞活化所致。
如实施例11展示的，IL-15 Fc VitoKine的固有基础活性可以通过掺入不同效力的IL-15部分来调节。同样，IL-15PD-1拮抗剂抗体VitoKine P-0875(SEQ ID NO:196和284)被构建为具有作为D2结构域的IL-15V63A/I68H变体。通过在处理人类PBMC后测量CD8+T细胞的Ki67表达来测试P-0875，与其IL-15/IL-15RαSushi抗体融合对应物P-0870(SEQ IDNO:196、297和5)和IL-15/IL-15RαSushi Fc融合对应物P-0773进行比较(图38B)。P-0773和P-0870在诱导Ki67表达方面同样有效，EC50为18.5nM，表明IL-15活性不受融合形式的影响。本发明的汇总数据表明，IL-15VitoKine平台通常产生～1000倍的活性减弱，P-0875诱导CD8 T细胞的EC50因此估计为18μM。这样的低效力的预测与P-0875甚至在最高测试浓度1μM没有活性一致(图38B)；然而，P-0875的效力不能用数据可靠地外推。然后在与人类PBMC相似制备的食蟹猴PBMC中进一步测试了P-0875和P-0773。两种化合物都显示出与人类细胞相比成比例增强的生物活性，并且在测试浓度范围内获得了活性曲线，以可靠地推断EC50值(图38C)。P-0773和P-0875诱导食蟹猴CD8+T细胞的Ki67表达，EC50分别为0.259nM和254nM。1000倍的效力降低是IL-15 VitoKine平台的特征，其对于基于典型的IL-15 VitoKine化合物的IL-15 Fc VitoKine P-0315对比P-0313一致地展示出(在诱导人类CD8+T细胞的Ki67表达方面，对于P-0313，EC50为18.6pM，并且对于P-0315，EC50为16.9pM，在图38A中图示出)。
总之，IL-15抗体VitoKine保留了该平台特征性的细胞因子效力减弱。此外，下调IL-15部分的效力可以使对应的Vitokine的固有基础活性最小化。
实施例20
Teff IL-2抗体VitoKine的离体功能生物活性评估
构建了示例性Teff IL-2抗体VitoKine，并评估其在每种IL-2突变背景下的D3结构域遮蔽效应。所有四种示例性IL-2抗体VitoKine P-0800、P-0830、P-0831和P-0802包含作为D1结构域的抗小鼠PD1抗体(SEQ ID NO:299和302)和作为D3结构域的IL-2RαSushiL42G变体。单价D2结构域(包括P-0800中的IL-2 P65R、P-0830中的IL-2 P65N、P-0831中的IL-2P65Q和P-0802中的IL-2野生型等效物)经由不可裂解的(G4S)3接头(SEQ ID NO:112)融合到异源二聚体HC链(SEQ ID NO:301)的C末端，并用MMP-2/9可裂解的接头(SEQ ID NO:94)连接到D3结构域的N末端。四种IL-2抗体VitoKine中的每一种还含有SEQ ID NO:300和302中列出的两种另外的多肽。四种VitoKine构建体中每一种的IL-2部分保留了完全的Teff效力，但与IL-2Rα具有不同水平的结合强度。如实施例16的表14所示，P65R突变消除了与IL-2Rα的结合，而P65N和P65Q分别使结合强度降低了8.6倍和43倍。
对四种示例性IL-2抗体VitoKine评估了它们在新鲜的人类PBMC中在诱导CD8+T细胞(图39A)和NK细胞(图39B)上剂量依赖性Ki67表达方面各自的效力。包含抗小鼠PD1抗体的P-0782与在异源二聚体重链的C末端融合的单价IL-2 P65R融合，被包括在内用于比较。P-0782、P-0800和P-0802分别是P-0704、P-0808和P-0755的抗体融合对应物。图34、图35和图39中所图示的确证数据清楚地展示出，D1结构域的形式，无论是作为Fc还是本文例示的抗体，都不影响D3结构域遮蔽细胞因子效力的效率。当D2结构域为野生型IL-2时，基于IL-2Rα的D3结构域致使～3-log活性减弱；当D2结构域为具有与IL-2Rα消除的结合的IL-2变体时，D3结构域仅贡献10至20倍的遮蔽效应。
有趣的是，当与IL-2Rα结合强度中等降低的IL-2变体(由P65N和P65Q例示的)被用作D2结构域时，D3结构域致使类似的遮蔽效率，因为它有利于野生型IL-2。在图39A中图示了在CD8+T细胞上Ki67表达的剂量依赖性诱导的数据，并且在图39B中图示了在NK细胞上Ki67表达的剂量依赖性诱导的数据。在表17中进一步总结了每种化合物诱导在NK细胞上Ki67表达的EC50值。
表17
各种IL-2抗体VitoKine构建体的活性比较
Figure BDA0003806684520001161
假设D2结构域和D3结构域之间的结合亲和力阈值和结合界面的最佳空间排列在决定Teff IL-2 VitoKine的遮蔽效率中发挥作用。由于IL-2 P65Q变体与IL-2Rα具有显著降低的结合强度，但仍可以被IL-2RαSushi L42G有效地遮蔽，以保持VitoKine的惰性。预计在体内蛋白酶裂解并完全恢复生物活性后，与野生型IL-2相比，IL-2 P65Q在刺激Treg细胞方面表现出明显受损的能力(数据未示出)。因此，IL-2 P65Q被选择为构建体Teff IL-2Vitokine的优选D2结构域。然而，可以采用其他突变的IL-2变体以在期望的抗肿瘤功效和不希望的全身性毒性之间实现最佳平衡。
通过改变细胞因子化合价和接头组合，构建了具有作为D1结构域的人类PD-1拮抗剂抗体、作为D2结构域的IL-2 P65Q变体和作为D3结构域的IL-2RαSushi L42G变体的各种IL-2抗体VitoKine。表18列出了示例性IL-2 PD-1抗体Vitokine。
表18
具有作为D2结构域的IL-2 P65Q和作为D3结构域的IL-2RαSushi L42G的示例性IL-2 PD-1抗体VitoKine
Figure BDA0003806684520001162
进一步评估了示例性IL-2 PD-1抗体VitoKine P-0872和P-0929在整体(bulk)D1结构域的背景下的蛋白酶裂解效率。P-0872含有单价IL-2部分和连接D2结构域和D3结构域的单个MMP-2/9可裂解接头(SEQ ID NO:94)。遵循实施例15中详述的方案用MMP-2蛋白酶消化P-0872。经消化的样品然后用蛋白A以结合洗脱模式纯化，并且洗脱的样品以还原性SDS-PAGE凝胶进行分析，并且其生物功能在离体功能测定中进行评估。
如图40A描绘的，P-0872的D3结构域被有效且完全地裂解，产生图2B中图示的活性形式2(P-0972-活性形式)，并且导致活性的完全恢复，如在诱导新鲜的人类PBMC中CD8+T细胞上的剂量依赖性Ki67表达方面与非VitoKine IL-2 PD-1抗体融合对应物P-0879(SEQ IDNO:285和296)相当的效力所例示的(图40B)。
另一种代表性IL-2抗体VitoKine P-0929含有二价IL-2部分和两个蛋白酶可裂解接头，包括连接D1结构域和D3结构域的MMP-2/9可裂解接头和连接D2结构域和D3结构域的MMP-14可裂解接头(SEQ ID NO:298)。遵循用于MMP-2消化的类似方案用MMP-14蛋白酶裂解P-0929。经消化的样品使用蛋白A纯化，并且流通和洗脱的样品以还原性SDS-PAGE凝胶进行分析。
图41A中图示的SDS-PAGE凝胶图展示出MMP-14蛋白酶可以识别并有效地裂解MMP-2/9和MMP-14底物肽两者，并且产生活性形式1和活性形式3，而不存在活性形式2。该观察结果与MMP底物对MMP家族的一个成员具有低特异性的事实相一致。样品中裂解的D3结构域的存在是由于纯化方案引起的，并不表明D3结构域在裂解后没有扩散。包含活性形式1和活性形式3的蛋白A流通样品(P-0929-活性形式)然后在人类PBMC中进行分析。如图41B描绘的，活化的P-0929比单价非VitoKine IL-2PD-1抗体融合对应物P-0879甚至更有效地诱导CD8+T细胞上的剂量依赖性Ki67表达。总之，与IL-2Rα结合受损的Teff IL-2的生物活性可以有效地在IL-2抗体VitoKine形式中遮蔽，并易于通过蛋白水解恢复。在两个蛋白酶可裂解接头的情况下，可以进一步优化两个可裂解接头的序列和选择，以适应不同的疾病适应症和/或阶段。
实施例21
对IL-15变体抗体VitoKine的体内评估
具有作为D2结构域的具有减弱的生物活性的IL-15变体的IL-15VitoKine的目标是调节VitoKine的固有基础活性，以便进一步使全身性中靶毒性和不希望的抗原吸收最小化，以提高生物利用度和增强治疗窗口。为了评估这一假设，体内测试IL-15抗体VitoKineP-0869。P-0869包含作为D1结构域的替代小鼠PD-1抗体(SEQ ID NO:299和302)、作为D2结构域的IL-15 V63A/I68H变体(SEQ ID NO:213)、不可裂解的L1接头和MMP-2/9可裂解的L2接头(SEQ ID NO:95)以及作为D3结构域的IL-15RαSushi(SEQ ID NO:5)。如实施例11和表13展示出的，IL-15氨基酸取代V63A/I68H在离体诱导CD8+T细胞上Ki67表达方面产生约2个对数的效力降低。此外，P-0869的人类PD-1抗体对应物P-0875在新鲜的人类PMBC中在最高测试浓度1μM未显示出可检测到的生物活性(图38B)，表明Vitokine的固有基础活性显著降低。
将P-0869以及作为阴性对照的媒介物(PBS)以1mg/kg、3mg/kg和5mg/kg的剂量单次i.p.注射到健康的BALB/c小鼠(8-10周龄，n＝6只/组)中来提供。IL-15 V63A/I68H变体/IL-15RαSushi Fc融合蛋白P-0773(SEQ ID NO:227和5)被包括在内作为阳性对照，并且以0.5mg/kg的单次i.p.注射给药。在给药前(第-1天)或给药后第3天、第5天和第7天采集血液样品用于免疫表型分析。基于IL-15的VitoKine平台，预计P-0869即使在非常高的给药水平也显示出极小的全身性活化和靶向淋巴细胞群体的扩增，因为基础活性显著降低。
P-0869在各种小鼠同系(syngeneic)模型(包括小鼠CT26肺转移模型、已建立的皮下CT26肿瘤模型和已建立的皮下MC38鼠结肠癌模型)中进一步测试。实验程序与实施例9和实施例10中描述的相似。预测具有减弱的D2结构域的IL-15抗体VitoKine显示出肿瘤生长抑制和在高剂量情况下极小的全身性细胞因子活化。VitoKine在意图治疗部位处活化之前的活性惰性使抗体VitoKine成为一种新型且创新形式的免疫细胞因子。通过下调D2结构域的效力来降低VitoKine的基础活性进一步有利于在细胞因子和靶向抗体之间建立化学计量平衡，以实现最佳给药。
实施例22
对Teff IL2抗体VitoKine的体内评估
Teff IL2抗体VitoKine P-0831与实施例21中描述相似地在体内评估。P-0831包含作为D1结构域的抗小鼠PD1抗体(SEQ ID NO:300、301和302)、作为D2结构域的IL-2P65Q/S125I变体(SEQ ID NO:240)和作为D3结构域的IL-2RαSushi L42G变体(SEQ ID NO:268)。P-0831还含有不可裂解的L1接头(SEQ ID NO:112)和15个氨基酸的MMP2/9可裂解接头(SEQ ID NO:94)。单体IL-2部分保留了完全的Teff效力，但与IL-2Rα的结合强度显著降低(43倍)，这预期会降低不希望的对Treg亚群的刺激。如实施例20描述的，P-0831中的D3结构域有效地遮蔽IL-2活性，并导致接近1000倍的效力降低(图39)。
P-0831在各种小鼠同系模型(包括小鼠CT26肺转移模型、已建立的皮下CT26肿瘤模型和已建立的皮下MC38鼠结肠癌模型)中进一步测试。实验程序与实施例9和实施例10中描述的相似。
几种IL-2抗体VitoKine P-0922A、P-0928A、P-0929A及它们的不可裂解的对应物P-0877在已建立的皮下MC38鼠结肠癌模型中进一步测试。所有四种VitoKine都包含作为D2结构域的IL-2 P65Q/S125I变体(SEQ ID NO:240)和作为D3结构域的IL-2RαSushi L42G变体(SEQ ID NO:268)。P-0922A和P-0929A含有作为D1结构域的具有同源二聚体重链(SEQ IDNO:299和302)的替代小鼠PD1拮抗剂抗体，而P-0928A和P-0877含有作为D1结构域的具有异源二聚体重链(SEQ ID NO:300、301和302)的替代小鼠PD1拮抗剂抗体。P-0922A中的L1接头和L2接头分别是不可裂解的(G4S)3(SEQ ID NO:112)和可裂解的(SEQ ID NO:94)。P-0928A中的L1接头和L2接头两者都是可裂解的，分别为SEQ ID NO:298和SEQ ID NO:94。P-0929A中的L1接头和L2接头两者都是可裂解的，分别为SEQ ID NO:94和SEQ ID NO:298。P-0877含有两个不可裂解的(G4S)3(SEQ ID NO:112)接头。简而言之，将5×105个MC38鼠结肠直肠癌细胞皮下植入7周龄雌性C57BL/6小鼠中。当肿瘤达到50-80mm3肿瘤体积时，开始用媒介物或VitoKine腹膜内给药。7只小鼠/组，并且每周两次监测肿瘤体积。图46示出了处理后第7天各组的个体小鼠的肿瘤体积。与媒介物相比，可裂解的IL-2抗体VitoKine P-0922A、P-0928A和P-0929A都展示出明显且相似的肿瘤生长抑制(TGI为75％-80％)，而它们的不可裂解的对应物P-0877展示出相对较弱但仍显著的抑制肿瘤生长的作用，TGI为47％，这可能是由于PD-1拮抗剂活性引起的。可裂解的VitoKine展示出显著优于P-0877的抗肿瘤功效的事实表明，在肿瘤或肿瘤微环境中或周围，可裂解的IL-2抗体VitoKine中的一个或两个接头的蛋白水解裂解以及随后释放活性形式的IL-2可能有助于可裂解的VitoKine优于不可裂解的对应物的体内功效。
VitoKine在意图治疗部位处活化之前的活性惰性使抗体VitoKine成为一种新型且创新形式的免疫细胞因子，并进一步有利于在细胞因子和靶向抗体之间建立化学计量平衡，以实现最佳给药。
P-0831在各种小鼠同系模型(包括小鼠CT26肺转移模型、已建立的皮下CT26肿瘤模型和已建立的皮下MC38鼠结肠癌模型)中进一步测试。实验程序与实施例9和实施例10中描述的相似。预测IL-2抗体VitoKine显示出肿瘤生长抑制和在高剂量情况下极小的全身性细胞因子活化。VitoKine在意图治疗部位处活化之前的活性惰性使抗体VitoKine成为一种新型且创新形式的免疫细胞因子，并进一步有利于在细胞因子和靶向抗体之间建立化学计量平衡，以实现最佳给药。
实施例23
基于IL-2Rβ的封闭肽产生蛋白酶可活化的惰性IL-15融合蛋白或IL-2融合蛋白
产生蛋白酶可活化的惰性IL-15融合蛋白或IL-2融合蛋白的不同方法是通过可裂解接头将封闭肽(例如基于IL-2Rβ的封闭肽)遗传融合到IL-15或IL-2。所探索的封闭肽基于包含与IL-15和IL-2直接接触的关键残基的两个IL-2Rβ环(SEQ ID NO:97和98)。表19中列出的肽基于这两个环的序列。
表19
Figure BDA0003806684520001211
合成表19中的五种肽L01至L05(SEQ ID NO:97-101)，并以ELISA形式评估它们与IL-15的结合。简而言之，将IL-15/IL-15RαSushi+Fc融合蛋白P-0153(SEQ ID NO:44和46)以1μg/孔的量包被在Nunc Maxisorp 96孔微孔板的孔上，并向每个孔中添加从100μM开始的3倍系列稀释的生物素化的肽。以制造商推荐的浓度添加链霉亲和素-HRP复合物，并通过TMB底物对信号进行显影。如图42中描绘的，观察到L03(SEQ ID NO:99)的特异性结合，L03是环化的环2(SEQ ID NO:98)。
采用基于环2的序列作为封闭肽，并掺入到IL-15融合蛋白中。包含通过可裂解接头和肽间隔物与IL-15融合的基于IL-2Rβ的封闭肽的融合蛋白的示例性序列(SEQ ID NO:102-106)在表19中示出，其中粗体表示基于IL-15Rβ的封闭肽，波浪下划线表示可裂解的接头，并且直下划线表示间隔肽。将IL-15αSushi+(SEQ ID NO:5)与含有封闭肽的IL-15融合蛋白共表达并形成非共价复合物。
基因合成、表达载体构建以及蛋白产生、纯化和表征遵循实施例1中详述的相同程序进行。首先将这些含有封闭肽的IL-15融合蛋白在ELISA测定中测试，以评估它们结合IL-2Rβ的能力。如图43中说明的，与P-0153相比，由于不同封闭肽的连接，结合亲和力存在中度降低。然而，评估免疫细胞(包括CD56+NK细胞或CD8+T细胞)被这些含有封闭肽的IL-15融合蛋白的活化的PBMC测定未展示出明显的活性降低(数据未示出)，表明测试的封闭肽的遮蔽效率不足。探索不同长度的封闭肽，包括IL-2Rβ的整个细胞外结构域，并且预计观察到较高的活性遮蔽效率。同样的方法可以以类似的方式应用于IL-2。
实施例24
VitoKine形式改进了融合蛋白的可开发性
本领域已知，天然存在的IL-2蛋白往往不太稳定并且易于聚集。这在我们的实验中得到了展示，野生型IL-2 Fc融合蛋白(P-0250)以低水平表达(在HEK-293F细胞中瞬时表达为约3mg/L)，具有高聚集倾向，通过图44A中描绘的SEC色谱图例示。将四种IL-2VitoKine分子P-0320、P-0382、P-0362和P-0379与P-0250进行比较。P-0320(SEQ ID NO:49)包含野生型IL-2结构域，该野生型IL-2结构域的N末端与Fc结构域融合，并且该野生型IL-2结构域的C末端与IL-2RαSushi结构域连接。连接Fc和IL-2的L1接头是包含uPA底物肽和侧翼间隔肽的可裂解的接头(SEQ ID NO:92)，并且IL-2和IL-2RαSushi之间的L2接头是柔性(GGGGS)3接头(SEQ ID NO:127)。P-0382(SEQ ID NO:51)与P-0320的差异仅在于接头序列；P-0382的L1接头是柔性(G3S)2接头(SEQ ID NO:115)，并且L2接头是MMP-2/9可裂解接头(SEQ ID NO:95)。P-0362(SEQ ID NO:53)和P-0379(SEQ ID NO:59)与P-382的差异在于单点突变。P-0362在IL-2RαSushi结构域中含有K38E突变，而P-0379在IL-2结构域中含有F42A取代。P-0250(SEQ ID NO:48)是一种IL-2 Fc融合蛋白，其IL-2使用柔性(G3S)2(SEQ ID NO:115)接头融合到Fc的C末端。
这5种分子的尺寸排阻图在图33A-图33E中示出。根据色谱图非常明显的是，所有四种IL-2 VitoKine构建体都具有优于IL-2 Fc融合蛋白的显著提高的纯度特性。P-0250含有超过25％的不期望的高分子量物质。相比之下，所有四种VitoKine分子都显示出尖锐的单体峰，具有超过96％的单体含量。接头变异、IL-2或IL-2RαSushi中的突变对质量没有明显影响。蛋白质量的这种显著提高明显归因于VitoKine中IL-2RαSushi结构域的融合。
除了蛋白质量之外，IL-2 VitoKine的表达水平也得到提高，特别是在Fc与IL-2之间具有GS接头和在IL-2与IL-2RαSushi之间具有10个氨基酸的MMP-2/9可活化接头的VitoKine形式。虽然由于细胞的生长条件，在不同批次之间蛋白表达水平可能不同，但明显的是，VitoKine的表达水平始终比IL-2 Fc融合蛋白高数倍。表20列出了蛋白表达滴度(mg/L)以及蛋白单体百分比。
表20
蛋白ID 表达滴度(mg/L) 通过SEC检测的单体
P-0250 3.1 74.3％
P-0320 9.3 96.2％
P-0382 23.3 97.8％
P-0362 18.1 100％
P-0379 16.6 99.1％
此外，本发明人对IL-2进行的工程化工作还鉴定了在位置125处用异亮氨酸取代丝氨酸的单个氨基酸取代，该取代导致了具有完全保留的生物活性的IL-2 Fc融合构建体的可开发性的普遍改进。在野生型IL-2和Fc融合形式中具有不同突变背景的IL-2变体的位置125处的异亮氨酸取代均导致表达水平的4至11倍提高，且聚集倾向一致地低。在表21中总结了通过SEC色谱术以示例性分子的聚集百分比评估的蛋白A纯化的物质的表达水平(mg/L)和纯度。表21同一行中的两种分子共有相同的其他氨基酸取代，并且差异仅在于残基125处为丝氨酸或异亮氨酸。作为实例，在图44F中进一步说明了P-250的IL-2-S125I对应分子P-0531的SEC色谱图。
表21
S125I取代改进了各种IL-2 Fc融合蛋白的可开发性谱
Figure BDA0003806684520001231
总之，VitoKine平台显著改进了蛋白的可开发性谱，这通过IL-2 Fc VitoKine构建体的蛋白表达增加和聚集倾向的大幅降低得到展示。此外，掺入有益的IL-2S125I氨基酸的IL-2(野生型或变体)VitoKine构建体可具有进一步增强的可开发性谱。
实施例25
VitoKine D3结构域的选择可以显著影响蛋白表达
还对VitoKine平台的D3结构域进行了探索，该D3结构域是D2结构域的同源受体的变体或不相关的蛋白结构域。基于晶体结构分析(Wang等人,Science 310:1159-1163,2005)，IL-2Rαsushi结构域1和2参与链交换事件，并且结果是IL-2Rα的残基1-19是sushi结构域2的一部分，并且残基102-122是sushi结构域1的一部分。在IL-2RαSushi变体(SEQ IDNO:147)中反映出这样的结构排列，该IL-2RαSushi变体在N末端含有IL-2Rα(SEQ ID NO:10)残基102-122并且在C末端含有IL-2Rα残基20-68。这样的IL-2RαSushi变体含有大部分与IL-2相互作用的残基，并且被推测以假定的结构完整性重现了大部分活性。用IL-2RαSushi变体替代P-0320(SEQ ID NO:49)中的IL-2RαSushi结构域产生了IL-2 VitoKine P-0321(SEQ ID NO:179)。出乎意料的是，包含IL-2RαSushi变体作为D3结构域的P-0321根本不表达或表达水平如此之低以至于没有物质可以被捕获和纯化。
类似地，用IL-2RαSushi(SEQ ID NO:10)替代VitoKine P-0315(SEQ ID NO:33)中的IL-15αSushi+结构域并且得到的蛋白是P-0389(SEQ ID NO:42)。与P-0315相比，P-0389以显著更低的水平表达。更值得注意的是，纯化的P-0389主要是高分子量聚集体，如图45A中描绘的SDS-PAGE凝胶图中展示的。为了比较的目的，对应分子P-0315的SDS-PAGE凝胶图以图45B示出。此外，尽管序列中存在MMP-2/9底物肽，但纯化的P-0389对MMP-2消化有抗性，表明分子未正确折叠，或者聚集限制了蛋白酶的接近。
总之，D3是VitoKine构建体的关键组分。除了作为遮蔽部分起作用之外，D3还能正面和负面地显著影响蛋白的可开发性谱。
本文公开的和要求保护的所有物品和方法鉴于本公开内容均可被制备和执行而无需过度实验。尽管已根据优选的实施方案描述本公开内容的物品和方法，但对于本领域技术人员将明显是，变化形式可被应用到所述物品和方法而不偏离本公开内容的精神和范围。对于本领域技术人员明显的是，所有这样的变化和等效物，无论是现存的或以后开发的，都被认为是在由所附权利要求限定的本公开内容的精神和范围之内。说明书中提及的所有专利、专利申请和出版物指示本公开内容所属领域的普通技术人员的水平。为了所有目的，所有专利、专利申请和出版物通过引用以其整体并入本文，并且其程度如同每个单独出版物单独地且具体地被指示为了任何和所有目的通过引用以其整体并入本文。本文例证地表述的公开内容可在本文未具体地公开的任何一个或更多个要素不存在的条件下被合适地实践。因此，应理解，尽管本公开内容已通过优选的实施方案和任选的特征被具体地公开，但是本领域技术人员可寻求本文公开的概念的改变和变化形式，并且认为此类改变和变化形式在由所附的权利要求限定的本公开内容的范围内。
序列表
在所附的序列表中列出的核酸序列和氨基酸序列按37C.F.R.1.822中规定的使用核苷酸碱基的标准字母缩写和氨基酸的单字母编码示出。
SEQ ID NO:1是人类IL-15前体氨基酸序列。
SEQ ID NO:2是人类IL-15成熟形式氨基酸序列。
SEQ ID NO:3是IL-15变体多肽的氨基酸序列。
SEQ ID NO:4是人类IL-15Rα氨基酸序列。
SEQ ID NO:5是人类IL-15Rα,sushi结构域+氨基酸序列。
SEQ ID NO:6是人类IL-2前体氨基酸序列。
SEQ ID NO:7是人类IL-2成熟形式天然存在的氨基酸序列。
SEQ ID NO:8是人类IL-2成熟形式野生型氨基酸序列。
SEQ ID NO:9是人类IL-2Rα(CD25)前体氨基酸序列。
SEQ ID NO:10是人类IL-2Rα、sushi结构域氨基酸序列。
SEQ ID NO:11是人类IL-2R S125I氨基酸序列。
SEQ ID NO:12是人类IL-2Rβ细胞外结构域氨基酸序列。
SEQ ID NO:13是人类IgG1-Fc氨基酸序列。
SEQ ID NO:14是具有减少/消除的效应子功能的人类IgG1-Fc的序列。
SEQ ID NO:15是Knob-Fc氨基酸序列。
SEQ ID NO:16是Hole-Fc氨基酸序列。
SEQ ID NO:17是人类IL-4成熟形式氨基酸序列。
SEQ ID NO:18是人类IL-7成熟形式氨基酸序列。
SEQ ID NO:19是人类IL-9成熟形式氨基酸序列。
SEQ ID NO:20是人类IL-10成熟形式氨基酸序列。
SEQ ID NO:21是人类IL-12亚基α成熟形式序列。
SEQ ID NO:22是人类IL-12亚基β成熟形式序列。
SEQ ID NO:23是人类IL-23亚基α成熟形式序列。
SEQ ID NO:24是人类IL-27亚基β成熟形式序列。
SEQ ID NO:25-43是各种IL-15 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:44是Hole-Fc-IL-15融合蛋白的氨基酸序列。
SEQ ID NO:45是Knob-Fc-IL-15融合蛋白的氨基酸序列。
SEQ ID NO:46是Knob-Fc-IL-15Rα-Sushi+融合蛋白的氨基酸序列。
SEQ ID NO:47是Fc-IL-15S58D融合蛋白的氨基酸序列。
SEQ ID NO:48是IL-2融合蛋白的氨基酸序列。
SEQ ID NO:49-65是各种IL-2 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:66-70是各种包含封闭肽的IL-15构建体的氨基酸序列。
SEQ ID NO:71-87和157-159是各种蛋白酶底物肽的氨基酸序列。
SEQ ID NO:88-96、160-161和298是各种蛋白酶可裂解接头的氨基酸序列，所述蛋白酶可裂解接头包含位于蛋白酶底物肽侧翼的各种间隔肽。
SEQ ID NO:97-106是各种封闭肽序列的氨基酸序列。
SEQ ID NO:107-127是各种不可裂解的接头序列的氨基酸序列。
SEQ ID NO:128-146是各种抗体VitoKine构建体的氨基酸序列。
SEQ ID NO:147是人类IL-2Rα变体序列。
SEQ ID NO:148-149是Hole-Fc-IL-15融合构建体的氨基酸序列。
SEQ ID NO:150-155是各种IL-2 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:156是具有减少/消除的效应子功能和延长的半衰期的人类IgG1-Fc序列。
SEQ ID NO:162-165是各种IL-15 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:166是具有减少/消除的效应子功能和延长的半衰期的人类IgG1-Fc的序列。
SEQ ID NO:167是具有延长的半衰期的Knob-Fc的氨基酸序列。
SEQ ID NO:168是具有延长的半衰期的Hole-Fc的氨基酸序列。
SEQ ID NO:169-174是各种IL-15 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:175-178是各种IL-15 Fc融合构建体的氨基酸序列。
SEQ ID NO:179是IL-2 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:180-181是抗体IL-15 VitoKine构建体的氨基酸序列。
SEQ ID NO:182-192是包含氨基酸缺失、插入和/或取代的各种IL-15变体多肽的氨基酸序列。
SEQ ID NO:193-194是人源化抗FAP抗体的重链和轻链的氨基酸序列。
SEQ ID NO:195-196是人源化PD-1拮抗剂抗体的重链和轻链的氨基酸序列。
SEQ ID NO:197-198是人类PD-1拮抗剂抗体的重链和轻链的氨基酸序列。
SEQ ID NO:199-215是各种IL-15变体多肽的氨基酸序列。
SEQ ID NO:216-229是各种IL-15变体Fc融合构建体的氨基酸序列。
SEQ ID NO:230-231是各种IL-15 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:232-247是各种IL-2变体多肽的氨基酸序列。
SEQ ID NO:248-266是各种IL-2变体Fc融合构建体的氨基酸序列。
SEQ ID NO:267-270是各种人类IL-2Rαsushi结构域变体多肽的氨基酸序列。
SEQ ID NO:271-274和292-295是各种IL-2 Fc VitoKine构建体的氨基酸序列。
SEQ ID NO:275-276是PD-1拮抗剂抗体的重链和轻链的氨基酸序列。
SEQ ID NO:277-278是PD-1拮抗剂抗体的重链和轻链的氨基酸序列。
SEQ ID NO:279-280是PD-L1拮抗剂抗体的重链和轻链的氨基酸序列。
SEQ ID NO:281-291是各种IL-15和IL-2抗体VitoKine构建体的氨基酸序列。
SEQ ID NO:296是与IL-2变体融合的PD-1拮抗剂抗体knob HC链的氨基酸序列。
SEQ ID NO:297是与IL-15变体融合的PD-1拮抗剂抗体HC的氨基酸序列。
SEQ ID NO:299-302是具有同源二聚体或异源二聚体重链的替代抗小鼠PD-1抗体的氨基酸序列。
SEQ ID NO:303-306是各种IL-15抗体VitoKine构建体的氨基酸序列。
序列表
人类IL-15前体序列
MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:1)
人类IL-15成熟形式序列
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:2)
人类IL-15 S58D突变蛋白
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:3)
人类IL-15Rα前体序列
MAPRRARGCRTLGLPALLLLLLLRPPATRGITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTAGVTPQPESLSPSGKEPAASSPSSNNTAATTAAIVPGSQLMPSKSPSTGTTEISSHESSHGTPSQTTAKNWELTASASHQPPGVYPQGHSDTTVAISTSTVLLCGLSAVSLLACYLKSRQTPPLASVEMEAMEALPVTWGTSSRDEDLENCSHHL(SEQ ID NO:4)
人类IL-15Rα,结构域+
ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:5)
人类IL-2前体序列
MYRMQLLSCIALSLALVTNSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT(SEQ ID NO:6)
人类IL-2成熟形式天然存在的序列
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT(SEQ ID NO:7)
人类IL-2成熟形式野生型序列
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLT(SEQ ID NO:8)
人类IL-2Rα(CD25)前体序列
MDSYLLMWGLLTFIMVPGCQAELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGEMETSQFPGEEKPQASPEGRPESETSCLVTTTDFQIQTEMAATMETSIFTTEYQVAVAGCVFLLISVLLLSGLTWQRRQRKSRRTI(SEQ ID NO:9)
人类IL-2RαSushi
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:10)
人类IL-2S125I序列
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:11)
人类IL-2Rβ细胞外结构域序列
AVNGTSQFTCFYNSRANISCVWSQDGALQDTSCQVHAWPDRRRWNQTCELLPVSQASWACNLILGAPDSQKLTTVDIVTLRVLCREGVRWRVMAIQDFKPFENLRLMAPISLQVVHVETHRCNISWEISQASHYFERHLEFEARTLSPGHTWEEAPLLTLKQKQEWICLETLTPDTQYEFQVRVKPLQGEFTTWSPWSQPLAFRTKPAALGKDT(SEQ IDNO:12)
人类IgG1-Fc
DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:13)
具有减少/消除的效应子功能的人类IgG1-Fc
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:14)
具有减少/消除的效应子功能的人类IgG1 Knob-Fc
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:15)
具有减少/消除的效应子功能的人类IgG1 Hole-Fc
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:16)
人类IL-4成熟形式序列
HKCDITLQEIIKTLNSLTEQKTLCTELTVTDIFAASKNTTEKETFCRAATVLRQFYSHHEKDTRCLGATAQQFHRHKQLIRFLKRLDRNLWGLAGLNSCPVKEANQSTLENFLERLKTIMREKYSKCSS(SEQ ID NO:17)
人类IL-7成熟形式序列
DCDIEGKDGKQYESVLMVSIDQLLDSMKEIGSNCLNNEFNFFKRHICDANKEGMFLFRAARKLRQFLKMNSTGDFDLHLLKVSEGTTILLNCTGQVKGRKPAALGEAQPTKSLEENKSLKEQKKLNDLCFLKRLLQEIKTCWNKILMGTKEH(SEQ ID NO:18)
人类IL-9成熟形式序列
QGCPTLAGILDINFLINKMQEDPASKCHCSANVTSCLCLGIPSDNCTRPCFSERLSQMTNTTMQTRYPLIFSRVKKSVEVLKNNKCPYFSCEQPCNQTTAGNALTFLKSLLEIFQKEKMRGMRGKI(SEQ ID NO:19)
人类IL-10成熟形式序列
SPGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN(SEQ ID NO:20)
人类IL-12亚基α成熟形式序列
RNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSYLNAS(SEQ ID NO:21)
人类IL-12亚基β成熟形式序列
IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRAQDRYYSSSWSEWASVPCS(SEQ ID NO:22)
人类IL-23亚基α成熟形式序列
RAVPGGSSPAWTQCQQLSQKLCTLAWSAHPLVGHMDLREEGDEETTNDVPHIQCGDGCDPQGLRDNSQFCLQRIHQGLIFYEKLLGSDIFTGEPSLLPDSPVGQLHASLLGLSQLLQPEGHHWETQQIPSLSPSQPWQRLLLRFKILRSLQAFVAVAARVFAHGAATLSP(SEQ ID NO:23)
人类TGFβ成熟形式序列
ALDTNYCFSSTEKNCCVRQLYIDFRKDLGWKWIHEPKGYHANFCLGPCPYIWSLDTQYSKVLALYNQHNPGASAAPCCVPQALEPLPIVYYVGRKPKVEQLSNMIVRSCKCS(SEQ ID NO:24)
P-0351(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:25)
P-0170(IL-15 Fc VitoKine)Hole链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGLGGSGRSANAILENWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:26)
P-0172(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGLGGSGRSANAILENWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:27)
P-0202(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSLSGRSDNHGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:28)
P-0203(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGSLGGSGRSANAILEGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:29)
P-0204(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSLGGSGRSANAILEGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:30)
P-0205(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSLGGSGRSANAILEGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:31)
P-0206(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSLGGSGRSANAILEGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:32)
P-0315(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:33)
P-0316(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGPLGMLSQGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:34)
P-0350(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:35)
P-0354(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGSGPLGMLSQGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGSGRSANAIITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:36)
P-0355(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGSGRSANAIITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:37)
P-0385(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGPLGMLSQITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:38)
P-0386(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYISNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKSIRDPALVHQRPAPP(SEQ ID NO:39)
P-0387(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSREEYICNSGFKEKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:40)
P-0388(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSDCGLPPDVPNAQPALEGRTSFPEDTVITYKCEESFVKIPGEKDSVICLKGSQWSDIEEFCNR(SEQ ID NO:41)
P-0389(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ IDNO:42)
P-0397(IL-15 Fc VitoKine)
ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPGGPLGMLSQSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSLGGSGRSANAILEGGSCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQID NO:43)
Hole-Fc-IL-15
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:44)
Knob-Fc-IL-15
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:45)
Knob-Fc-IL-15Rα-Sushi+
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:46)
Fc-IL-15S58D
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:47)
P-0250(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLT(SEQID NO:48)
P-0320(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGGGSGGGGSGGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:49)
P-0352(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGGGSGGGGSGGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:50)
P-0382(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:51)
P-0398(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:52)
P-0362(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:53)
P-0380(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLAMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:54)
P-0384(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLAMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:55)
P-0400(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLNDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:56)
P-0404(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLELQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:57)
P-0399(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:58)
P-0379(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:59)
P-0381(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTEMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:60)
P-0383(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTEMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:61)
P-0329(IL-2 Fc VitoKine)
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGGGSLGGSGRSANAILEGGSCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:62)
P-0401(IL-2 Fc VitoKine)
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGGGPLGMLSQSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGGGSGGGGSGGGGSGGGGSCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:63)
P-0402(IL-2 Fc VitoKine)
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGGGSGPLGMLSQGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGGGSGGGGSGGGGSGGGGSCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:64)
P-0403(IL-2 Fc VitoKine)
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGGGPLGMLSQSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTAEAAAKEAAAKEAAAKACPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:65)
Hole-Fc-15p1
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGSLGGSGRSANAILEGGGSGGGSGGGSIYNCEISQASHYFERHLCYSI(SEQ ID NO:66)
Hole-Fc-15p2
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGSLGGSGRSANAILEGGGSGGGSGGGSIYNCELHREFYHSAQSIEWCYSI(SEQ ID NO:67)
Hole-Fc-15p3
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGSLGGSGRSANAILEGGGSGGGSGGGSETHRCNISWEISQASHYFERHLEFEARTLCPGH(SEQ ID NO:68)
p1’-15-Fc
QGQSGQCEISQASHYFERHLCYSIGSSGGSGGSGGSGLSGRSDNHGSSGTNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:69)
p3’-15-Fc
QGQSGQCNISWEISQASHYFERHLEFEARTLCPGHGSSGGSGGSGGSGLSGRSDNHGSSGTNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:70)
蛋白酶底物肽序列
SPLGLAGS(SEQ ID NO:71)
蛋白酶底物肽序列
EPLELRAG(SEQ ID NO:72)
蛋白酶底物肽序列
LSGRSDNH(SEQ ID NO:73)
蛋白酶底物肽序列
GPLGIAGQ(SEQ ID NO:74)
蛋白酶底物肽序列
GTAHLMGG(SEQ ID NO:75)
蛋白酶底物肽序列
RIGSLRTA(SEQ ID NO:76)
蛋白酶底物肽序列
GPLGMLSQ(SEQ ID NO:77)
蛋白酶底物肽序列
RPSASRSA(SEQ ID NO:78)
蛋白酶底物肽序列
PLGLAG(SEQ ID NO:79)
蛋白酶底物肽序列
LGGSGRSANAILE(SEQ ID NO:80)
蛋白酶底物肽序列
GGSGRSANAI(SEQ ID NO:81)
蛋白酶底物肽序列
SGRSA(SEQ ID NO:82)
蛋白酶底物肽序列
AANL(SEQ ID NO:83)
蛋白酶底物肽序列
GFFY(SEQ ID NO:84)
蛋白酶底物肽序列
GPICFRLG(SEQ ID NO:85)
蛋白酶底物肽序列
RQAGFSL(SEQ ID NO:86)
蛋白酶底物肽序列
HSSKLQ(SEQ ID NO:87)
蛋白酶可裂解的接头序列
GGGSGGGGSGGGGSLSGRSDNHGGSGGGGS(SEQ ID NO:88)
蛋白酶可裂解的接头序列
GSSSGRSENIRTAGT(SEQ ID NO:89)
蛋白酶可裂解的接头序列
GGGGSGGGGSGGGSLGGSGRSANAILEGGSGGGGS(SEQ ID NO:90)
蛋白酶可裂解的接头序列
GGGGSGGGGSLGGSGRSANAILEGGGGS(SEQ ID NO:91)
蛋白酶可裂解的接头序列
GGGGSLGGSGRSANAILEGGS(SEQ ID NO:92)
蛋白酶可裂解的接头序列
GGGSGPTNKVRGGS(SEQ ID NO:93)
蛋白酶可裂解的接头序列
GGSGPLGMLSQGGGS(SEQ ID NO:94)
蛋白酶可裂解的接头序列
GGPLGMLSQS(SEQ ID NO:95)
蛋白酶可裂解的接头序列
GGGPLGMLSQGGS(SEQ ID NO:96)
基于IL-2Rβ的封闭肽序列
LGAPDSQKLTTVDIV(SEQ ID NO:97)
基于IL-2Rβ的封闭肽序列
EISQASHYFERHL(SEQ ID NO:98)
基于IL-2Rβ的封闭肽序列
CEISQASHYFERHLC(SEQ ID NO:99)
基于IL-2Rβ的封闭肽序列
LGAPDSQKLTTVDIVGGGGGGGGEISQASHYFERHL(SEQ ID NO:100)
基于IL-2Rβ的封闭肽序列
KPFENLRLMAPIS(SEQ ID NO:101)
基于IL-2Rβ的封闭肽序列
GGGSLGGSGRSANAILEGGGSGGGSGGGSIYNCEISQASHYFERHLCYSI(SEQ ID NO:102)
基于IL-2Rβ的封闭肽序列
GGGSLGGSGRSANAILEGGGSGGGSGGGSIYNCELHREFYHSAQSIEWCYSI(SEQ ID NO:103)
基于IL-2Rβ的封闭肽序列
GGGSLGGSGRSANAILEGGGSGGGSGGGSETHRCNISWEISQASHYFERHLEFEARTLCPGH(SEQ IDNO:104)
基于IL-2Rβ的封闭肽序列
QGQSGQCEISQASHYFERHLCYSIGSSGGSGGSGGSGLSGRSDNHGSSGT(SEQ ID NO:105)
基于IL-2Rβ的封闭肽序列
QGQSGQCNISWEISQASHYFERHLEFEARTLCPGHGSSGGSGGSGGSGLSGRSDNHGSSGT(SEQ IDNO:106)
不可裂解的接头序列
EPKSSDKTHTSPPS(SEQ ID NO:107)
不可裂解的接头序列
GGGSGGGSGGGS(SEQ ID NO:108)
不可裂解的接头序列
GGGS(SEQ ID NO:109)
不可裂解的接头序列
GSSGGSGGS(SEQ ID NO:110)
不可裂解的接头序列
GSSGT(SEQ ID NO:111)
不可裂解的接头序列
GGGGSGGGGSGGGGS(SEQ ID NO:112)
不可裂解的接头序列
AEAAAKEAAAKEAAAKA(SEQ ID NO:113)
不可裂解的接头序列
GGGGSGGGGSGGGGSGGGGS(SEQ ID NO:114)
不可裂解的接头序列
GGGSGGGS(SEQ ID NO:115)
不可裂解的接头序列
GSGS(SEQ ID NO:116)
不可裂解的接头序列
GGS(SEQ ID NO:117)
不可裂解的接头序列
GGGGS(SEQ ID NO:118)
不可裂解的接头序列
GGSGG(SEQ ID NO:119)
不可裂解的接头序列
SGGG(SEQ ID NO:120)
不可裂解的接头序列
GSGS(SEQ ID NO:121)
不可裂解的接头序列
GSGSGS(SEQ ID NO:122)
不可裂解的接头序列
GSGSGSGS(SEQ ID NO:123)
不可裂解的接头序列
GSGSGSGSGS(SEQ ID NO:124)
不可裂解的接头序列
GSGSGSGSGSGS(SEQ ID NO:125)
不可裂解的接头序列
GGGGSGGGGS(SEQ ID NO:126)
不可裂解的接头序列
GGGGSGGGGSGGGGS(SEQ ID NO:127)
人源化PD-1拮抗剂抗体-IL-15-VitoKine-HC
QGQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQAPGQGLEWMGVIESETGGTAYNQKFKGRAKITADKSTSTAYMELSSLRSEDTAVYYCTREGITTVATTYYWYFDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:128)
人源化PD-1拮抗剂抗体-Lκ
DVVMTQSPLSLPVTLGQPASISCRSSQSIVHSNGNTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:129)
CTLA-4拮抗剂抗体-IL-15-VitoKine-HC
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEWVTFISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:130)
CTLA-4拮抗剂抗体-Lκ
EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGAFSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQID NO:131)
CD40激动剂抗体-IL-15-VitoKine-HC
QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:132)
CD40激动剂抗体-Lκ
DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:133)
人源化抗纤连蛋白抗体-IL-15-VitoKine-HC
EVQLLESGGGLVQPGGSLRLSCAASGFTFSSFSMSWVRQAPGKGLEWVSSISGSSGTTYYADSVKGRFTISRDSKNTLYLQMNSLRAEDTAVYYCAKPFPYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:134)
人源化抗纤连蛋白抗体-Lκ
