CN114748618A - 包含针对狂犬病病毒g蛋白的双特异性抗体的药物制剂及其制备方法 - Google Patents

包含针对狂犬病病毒g蛋白的双特异性抗体的药物制剂及其制备方法 Download PDF

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CN114748618A
CN114748618A CN202210357290.XA CN202210357290A CN114748618A CN 114748618 A CN114748618 A CN 114748618A CN 202210357290 A CN202210357290 A CN 202210357290A CN 114748618 A CN114748618 A CN 114748618A
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常志远
邹志通
张鑫
王威
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Genrix Shanghai Biopharmaceutical Co ltd
Chongqing Zhixiang Jintai Biopharmaceutical Co ltd
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Abstract

本申请提供了包含针对狂犬病病毒G蛋白的双特异性抗体和抗氧化剂的药物制剂，如液体制剂。所述双特异性抗体包含分别结合狂犬病病毒G蛋白的表位I和表位III的两个抗原结合片段，并且具有中和狂犬病病毒的活性。所述制剂可用于疑似狂犬病病毒暴露后的预防中，并通过注射给药。

Description

包含针对狂犬病病毒G蛋白的双特异性抗体的药物制剂及其 制备方法
技术领域
本申请大体涉及包含抗体的药物制剂领域。具体而言，本申请涉及包含针对狂犬病病毒G蛋白的双特异性抗体的稳定的药物制剂及其制备方法。
背景技术
狂犬病病毒是一种子弹形状的、有包膜的、单链RNA病毒，属于弹状病毒科，狂犬病病毒属。狂犬病病毒的基因组编码5种病毒蛋白质：RNA依赖性RNA聚合酶(L)、核蛋白(N)、磷酸化蛋白质(P)、位于病毒蛋白包膜内侧的基质蛋白(M)及外表面糖蛋白(G)。狂犬病病毒的糖蛋白(G蛋白)能与乙酰胆碱结合，决定了狂犬病病毒的噬神经性。G蛋白(62-67kDa)是由505个氨基酸组成的一种I型糖蛋白，G蛋白形成覆盖病毒粒包膜外表面的突起，研究证实G蛋白能够诱导病毒中和抗体。G蛋白至少有5个中和表位，其中表位II是不连续空间表位，包括34-42位氨基酸残基和198-200位氨基酸残基，表位III位于330-338位，是线性表位，报道的抗体中约97％识别的都是表位II和表位III，狂犬病病毒中和抗体CR4098结合表位III。识别表位I和表位IV的抗体很少，狂犬病病毒中和抗体CR57识别线性表位I，即218-240位，核心结合域是226-231位的KLCGVL。表位IV含251和264位残基。还有一个表位是与表位III间隔3个氨基酸残基与表位III不重叠的微表位a，只有两个氨基酸残基342-343。
针对狂犬病病毒的预防和治疗，WHO建议：III类暴露以及野生动物咬伤的II类以上的暴露，应同时进行主动和被动免疫治疗，以获得快速的保护作用。目前已开发出用于被动免疫治疗的针对狂犬病病毒G蛋白的双特异性抗体。
与任何蛋白质一样，抗体的生物学活性依赖于至少核心氨基酸序列的构象完整性保持完整同时保护蛋白质的多个功能基团免于降解。化学和物理不稳定性均可导致抗体降解。因为抗体比传统的有机和无机药物更大且更复杂，因此这种抗体的制剂会出现特殊问题。抗体稳定性可以受到多种因素影响，例如，pH、温度、重复冷冻/解冻循环和剪切力等等。活性抗体可以由于物理不稳定性和化学不稳定性的结果而丧失，所述物理不稳定性包括变性、聚集(可溶及不可溶聚集体形成)、沉淀与吸附，所述化学不稳定性包括例如外消旋化、水解和脱酰胺作用等等。任何这些不稳定性均可以潜在地导致具有降低的生物学活性、增加的毒性和/或增加的免疫原性的抗体副产物或者衍生物的形成。
虽然现有技术领域指出了可适用于产生特定抗体的抗体制剂的许多赋形剂的例子，但是不能预测为了克服特定抗体可能具有的特定的不稳定性问题而应该加入哪些赋形剂以及应该加入的量。此外，难以发现在特定制剂内保持特定抗体的化学和生物学稳定性的最佳条件。鉴于可以变化的所有因素，找寻用于配制抗体的合适赋形剂和最佳条件孕育着挑战。
因此，本领域依然需要能长期储存的稳定的包含针对狂犬病病毒G蛋白的抗体的药物制剂。
发明概述
一方面，本申请提供了药物组合物，其包含约0.2mg/ml～约20mg/ml的针对狂犬病病毒G蛋白的双特异性抗体(下文简称为“双特异性抗体”)、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2，其中所述双特异性抗体包含结合狂犬病病毒G蛋白的表位I的抗原结合片段和结合狂犬病病毒G蛋白的表位III的抗原结合片段，并且所述双特异性抗体具有中和狂犬病病毒的活性。
一方面，本申请提供了药物组合物，其包含约0.2mg/ml～约20mg/ml的针对狂犬病病毒G蛋白的双特异性抗体(下文简称为“双特异性抗体”)、缓冲盐、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2，其中所述双特异性抗体包含结合狂犬病病毒G蛋白的表位I的抗原结合片段和结合狂犬病病毒G蛋白的表位III的抗原结合片段，并且所述双特异性抗体具有中和狂犬病病毒的活性。
在一些实施方案中，本申请提供了药物组合物，其包含约0.2mg/ml～约20mg/ml的所述双特异性抗体、约10mM-约50mM的缓冲盐、约100mM-约400mM的选自海藻糖和甘露醇的渗透压调节剂、约0.2mM-约5mM的金属离子螯合剂型抗氧化剂、约0.05mg/ml-约1.0mg/ml的非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
另一方面，本申请提供了药物组合物在制备用于预防或治疗狂犬病的药物中的用途，其中所述药物组合物包含约0.2mg/ml～约20mg/ml的所述双特异性抗体、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
另一方面，本申请提供了用于预防或治疗狂犬病的方法，其包括向有需要的个体施用药物组合物，所述药物组合物包含约0.2mg/ml～约20mg/ml的所述双特异性抗体、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
另一方面，本申请提供了用于预防或治疗狂犬病的药物组合物，其包含约0.2mg/ml～约20mg/ml的所述双特异性抗体、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
在本申请的一个实施方案中，所述药物组合物还包含缓冲盐，例如约10mM-约50mM的缓冲盐。
在本申请的一个实施方案中，所述金属离子螯合剂型抗氧化剂为乙二胺四乙酸盐。
在一些实施方案中，本申请的药物组合物通过实施疑似狂犬病病毒暴露后的被动免疫来预防或治疗狂犬病。
附图说明
图1A-1F示例性说明了渗透压调节剂对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图2A-2F示例性说明了抗氧化剂对包含低浓度的S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图3A-3F示例性说明了抗氧化剂对包含高浓度的S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图4A-4F示例性说明了在不存在抗氧化剂的情况下，pH值对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图5A-5F示例性说明了在存在抗氧化剂的情况下，pH值对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图6A-6F示例性说明了缓冲盐对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图7A-7E示例性说明了在振摇实验中，聚山梨酯80的浓度对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂中不溶性微粒数量和SEC聚体的水平的影响。
图8A-8F示例性说明了聚山梨酯80的存在对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图9A-9F示例性说明了在加速稳定性实验中，S2E3-SCFV-FCH+C34M-IGG1K抗体的浓度对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