EIVLTQSPGTLSLSPGERATLSCRASQSVSSSFLAWYQQKPGQAPRLLIYYASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQTGRIPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQID NO:135)
利妥昔单抗-IL-2-VitoKine-HC
QVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKAEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:136)
利妥昔单抗-Lκ
QIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:137)
赫塞汀-IL-2-VitoKine-HC
EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:138)
赫塞汀-Lκ
DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:139)
西妥昔单抗-IL-2-VitoKine-HC
QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:140)
西妥昔单抗-Lκ
DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:141)
人源化PD-1拮抗剂抗体-IL-2-VitoKine-HC
QGQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQAPGQGLEWMGVIESETGGTAYNQKFKGRAKITADKSTSTAYMELSSLRSEDTAVYYCTREGITTVATTYYWYFDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:142)
维多珠单抗-IL-2-VitoKine-HC
QVQLVQSGAEVKKPGASVKVSCKGSGYTFTSYWMHWVRQAPGQRLEWIGEIDPSESNTNYNQKFKGRVTLTVDISASTAYMELSSLRSEDTAVYYCARGGYDGWDYAIDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLTLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:143)
维多珠单抗-Lκ
DVVMTQSPLSLPVTPGEPASISCRSSQSLAKSYGNTYLSWYLQKPGQSPQLLIYGISNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCLQGTHQPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:144)
修美乐-IL-2-VitoKine-HC
EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSAITWNSGHIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKVSYLSTASSLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLTLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:145)
修美乐-Lκ
DIQMTQSPSSLSASVGDRVTITCRASQGIRNYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQRYNRAPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:146)
IL-2Rα结构域调换的Sushi
GHCREPPPWENEATERIYHFVYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRN(SEQ ID NO:147)
Hole-Fc-IL-15 2
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:148)
Hole-Fc-IL-15 3
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:149)
P-0420(IL-2 Treg-偏好性(biased)变体Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISRINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:150)
P-0421(IL-2 Treg-偏好性变体Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLTLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:151)
P-0423(IL-2 Treg-偏好性变体Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSESIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:152)
P-0424(IL-2 Treg-偏好性变体Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLNDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSESIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:153)
P-0425(IL-2 Treg-偏好性变体Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLRDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSESIISTLTGGPLGMLSQSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:154)
P-0426(IL-2 Treg-偏好性变体Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLHDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIESIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:155)
具有减少/消除的效应子功能和延长的半衰期的人类IgG1-Fc
DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:156)
蛋白酶底物肽序列
SGRSENIRTA(SEQ ID NO:157)
蛋白酶底物肽序列
GPTNKVR(SEQ ID NO:158)
蛋白酶底物肽序列
RQARAVGG(SEQ ID NO:159)
蛋白酶可裂解的接头序列
GGPTNKVRGS(SEQ ID NO:160)
蛋白酶可裂解的接头序列
GRQARAVGGS(SEQ ID NO:161)
P-0660(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGSSSGRSENIRTAGTNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:162)
P-0488(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSSGRSENIRTAITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:163)
P-0489(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPTNKVRGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:164)
P-0661(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGPTNKVRGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:165)
具有减少/消除的效应子功能和延长的体内半衰期的人类IgG1-Fc
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPG(SEQ ID NO:166)
具有延长的体内半衰期的人类IgG1 Knob-Fc
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPG(SEQ ID NO:167)
具有延长的体内半衰期的人类IgG1 Hole-Fc
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPG(SEQ ID NO:168)
P-0650(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:169)
P-0651(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:170)
P-0662(IL-15 Fc VitoKine)Hole链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:171)
具有延长的半衰期的P-0663(IL-15 Fc VitoKine)Hole链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:172)
具有延长的半衰期的P-0664(IL-15 Fc VitoKine)Hole链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:173)
具有延长的半衰期的P-0665(IL-15 Fc VitoKine)Hole链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGSSSGRSENIRTAGTNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:174)
P-0156链1
ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:175)
P-0156链2
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:176)
Benchmark链1
NWVNVISDLKKIEDLIQSMHIDATLYTESNVHPSCKVTAMKCFLLELQVISLESGDASIHDTVQDLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSEPKSSDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHEALHSHYTQKSLSLSPGK(SEQ ID NO:177)
Benchmark链2
ITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSEPKSSDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK(SEQ ID NO:178)
P-0321(IL-2 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFSQSIISTLTGGGGSGGGGSGGGGSGHCREPPPWENEATERIYHFVYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRN(SEQ ID NO:179)
PD-L1阻断抗体-IL-15-VitoKine-HC
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDADIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:180)
PD-L1阻断抗体-Lκ
DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:181)
缺失111-114的人类IL-15变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMF(SEQ ID NO:182)
缺失109-114的人类IL-15变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQ(SEQ ID NO:183)
缺失108-114的人类IL-15变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIV(SEQ ID NO:184)
N95后插入‘GS’的人类IL-15变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNGSIKEFLQSFVHIVQMFINTS(SEQ ID NO:185)
N95后插入‘GGSGG’的人类IL-15变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNGGSGGIKEFLQSFVHIVQMFINTS(SEQ ID NO:186)
在N末端处具有1个氨基酸缺失的人类IL-15变体
WVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:187)
在N末端处具有2个氨基酸缺失的人类IL-15变体
VNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:188)
在N末端处具有3个氨基酸缺失的人类IL-15变体
NVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:189)
在N末端处具有4个氨基酸缺失的人类IL-15变体
VISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:190)
在N末端处具有2个氨基酸缺失的人类IL-15 V63A变体
VNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:191)
在N末端处具有2个氨基酸缺失的人类IL-15 I68H变体
VNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:192)
人源化抗FAP抗体重链
QVQLVQSGAEVKKPGASVKVSCKASGYTFTENIIHWVRQAPGQGLEWMGWFHPGSGSIKYAQKFQGRVTMTADKSTSTVYMELSSLRSEDTAVYYCARHGGTGRGAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:193)
人源化抗FAP抗体κ轻链
DIQMTQSPSSLSASVGDRVTITCRASRSISTSAYSYMHWYQQKPGKAPKLLIYLASNLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHSRELPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:194)
人源化PD-1拮抗剂抗体重链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:195)
人源化PD-1拮抗剂抗体κ轻链
DIVMTQSPLSLPVTPGEPASITCKASQDVETVVAWYLQKPGQSPRLLIYWASTRHTGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQYSRYPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:196)
人类PD-1拮抗剂抗体重链
EVQLVQSGAEVKKPGASVKVSCKASGYRFTSYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDADYSSGSGYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:197)
人类PD-1拮抗剂抗体λ轻链
SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSADNSITYRVFGGGTKVTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS(SEQ IDNO:198)
人类IL-15 V63A变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:199)
人类IL-15 V63K变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTKENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:200)
人类IL-15 V63R变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTRENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:201)
人类IL-15 I68H变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:202)
人类IL-15 I68F变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIFLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:203)
人类IL-15 I68Q变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIQLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:204)
人类IL-15 I68G变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIGLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:205)
人类IL-15 I68K变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIKLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:206)
人类IL-15 I68D变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIDLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:207)
人类IL-15 Q108S变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVSMFINTS(SEQ ID NO:208)
人类IL-15 Q108A变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVAMFINTS(SEQ ID NO:209)
人类IL-15 Q108E变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVEMFINTS(SEQ ID NO:210)
人类IL-15 Q108K变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVKMFINTS(SEQ ID NO:211)
人类IL-15 Q108M变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVMMFINTS(SEQ ID NO:212)
人类IL-15 V63A/I68H变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:213)
人类IL-15 V63A/I68Q变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIQLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:214)
人类IL-15 V63A/I68G变体多肽
NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIGLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:215)
Fc-IL-15 V63K链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTKENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:216)
Fc-IL-15 V63R链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTRENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:217)
Fc-IL-15 V63A链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:218)
Fc-IL-15 I68H链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:219)
Fc-IL-15 I68F链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIFLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:220)
Fc-IL-15 I68Q链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIQLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:221)
Fc-IL-15 I68G链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIGLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:222)
Fc-IL-15 I68K链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIKLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:223)
Fc-IL-15 I68D链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIDLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:224)
Hole Fc-IL-15 Q108S链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVSMFINTS(SEQ ID NO:225)
Hole Fc-IL-15 Q108A链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGEPKSSDKTHTSPPSPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVAMFINTS(SEQ ID NO:226)
Fc-IL-15 V63A/I68H链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:227)
Fc-IL-15 V63A/I68Q链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIQLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:228)
Fc-IL-15 V63A/I68G链
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIGLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:229)
P-0682(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVSMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:230)
P-0806(IL-15 Fc VitoKine)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:231)
人类IL-2R38A/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:232)
人类IL-2 F42A/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:233)
人类IL-2E62A/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEALKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:234)
人类IL-2E62F/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEFLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:235)
人类IL-2E62H/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEHLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:236)
人类IL-2E62L/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEELLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:237)
人类IL-2 P65H/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKHLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:238)
人类IL-2 P65N/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKNLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:239)
人类IL-2 P65Q/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:240)
人类IL-2 P65R/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:241)
人类IL-2 P65K/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKKLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:242)
人类IL-2 F42A/E62F/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEFLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:243)
人类IL-2 F42A/E62A/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEALKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:244)
人类IL-2 F42A/E62H/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEHLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:245)
人类IL-2 F42A/P65H/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKHLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:246)
人类IL-2 F42A/P65R/S125I变体多肽
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ ID NO:247)
P-0531(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:248)
P-0689(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:249)
P-0602(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:250)
P-0613(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:251)
P-0624(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEALKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:252)
P-0625(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEFLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:253)
P-0626(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEHLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:254)
P-0627(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEELLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:255)
P-0634(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKHLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:256)
P-0708(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKNLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:257)
P-0709(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:258)
P-0635(IL-2 Fc融合体)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:259)
P-0704(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:260)
P-0707(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKKLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:261)
P-0702(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEFLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:262)
P-0766(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEALKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:263)
P-0767(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEHLKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:264)
P-0703(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKHLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:265)
P-0705(IL-2 Fc融合体knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQID NO:266)
人类IL-2RαSushi Y43A变体
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLAMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:267)
人类IL-2RαSushi L42G变体
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:268)
人类IL-2RαSushi R36A变体
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRAIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:269)
人类IL-2RαSushi K38E变体
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIESGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:270)
P-0754(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLAMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:271)
P-0755(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:272)
P-0756(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRAIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:273)
P-0745(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:274)
人源化PD-1拮抗剂抗体-HC
QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQ ID NO:275)
人源化PD-1拮抗剂抗体-Lκ
EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:276)
人类PD-1拮抗剂抗体-HC
QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQ IDNO:277)
人类PD-1拮抗剂抗体-Lκ
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:278)
人源化PD-L1拮抗剂抗体-HC
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO:279)
人源化PD-L1拮抗剂抗体-Lκ
DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ IDNO:280)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVSMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:281)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIGLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:282)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIGLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:283)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ ID NO:284)
人源化PD-1拮抗剂抗体-HC-Hole链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:285)
人源化PD-1拮抗剂抗体IL-2 VitoKine HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:286)
人源化PD-1拮抗剂抗体IL-2 VitoKine HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRAIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:287)
人源化PD-1拮抗剂抗体IL-2 VitoKine HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:288)
人源化PD-1拮抗剂抗体IL-2 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:289)
人源化PD-1拮抗剂抗体IL-2 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGRSENIRTAGGAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:290)
人源化PD-1拮抗剂抗体IL-2 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGSGPLGMLSQGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGGSGRSENIRTAGGELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:291)
P-0701(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:292)
P-0807(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLAMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:293)
P-0708(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSGYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:294)
P-0809(IL-2 Fc VitoKine knob链)
DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHAHYTQKSLSLSPGGGGSGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKRLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLTGGSGPLGMLSQGGGSELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRAIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG(SEQ ID NO:295)
人源化PD-1拮抗剂抗体IL-2HC knob链
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKQLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFIQSIISTLT(SEQ IDNO:296)
人源化PD-1拮抗剂抗体IL-15HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTAENLIHLANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS(SEQ ID NO:297)
蛋白酶可裂解的接头序列
GGGSGRSENIRTAGG(SEQ ID NO:298)
替代小鼠PD-1拮抗剂抗体HC链
EVQLQESGPGLVKPSQSLSLTCSVTGYSITSSYRWNWIRKFPGNRLEWMGYINSAGISNYNPSLKRRISITRDTSKNQFFLQVNSVTTEDAATYYCARSDNMGTTPFTYWGQGTLVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVAISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFGAPIEKTISKTKGGRPKAPQVYTIPPPKEQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG(SEQID NO:299)
替代小鼠PD-1拮抗剂抗体异源二聚体HC链1
EVQLQESGPGLVKPSQSLSLTCSVTGYSITSSYRWNWIRKFPGNRLEWMGYINSAGISNYNPSLKRRISITRDTSKNQFFLQVNSVTTEDAATYYCARSDNMGTTPFTYWGQGTLVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVAISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFGAPIEKTISKTKGRPKAPQVYTIPPPKKQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYKNTQPIMKTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG(SEQID NO:300)
替代小鼠PD-1拮抗剂抗体异源二聚体HC链2
EVQLQESGPGLVKPSQSLSLTCSVTGYSITSSYRWNWIRKFPGNRLEWMGYINSAGISNYNPSLKRRISITRDTSKNQFFLQVNSVTTEDAATYYCARSDNMGTTPFTYWGQGTLVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVAISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFGAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYDNTQPIMDTDGSYFVYSDLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG(SEQID NO:301)
替代小鼠PD-1拮抗剂抗体LC
DIVMTQGTLPNPVPSGESVSITCRSSKSLLYSDGKTYLNWYLQRPGQSPQLLIYWMSTRASGVSDRFSGSGSGTDFTLKISGVEAEDVGIYYCQQGLEFPTFGGGTKLELKRTDAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPRDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC(SEQ ID NO:302)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGPLGMLSQSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:303)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSLGGSGRSANAILEGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGSGPLGMLSQGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQID NO:304)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGRSENIRTAGGNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGSGPLGMLSQGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:305)
人源化PD-1拮抗剂抗体IL-15 VitoKine HC
EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVATISGGGSYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCASPDSSGVAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGSGPLGMLSQGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP(SEQ IDNO:306)。
序列表
<110> 科优基因公司
<120> 基于细胞因子的可生物活化药物及其使用方法
<130> CACCG1.0002WO3
<160> 306
<170> PatentIn version 3.5
<210> 1
<211> 162
<212> PRT
<213> 智人(Homo sapiens)
<400> 1
Met Arg Ile Ser Lys Pro His Leu Arg Ser Ile Ser Ile Gln Cys Tyr
1 5 10 15
Leu Cys Leu Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly Ile His
20 25 30
Val Phe Ile Leu Gly Cys Phe Ser Ala Gly Leu Pro Lys Thr Glu Ala
35 40 45
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
50 55 60
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
65 70 75 80
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
85 90 95
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
100 105 110
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
115 120 125
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
130 135 140
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
145 150 155 160
Thr Ser
<210> 2
<211> 114
<212> PRT
<213> 智人(Homo sapiens)
<400> 2
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 3
<211> 114
<212> PRT
<213> 智人(Homo sapiens)
<400> 3
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 4
<211> 267
<212> PRT
<213> 智人(Homo sapiens)
<400> 4
Met Ala Pro Arg Arg Ala Arg Gly Cys Arg Thr Leu Gly Leu Pro Ala
1 5 10 15
Leu Leu Leu Leu Leu Leu Leu Arg Pro Pro Ala Thr Arg Gly Ile Thr
20 25 30
Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser
35 40 45
Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys
50 55 60
Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala
65 70 75 80
Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp
85 90 95
Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro Ser Thr Val Thr Thr
100 105 110
Ala Gly Val Thr Pro Gln Pro Glu Ser Leu Ser Pro Ser Gly Lys Glu
115 120 125
Pro Ala Ala Ser Ser Pro Ser Ser Asn Asn Thr Ala Ala Thr Thr Ala
130 135 140
Ala Ile Val Pro Gly Ser Gln Leu Met Pro Ser Lys Ser Pro Ser Thr
145 150 155 160
Gly Thr Thr Glu Ile Ser Ser His Glu Ser Ser His Gly Thr Pro Ser
165 170 175
Gln Thr Thr Ala Lys Asn Trp Glu Leu Thr Ala Ser Ala Ser His Gln
180 185 190
Pro Pro Gly Val Tyr Pro Gln Gly His Ser Asp Thr Thr Val Ala Ile
195 200 205
Ser Thr Ser Thr Val Leu Leu Cys Gly Leu Ser Ala Val Ser Leu Leu
210 215 220
Ala Cys Tyr Leu Lys Ser Arg Gln Thr Pro Pro Leu Ala Ser Val Glu
225 230 235 240
Met Glu Ala Met Glu Ala Leu Pro Val Thr Trp Gly Thr Ser Ser Arg
245 250 255
Asp Glu Asp Leu Glu Asn Cys Ser His His Leu
260 265
<210> 5
<211> 77
<212> PRT
<213> 智人(Homo sapiens)
<400> 5
Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val
1 5 10 15
Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly
20 25 30
Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn
35 40 45
Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile
50 55 60
Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
65 70 75
<210> 6
<211> 153
<212> PRT
<213> 智人(Homo sapiens)
<400> 6
Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu
1 5 10 15
Val Thr Asn Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu
20 25 30
Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile
35 40 45
Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe
50 55 60
Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu
65 70 75 80
Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys
85 90 95
Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile
100 105 110
Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala
115 120 125
Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe
130 135 140
Cys Gln Ser Ile Ile Ser Thr Leu Thr
145 150
<210> 7
<211> 133
<212> PRT
<213> 智人(Homo sapiens)
<400> 7
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 8
<211> 133
<212> PRT
<213> 智人(Homo sapiens)
<400> 8
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 9
<211> 272
<212> PRT
<213> 智人(Homo sapiens)
<400> 9
Met Asp Ser Tyr Leu Leu Met Trp Gly Leu Leu Thr Phe Ile Met Val
1 5 10 15
Pro Gly Cys Gln Ala Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro
20 25 30
His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn
35 40 45
Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr
50 55 60
Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys
65 70 75 80
Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro
85 90 95
Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro
100 105 110
Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro
115 120 125
Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val
130 135 140
Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His
145 150 155 160
Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg
165 170 175
Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu Thr Ser Gln
180 185 190
Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro Glu
195 200 205
Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln Ile Gln Thr
210 215 220
Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr Gln
225 230 235 240
Val Ala Val Ala Gly Cys Val Phe Leu Leu Ile Ser Val Leu Leu Leu
245 250 255
Ser Gly Leu Thr Trp Gln Arg Arg Gln Arg Lys Ser Arg Arg Thr Ile
260 265 270
<210> 10
<211> 165
<212> PRT
<213> 智人(Homo sapiens)
<400> 10
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 11
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 S125I序列
<400> 11
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 12
<211> 214
<212> PRT
<213> 智人(Homo sapiens)
<400> 12
Ala Val Asn Gly Thr Ser Gln Phe Thr Cys Phe Tyr Asn Ser Arg Ala
1 5 10 15
Asn Ile Ser Cys Val Trp Ser Gln Asp Gly Ala Leu Gln Asp Thr Ser
20 25 30
Cys Gln Val His Ala Trp Pro Asp Arg Arg Arg Trp Asn Gln Thr Cys
35 40 45
Glu Leu Leu Pro Val Ser Gln Ala Ser Trp Ala Cys Asn Leu Ile Leu
50 55 60
Gly Ala Pro Asp Ser Gln Lys Leu Thr Thr Val Asp Ile Val Thr Leu
65 70 75 80
Arg Val Leu Cys Arg Glu Gly Val Arg Trp Arg Val Met Ala Ile Gln
85 90 95
Asp Phe Lys Pro Phe Glu Asn Leu Arg Leu Met Ala Pro Ile Ser Leu
100 105 110
Gln Val Val His Val Glu Thr His Arg Cys Asn Ile Ser Trp Glu Ile
115 120 125
Ser Gln Ala Ser His Tyr Phe Glu Arg His Leu Glu Phe Glu Ala Arg
130 135 140
Thr Leu Ser Pro Gly His Thr Trp Glu Glu Ala Pro Leu Leu Thr Leu
145 150 155 160
Lys Gln Lys Gln Glu Trp Ile Cys Leu Glu Thr Leu Thr Pro Asp Thr
165 170 175
Gln Tyr Glu Phe Gln Val Arg Val Lys Pro Leu Gln Gly Glu Phe Thr
180 185 190
Thr Trp Ser Pro Trp Ser Gln Pro Leu Ala Phe Arg Thr Lys Pro Ala
195 200 205
Ala Leu Gly Lys Asp Thr
210
<210> 13
<211> 226
<212> PRT
<213> 智人(Homo sapiens)
<400> 13
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly
1 5 10 15
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 14
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有减少/消除的效应子功能的人类IgG1-Fc
<400> 14
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 15
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有减少/消除的效应子功能的人类IgG1 Knob-Fc
<400> 15
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 16
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有减少/消除的效应子功能的人类IgG1 Hole-Fc
<400> 16
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 17
<211> 129
<212> PRT
<213> 智人(Homo sapiens)
<400> 17
His Lys Cys Asp Ile Thr Leu Gln Glu Ile Ile Lys Thr Leu Asn Ser
1 5 10 15
Leu Thr Glu Gln Lys Thr Leu Cys Thr Glu Leu Thr Val Thr Asp Ile
20 25 30
Phe Ala Ala Ser Lys Asn Thr Thr Glu Lys Glu Thr Phe Cys Arg Ala
35 40 45
Ala Thr Val Leu Arg Gln Phe Tyr Ser His His Glu Lys Asp Thr Arg
50 55 60
Cys Leu Gly Ala Thr Ala Gln Gln Phe His Arg His Lys Gln Leu Ile
65 70 75 80
Arg Phe Leu Lys Arg Leu Asp Arg Asn Leu Trp Gly Leu Ala Gly Leu
85 90 95
Asn Ser Cys Pro Val Lys Glu Ala Asn Gln Ser Thr Leu Glu Asn Phe
100 105 110
Leu Glu Arg Leu Lys Thr Ile Met Arg Glu Lys Tyr Ser Lys Cys Ser
115 120 125
Ser
<210> 18
<211> 152
<212> PRT
<213> 智人(Homo sapiens)
<400> 18
Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr Glu Ser Val Leu
1 5 10 15
Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys Glu Ile Gly Ser
20 25 30
Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg His Ile Cys Asp
35 40 45
Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala Arg Lys Leu Arg
50 55 60
Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp Leu His Leu Leu
65 70 75 80
Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys Thr Gly Gln Val
85 90 95
Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln Pro Thr Lys Ser
100 105 110
Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys Leu Asn Asp Leu
115 120 125
Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr Cys Trp Asn Lys
130 135 140
Ile Leu Met Gly Thr Lys Glu His
145 150
<210> 19
<211> 126
<212> PRT
<213> 智人(Homo sapiens)
<400> 19
Gln Gly Cys Pro Thr Leu Ala Gly Ile Leu Asp Ile Asn Phe Leu Ile
1 5 10 15
Asn Lys Met Gln Glu Asp Pro Ala Ser Lys Cys His Cys Ser Ala Asn
20 25 30
Val Thr Ser Cys Leu Cys Leu Gly Ile Pro Ser Asp Asn Cys Thr Arg
35 40 45
Pro Cys Phe Ser Glu Arg Leu Ser Gln Met Thr Asn Thr Thr Met Gln
50 55 60
Thr Arg Tyr Pro Leu Ile Phe Ser Arg Val Lys Lys Ser Val Glu Val
65 70 75 80
Leu Lys Asn Asn Lys Cys Pro Tyr Phe Ser Cys Glu Gln Pro Cys Asn
85 90 95
Gln Thr Thr Ala Gly Asn Ala Leu Thr Phe Leu Lys Ser Leu Leu Glu
100 105 110
Ile Phe Gln Lys Glu Lys Met Arg Gly Met Arg Gly Lys Ile
115 120 125
<210> 20
<211> 160
<212> PRT
<213> 智人(Homo sapiens)
<400> 20
Ser Pro Gly Gln Gly Thr Gln Ser Glu Asn Ser Cys Thr His Phe Pro
1 5 10 15
Gly Asn Leu Pro Asn Met Leu Arg Asp Leu Arg Asp Ala Phe Ser Arg
20 25 30
Val Lys Thr Phe Phe Gln Met Lys Asp Gln Leu Asp Asn Leu Leu Leu
35 40 45
Lys Glu Ser Leu Leu Glu Asp Phe Lys Gly Tyr Leu Gly Cys Gln Ala
50 55 60
Leu Ser Glu Met Ile Gln Phe Tyr Leu Glu Glu Val Met Pro Gln Ala
65 70 75 80
Glu Asn Gln Asp Pro Asp Ile Lys Ala His Val Asn Ser Leu Gly Glu
85 90 95
Asn Leu Lys Thr Leu Arg Leu Arg Leu Arg Arg Cys His Arg Phe Leu
100 105 110
Pro Cys Glu Asn Lys Ser Lys Ala Val Glu Gln Val Lys Asn Ala Phe
115 120 125
Asn Lys Leu Gln Glu Lys Gly Ile Tyr Lys Ala Met Ser Glu Phe Asp
130 135 140
Ile Phe Ile Asn Tyr Ile Glu Ala Tyr Met Thr Met Lys Ile Arg Asn
145 150 155 160
<210> 21
<211> 197
<212> PRT
<213> 智人(Homo sapiens)
<400> 21
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser
195
<210> 22
<211> 306
<212> PRT
<213> 智人(Homo sapiens)
<400> 22
Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr
1 5 10 15
Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys
85 90 95
Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys
100 105 110
Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr
115 120 125
Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln
130 135 140
Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly
145 150 155 160
Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala
165 170 175
Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala
180 185 190
Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg
195 200 205
Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu
210 215 220
Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp
225 230 235 240
Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln
245 250 255
Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr
260 265 270
Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala
275 280 285
Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro
290 295 300
Cys Ser
305
<210> 23
<211> 170
<212> PRT
<213> 智人(Homo sapiens)
<400> 23
Arg Ala Val Pro Gly Gly Ser Ser Pro Ala Trp Thr Gln Cys Gln Gln
1 5 10 15
Leu Ser Gln Lys Leu Cys Thr Leu Ala Trp Ser Ala His Pro Leu Val
20 25 30
Gly His Met Asp Leu Arg Glu Glu Gly Asp Glu Glu Thr Thr Asn Asp
35 40 45
Val Pro His Ile Gln Cys Gly Asp Gly Cys Asp Pro Gln Gly Leu Arg
50 55 60
Asp Asn Ser Gln Phe Cys Leu Gln Arg Ile His Gln Gly Leu Ile Phe
65 70 75 80
Tyr Glu Lys Leu Leu Gly Ser Asp Ile Phe Thr Gly Glu Pro Ser Leu
85 90 95
Leu Pro Asp Ser Pro Val Gly Gln Leu His Ala Ser Leu Leu Gly Leu
100 105 110
Ser Gln Leu Leu Gln Pro Glu Gly His His Trp Glu Thr Gln Gln Ile
115 120 125
Pro Ser Leu Ser Pro Ser Gln Pro Trp Gln Arg Leu Leu Leu Arg Phe
130 135 140
Lys Ile Leu Arg Ser Leu Gln Ala Phe Val Ala Val Ala Ala Arg Val
145 150 155 160
Phe Ala His Gly Ala Ala Thr Leu Ser Pro
165 170
<210> 24
<211> 112
<212> PRT
<213> 智人(Homo sapiens)
<400> 24
Ala Leu Asp Thr Asn Tyr Cys Phe Ser Ser Thr Glu Lys Asn Cys Cys
1 5 10 15
Val Arg Gln Leu Tyr Ile Asp Phe Arg Lys Asp Leu Gly Trp Lys Trp
20 25 30
Ile His Glu Pro Lys Gly Tyr His Ala Asn Phe Cys Leu Gly Pro Cys
35 40 45
Pro Tyr Ile Trp Ser Leu Asp Thr Gln Tyr Ser Lys Val Leu Ala Leu
50 55 60
Tyr Asn Gln His Asn Pro Gly Ala Ser Ala Ala Pro Cys Cys Val Pro
65 70 75 80
Gln Ala Leu Glu Pro Leu Pro Ile Val Tyr Tyr Val Gly Arg Lys Pro
85 90 95
Lys Val Glu Gln Leu Ser Asn Met Ile Val Arg Ser Cys Lys Cys Ser
100 105 110
<210> 25
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0351 (IL-15 Fc VitoKine)
<400> 25
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 26
<211> 432
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0170 (IL-15 Fc VitoKine) Hole链
<400> 26
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Asn
225 230 235 240
Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln
245 250 255
Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro
260 265 270
Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val
275 280 285
Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn
290 295 300
Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr
305 310 315 320
Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys
325 330 335
Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr
340 345 350
Ser Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
355 360 365
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys
370 375 380
Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
385 390 395 400
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
405 410 415
Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
420 425 430
<210> 27
<211> 432
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0172 (IL-15 Fc VitoKine)
<400> 27
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Asn
225 230 235 240
Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln
245 250 255
Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro
260 265 270
Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val
275 280 285
Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn
290 295 300
Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr
305 310 315 320
Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys
325 330 335
Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr
340 345 350
Ser Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
355 360 365
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys
370 375 380
Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
385 390 395 400
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
405 410 415
Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
420 425 430
<210> 28
<211> 450
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0202 (IL-15 Fc VitoKine)
<400> 28
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Leu Ser Gly Arg Ser Asp Asn His Gly Gly Ser Gly Gly Gly Gly
245 250 255
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
260 265 270
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
275 280 285
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
290 295 300
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
305 310 315 320
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
325 330 335
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
340 345 350
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
355 360 365
Asn Thr Ser Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His
370 375 380
Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr
385 390 395 400
Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr
405 410 415
Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro
420 425 430
Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala
435 440 445
Pro Pro
450
<210> 29
<211> 467
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0203 (IL-15 Fc VitoKine)
<400> 29
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser
225 230 235 240
Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Ser
245 250 255
Gly Gly Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys
260 265 270
Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr
275 280 285
Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe
290 295 300
Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile
305 310 315 320
His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser
325 330 335
Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu
340 345 350
Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val
355 360 365
Gln Met Phe Ile Asn Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
370 375 380
Ser Gly Gly Gly Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu
385 390 395 400
His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg
405 410 415
Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu
420 425 430
Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr
435 440 445
Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro
450 455 460
Ala Pro Pro
465
<210> 30
<211> 460
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0204 (IL-15 Fc VitoKine)
<400> 30
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Gly Gly Ser
225 230 235 240
Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Gly Gly Ser Asn Trp
245 250 255
Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser
260 265 270
Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser
275 280 285
Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile
290 295 300
Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu
305 310 315 320
Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu
325 330 335
Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu
340 345 350
Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
355 360 365
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ile
370 375 380
Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys
385 390 395 400
Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe
405 410 415
Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys
420 425 430
Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg
435 440 445
Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
450 455 460
<210> 31
<211> 455
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0205 (IL-15 Fc VitoKine)
<400> 31
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Gly Gly Ser
225 230 235 240
Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Gly Gly Ser Asn Trp
245 250 255
Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser
260 265 270
Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser
275 280 285
Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile
290 295 300
Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu
305 310 315 320
Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu
325 330 335
Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu
340 345 350
Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
355 360 365
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ile Thr Cys Pro Pro Pro
370 375 380
Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr
385 390 395 400
Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly
405 410 415
Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala
420 425 430
His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val
435 440 445
His Gln Arg Pro Ala Pro Pro
450 455
<210> 32
<211> 450
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0206 (IL-15 Fc VitoKine)
<400> 32
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Gly Gly Ser
225 230 235 240
Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Gly Gly Ser Asn Trp
245 250 255
Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser
260 265 270
Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser
275 280 285
Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile
290 295 300
Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu
305 310 315 320
Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu
325 330 335
Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu
340 345 350
Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
355 360 365
Gly Gly Gly Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His
370 375 380
Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr
385 390 395 400
Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr
405 410 415
Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro
420 425 430
Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala
435 440 445
Pro Pro
450
<210> 33
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0315 (IL-15 Fc VitoKine)
<400> 33
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
370 375 380
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys
385 390 395 400
Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
405 410 415
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
420 425 430
Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440 445
<210> 34
<211> 451
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0316 (IL-15 Fc VitoKine)
<400> 34
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Gly Pro Leu Gly Met
355 360 365
Leu Ser Gln Gly Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu
370 375 380
His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg
385 390 395 400
Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu
405 410 415
Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr
420 425 430
Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro
435 440 445
Ala Pro Pro
450
<210> 35
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0350 (IL-15 Fc VitoKine)
<400> 35
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 36
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0354 (IL-15 Fc VitoKine)
<400> 36
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 37
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0355 (IL-15 Fc VitoKine)
<400> 37
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 38
<211> 446
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0385 (IL-15 Fc VitoKine)
<400> 38
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Pro Leu Gly Met Leu Ser
355 360 365
Gln Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp
370 375 380
Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser
385 390 395 400
Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu
405 410 415
Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys
420 425 430
Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440 445
<210> 39
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0386 (IL-15 Fc VitoKine)
<400> 39
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
370 375 380
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Ser
385 390 395 400
Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
405 410 415
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
420 425 430
Lys Ser Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440 445
<210> 40
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0387 (IL-15 Fc VitoKine)
<400> 40
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
370 375 380
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Glu Tyr Ile Cys
385 390 395 400
Asn Ser Gly Phe Lys Glu Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
405 410 415
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
420 425 430
Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440 445
<210> 41
<211> 433
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0388 (IL-15 Fc VitoKine)
<400> 41
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Asp Cys Gly Leu Pro Pro Asp Val Pro Asn Ala Gln Pro
370 375 380
Ala Leu Glu Gly Arg Thr Ser Phe Pro Glu Asp Thr Val Ile Thr Tyr
385 390 395 400
Lys Cys Glu Glu Ser Phe Val Lys Ile Pro Gly Glu Lys Asp Ser Val
405 410 415
Ile Cys Leu Lys Gly Ser Gln Trp Ser Asp Ile Glu Glu Phe Cys Asn
420 425 430
Arg
<210> 42
<211> 536
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0389 (IL-15 Fc VitoKine)
<400> 42
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala
370 375 380
Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu
385 390 395 400
Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu
405 410 415
Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys
420 425 430
Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro
435 440 445
Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln
450 455 460
Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro
465 470 475 480
Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln
485 490 495
Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly
500 505 510
Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr
515 520 525
Gln Pro Gln Leu Ile Cys Thr Gly
530 535
<210> 43
<211> 443
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0397 (IL-15 Fc VitoKine)
<400> 43
Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val
1 5 10 15
Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly
20 25 30
Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn
35 40 45
Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile
50 55 60
Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro Gly Gly Pro
65 70 75 80
Leu Gly Met Leu Ser Gln Ser Asn Trp Val Asn Val Ile Ser Asp Leu
85 90 95
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
100 105 110
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
115 120 125
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
130 135 140
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
145 150 155 160
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
165 170 175
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
180 185 190
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Gly Gly Ser Leu Gly
195 200 205
Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Ser Cys Pro
210 215 220
Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
245 250 255
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
260 265 270
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
275 280 285
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
290 295 300
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
305 310 315 320
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
325 330 335
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
340 345 350
Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
355 360 365
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
370 375 380
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
385 390 395 400
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
405 410 415
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
420 425 430
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440
<210> 44
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole-Fc-IL-15
<400> 44
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 45
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Knob-Fc-IL-15
<400> 45
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 46
<211> 318
<212> PRT
<213> 人工(Artificial)
<220>
<223> Knob-Fc-IL-15α-Sushi+
<400> 46
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp
245 250 255
Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser
260 265 270
Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu
275 280 285
Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys
290 295 300
Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
305 310 315
<210> 47
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 S58D
<400> 47
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 48
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0250 (IL-2 Fc融合体)
<400> 48
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 49
<211> 560
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0320 (IL-2 Fc VitoKine)
<400> 49
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys
245 250 255
Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu
260 265 270
Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr
275 280 285
Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln
290 295 300
Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala
305 310 315 320
Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile
325 330 335
Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys
340 345 350
Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp
355 360 365
Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly
370 375 380
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp
385 390 395 400
Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys
405 410 415
Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile
420 425 430
Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser
435 440 445
Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr
450 455 460
Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr
465 470 475 480
Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro
485 490 495
Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg
500 505 510
Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln
515 520 525
Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met
530 535 540
Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
545 550 555 560
<210> 50
<211> 560
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0352 (IL-2 Fc VitoKine)
<400> 50
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys
245 250 255
Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu
260 265 270
Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr
275 280 285
Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln
290 295 300
Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala
305 310 315 320
Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile
325 330 335
Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys
340 345 350
Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp
355 360 365
Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly
370 375 380
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Leu Cys Asp Asp
385 390 395 400
Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys
405 410 415
Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile
420 425 430
Glu Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser
435 440 445
Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr
450 455 460
Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr
465 470 475 480
Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro
485 490 495
Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg
500 505 510
Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln
515 520 525
Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met
530 535 540
Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
545 550 555 560
<210> 51
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0382 (IL-2 Fc VitoKine)
<400> 51
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 52
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0398 (IL-2 Fc VitoKine)
<400> 52
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 53
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0362 (IL-2 Fc VitoKine)
<400> 53
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 54
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0380 (IL-2 Fc VitoKine)
<400> 54
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Ala Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 55
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0384 (IL-2 Fc VitoKine)
<400> 55
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
405 410 415
Gly Ser Leu Ala Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 56
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0400 (IL-2 Fc VitoKine)
<400> 56
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Asn Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 57
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0404 (IL-2 Fc VitoKine)
<400> 57
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Glu Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 58
<211> 454
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0399 (IL-2 Fc VitoKine)
<400> 58
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met
370 375 380
Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser
385 390 395 400
Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr
405 410 415
Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His
420 425 430
Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His
435 440 445
Gln Arg Pro Ala Pro Pro
450
<210> 59
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0379 (IL-2 Fc VitoKine)
<400> 59
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 60
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0381 (IL-2 Fc VitoKine)
<400> 60
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Glu
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 61
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0383 (IL-2 Fc VitoKine)
<400> 61
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Glu
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 62
<211> 555
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0329 (IL-2 Fc VitoKine)
<400> 62
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
165 170 175
Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu
180 185 190
Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile
195 200 205
Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe
210 215 220
Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu
225 230 235 240
Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys
245 250 255
Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile
260 265 270
Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala
275 280 285
Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe
290 295 300
Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Leu Gly
305 310 315 320
Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Ser Cys Pro
325 330 335
Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe
340 345 350
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
355 360 365
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
370 375 380
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
385 390 395 400
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
405 410 415
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
420 425 430
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
435 440 445
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
450 455 460
Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
465 470 475 480
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
485 490 495
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
500 505 510
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
515 520 525
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
530 535 540
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
545 550 555
<210> 63
<211> 549
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0401 (IL-2 Fc VitoKine)
<400> 63
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ala
165 170 175
Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu
180 185 190
Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn
195 200 205
Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys
210 215 220
Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro
225 230 235 240
Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg
245 250 255
Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys
260 265 270
Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr
275 280 285
Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile
290 295 300
Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Gly Ser Gly Gly Gly Gly Ser Cys Pro Pro Cys Pro Ala Pro Glu
325 330 335
Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
340 345 350
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
355 360 365
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
370 375 380
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
385 390 395 400
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
405 410 415
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
420 425 430
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
435 440 445
Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn
450 455 460
Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
465 470 475 480
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
485 490 495
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
500 505 510
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
515 520 525
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
530 535 540
Ser Leu Ser Pro Gly
545
<210> 64
<211> 554
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0402 (IL-2 Fc VitoKine)
<400> 64
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly Gly Gly Ser Gly Pro Leu Gly Met Leu Ser Gln
165 170 175
Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu
180 185 190
Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile
195 200 205
Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe
210 215 220
Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu
225 230 235 240
Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys
245 250 255
Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile
260 265 270
Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala
275 280 285
Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe
290 295 300
Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys Pro Pro
325 330 335
Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro
340 345 350
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
355 360 365
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
370 375 380
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
385 390 395 400
Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
405 410 415
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
420 425 430
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
435 440 445
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
450 455 460
Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
465 470 475 480
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
485 490 495
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
500 505 510
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
515 520 525
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
530 535 540
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
545 550
<210> 65
<211> 546
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0403 (IL-2 Fc VitoKine)
<400> 65
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ala
165 170 175
Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu
180 185 190
Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn
195 200 205
Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys
210 215 220
Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro
225 230 235 240
Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg
245 250 255
Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys
260 265 270
Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr
275 280 285
Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile
290 295 300
Ser Thr Leu Thr Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu
305 310 315 320
Ala Ala Ala Lys Ala Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
325 330 335
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
340 345 350
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
355 360 365
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
370 375 380
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
385 390 395 400
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
405 410 415
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
420 425 430
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
435 440 445
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
450 455 460
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
465 470 475 480
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
485 490 495
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
500 505 510
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
515 520 525
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
530 535 540
Pro Gly
545
<210> 66
<211> 405
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole-Fc-15p1
<400> 66
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
355 360 365
Ala Ile Leu Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
370 375 380
Ile Tyr Asn Cys Glu Ile Ser Gln Ala Ser His Tyr Phe Glu Arg His
385 390 395 400
Leu Cys Tyr Ser Ile
405
<210> 67
<211> 407
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole-Fc-15p2
<400> 67
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
355 360 365
Ala Ile Leu Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
370 375 380
Ile Tyr Asn Cys Glu Leu His Arg Glu Phe Tyr His Ser Ala Gln Ser
385 390 395 400
Ile Glu Trp Cys Tyr Ser Ile
405
<210> 68
<211> 417
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole-Fc-15p3
<400> 68
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
355 360 365
Ala Ile Leu Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
370 375 380
Glu Thr His Arg Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala Ser His
385 390 395 400
Tyr Phe Glu Arg His Leu Glu Phe Glu Ala Arg Thr Leu Cys Pro Gly
405 410 415
His
<210> 69
<211> 386
<212> PRT
<213> 人工(Artificial)
<220>
<223> p1'-15-Fc
<400> 69
Gln Gly Gln Ser Gly Gln Cys Glu Ile Ser Gln Ala Ser His Tyr Phe
1 5 10 15
Glu Arg His Leu Cys Tyr Ser Ile Gly Ser Ser Gly Gly Ser Gly Gly
20 25 30
Ser Gly Gly Ser Gly Leu Ser Gly Arg Ser Asp Asn His Gly Ser Ser
35 40 45
Gly Thr Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp
50 55 60
Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp
65 70 75 80
Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu
85 90 95
Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr
100 105 110
Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly
115 120 125
Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys
130 135 140
Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe
145 150 155 160
Ile Asn Thr Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
165 170 175
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
180 185 190
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
195 200 205
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
210 215 220
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
225 230 235 240
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
245 250 255
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
260 265 270
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
275 280 285
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
290 295 300
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
305 310 315 320
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
325 330 335
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
340 345 350
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
355 360 365
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
370 375 380
Pro Gly
385
<210> 70
<211> 397
<212> PRT
<213> 人工(Artificial)
<220>
<223> p3'-15-Fc
<400> 70
Gln Gly Gln Ser Gly Gln Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala
1 5 10 15
Ser His Tyr Phe Glu Arg His Leu Glu Phe Glu Ala Arg Thr Leu Cys
20 25 30
Pro Gly His Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly
35 40 45
Leu Ser Gly Arg Ser Asp Asn His Gly Ser Ser Gly Thr Asn Trp Val
50 55 60
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
65 70 75 80
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
85 90 95
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
100 105 110
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile
115 120 125
Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
130 135 140
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
145 150 155 160
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly
165 170 175
Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe
180 185 190
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
195 200 205
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
210 215 220
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
225 230 235 240
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
245 250 255
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
260 265 270
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
275 280 285
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
290 295 300
Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val
305 310 315 320
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
325 330 335
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
340 345 350
Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
355 360 365
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
370 375 380
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
385 390 395
<210> 71
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 71
Ser Pro Leu Gly Leu Ala Gly Ser
1 5
<210> 72
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 72
Glu Pro Leu Glu Leu Arg Ala Gly
1 5
<210> 73
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 73
Leu Ser Gly Arg Ser Asp Asn His
1 5
<210> 74
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 74
Gly Pro Leu Gly Ile Ala Gly Gln
1 5
<210> 75
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 75
Gly Thr Ala His Leu Met Gly Gly
1 5
<210> 76
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 76
Arg Ile Gly Ser Leu Arg Thr Ala
1 5
<210> 77
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 77
Gly Pro Leu Gly Met Leu Ser Gln
1 5
<210> 78
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 78
Arg Pro Ser Ala Ser Arg Ser Ala
1 5
<210> 79
<211> 6
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 79
Pro Leu Gly Leu Ala Gly
1 5
<210> 80
<211> 13
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 80
Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu
1 5 10
<210> 81
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 81
Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile
1 5 10
<210> 82
<211> 5
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 82
Ser Gly Arg Ser Ala
1 5
<210> 83
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 83
Ala Ala Asn Leu
1
<210> 84
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 84
Gly Phe Phe Tyr
1
<210> 85
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 85
Gly Pro Ile Cys Phe Arg Leu Gly
1 5
<210> 86
<211> 7
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 86
Arg Gln Ala Gly Phe Ser Leu
1 5
<210> 87
<211> 6
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 87
His Ser Ser Lys Leu Gln
1 5
<210> 88
<211> 30
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 88
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Ser
1 5 10 15
Gly Arg Ser Asp Asn His Gly Gly Ser Gly Gly Gly Gly Ser
20 25 30
<210> 89
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 89
Gly Ser Ser Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly Thr
1 5 10 15
<210> 90
<211> 35
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 90
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Ser Leu Gly
1 5 10 15
Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Ser Gly Gly
20 25 30
Gly Gly Ser
35
<210> 91
<211> 28
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 91
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg
1 5 10 15
Ser Ala Asn Ala Ile Leu Glu Gly Gly Gly Gly Ser
20 25
<210> 92
<211> 21
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 92
Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile
1 5 10 15
Leu Glu Gly Gly Ser
20
<210> 93
<211> 14
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 93
Gly Gly Gly Ser Gly Pro Thr Asn Lys Val Arg Gly Gly Ser
1 5 10
<210> 94
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 94
Gly Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser
1 5 10 15
<210> 95
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 95
Gly Gly Pro Leu Gly Met Leu Ser Gln Ser
1 5 10
<210> 96
<211> 13
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 96
Gly Gly Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Ser
1 5 10
<210> 97
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 97
Leu Gly Ala Pro Asp Ser Gln Lys Leu Thr Thr Val Asp Ile Val
1 5 10 15
<210> 98
<211> 13
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 98
Glu Ile Ser Gln Ala Ser His Tyr Phe Glu Arg His Leu
1 5 10
<210> 99
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 99
Cys Glu Ile Ser Gln Ala Ser His Tyr Phe Glu Arg His Leu Cys
1 5 10 15
<210> 100
<211> 36
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 100
Leu Gly Ala Pro Asp Ser Gln Lys Leu Thr Thr Val Asp Ile Val Gly
1 5 10 15
Gly Gly Gly Gly Gly Gly Gly Glu Ile Ser Gln Ala Ser His Tyr Phe
20 25 30
Glu Arg His Leu
35
<210> 101
<211> 13
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 101
Lys Pro Phe Glu Asn Leu Arg Leu Met Ala Pro Ile Ser
1 5 10
<210> 102
<211> 50
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 102
Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu
1 5 10 15
Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Ile Tyr Asn
20 25 30
Cys Glu Ile Ser Gln Ala Ser His Tyr Phe Glu Arg His Leu Cys Tyr
35 40 45
Ser Ile
50
<210> 103
<211> 52
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 103
Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu
1 5 10 15
Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Ile Tyr Asn
20 25 30
Cys Glu Leu His Arg Glu Phe Tyr His Ser Ala Gln Ser Ile Glu Trp
35 40 45
Cys Tyr Ser Ile
50
<210> 104
<211> 62
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 104
Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu
1 5 10 15
Glu Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Glu Thr His
20 25 30
Arg Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala Ser His Tyr Phe Glu
35 40 45
Arg His Leu Glu Phe Glu Ala Arg Thr Leu Cys Pro Gly His
50 55 60
<210> 105
<211> 50
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 105
Gln Gly Gln Ser Gly Gln Cys Glu Ile Ser Gln Ala Ser His Tyr Phe
1 5 10 15
Glu Arg His Leu Cys Tyr Ser Ile Gly Ser Ser Gly Gly Ser Gly Gly
20 25 30
Ser Gly Gly Ser Gly Leu Ser Gly Arg Ser Asp Asn His Gly Ser Ser
35 40 45
Gly Thr
50
<210> 106
<211> 61
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基于IL-2Rβ的封闭肽序列
<400> 106
Gln Gly Gln Ser Gly Gln Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala
1 5 10 15
Ser His Tyr Phe Glu Arg His Leu Glu Phe Glu Ala Arg Thr Leu Cys
20 25 30
Pro Gly His Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly
35 40 45
Leu Ser Gly Arg Ser Asp Asn His Gly Ser Ser Gly Thr
50 55 60
<210> 107
<211> 14
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 107
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
1 5 10
<210> 108
<211> 12
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 108
Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser
1 5 10
<210> 109
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 109
Gly Gly Gly Ser
1
<210> 110
<211> 9
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 110
Gly Ser Ser Gly Gly Ser Gly Gly Ser
1 5
<210> 111
<211> 5
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 111
Gly Ser Ser Gly Thr
1 5
<210> 112
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 112
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 113
<211> 17
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 113
Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys
1 5 10 15
Ala
<210> 114
<211> 20
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 114
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1 5 10 15
Gly Gly Gly Ser
20
<210> 115
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 115
Gly Gly Gly Ser Gly Gly Gly Ser
1 5
<210> 116
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 116
Gly Ser Gly Ser
1
<210> 117
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 117
Gly Gly Ser Ser
1
<210> 118
<211> 5
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 118
Gly Gly Gly Gly Ser
1 5
<210> 119
<211> 5
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 119
Gly Gly Ser Gly Gly
1 5
<210> 120
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 120
Ser Gly Gly Gly
1
<210> 121
<211> 4
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 121
Gly Ser Gly Ser
1
<210> 122
<211> 6
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 122
Gly Ser Gly Ser Gly Ser
1 5
<210> 123
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 123
Gly Ser Gly Ser Gly Ser Gly Ser
1 5
<210> 124
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 124
Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser
1 5 10
<210> 125
<211> 12
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 125
Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser
1 5 10
<210> 126
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 126
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10
<210> 127
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 不可裂解的接头序列
<400> 127
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 128
<211> 676
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体-IL-15-VitoKine-HC
<400> 128
Gln Gly Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Glu Ser Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Arg Ala Lys Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Thr Arg Glu Gly Ile Thr Thr Val Ala Thr Thr Tyr Tyr Trp Tyr Phe
100 105 110
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
225 230 235 240
Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
245 250 255
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
260 265 270
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
305 310 315 320
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
325 330 335
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
340 345 350
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
355 360 365
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
370 375 380
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
385 390 395 400
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
420 425 430
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
435 440 445
Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly
450 455 460
Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val
465 470 475 480
Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile
485 490 495
Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val
500 505 510
Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu
515 520 525
Ser Gly Asp Ala Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu
530 535 540
Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys
545 550 555 560
Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln
565 570 575
Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro
580 585 590
Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser Val
595 600 605
Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu
610 615 620
Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser
625 630 635 640
Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr
645 650 655
Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg
660 665 670
Pro Ala Pro Pro
675
<210> 129
<211> 219
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体-LC
<400> 129
Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
1 5 10 15
Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser
20 25 30
Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly
85 90 95
Ser His Val Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110
Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
115 120 125
Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