图10A-10F示例性说明了在长期稳定性实验中，S2E3-SCFV-FCH+C34M-IGG1K抗体的浓度对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。
发明详述
针对狂犬病病毒G蛋白的双特异性抗体
本申请的针对狂犬病病毒G蛋白的双特异性抗体包含结合狂犬病病毒G蛋白不同表位的两个抗原结合片段，特别包含结合狂犬病病毒G蛋白的表位I的抗原结合片段和结合狂犬病病毒G蛋白的表位III的抗原结合片段，并且所述双特异性抗体具有中和狂犬病病毒的活性。中国专利申请第201910706710.9号已描述了本申请的针对狂犬病病毒G蛋白的双特异性抗体的结构、氨基酸序列、制备方法和生物活性，特此将其全部内容以其整体通过引用并入本文中。
在一些实施方案中，所述结合狂犬病病毒G蛋白的表位I的抗原结合片段包含：
氨基酸序列为RYTIN的HCDR1，氨基酸序列为GIIPIFGTANYAQRFQG的HCDR2，氨基酸序列为ENLDNSGTYYYFSGWFDP的HCDR3，氨基酸序列为TGTSSDIGAYDYVS的LCDR1，氨基酸序列为DATKRPS的LCDR2，氨基酸序列为CSYAGDYTPGVV的LCDR3；或者
氨基酸序列为RYSIN的HCDR1，氨基酸序列为GIIPIFGTANYAQRFQG的HCDR2，氨基酸序列为ENLDNSGTYYYFSGWFDP的HCDR3，氨基酸序列为TGTSSDIDGYDFVS的LCDR1，氨基酸序列为DATKRPS的LCDR2，氨基酸序列为CSYAGDYTPGVV的LCDR3；或者
氨基酸序列为GYTIN的HCDR1，氨基酸序列为GIIPIFGTANYAQRFQG的HCDR2，氨基酸序列为ENLDNSGTYYYFSGWFDP的HCDR3，氨基酸序列为TGTSSDLGGYDFVS的LCDR1，氨基酸序列为DATKRPS的LCDR2，氨基酸序列为CSYAGDYTPGVV的LCDR3；
其中，HCDR和LCDR氨基酸序列根据Kabat定义。
在一些实施方案中，所述结合狂犬病病毒G蛋白的表位III的抗原结合片段包含：
氨基酸序列为SYGMH的HCDR1，氨基酸序列为TISYDGSIKDYADSVKG的HCDR2，氨基酸序列为GDRTGNLDY的HCDR3，氨基酸序列为RASQNIRNALN的LCDR1，氨基酸序列为DASTRQS的LCDR2，氨基酸序列为QQNSEFPPT的LCDR3；
其中，HCDR和LCDR氨基酸序列根据Kabat定义。
在一些实施方案中，所述结合狂犬病病毒G蛋白的表位I的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：24所示，轻链可变区的氨基酸序列如SEQ ID NO：25所示；或者
所述结合狂犬病病毒G蛋白的表位I的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：26所示，轻链可变区的氨基酸序列如SEQ ID NO：27所示；或者
所述结合狂犬病病毒G蛋白的表位I的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：28所示，轻链可变区的氨基酸序列如SEQ ID NO：29所示。
在一些实施方案中，所述结合狂犬病病毒G蛋白的表位III的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：1所示，轻链可变区的氨基酸序列如SEQ ID NO：3所示。
在一些实施方案中，所述结合狂犬病病毒G蛋白的表位I的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：24所示，轻链可变区的氨基酸序列如SEQ ID NO：25所示，所述结合狂犬病病毒G蛋白的表位III的抗原结合片段的重链可变区的氨基酸序列如SEQID NO：1所示，轻链可变区的氨基酸序列如SEQ ID NO：3所示；或者
所述结合狂犬病病毒G蛋白的表位I的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：26所示，轻链可变区的氨基酸序列如SEQ ID NO：27所示，所述结合狂犬病病毒G蛋白的表位III的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：1所示，轻链可变区的氨基酸序列如SEQ ID NO：3所示；或者