130 135 140
Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
145 150 155 160
Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
165 170 175
Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
180 185 190
Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
195 200 205
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 130
<211> 669
<212> PRT
<213> 人工(Artificial)
<220>
<223> CTLA-4拮抗剂抗体-IL-15-VitoKine-HC
<400> 130
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Ile Tyr Tyr Cys
85 90 95
Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly
435 440 445
Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu
450 455 460
Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile
465 470 475 480
Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu
485 490 495
Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu
500 505 510
Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His
515 520 525
Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser
530 535 540
Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu
545 550 555 560
Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln
565 570 575
Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser
580 585 590
Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val
595 600 605
Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly
610 615 620
Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn
625 630 635 640
Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile
645 650 655
Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
660 665
<210> 131
<211> 215
<212> PRT
<213> 人工(Artificial)
<220>
<223> CTLA-4拮抗剂抗体-LC
<400> 131
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Phe Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 132
<211> 677
<212> PRT
<213> 人工(Artificial)
<220>
<223> CD40激动剂抗体-IL-15-VitoKine-HC
<400> 132
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr
20 25 30
Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Trp Ile Asn Pro Asp Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Asn Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Gln Pro Leu Gly Tyr Cys Thr Asn Gly Val Cys Ser Tyr
100 105 110
Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser
115 120 125
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
130 135 140
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
145 150 155 160
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
165 170 175
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
180 185 190
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
195 200 205
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
210 215 220
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
225 230 235 240
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
245 250 255
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
260 265 270
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
275 280 285
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
290 295 300
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
305 310 315 320
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
325 330 335
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
340 345 350
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
355 360 365
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
370 375 380
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
385 390 395 400
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
405 410 415
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
420 425 430
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
435 440 445
Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser
450 455 460
Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn
465 470 475 480
Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His
485 490 495
Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys
500 505 510
Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu
515 520 525
Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile
530 535 540
Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly
545 550 555 560
Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu
565 570 575
Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly
580 585 590
Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser
595 600 605
Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg
610 615 620
Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser
625 630 635 640
Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp
645 650 655
Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln
660 665 670
Arg Pro Ala Pro Pro
675
<210> 133
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> CD40激动剂抗体-LC
<400> 133
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Tyr Ser Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile
35 40 45
Tyr Thr Ala Ser Thr Leu Gln 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 Gln Ala Asn Ile Phe Pro Leu
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 134
<211> 666
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化抗纤连蛋白抗体-IL-15-VitoKine-HC
<400> 134
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 Phe
20 25 30
Ser Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Ser Lys Asn Thr Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
115 120 125
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
130 135 140
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
145 150 155 160
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
165 170 175
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
180 185 190
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
195 200 205
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
210 215 220
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
225 230 235 240
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
245 250 255
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
260 265 270
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
275 280 285
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
290 295 300
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
305 310 315 320
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
325 330 335
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
340 345 350
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
355 360 365
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
370 375 380
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
385 390 395 400
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
405 410 415
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
420 425 430
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly Gly
435 440 445
Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu Gly Gly
450 455 460
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
465 470 475 480
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
485 490 495
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
500 505 510
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
515 520 525
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
530 535 540
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
545 550 555 560
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
565 570 575
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
580 585 590
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
595 600 605
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
610 615 620
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
625 630 635 640
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
645 650 655
Ala Leu Val His Gln Arg Pro Ala Pro Pro
660 665
<210> 135
<211> 215
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化抗纤连蛋白抗体-LC
<400> 135
Glu Ile Val Leu Thr Gln Ser Pro Gly 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 Ser
20 25 30
Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Tyr Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Thr Gly Arg Ile Pro
85 90 95
Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
100 105 110
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140
Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
145 150 155 160
Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190
Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
195 200 205
Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 136
<211> 766
<212> PRT
<213> 人工(Artificial)
<220>
<223> 利妥昔单抗-IL-2-VitoKine-HC
<400> 136
Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile
35 40 45
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly
100 105 110
Ala Gly Thr Thr Val Thr Val Ser Ala 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 Ala 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 Asn 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 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr 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 Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
450 455 460
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
465 470 475 480
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
485 490 495
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
500 505 510
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
515 520 525
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
530 535 540
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
545 550 555 560
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
565 570 575
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
580 585 590
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
595 600 605
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
610 615 620
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
625 630 635 640
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
645 650 655
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
660 665 670
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
675 680 685
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
690 695 700
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
705 710 715 720
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
725 730 735
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
740 745 750
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 137
<211> 213
<212> PRT
<213> 人工(Artificial)
<220>
<223> 利妥昔单抗-LC
<400> 137
Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly
1 5 10 15
Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile
20 25 30
His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr
35 40 45
Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu
65 70 75 80
Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr
85 90 95
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro
100 105 110
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
115 120 125
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
130 135 140
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
145 150 155 160
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
165 170 175
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
180 185 190
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
195 200 205
Asn Arg Gly Glu Cys
210
<210> 138
<211> 766
<212> PRT
<213> 人工(Artificial)
<220>
<223> 赫塞汀-IL-2-VitoKine-HC
<400> 138
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 Asn Ile Lys Asp Thr
20 25 30
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val
50 55 60
Lys Gly 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
Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met 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 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 Asn 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 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr 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 Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
450 455 460
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
465 470 475 480
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
485 490 495
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
500 505 510
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
515 520 525
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
530 535 540
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
545 550 555 560
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
565 570 575
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
580 585 590
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
595 600 605
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
610 615 620
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser
625 630 635 640
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
645 650 655
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
660 665 670
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
675 680 685
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
690 695 700
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
705 710 715 720
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
725 730 735
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
740 745 750
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 139
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 赫塞汀-LC
<400> 139
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Arg 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 Gln His Tyr Thr Thr Pro Pro
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 140
<211> 764
<212> PRT
<213> 人工(Artificial)
<220>
<223> 西妥昔单抗-IL-2-VitoKine-HC
<400> 140
Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln
1 5 10 15
Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr
20 25 30
Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu
35 40 45
Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr
50 55 60
Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe
65 70 75 80
Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala
85 90 95
Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys
450 455 460
Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile
465 470 475 480
Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu
485 490 495
Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu
500 505 510
Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu
515 520 525
Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn
530 535 540
Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met
545 550 555 560
Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg
565 570 575
Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Pro
580 585 590
Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro Pro Glu
595 600 605
Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met
610 615 620
Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser
625 630 635 640
Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn
645 650 655
Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val
660 665 670
Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln
675 680 685
Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg
690 695 700
Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe
705 710 715 720
Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala
725 730 735
Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys
740 745 750
Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760
<210> 141
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 西妥昔单抗-LC
<400> 141
Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly
1 5 10 15
Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn
20 25 30
Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile
35 40 45
Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser
65 70 75 80
Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr
85 90 95
Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 142
<211> 770
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体-IL-2-VitoKine-HC
<400> 142
Gln Gly Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Val Ile Glu Ser Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Arg Ala Lys Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Thr Arg Glu Gly Ile Thr Thr Val Ala Thr Thr Tyr Tyr Trp Tyr Phe
100 105 110
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr
115 120 125
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
130 135 140
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
145 150 155 160
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
195 200 205
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
210 215 220
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
225 230 235 240
Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
245 250 255
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
260 265 270
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
305 310 315 320
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
325 330 335
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
340 345 350
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
355 360 365
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
370 375 380
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
385 390 395 400
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
420 425 430
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
435 440 445
Leu Ser Leu Ser Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro
450 455 460
Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu
465 470 475 480
Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro
485 490 495
Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala
500 505 510
Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu
515 520 525
Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro
530 535 540
Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly
545 550 555 560
Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile
565 570 575
Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser
580 585 590
Thr Leu Thr Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys
595 600 605
Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala
610 615 620
Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg
625 630 635 640
Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser
645 650 655
His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg
660 665 670
Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg
675 680 685
Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser
690 695 700
Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr
705 710 715 720
Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys
725 730 735
Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys
740 745 750
Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys
755 760 765
Thr Gly
770
<210> 143
<211> 766
<212> PRT
<213> 人工(Artificial)
<220>
<223> 维多珠单抗-IL-2-VitoKine-HC
<400> 143
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Ser Tyr
20 25 30
Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Ile
35 40 45
Gly Glu Ile Asp Pro Ser Glu Ser Asn Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Arg Val Thr Leu Thr Val Asp Ile Ser Ala Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Gly Gly Tyr Asp Gly Trp Asp Tyr Ala Ile 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 Ala Gly
225 230 235 240
Ala 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 Asn 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 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr 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 Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
450 455 460
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Thr Leu Gln
465 470 475 480
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
485 490 495
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
500 505 510
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
515 520 525
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
530 535 540
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
545 550 555 560
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
565 570 575
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
580 585 590
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
595 600 605
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
610 615 620
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
625 630 635 640
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
645 650 655
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
660 665 670
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
675 680 685
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
690 695 700
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
705 710 715 720
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
725 730 735
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
740 745 750
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 144
<211> 219
<212> PRT
<213> 人工(Artificial)
<220>
<223> 维多珠单抗-LC
<400> 144
Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Ala Lys Ser
20 25 30
Tyr Gly Asn Thr Tyr Leu Ser Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Gly Ile Ser Asn Arg Phe Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Leu Gln Gly
85 90 95
Thr His Gln Pro Tyr Thr 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> 145
<211> 766
<212> PRT
<213> 人工(Artificial)
<220>
<223> 修美乐-IL-2-VitoKine-HC
<400> 145
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 Phe Asp Asp Tyr
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Thr Trp Asn Ser Gly His Ile Asp Tyr Ala Asp Ser Val
50 55 60
Glu Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Val Ser Tyr Leu Ser Thr Ala Ser Ser Leu 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 Asn 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 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365
Leu Thr 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 Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
450 455 460
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Thr Leu Gln
465 470 475 480
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
485 490 495
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
500 505 510
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
515 520 525
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
530 535 540
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
545 550 555 560
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
565 570 575
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
580 585 590
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
595 600 605
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
610 615 620
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
625 630 635 640
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
645 650 655
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
660 665 670
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
675 680 685
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
690 695 700
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
705 710 715 720
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
725 730 735
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
740 745 750
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 146
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 修美乐-LC
<400> 146
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Thr Leu Gln 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 Val Ala Thr Tyr Tyr Cys Gln Arg Tyr Asn Arg Ala Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 147
<211> 70
<212> PRT
<213> 人工(Artificial)
<220>
<223> IL-2α结构域调换的Sushi
<400> 147
Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg
1 5 10 15
Ile Tyr His Phe Val Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys
20 25 30
Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys
35 40 45
Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr
50 55 60
Ser Ser Ala Thr Arg Asn
65 70
<210> 148
<211> 350
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole-Fc-IL-15 2
<400> 148
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val Asn
225 230 235 240
Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His
245 250 255
Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys
260 265 270
Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu
275 280 285
Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile
290 295 300
Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly
305 310 315 320
Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu
325 330 335
Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
340 345 350
<210> 149
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole-Fc-IL-15 3
<400> 149
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 150
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0420 (IL-2 Treg-偏好性(biased)变体Fc VitoKine)
<400> 150
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Arg Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 151
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0421 (IL-2 Treg-偏好性变体Fc VitoKine)
<400> 151
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Thr Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 152
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0423 (IL-2 Treg-偏好性变体Fc VitoKine)
<400> 152
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Glu Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 153
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0424 (IL-2 Treg-偏好性变体Fc VitoKine)
<400> 153
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Asn Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Glu Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 154
<211> 542