所述结合狂犬病病毒G蛋白的表位I的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：28所示，轻链可变区的氨基酸序列如SEQ ID NO：29所示，所述结合狂犬病病毒G蛋白的表位III的抗原结合片段的重链可变区的氨基酸序列如SEQ ID NO：1所示，轻链可变区的氨基酸序列如SEQ ID NO：3所示。
在一些实施方案中，两个抗原结合片段的形式独立地选自单链抗体(scFv)或Fab片段。
在一些实施方案中，结合狂犬病病毒G蛋白的表位I的抗原结合片段为单链抗体(scFv)，结合狂犬病病毒G蛋白的表位III的抗原结合片段为Fab片段。
在一些实施方案中，所述双特异性抗体包含SEQ ID NO:32、33、34之一所示的氨基酸序列。在一些实施方案中，所述双特异性抗体包含SEQ ID NO:30和SEQ ID NO:31所示的氨基酸序列。
药物组合物
一方面，本申请提供了药物组合物，其包含约0.2mg/ml-约20mg/ml的所述双特异性抗体、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
一方面，本申请提供了药物组合物，其包含约0.2mg/ml-约20mg/ml的所述双特异性抗体、缓冲盐、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
在一些实施方案中，本申请提供了药物组合物，其包含约0.2mg/ml-约20mg/ml的所述双特异性抗体、约10mM-约50mM的缓冲盐、约100mM-约400mM的选自海藻糖和甘露醇的渗透压调节剂、约0.2mM-约5mM的金属离子螯合剂型抗氧化剂、约0.05mg/ml-约1.0mg/ml的非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2。
在一些实施方案中，本申请提供了药物组合物，其包含约0.5mg/ml-约2.0mg/ml的所述双特异性抗体、约10mM-约50mM的缓冲盐、约100mM-约400mM的选自海藻糖和甘露醇的渗透压调节剂、约0.2mM-约5mM的金属离子螯合剂型抗氧化剂、约0.05mg/ml-约1.0mg/ml的非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.2-约6.2。
在一些实施方案中，在所述药物组合物中，所述双特异性抗体的含量为约0.2mg/ml～约20mg/ml、约0.5mg/ml～约2.0mg/ml、约0.5mg/ml-约1.0mg/ml、约1.0mg/ml-约2.0mg/ml、约0.5mg/ml、约1.0mg/ml、约1.5mg/ml或约2.0mg/ml。或者，本领域技术人员可以根据实践的需要选择合适含量的所述双特异性抗体。
在一些实施方案中，所述缓冲盐可以是本领域中用于制备液体制剂的常用的缓冲盐，其实例包括但不限于磷酸/磷酸盐缓冲对、组氨酸/组氨酸的无机盐缓冲对、醋酸/醋酸盐缓冲对、柠檬酸/柠檬酸盐缓冲对等等。在一些实施方案中，所述缓冲盐是组氨酸/组氨酸的无机盐缓冲对或醋酸/醋酸盐缓冲对，例如组氨酸/盐酸组氨酸缓冲对或醋酸/醋酸钠缓冲对。在一些实施方案中，所述缓冲盐是组氨酸/盐酸组氨酸缓冲对。
在一些实施方案中，在所述药物组合物中所述缓冲盐的浓度为约10mM-约50mM、约15mM-约30mM、约15mM-约25mM、约15mM-约20mM、约20mM-约25mM或约20mM。在一些实施方案中，所述缓冲盐为浓度为约10mM-约50mM、约15mM-约30mM、约15mM-约25mM、约15mM-约20mM、约20mM-约25mM或约20mM的组氨酸/盐酸组氨酸缓冲对或醋酸/醋酸钠缓冲对。或者，本领域技术人员可以根据实践的需要选择合适浓度的组氨酸/盐酸组氨酸缓冲对或醋酸/醋酸钠缓冲对。选用组氨酸/盐酸组氨酸缓冲对和醋酸/醋酸钠缓冲对时，药物组合物中的所述双特异性抗体聚集速度较慢，并且该缓冲盐既能维持药物组合物的pH，又可以在临床使用进入人体内时被迅速中和至生理性pH。
在一些实施方案中，在所述药物组合物中所述渗透压调节剂的浓度为约100mM-约400mM、约200mM-约300mM、约200mM-约250mM、约230mM-约260mM或约250mM-约300mM，例如，约200、210、220、230、240、250、260、270、280、290或300mM。所述渗透压调节剂选自海藻糖和甘露醇，优选海藻糖。海藻糖和甘露醇在本申请的包含所述双特异性抗体的药物制剂中具有溶剂排阻效应，会提高所述双特异性抗体在水溶液中的稳定性，抑制所述双特异性抗体的聚集。
在一些实施方案中，在所述药物组合物中，所述金属离子螯合剂型抗氧化剂的实例包括但不限于乙二胺四乙酸盐、乙二醇四乙酸(EGTA)、硫胺四氢糖基二硫化物(TTFD)、2,3-二巯基乙二酸(DMSA)、二乙基三胺五醋酸或二巯基丙醇。在一些实施方案中，所述乙二胺四乙酸盐可以为乙二胺四乙酸的碱金属盐或碱土金属盐，例如乙二胺四乙酸二钠或乙二胺四乙酸钙钠等。在一些实施方案中，所述金属离子螯合剂型抗氧化剂(例如，乙二胺四乙酸盐)的浓度为约0.2mM-约5mM、约0.5mM-约5mM、约0.5mM-约2.0mM、0.5mM-约1.5mM、约0.5mM-约1.0mM、约1.0mM-约1.5mM或约1.0mM。在一些实施方案中，金属离子螯合剂型抗氧化剂(例如，乙二胺四乙酸盐)的浓度为约0.5、约1.0、约1.5、约2.0、约2.5、约3.0、约3.5、约4.0、约4.5或5.0mM。或者，本领域技术人员可以根据实践的需要选择合适浓度的金属离子螯合剂型抗氧化剂(例如，乙二胺四乙酸盐)。
在一些实施方案中，在所述药物组合物中，所述非离子型表面活性剂可以为聚山梨酯型表面活性剂，例如聚山梨酯20或聚山梨酯80。在一些实施方案中，非离子型表面活性剂(例如，聚山梨酯型表面活性剂，例如聚山梨酯20或聚山梨酯80)的含量为约0.05mg/ml-约1.0mg/ml、约0.1mg/ml-约1.0mg/ml、约0.2mg/ml-约0.5mg/ml、约0.3mg/ml-约1.0mg/ml或约0.5mg/ml-约1.0mg/ml，例如，约0.3mg/ml、0.34mg/ml、约0.4mg/ml、0.5mg/ml、0.6mg/ml、0.7mg/ml、0.8mg/ml、0.9mg/ml或1.0mg/ml。所述双特异性抗体作为蛋白质可能会在界面(空气/液体界面、液体/玻璃界面等)处由于疏水作用而发生变性，进而发生蛋白聚集甚至沉淀。选用非离子型表面活性剂(例如，聚山梨酯型表面活性剂，例如聚山梨酯20或聚山梨酯80)可以防止在振摇、搅动或冻融时出现蛋白聚集沉淀。
在一些实施方案中，所述药物组合物的pH值为约5.0-约6.2、约5.0-约6.0、约5.2-约6.2、约5.5-约6.0或约5.5-约5.8，例如约5.0、5.1、5.2、5.3、5.4、5.5、5.6、5.7、5.8、5.9或6.0。
在一些实施方案中，所述药物组合物中的水可以是无菌无热源的水，例如注射用水。
在一些实施方案中，本申请提供了药物组合物，其包含约0.5mg/ml～约2.0mg/ml的所述双特异性抗体、约10mM-约50mM的组氨酸/盐酸组氨酸缓冲对或醋酸/醋酸钠缓冲对、约100mM-约400mM的选自海藻糖和甘露醇的渗透压调节剂、约0.2mM-约5mM的乙二胺四乙酸盐、约0.05mg/ml-约1.0mg/ml的聚山梨酯80和余量的水，其中所述药物组合物的pH值为约5.5-约6.0。
在一些实施方案中，本申请提供了药物组合物，其包含约0.5mg/ml～约2.0mg/ml的所述双特异性抗体、约15mM-约25mM的组氨酸/盐酸组氨酸缓冲对或醋酸/醋酸钠缓冲对、约200mM-约300mM的选自海藻糖和甘露醇的渗透压调节剂、约0.5mM-约5mM的乙二胺四乙酸盐、约0.3mg/ml-约1.0mg/ml的聚山梨酯80和余量的水，其中所述药物组合物的pH值为约5.5-约6.0。
在一些实施方案中，本申请提供了药物组合物，其包含约0.5mg/ml～约2.0mg/ml的所述双特异性抗体、约10mM-约50mM的组氨酸/盐酸组氨酸缓冲对、约100mM-约400mM的海藻糖、约0.2mM-约5mM的乙二胺四乙酸盐、约0.05mg/ml-约1.0mg/ml的聚山梨酯80和余量的水，其中所述药物组合物的pH值为约5.5-约6.0。
在一些实施方案中，本申请提供了药物组合物，其包含约0.5mg/ml～约2.0mg/ml的所述双特异性抗体、约15mM-约25mM的组氨酸/盐酸组氨酸缓冲对、约200mM-约300mM的海藻糖、约0.5mM-约5mM的乙二胺四乙酸盐、约0.3mg/ml-约1.0mg/ml的聚山梨酯80和余量的水，其中所述药物组合物的pH值为约5.5-约6.0。
在一些实施方案中，本申请提供了药物组合物，其包含约0.5mg/ml～约2.0mg/ml的所述双特异性抗体、约20mM的组氨酸/盐酸组氨酸缓冲对、约240mM的海藻糖、约1.0mM的乙二胺四乙酸盐、约0.5mg/ml-约1.0mg/ml的聚山梨酯80和余量的水，其中所述药物组合物的pH值为约5.5-约5.8。