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0425 (IL-2 Treg-偏好性变体Fc VitoKine)
<400> 154
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Arg Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ser Glu Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Pro Leu Gly Met Leu Ser Gln Ser Glu Leu Cys Asp Asp Asp Pro
370 375 380
Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly
385 390 395 400
Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Glu Ser
405 410 415
Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp
420 425 430
Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys
435 440 445
Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu
450 455 460
Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His
465 470 475 480
Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr
485 490 495
His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr
500 505 510
Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His
515 520 525
Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
530 535 540
<210> 155
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0426 (IL-2 Treg-偏好性变体Fc VitoKine)
<400> 155
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu His Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Glu Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Glu Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 156
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有减少/消除的效应子功能和延长的半衰期的人类IgG1-Fc
<400> 156
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr
20 25 30
Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 157
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 157
Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala
1 5 10
<210> 158
<211> 7
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 158
Gly Pro Thr Asn Lys Val Arg
1 5
<210> 159
<211> 8
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶底物肽序列
<400> 159
Arg Gln Ala Arg Ala Val Gly Gly
1 5
<210> 160
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 160
Gly Gly Pro Thr Asn Lys Val Arg Gly Ser
1 5 10
<210> 161
<211> 10
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 161
Gly Arg Gln Ala Arg Ala Val Gly Gly Ser
1 5 10
<210> 162
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0660 (IL-15 Fc VitoKine)
<400> 162
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Ser Ser Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly
225 230 235 240
Thr Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 163
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0488 (IL-15 Fc VitoKine)
<400> 163
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 164
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0489 (IL-15 Fc VitoKine)
<400> 164
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Thr Asn Lys Val Arg Gly Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 165
<211> 441
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0661 (IL-15 Fc VitoKine)
<400> 165
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Pro Thr Asn Lys Val Arg Gly Gly Ser
225 230 235 240
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
245 250 255
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
260 265 270
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
275 280 285
Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val Glu
290 295 300
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
305 310 315 320
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
325 330 335
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
340 345 350
Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro
355 360 365
Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser
370 375 380
Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys
385 390 395 400
Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn
405 410 415
Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala
420 425 430
Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 166
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有减少/消除的效应子功能和延长的体内半衰期的人类IgG1-Fc
<400> 166
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 167
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有延长的体内半衰期的人类IgG1 Knob-Fc
<400> 167
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 168
<211> 226
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有延长的体内半衰期的人类IgG1 Hole-Fc
<400> 168
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly
225
<210> 169
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0650 (IL-15 Fc VitoKine)
<400> 169
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 170
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0651 (IL-15 Fc VitoKine)
<400> 170
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 171
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0662 (IL-15 Fc VitoKine) Hole链
<400> 171
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
370 375 380
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys
385 390 395 400
Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
405 410 415
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
420 425 430
Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440 445
<210> 172
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有延长的半衰期的P-0663 (IL-15 Fc VitoKine) Hole链
<400> 172
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu
245 250 255
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
260 265 270
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
275 280 285
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
290 295 300
Asp Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
305 310 315 320
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
325 330 335
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
340 345 350
Ile Val Gln Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu
355 360 365
Ser Gln Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp
370 375 380
Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys
385 390 395 400
Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys
405 410 415
Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu
420 425 430
Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440 445
<210> 173
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有延长的半衰期的P-0664 (IL-15 Fc VitoKine) Hole链
<400> 173
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 174
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> 具有延长的半衰期的P-0665 (IL-15 Fc VitoKine) Hole链
<400> 174
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Ser Ser Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly
225 230 235 240
Thr Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 175
<211> 299
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0156链1
<400> 175
Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val
1 5 10 15
Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly
20 25 30
Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn
35 40 45
Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile
50 55 60
Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro Gly Cys Pro
65 70 75 80
Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe
85 90 95
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
100 105 110
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
115 120 125
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
130 135 140
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
145 150 155 160
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
165 170 175
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
180 185 190
Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg
195 200 205
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly
210 215 220
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
225 230 235 240
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
245 250 255
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
260 265 270
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
275 280 285
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
290 295
<210> 176
<211> 336
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0156链2
<400> 176
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro
115 120 125
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
130 135 140
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
145 150 155 160
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
165 170 175
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
180 185 190
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
195 200 205
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
210 215 220
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
225 230 235 240
Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser
245 250 255
Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
260 265 270
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
275 280 285
Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys
290 295 300
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
305 310 315 320
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
325 330 335
<210> 177
<211> 350
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基准链1
<400> 177
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asn Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Gln
50 55 60
Asp Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser Gly Gly Gly Gly Ser Glu Pro Lys Ser Ser Asp Lys Thr His
115 120 125
Thr Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe
130 135 140
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
145 150 155 160
Glu Val Thr Cys Val Val Val Asp Val Lys His Glu Asp Pro Glu Val
165 170 175
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
180 185 190
Lys Pro Arg Glu Glu Glu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
195 200 205
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
210 215 220
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
225 230 235 240
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
245 250 255
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Asp Val
260 265 270
Ser Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asp Gly
275 280 285
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
290 295 300
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
305 310 315 320
Glu Gln Gly Asp Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His
325 330 335
Ser His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
340 345 350
<210> 178
<211> 301
<212> PRT
<213> 人工(Artificial)
<220>
<223> 基准链2
<400> 178
Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val
1 5 10 15
Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly
20 25 30
Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn
35 40 45
Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile
50 55 60
Arg Gly Gly Gly Gly Ser Glu Pro Lys Ser Ser Asp Lys Thr His Thr
65 70 75 80
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu
85 90 95
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
100 105 110
Val Thr Cys Val Val Val Asp Val Lys His Glu Asp Pro Glu Val Lys
115 120 125
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
130 135 140
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
145 150 155 160
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
165 170 175
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
180 185 190
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
195 200 205
Arg Glu Gln Met Thr Lys Asn Gln Val Lys Leu Thr Cys Leu Val Lys
210 215 220
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
225 230 235 240
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
245 250 255
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
260 265 270
Gln Gly Asn Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ser
275 280 285
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
290 295 300
<210> 179
<211> 465
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0321 (IL-2 Fc VitoKine)
<400> 179
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn
225 230 235 240
Ala Ile Leu Glu Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys
245 250 255
Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu
260 265 270
Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr
275 280 285
Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln
290 295 300
Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala
305 310 315 320
Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile
325 330 335
Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys
340 345 350
Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp
355 360 365
Ile Thr Phe Ser Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Gly Gly
370 375 380
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly His Cys Arg Glu
385 390 395 400
Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val
405 410 415
Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg
420 425 430
Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser
435 440 445
His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg
450 455 460
Asn
465
<210> 180
<211> 669
<212> PRT
<213> 人工(Artificial)
<220>
<223> PD-L1阻断抗体-IL-15-VitoKine-HC
<400> 180
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 Thr Phe Ser Asp Ser
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly 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 Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly
435 440 445
Gly Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu
450 455 460
Glu Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile
465 470 475 480
Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu
485 490 495
Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu
500 505 510
Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Asp Ile His
515 520 525
Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser
530 535 540
Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu
545 550 555 560
Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln
565 570 575
Met Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser
580 585 590
Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val
595 600 605
Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly
610 615 620
Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn
625 630 635 640
Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile
645 650 655
Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
660 665
<210> 181
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> PD-L1阻断抗体-LC
<400> 181
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr 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 Gln Tyr Leu Tyr His Pro Ala
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 182
<211> 110
<212> PRT
<213> 人工(Artificial)
<220>
<223> 缺失111-114的人类IL-15变体多肽
<400> 182
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe
100 105 110
<210> 183
<211> 108
<212> PRT
<213> 人工(Artificial)
<220>
<223> 缺失109-114的人类IL-15变体多肽
<400> 183
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln
100 105
<210> 184
<211> 107
<212> PRT
<213> 人工(Artificial)
<220>
<223> 缺失108-114的人类IL-15变体多肽
<400> 184
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val
100 105
<210> 185
<211> 116
<212> PRT
<213> 人工(Artificial)
<220>
<223> N95后插入'GS'的人类IL-15变体多肽
<400> 185
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Gly
85 90 95
Ser Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe
100 105 110
Ile Asn Thr Ser
115
<210> 186
<211> 119
<212> PRT
<213> 人工(Artificial)
<220>
<223> N95后插入'GGSGG'的人类IL-15变体多肽
<400> 186
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Gly
85 90 95
Gly Ser Gly Gly Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val
100 105 110
Gln Met Phe Ile Asn Thr Ser
115
<210> 187
<211> 113
<212> PRT
<213> 人工(Artificial)
<220>
<223> 在N末端处具有1个氨基酸缺失的人类IL-15变体
<400> 187
Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln
1 5 10 15
Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro
20 25 30
Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val
35 40 45
Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn
50 55 60
Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr
65 70 75 80
Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys
85 90 95
Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr
100 105 110
Ser
<210> 188
<211> 112
<212> PRT
<213> 人工(Artificial)
<220>
<223> 在N末端处具有2个氨基酸缺失的人类IL-15变体
<400> 188
Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser
1 5 10 15
Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser
20 25 30
Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile
35 40 45
Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu
50 55 60
Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu
65 70 75 80
Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu
85 90 95
Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
100 105 110
<210> 189
<211> 111
<212> PRT
<213> 人工(Artificial)
<220>
<223> 在N末端处具有3个氨基酸缺失的人类IL-15变体
<400> 189
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
1 5 10 15
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
20 25 30
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
35 40 45
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile
50 55 60
Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
65 70 75 80
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
85 90 95
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
100 105 110
<210> 190
<211> 110
<212> PRT
<213> 人工(Artificial)
<220>
<223> 在N末端处具有4个氨基酸缺失的人类IL-15变体
<400> 190
Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His
1 5 10 15
Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys
20 25 30
Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu
35 40 45
Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile
50 55 60
Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly
65 70 75 80
Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu
85 90 95
Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
100 105 110
<210> 191
<211> 112
<212> PRT
<213> 人工(Artificial)
<220>
<223> 在N末端处具有2个氨基酸缺失的人类IL-15 V63A变体
<400> 191
Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser
1 5 10 15
Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser
20 25 30
Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile
35 40 45
Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu Asn Leu
50 55 60
Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu
65 70 75 80
Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu
85 90 95
Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
100 105 110
<210> 192
<211> 112
<212> PRT
<213> 人工(Artificial)
<220>
<223> 在N末端处具有2个氨基酸缺失的人类IL-15 I68H变体
<400> 192
Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser
1 5 10 15
Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser
20 25 30
Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile
35 40 45
Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu
50 55 60
Ile His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu
65 70 75 80
Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu
85 90 95
Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
100 105 110
<210> 193
<211> 449
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化抗FAP抗体重链
<400> 193
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Asn
20 25 30
Ile Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Trp Phe His Pro Gly Ser Gly Ser Ile Lys Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Val 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 His Gly Gly Thr Gly Arg Gly Ala Met 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 Ala Ala Gly Ala
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 Asn Ser Thr Tyr Arg
290 295 300
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
305 310 315 320
Glu Tyr Lys Cys Lys Val Ser Asn Lys 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 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
355 360 365
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
385 390 395 400
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415
Lys Ser Arg Trp Gln 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
<210> 194
<211> 218
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化抗FAP抗体κ轻链
<400> 194
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Arg Ser Ile Ser Thr Ser
20 25 30
Ala Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ser
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
65 70 75 80
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Ser Arg
85 90 95
Glu Leu Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105 110
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 195
<211> 446
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体重链
<400> 195
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 196
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体κ轻链
<400> 196
Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
1 5 10 15
Glu Pro Ala Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Glu Thr Val
20 25 30
Val Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Arg Leu Leu Ile
35 40 45
Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala
65 70 75 80
Glu Asp Val Gly Val Tyr Tyr Cys Gln Gln Tyr Ser Arg Tyr Pro Trp
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 197
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类PD-1拮抗剂抗体重链
<400> 197
Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Arg Phe Thr Ser Tyr
20 25 30
Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu
50 55 60
Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Asn Thr Ala Tyr
65 70 75 80
Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Ala Asp Tyr Ser Ser Gly Ser Gly Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
115 120 125
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
145 150 155 160
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
165 170 175
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
180 185 190
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
195 200 205
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
210 215 220
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro
225 230 235 240
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
290 295 300
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
305 310 315 320
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
325 330 335
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
355 360 365
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
385 390 395 400
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
405 410 415
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
420 425 430
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 198
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类PD-1拮抗剂抗体λ轻链
<400> 198
Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser Pro Gly Gln
1 5 10 15
Thr Ala Arg Ile Thr Cys Ser Gly Asp Ala Leu Pro Lys Gln Tyr Ala
20 25 30
Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Met Val Ile Tyr
35 40 45
Lys Asp Thr Glu Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser
50 55 60
Ser Ser Gly Thr Lys Val Thr Leu Thr Ile Ser Gly Val Gln Ala Glu
65 70 75 80
Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Ala Asp Asn Ser Ile Thr Tyr
85 90 95
Arg Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gln Pro Lys
100 105 110
Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln
115 120 125
Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly
130 135 140
Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly
145 150 155 160
Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala
165 170 175
Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser
180 185 190
Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val
195 200 205
Ala Pro Thr Glu Cys Ser
210
<210> 199
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 V63A变体多肽
<400> 199
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 200
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 V63K变体多肽
<400> 200
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Lys Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 201
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 V63R变体多肽
<400> 201
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Arg Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 202
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 I68H变体多肽
<400> 202
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 203
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 I68F变体多肽
<400> 203
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Phe Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 204
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 I68Q变体多肽
<400> 204
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Gln Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 205
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 I68G变体多肽
<400> 205
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Gly Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 206
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 I68K变体多肽
<400> 206
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Lys Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 207
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 I68D变体多肽
<400> 207
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Asp Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 208
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 Q108S变体多肽
<400> 208
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Ser Met Phe Ile Asn
100 105 110
Thr Ser
<210> 209
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 Q108A变体多肽
<400> 209