本申请的药物组合物为液体制剂，其可以为溶液剂、乳剂或混悬剂的形式，优选为溶液剂。本申请的药物组合物可通过肠胃外给药途径，如注射给药途径施用于有需要的个体。所述注射给药途径包括但不限于皮下注射(例如，伤口皮下浸润注射)、肌肉注射和皮内注射等。
本申请的包含所述双特异性抗体的药物组合物在5℃±3℃下放置6个月后，SEC-HPLC单体峰面积％、CEX-HPLC酸性峰面积％、CE-SDS(非还原法)和活性水平均基本没有发生变化。本申请的包含所述双特异性抗体的药物组合物可以在5℃±3℃的避光条件下长期贮存，并且能够保持质量稳定，保证药品的安全性、有效性和均一性。
应当理解，以上详细描述仅为了使本领域技术人员更清楚地了解本申请的内容，而并非意图在任何方面加以限制。本领域技术人员能够对所述实施方案进行各种改动和变化。
以下实施例仅用于说明而非限制本申请范围的目的。在以下实施例中，以CN201910706710.9中公开的S2E3-scFv-FcH+C34m-IgG1K抗体为本申请的所述双特异性抗体的实例。
实施例
实施例1：渗透压调节剂对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响
取适量S2E3-SCFV-FCH+C34M-IGG1K抗体样品，超滤换液置换至纯水中，并用纯化水稀释至2mg/ml，即得S2E3-SCFV-FCH+C34M-IGG1K抗体水溶液。配制表1所示的各配方的2×制剂buffer母液(包含2倍浓度的缓冲盐和渗透压调节剂，pH约为6.0)。然后，取7mlS2E3-SCFV-FCH+C34M-IGG1K抗体水溶液、7ml 2×制剂buffer母液和28μL的10％聚山梨酯80溶液，混匀，分别制得渗透压调节剂为海藻糖、甘露醇、蔗糖、盐酸精氨酸、甘氨酸和脯氨酸，以及不含渗透压调节剂的制剂样品。
表1液体制剂的配方组成
Figure BDA0003580083210000071
√：表示处方中含有所对应的物质；-：表示处方中未含有所对应的物质；
20mM组氨酸盐为组氨酸和盐酸组氨酸组成的包含20mM组氨酸/组氨酸离子的缓冲体系
将各个处方的样品置于高温(37±2℃)条件下储存以进行稳定性试验以加速S2E3-SCFV-FCH+C34M-IGG1K抗体的降解，并在第0天、2周、4周、6周和8周进行取样，采用如下所述的SEC-HPLC、CEX-HPLC、非还原CE-SDS和还原CE-SDS方法测定各个时间点制剂样品的纯度。使用DSF方法测定各个处方样品的Tm值，以确定其构象稳定性性质。检测结果显示于图1A至图1F中。
SEC-HPLC(分子排阻色谱法)纯度测定采用TSKgel G3000SWXL色谱柱和高效液相色谱仪，流动相为50mM PB/300mM NaCl，pH 7.0溶液，将样品稀释后进样，于1.0ml/min的流速下进行等度洗脱，洗脱15min，280nm紫外波长下进行检测，采用峰面积归一化法得到单体和聚体的峰面积百分比。
CEX-HPLC(阳离子交换色谱法)纯度测定采用MabPacTMSCX-10BioLCTM Analytical色谱柱和高效液相色谱仪，流动相A为20mM MES，pH 5.6溶液，流动相B为20mM MES+0.5MNaCl，pH5.6溶液，将样品稀释后进样，于0.5ml/min的流速下进行梯度洗脱，洗脱50min，280nm紫外波长下进行检测，采用峰面积归一化法得到酸性峰、主峰和碱性峰的峰面积百分比。
非还原CE-SDS(十二烷基硫酸钠毛细管电泳)和还原CE-SDS方法采用毛细管电泳仪进行测定，采用有效长度为20cm的无涂层毛细管柱和SDS-MW Gel分离胶，柱温25℃，分离电压15kV，检测波长214nm，分别在非还原和还原条件下测定样品的纯度。
DSF(差式扫描荧光)使用Real-Time PCR仪进行测定，将样品稀释后与荧光染料(
Figure BDA0003580083210000081
Orange Protein gel stain，Life)混合后转移至PCR管内，设置升温程序(25℃到95℃，1℃/min)，实时收集荧光并进行数据记录，对荧光-温度曲线求导，求导后曲线最高峰即为Tm值。
表2渗透压调节剂对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的Tm值的影响
制剂编号 H60T H60 H60M H60S H60R H60G H60P