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Ala Met Phe Ile Asn
100 105 110
Thr Ser
<210> 210
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 Q108E变体多肽
<400> 210
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Glu Met Phe Ile Asn
100 105 110
Thr Ser
<210> 211
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 Q108K变体多肽
<400> 211
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Lys Met Phe Ile Asn
100 105 110
Thr Ser
<210> 212
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 Q108M变体多肽
<400> 212
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Met Met Phe Ile Asn
100 105 110
Thr Ser
<210> 213
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 V63A/I68H变体多肽
<400> 213
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu
50 55 60
Asn Leu Ile His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 214
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 V63A/I68Q变体多肽
<400> 214
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu
50 55 60
Asn Leu Ile Gln Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 215
<211> 114
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-15 V63A/I68G变体多肽
<400> 215
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu
50 55 60
Asn Leu Ile Gly Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 216
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 V63K链
<400> 216
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Lys
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 217
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 V63R链
<400> 217
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Arg
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 218
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 V63A链
<400> 218
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 219
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 I68H链
<400> 219
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 220
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 I68F链
<400> 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Phe Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 221
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 I68Q链
<400> 221
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Gln Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 222
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 I68G链
<400> 222
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Gly Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 223
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 I68K链
<400> 223
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Lys Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 224
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 I68D链
<400> 224
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Asp Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 225
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole Fc-IL-15 Q108S链
<400> 225
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Ser Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 226
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Hole Fc-IL-15 Q108A链
<400> 226
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser
225 230 235 240
Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Ala Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 227
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 V63A/I68H链
<400> 227
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala
290 295 300
Glu Asn Leu Ile His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 228
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 V63A/I68Q链
<400> 228
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala
290 295 300
Glu Asn Leu Ile Gln Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 229
<211> 355
<212> PRT
<213> 人工(Artificial)
<220>
<223> Fc-IL-15 V63A/I68G链
<400> 229
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala
290 295 300
Glu Asn Leu Ile Gly Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser
355
<210> 230
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0682 (IL-15 Fc VitoKine)
<400> 230
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val
290 295 300
Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Ser Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 231
<211> 442
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0806 (IL-15 Fc VitoKine)
<400> 231
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
225 230 235 240
Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu
245 250 255
Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val
260 265 270
His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu
275 280 285
Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala
290 295 300
Glu Asn Leu Ile His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn
305 310 315 320
Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn
325 330 335
Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile
340 345 350
Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys
355 360 365
Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr
370 375 380
Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg
385 390 395 400
Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr
405 410 415
Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro
420 425 430
Ala Leu Val His Gln Arg Pro Ala Pro Pro
435 440
<210> 232
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 R38A/S125I变体多肽
<400> 232
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Ala Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 233
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 F42A/S125I变体多肽
<400> 233
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 234
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 E62A/S125I变体多肽
<400> 234
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Ala Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 235
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 E62F/S125I变体多肽
<400> 235
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Phe Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 236
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 E62H/S125I变体多肽
<400> 236
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu His Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 237
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 E62L/S125I变体多肽
<400> 237
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Leu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 238
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 P65H/S125I变体多肽
<400> 238
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
His Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 239
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 P65N/S125I变体多肽
<400> 239
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Asn Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 240
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 P65Q/S125I变体多肽
<400> 240
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Gln Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 241
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 P65R/S125I变体多肽
<400> 241
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Arg Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 242
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 P65K/S125I变体多肽
<400> 242
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Lys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 243
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 F42A/E62F/S125I变体多肽
<400> 243
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Phe Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 244
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 F42A/E62A/S125I变体多肽
<400> 244
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Ala Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 245
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 F42A/E62H/S125I变体多肽
<400> 245
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu His Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 246
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 F42A/P65H/S125I变体多肽
<400> 246
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
His Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 247
<211> 133
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2 F42A/P65R/S125I变体多肽
<400> 247
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Arg Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 248
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0531 (IL-2 Fc融合体)
<400> 248
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 249
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0689 (IL-2 Fc融合体knob链)
<400> 249
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 250
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0602 (IL-2 Fc融合体)
<400> 250
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Ala
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 251
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0613 (IL-2 Fc融合体)
<400> 251
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 252
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0624 (IL-2 Fc融合体)
<400> 252
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Ala Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 253
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0625 (IL-2 Fc融合体)
<400> 253
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Phe Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 254
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0626 (IL-2 Fc融合体)
<400> 254
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu His Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 255
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0627 (IL-2 Fc融合体)
<400> 255
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Leu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 256
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0634 (IL-2 Fc融合体)
<400> 256
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys His Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 257
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0708 (IL-2 Fc融合体knob链)
<400> 257
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Asn Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 258
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0709 (IL-2 Fc融合体knob链)
<400> 258
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Gln Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 259
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0635 (IL-2 Fc融合体)
<400> 259
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
130 135 140
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 260
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0704 (IL-2 Fc融合体knob链)
<400> 260
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 261
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0707 (IL-2 Fc融合体knob链)
<400> 261
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Lys Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 262
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0702 (IL-2 Fc融合体knob链)
<400> 262
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Phe Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 263
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0766 (IL-2 Fc融合体knob链)
<400> 263
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Ala Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 264
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0767 (IL-2 Fc融合体knob链)
<400> 264
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu His Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 265
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0703 (IL-2 Fc融合体knob链)
<400> 265
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys His Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 266
<211> 367
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0705 (IL-2 Fc融合体knob链)
<400> 266
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Ala Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr
355 360 365
<210> 267
<211> 165
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2α Sushi Y43A变体
<400> 267
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Ala Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 268
<211> 165
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2α Sushi L42G变体
<400> 268
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Gly Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 269
<211> 165
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2α Sushi R36A变体
<400> 269
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Ala Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 270
<211> 165
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类IL-2α Sushi K38E变体
<400> 270
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Glu Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 271
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0754 (IL-2 Fc VitoKine knob链)
<400> 271
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Leu Ala Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 272
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0755 (IL-2 Fc VitoKine knob链)
<400> 272
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Gly Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 273
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0756 (IL-2 Fc VitoKine knob链)
<400> 273
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Ala Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 274
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0745 (IL-2 Fc VitoKine knob链)
<400> 274
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 275
<211> 447
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体-HC
<400> 275
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 Ala Ser Thr Lys Gly Pro Ser Val
115 120 125
Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 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 Lys Thr Tyr Thr Cys Asn Val Asp His Lys
195 200 205
Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro
210 215 220
Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu
260 265 270
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
435 440 445
<210> 276
<211> 218
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体-LC
<400> 276
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
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
115 120 125
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
145 150 155 160
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
180 185 190
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
195 200 205
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 277
<211> 440
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类PD-1拮抗剂抗体-HC
<400> 277
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 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
115 120 125
Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
130 135 140
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
145 150 155 160
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
165 170 175
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys
180 185 190
Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp
195 200 205
Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala
210 215 220
Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
225 230 235 240
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
245 250 255
Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val
260 265 270
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
275 280 285
Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
290 295 300
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly
305 310 315 320
Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
325 330 335
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr
340 345 350
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
355 360 365
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
370 375 380
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
385 390 395 400
Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe
405 410 415
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
420 425 430
Ser Leu Ser Leu Ser Leu Gly Lys
435 440
<210> 278
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人类PD-1拮抗剂抗体-LC
<400> 278
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 Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 279
<211> 448
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-L1拮抗剂抗体-HC
<400> 279
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 Thr Phe Ser Asp Ser
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly 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 Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val
290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
305 310 315 320
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr
325 330 335
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
355 360 365
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
385 390 395 400
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
405 410 415
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
420 425 430
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 280
<211> 214
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-L1拮抗剂抗体-LC
<400> 280
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr 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 Gln Tyr Leu Tyr His Pro Ala
85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 281
<211> 662
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC knob链
<400> 281
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile
515 520 525
Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Ser Met Phe Ile Asn Thr Ser Gly
565 570 575
Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met
580 585 590
Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser
595 600 605
Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr
610 615 620
Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His
625 630 635 640
Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His
645 650 655
Gln Arg Pro Ala Pro Pro
660
<210> 282
<211> 662
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC knob链
<400> 282
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu Asn Leu Ile
515 520 525
Gly Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly
565 570 575
Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met
580 585 590
Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser
595 600 605
Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr
610 615 620
Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His
625 630 635 640
Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His
645 650 655
Gln Arg Pro Ala Pro Pro
660
<210> 283
<211> 662
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC knob链
<400> 283
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile
515 520 525
Gly Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly
565 570 575
Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met
580 585 590
Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser
595 600 605
Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr
610 615 620
Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His
625 630 635 640
Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His
645 650 655
Gln Arg Pro Ala Pro Pro
660
<210> 284
<211> 662
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC
<400> 284
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu Asn Leu Ile
515 520 525
His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly
565 570 575
Gly Pro Leu Gly Met Leu Ser Gln Ser Ile Thr Cys Pro Pro Pro Met
580 585 590
Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser
595 600 605
Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr
610 615 620
Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala His
625 630 635 640
Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His
645 650 655
Gln Arg Pro Ala Pro Pro
660
<210> 285
<211> 446
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体-HC-Hole链
<400> 285
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 286
<211> 767
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 VitoKine HC knob链
<400> 286
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr
450 455 460
Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn
465 470 475 480
Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe
485 490 495
Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys
500 505 510
Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu Asn Leu Ala Gln
515 520 525
Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn
530 535 540
Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu
545 550 555 560
Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile
565 570 575
Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Ser Gly Pro
580 585 590
Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp
595 600 605
Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu
610 615 620
Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys
625 630 635 640
Ser Gly Ser Gly Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser
645 650 655
Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr
660 665 670
Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr
675 680 685
Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly
690 695 700
His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile
705 710 715 720
Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly
725 730 735
Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr
740 745 750
His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 287
<211> 767
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 VitoKine HC knob链
<400> 287
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr
450 455 460
Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn
465 470 475 480
Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe
485 490 495
Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys
500 505 510
Leu Glu Glu Glu Leu Lys Gln Leu Glu Glu Val Leu Asn Leu Ala Gln
515 520 525
Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn
530 535 540
Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu
545 550 555 560
Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile
565 570 575
Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Ser Gly Pro
580 585 590
Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp
595 600 605
Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu
610 615 620
Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Ala Ile Lys
625 630 635 640
Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser
645 650 655
Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr
660 665 670
Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr
675 680 685
Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly
690 695 700
His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile
705 710 715 720
Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly
725 730 735
Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr
740 745 750
His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 288
<211> 767
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 VitoKine HC knob链
<400> 288
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr
450 455 460
Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn
465 470 475 480
Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe
485 490 495
Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys
500 505 510
Leu Glu Glu Glu Leu Lys Gln Leu Glu Glu Val Leu Asn Leu Ala Gln
515 520 525
Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn
530 535 540
Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu
545 550 555 560
Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile
565 570 575
Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Ser Gly Pro
580 585 590
Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp
595 600 605
Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu
610 615 620
Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys
625 630 635 640
Ser Gly Ser Gly Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser
645 650 655
Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr
660 665 670
Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr
675 680 685
Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly
690 695 700
His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile
705 710 715 720
Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly
725 730 735
Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr
740 745 750
His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 289
<211> 767
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 VitoKine HC
<400> 289
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr
450 455 460
Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn
465 470 475 480
Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe
485 490 495
Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys
500 505 510
Leu Glu Glu Glu Leu Lys Gln Leu Glu Glu Val Leu Asn Leu Ala Gln
515 520 525
Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn
530 535 540
Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu
545 550 555 560
Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile
565 570 575
Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly Gly Ser Gly Pro
580 585 590
Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu Cys Asp Asp Asp
595 600 605
Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu
610 615 620
Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys
625 630 635 640
Ser Gly Ser Gly Tyr Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser
645 650 655
Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr
660 665 670
Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr
675 680 685
Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly
690 695 700
His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile
705 710 715 720
Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly
725 730 735
Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr
740 745 750
His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly
755 760 765
<210> 290
<211> 774
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 VitoKine HC
<400> 290
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly Gly Ala Pro Thr
450 455 460
Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu
465 470 475 480
Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys
485 490 495
Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr
500 505 510
Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Gln Leu Glu
515 520 525
Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg
530 535 540
Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser
545 550 555 560
Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val
565 570 575
Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr
580 585 590
Leu Thr Gly Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly
595 600 605
Ser Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe
610 615 620
Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys
625 630 635 640
Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Gly Tyr Met Leu Cys Thr
645 650 655
Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser
660 665 670
Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu
675 680 685
Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val
690 695 700
Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu
705 710 715 720
Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val
725 730 735
Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala
740 745 750
Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro
755 760 765
Gln Leu Ile Cys Thr Gly
770
<210> 291
<211> 774
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 VitoKine HC
<400> 291
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Ala Pro Thr
450 455 460
Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu
465 470 475 480
Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys
485 490 495
Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr
500 505 510
Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Gln Leu Glu
515 520 525
Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg
530 535 540
Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser
545 550 555 560
Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val
565 570 575
Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr
580 585 590
Leu Thr Gly Gly Gly Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly
595 600 605
Gly Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe
610 615 620
Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys
625 630 635 640
Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Gly Tyr Met Leu Cys Thr
645 650 655
Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser
660 665 670
Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu
675 680 685
Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val
690 695 700
Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu
705 710 715 720
Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val
725 730 735
Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala
740 745 750
Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro
755 760 765
Gln Leu Ile Cys Thr Gly
770
<210> 292
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0701 (IL-2 Fc VitoKine knob链)
<400> 292
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 293
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0807 (IL-2 Fc VitoKine knob链)
<400> 293
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Leu Ala Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 294
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0708 (IL-2 Fc VitoKine knob链)
<400> 294
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Arg Ile Lys Ser Gly Ser Gly Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 295
<211> 547
<212> PRT
<213> 人工(Artificial)
<220>
<223> P-0809 (IL-2 Fc VitoKine knob链)
<400> 295
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly
1 5 10 15
Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
20 25 30
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
35 40 45
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val
50 55 60
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
65 70 75 80
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
85 90 95
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
100 105 110
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
115 120 125
Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
130 135 140
Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
145 150 155 160
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
165 170 175
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
180 185 190
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
195 200 205
His Glu Ala Leu His Ala His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
210 215 220
Pro Gly Gly Gly Gly Ser Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser
225 230 235 240
Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu Asp Leu Gln
245 250 255
Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg
260 265 270
Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys
275 280 285
His Leu Gln Cys Leu Glu Glu Glu Leu Lys Arg Leu Glu Glu Val Leu
290 295 300
Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu Ile
305 310 315 320
Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser Glu Thr Thr
325 330 335
Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val Glu Phe Leu
340 345 350
Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr Leu Thr Gly
355 360 365
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Glu Leu
370 375 380
Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys Ala Met
385 390 395 400
Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg Gly Phe
405 410 415
Arg Ala Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly Asn Ser
420 425 430
Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser Ala Thr
435 440 445
Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln Lys Glu
450 455 460
Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp Gln Ala
465 470 475 480
Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn Glu Ala
485 490 495
Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr Tyr Gln
500 505 510
Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu Ser Val
515 520 525
Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln Leu Ile
530 535 540
Cys Thr Gly
545
<210> 296
<211> 594
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-2 HC knob链
<400> 296
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro
340 345 350
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr
450 455 460
Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His Leu Leu Leu
465 470 475 480
Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys Asn Pro Lys
485 490 495
Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr
500 505 510
Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys Gln Leu Glu
515 520 525
Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu Arg Pro Arg
530 535 540
Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu Lys Gly Ser
545 550 555 560
Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr Ile Val
565 570 575
Glu Phe Leu Asn Arg Trp Ile Thr Phe Ile Gln Ser Ile Ile Ser Thr
580 585 590
Leu Thr
<210> 297
<211> 575
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 HC
<400> 297
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Ala Glu Asn Leu Ile
515 520 525
His Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser
565 570 575
<210> 298
<211> 15
<212> PRT
<213> 人工(Artificial)
<220>
<223> 蛋白酶可裂解的接头序列
<400> 298
Gly Gly Gly Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly Gly
1 5 10 15
<210> 299
<211> 444
<212> PRT
<213> 人工(Artificial)
<220>
<223> 替代小鼠PD-1拮抗剂抗体HC链
<400> 299
Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Ser Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Ser
20 25 30
Tyr Arg Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Arg Leu Glu Trp
35 40 45
Met Gly Tyr Ile Asn Ser Ala Gly Ile Ser Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Arg Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe
65 70 75 80
Leu Gln Val Asn Ser Val Thr Thr Glu Asp Ala Ala Thr Tyr Tyr Cys
85 90 95
Ala Arg Ser Asp Asn Met Gly Thr Thr Pro Phe Thr Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val
115 120 125
Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr
130 135 140
Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr
145 150 155 160
Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser
180 185 190
Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala
195 200 205
Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys
210 215 220
Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val
245 250 255
Thr Cys Val Val Val Ala Ile Ser Lys Asp Asp Pro Glu Val Gln Phe
260 265 270
Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Lys Pro
275 280 285
Arg Glu Glu Gln Ile Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro
290 295 300
Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val
305 310 315 320
Asn Ser Ala Ala Phe Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr
325 330 335
Lys Gly Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro
340 345 350
Lys Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr
355 360 365
Asn Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln
370 375 380
Pro Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly
385 390 395 400
Ser Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu
405 410 415
Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn
420 425 430
His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly
435 440
<210> 300
<211> 443
<212> PRT
<213> 人工(Artificial)
<220>
<223> 替代小鼠PD-1拮抗剂抗体异源二聚体HC链1
<400> 300
Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Ser Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Ser
20 25 30
Tyr Arg Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Arg Leu Glu Trp
35 40 45
Met Gly Tyr Ile Asn Ser Ala Gly Ile Ser Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Arg Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe
65 70 75 80
Leu Gln Val Asn Ser Val Thr Thr Glu Asp Ala Ala Thr Tyr Tyr Cys
85 90 95
Ala Arg Ser Asp Asn Met Gly Thr Thr Pro Phe Thr Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val
115 120 125
Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr
130 135 140
Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr
145 150 155 160
Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser
180 185 190
Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala
195 200 205
Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys
210 215 220
Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val
245 250 255
Thr Cys Val Val Val Ala Ile Ser Lys Asp Asp Pro Glu Val Gln Phe
260 265 270
Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Lys Pro
275 280 285
Arg Glu Glu Gln Ile Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro
290 295 300
Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val
305 310 315 320
Asn Ser Ala Ala Phe Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr
325 330 335
Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys
340 345 350
Lys Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asn
355 360 365
Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro
370 375 380
Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Lys Thr Asp Gly Ser
385 390 395 400
Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala
405 410 415
Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His
420 425 430
His Thr Glu Lys Ser Leu Ser His Ser Pro Gly
435 440
<210> 301
<211> 443
<212> PRT
<213> 人工(Artificial)
<220>
<223> 替代小鼠PD-1拮抗剂抗体异源二聚体HC链2
<400> 301
Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln
1 5 10 15
Ser Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Ser
20 25 30
Tyr Arg Trp Asn Trp Ile Arg Lys Phe Pro Gly Asn Arg Leu Glu Trp
35 40 45
Met Gly Tyr Ile Asn Ser Ala Gly Ile Ser Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Arg Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe
65 70 75 80
Leu Gln Val Asn Ser Val Thr Thr Glu Asp Ala Ala Thr Tyr Tyr Cys
85 90 95
Ala Arg Ser Asp Asn Met Gly Thr Thr Pro Phe Thr Tyr Trp Gly Gln
100 105 110
Gly Thr Leu Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val
115 120 125
Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr
130 135 140
Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr
145 150 155 160
Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val
165 170 175
Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser
180 185 190
Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala
195 200 205
Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys
210 215 220
Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe
225 230 235 240
Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val
245 250 255
Thr Cys Val Val Val Ala Ile Ser Lys Asp Asp Pro Glu Val Gln Phe
260 265 270
Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Lys Pro
275 280 285
Arg Glu Glu Gln Ile Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro
290 295 300
Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val
305 310 315 320
Asn Ser Ala Ala Phe Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr
325 330 335
Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys
340 345 350
Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asn
355 360 365
Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro
370 375 380
Ala Glu Asn Tyr Asp Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser
385 390 395 400
Tyr Phe Val Tyr Ser Asp Leu Asn Val Gln Lys Ser Asn Trp Glu Ala
405 410 415
Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His
420 425 430
His Thr Glu Lys Ser Leu Ser His Ser Pro Gly
435 440
<210> 302
<211> 218
<212> PRT
<213> 人工(Artificial)
<220>
<223> 替代小鼠PD-1拮抗剂抗体LC
<400> 302
Asp Ile Val Met Thr Gln Gly Thr Leu Pro Asn Pro Val Pro Ser Gly
1 5 10 15
Glu Ser Val Ser Ile Thr Cys Arg Ser Ser Lys Ser Leu Leu Tyr Ser
20 25 30
Asp Gly Lys Thr Tyr Leu Asn Trp Tyr Leu Gln Arg Pro Gly Gln Ser
35 40 45
Pro Gln Leu Leu Ile Tyr Trp Met Ser Thr Arg Ala Ser Gly Val Ser
50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
65 70 75 80
Ser Gly Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Gln Gln Gly
85 90 95
Leu Glu Phe Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys Arg
100 105 110
Thr Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln
115 120 125
Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr
130 135 140
Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln
145 150 155 160
Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr
165 170 175
Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg
180 185 190
His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro
195 200 205
Ile Val Lys Ser Phe Asn Arg Asn Glu Cys
210 215
<210> 303
<211> 668
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC
<400> 303
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu
450 455 460
Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu
465 470 475 480
Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser
485 490 495
Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu
500 505 510
Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp
515 520 525
Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn
530 535 540
Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu
545 550 555 560
Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met
565 570 575
Phe Ile Asn Thr Ser Gly Gly Pro Leu Gly Met Leu Ser Gln Ser Ile
580 585 590
Thr Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys
595 600 605
Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe
610 615 620
Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys
625 630 635 640
Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg
645 650 655
Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
660 665
<210> 304
<211> 673
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC
<400> 304
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Leu Gly Gly Ser Gly Arg Ser Ala Asn Ala Ile Leu Glu
450 455 460
Gly Gly Ser Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu
465 470 475 480
Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser
485 490 495
Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu
500 505 510
Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp
515 520 525
Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn
530 535 540
Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu
545 550 555 560
Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met
565 570 575
Phe Ile Asn Thr Ser Gly Gly Ser Gly Pro Leu Gly Met Leu Ser Gln
580 585 590
Gly Gly Gly Ser Ile Thr Cys Pro Pro Pro Met Ser Val Glu His Ala
595 600 605
Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile
610 615 620
Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu
625 630 635 640
Cys Val Leu Asn Lys Ala Thr Asn Val Ala His Trp Thr Thr Pro Ser
645 650 655
Leu Lys Cys Ile Arg Asp Pro Ala Leu Val His Gln Arg Pro Ala Pro
660 665 670
Pro
<210> 305
<211> 667
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC
<400> 305
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Ser Gly Arg Ser Glu Asn Ile Arg Thr Ala Gly Gly Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile
515 520 525
Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly
565 570 575
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Ile Thr
580 585 590
Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser
595 600 605
Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys
610 615 620
Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala
625 630 635 640
Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp
645 650 655
Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
660 665
<210> 306
<211> 667
<212> PRT
<213> 人工(Artificial)
<220>
<223> 人源化PD-1拮抗剂抗体IL-15 VitoKine HC
<400> 306
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Gly Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Ser Pro Asp Ser Ser Gly Val Ala Tyr Trp Gly Gln Gly Thr Leu
100 105 110
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
145 150 155 160
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
195 200 205
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220
Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val
225 230 235 240
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
245 250 255
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
305 310 315 320
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
385 390 395 400
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly
435 440 445
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Trp Val
450 455 460
Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met
465 470 475 480
His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys
485 490 495
Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser
500 505 510
Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile
515 520 525
Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser
530 535 540
Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe
545 550 555 560
Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser Gly
565 570 575
Gly Ser Gly Pro Leu Gly Met Leu Ser Gln Gly Gly Gly Ser Ile Thr
580 585 590
Cys Pro Pro Pro Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser
595 600 605
Tyr Ser Leu Tyr Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys
610 615 620
Arg Lys Ala Gly Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala
625 630 635 640
Thr Asn Val Ala His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp
645 650 655
Pro Ala Leu Val His Gln Arg Pro Ala Pro Pro
660 665

Claims (48)

1.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D1-D2-D3)包含：1)组织或疾病部位靶向部分D1结构域(“D1”)、2)可生物活化部分D2结构域(“D2”)和3)遮蔽部分D3结构域(“D3”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2是选自具有SEQ ID NO:182-192和199-215中列出的氨基酸序列的多肽的组的IL-15变体多肽。
2.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D1-D2-D3)包含：1)双重功能部分D1结构域(“D1”)、2)可生物活化部分D2结构域(“D2”)和3)遮蔽部分D3结构域(“D3”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位和保持在意图的治疗部位处；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2是选自具有SEQ ID NO:182-192和199-215中列出的氨基酸序列的多肽的组的IL-15变体多肽。
3.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D3-D2-D1)包含：1)遮蔽部分D3结构域(“D3”)、2)可生物活化部分D2结构域(“D2”)和3)组织或疾病部位靶向部分D1结构域(“D1”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2是选自具有SEQ ID NO:182-192和199-215中列出的氨基酸序列的多肽的组的IL-15变体多肽。
4.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D3-D2-D1)包含：1)遮蔽部分D3结构域(“D3”)、2)可生物活化部分D2结构域(“D2”)和3)双重功能部分D1结构域(“D1”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位和保持在意图的治疗部位处；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2是选自具有SEQ ID NO:182-192和199-215中列出的氨基酸序列的多肽的组的IL-15变体多肽。
5.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D1-D2-D3)包含：1)组织或疾病部位靶向部分D1结构域(“D1”)、2)可生物活化部分D2结构域(“D2”)和3)遮蔽部分D3结构域(“D3”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2结构域是选自具有SEQ ID NO:232-247中列出的氨基酸序列的多肽的组的IL-2变体多肽。
6.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D1-D2-D3)包含：1)双重功能部分D1结构域(“D1”)、2)可生物活化部分D2结构域(“D2”)和3)遮蔽部分D3结构域(“D3”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位和保持在意图的治疗部位处；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2结构域是选自具有SEQ ID NO:232-247中列出的氨基酸序列的多肽的组的IL-2变体多肽。
7.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D3-D2-D1)包含：1)遮蔽部分D3结构域(“D3”)、2)可生物活化部分D2结构域(“D2”)和3)组织或疾病部位靶向部分D1结构域(“D1”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2结构域是选自具有SEQ ID NO:232-247中列出的氨基酸序列的多肽的组的IL-2变体多肽。
8.一种可生物活化的多肽药物构建体，所述构建体在N末端至C末端方向(D3-D2-D1)包含：1)遮蔽部分D3结构域(“D3”)、2)可生物活化部分D2结构域(“D2”)和3)双重功能部分D1结构域(“D1”)；其中D1功能为将所述可生物活化部分靶向到意图的治疗部位和保持在意图的治疗部位处；其中D3能够遮蔽D2的功能活性，直到D2在所述意图的治疗部位处被活化；并且其中D2结构域是选自具有SEQ ID NO:232-247中列出的氨基酸序列的多肽的组的IL-2变体多肽。
9.根据权利要求1至8中任一项所述的构建体，其中所述构建体的D1结构域、D2结构域和D3结构域各自为单体形式，各自为二聚体形式，或者共同为二聚体和单体的组合形式。
10.根据权利要求1至9中任一项所述的构建体，其中所述D1结构域选自由以下组成的组：能够与肿瘤相关抗原(TAA)或组织特异性抗原或靶、细胞表面分子或细胞外基质蛋白、蛋白酶和任何翻译后修饰残基结合的抗体，或抗体片段、或配体或其变体、或受体或其变体。
11.根据权利要求10所述的构建体，其中所述D1结构域是能够与肿瘤相关抗原(TAA)或免疫检查点调节剂结合的抗体或抗体片段。
12.根据权利要求11所述的构建体，其中所述抗体或抗体片段选自由以下组成的组：PD-1拮抗性抗体；PD-L1拮抗性抗体；CTLA-4拮抗性抗体；CD20拮抗性抗体；Her-2/neu拮抗性抗体；EGFR拮抗性抗体；FAP拮抗性抗体；抗整联蛋白α4β7的抗炎抗体；TNFα拮抗性抗体；和激动性CD40抗体。
13.根据权利要求12所述的构建体，其中所述抗体是拮抗性成纤维细胞活化蛋白(FAP)抗体或抗体片段。
14.根据权利要求13所述的构建体，其中所述抗体是包含SEQ ID NO:193和194中列出的重链和轻链氨基酸序列的人源化抗FAP抗体。
15.根据权利要求12所述的构建体，其中所述抗体是拮抗性程序性死亡-1(PD-1)抗体或抗体片段。
16.根据权利要求15所述的构建体，其中所述抗体是选自包含以下的抗体的拮抗性人源化PD-1抗体：SEQ ID NO:195和196中列出的重链和轻链氨基酸序列；SEQ ID NO:197和198中列出的重链和轻链氨基酸序列；SEQ ID NO:275和276中列出的重链和轻链氨基酸序列；或SEQ ID NO:277和278中列出的重链和轻链氨基酸序列。
17.根据权利要求1至10中任一项所述的构建体，其中所述D1结构域是选自由以下组成的组的Fc结构域：人类IgG1 Fc结构域、人类IgG2 Fc结构域、人类IgG3 Fc结构域、人类IgG4Fc结构域、IgA Fc结构域、IgD Fc结构域、IgE Fc结构域、IgG Fc结构域和IgM Fc结构域。
18.根据权利要求17所述的构建体，其中所述Fc结构域是具有沉默的效应子功能和/或具有半衰期延长功能的Fc结构域。
19.根据权利要求17至18中任一项所述的构建体，其中所述Fc结构域是具有选自由SEQID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:156和SEQ ID NO:166-168中列出的氨基酸序列组成的组的氨基酸序列的Fc结构域。
20.根据权利要求1至19中任一项所述的构建体，其中D2通过选自由蛋白酶可裂解的肽接头和不可裂解的肽接头组成的组的肽接头(“L1”)附接至D1。
21.根据权利要求20所述的构建体，其中所述蛋白酶可裂解的肽接头选自SEQ ID NO:71-96、157-161和298中列出的序列的组。
22.根据权利要求20所述的构建体，其中所述不可裂解的肽接头选自SEQ ID NO:107-127中列出的序列的组。
23.根据权利要求1至22中任一项所述的构建体，其中所述D3结构域选自由以下组成的组：针对D2鉴定并能够遮蔽D2的活性的同源受体/结合配偶体(或其变体)和任何结合配偶体。
24.根据权利要求1至23中任一项所述的构建体，其中所述D3结构域选自由以下组成的组：能够遮蔽D2的活性的蛋白、肽、DNA片段、RNA片段、聚合物、抗体和抗体片段。
25.根据权利要求23所述的构建体，其中所述D3结构域是IL-15的同源受体/结合配偶体(或其变体)，并且包含SEQ ID NO:4中列出的氨基酸序列或其任何功能片段。
26.根据权利要求25所述的构建体，其中所述D3结构域是IL-15的同源受体/结合配偶体(或其变体)，并且包含SEQ ID NO:5中列出的氨基酸序列。
27.根据权利要求26所述的构建体，其中所述D3结构域是IL-2的同源受体/结合配偶体(或其变体)，并且包含SEQ ID NO:10和267-270中列出的氨基酸序列。
28.根据权利要求1至27中任一项所述的构建体，其中D2通过选自由蛋白酶可裂解的肽接头和不可裂解的肽接头组成的组的肽接头(“L2”)附接至D3。
29.根据权利要求28所述的构建体，其中所述蛋白酶可裂解的肽接头选自SEQ ID NO:71-96、157-161和298中列出的序列的组。
30.根据权利要求28所述的构建体，其中所述不可裂解的肽接头选自SEQ ID NO:107-127中列出的序列的组。
31.根据权利要求1至30中任一项所述的构建体，其中L1和L2两者都是蛋白酶可裂解的肽接头。
32.根据权利要求1至30中任一项所述的构建体，其中L1和L2两者都是不可裂解的肽接头。
33.根据权利要求1至30中任一项所述的构建体，其中L1是蛋白酶可裂解的肽接头，并且L2是不可裂解的肽接头。
34.根据权利要求1至30中任一项所述的构建体，其中L1是不可裂解的肽接头，并且L2是蛋白酶可裂解的肽接头。
35.一种药物组合物，所述药物组合物包含与药学上可接受的载体混合的根据权利要求1至34中任一项所述的构建体。
36.一种治疗受试者的癌症或癌症转移的方法，所述方法包括向所述受试者施用治疗有效量的根据权利要求35所述的药物组合物。
37.根据权利要求36所述的方法，其中所述方法还包括能够治疗受试者的癌症或癌症转移的第二治疗剂或疗法。
38.一种治疗受试者的自身免疫性疾病的方法，所述方法包括向所述受试者施用治疗有效量的根据权利要求35所述的药物组合物。
39.根据权利要求38所述的方法，其中所述方法还包括能够治疗受试者的自身免疫性疾病的第二治疗剂或疗法。
40.一种治疗受试者的炎性疾病的方法，所述方法包括向所述受试者施用治疗有效量的根据权利要求35所述的药物组合物。
41.根据权利要求40所述的方法，其中所述方法还包括能够治疗受试者的炎性疾病的第二治疗剂或疗法。
42.一种用于治疗受试者的病毒感染的方法，所述方法包括施用治疗有效量的根据权利要求35所述的药物组合物。
43.根据权利要求42所述的方法，其中所述方法还包括能够治疗受试者的病毒感染的第二治疗剂或疗法。
44.一种核酸分子，所述核酸分子编码根据权利要求1至34中任一项所述的构建体。
45.一种表达载体，所述表达载体包含根据权利要求44所述的核酸分子。
46.一种宿主细胞，所述宿主细胞包含根据权利要求45所述的表达载体。
47.一种产生根据权利要求1至34中任一项所述的可生物活化的多肽药物构建体的方法，所述方法包括在促进所述可生物活化的多肽药物构建体的表达的条件下培养根据权利要求46所述的宿主细胞，并回收可生物活化的多肽药物构建体蛋白。
48.一种分离的可生物活化的多肽药物构建体蛋白，所述分离的可生物活化的多肽药物构建体蛋白通过根据权利要求46所述的方法产生。
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