Tm值(℃) 61.1 59.9 60.6 60.9 58.8 60.6 60.2
从上表和图1A至图1F中可以看出，不同的渗透压调节剂对S2E3-SCFV-FCH+C34M-IGG1K抗体的Tm值影响较小，其变化小于2℃，均在60℃左右，其中使用盐酸精氨酸的制剂Tm值稍低。在高温储存条件下，蔗糖、盐酸精氨酸、甘氨酸和脯氨酸使包含S2E3-SCFV-FCH+C34M-IGG1K抗体的液体制剂中的SEC-HPLC聚体和CEX-HPLC碱性峰的水平增加的速度比较快，非还原CE-SDS纯度的水平下降的速度也比较快，因此，这些渗透压调节剂不适合用于制备包含S2E3-SCFV-FCH+C34M-IGG1K抗体的液体制剂。使用海藻糖和甘露醇的制剂样品在高温储存条件下比较稳定。
实施例2.抗氧化剂对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响
除了还加入作为抗氧化剂的1mM乙二胺四乙酸二钠和1mM甲硫氨酸以外，采用类似于实施例1中所述的制备方法并根据下表中所示的配方组成制备液体制剂H60T、H60TE和H60TM。
表3液体制剂的配方组成
Figure BDA0003580083210000082
√：表示处方中含有所对应的物质；-：表示处方中未含有所对应的物质；
20mM组氨酸盐为组氨酸和盐酸组氨酸组成的包含20mM组氨酸/组氨酸离子的缓冲体系
采用类似于实施例1中所述的储存条件、采样时间点和检测项目对各个制剂样品的纯度进行检测。检测结果显示于下表和图2A至图2F中。
表4抗氧化剂对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的Tm值的影响
制剂编号 H60T H60TE H60TM
Tm值(℃) 61.1 61.1 61.2
从上表和图2A至图2F中可以看出，抗氧化剂的添加不影响S2E3-SCFV-FCH+C34M-IGG1K抗体的Tm值。乙二胺四乙酸二钠和甲硫氨酸均可以降低SEC聚体增加的速度，CEX纯度下降趋势不会发生变化，但是甲硫氨酸会使非还原CE-SDS纯度下降的速度加快。
采用以下配方组成，重复实施上述实验步骤，进一步考察抗氧化剂对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。检测结果显示于图3A至图3F中。
表5液体制剂的配方组成
Figure BDA0003580083210000091
√：表示处方中含有所对应的物质；
20mM组氨酸盐为组氨酸和盐酸组氨酸组成的包含20mM组氨酸/组氨酸离子的缓冲体系
从图3A至图3F中可知，在包含较高浓度的S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂中，使用L-甲硫氨酸作为抗氧化剂时，SEC-HPLC、CEX-HPLC和非还原CE-SDS的纯度在高温(37±2℃)条件下下降速度均加快。
由此可见，乙二胺四乙酸二钠比L-甲硫氨酸更适合作为包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂中的抗氧化剂。
实施例3.pH值对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响
除了将制剂的最终pH值调整为5.5-6.2以外，采用类似于实施例1中所述的制备方法并根据下表中所示的配方组成制备液体制剂H55T、H58T、H60T和H62T。
表6液体制剂的配方组成
Figure BDA0003580083210000092
√：表示处方中含有所对应的物质；
20mM组氨酸盐为组氨酸和盐酸组氨酸组成的包含20mM组氨酸/组氨酸离子的缓冲体系
采用类似于实施例1中所述的储存条件、采样时间点和检测项目对各个制剂样品的纯度进行检测。检测结果显示于下表和图4A至图4F中。
表7pH值对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的Tm值的影响
制剂编号 H55T H58T H60T H62T
Tm值(℃) 60.5 60.9 61.1 60.9
从上表和图4A至图4F中可以看出，S2E3-SCFV-FCH+C34M-IGG1K抗体的Tm值基本不受制剂的pH值变化的影响。在pH为5.5～6.2范围内，在高温储存条件下，除pH为6.2的制剂的纯度下降较快以外，pH为5.8的制剂的聚体增加的速度最慢，而pH为5.5、5.8和6.0的制剂对CEX-HPLC和非还原的CE-SDS纯度下降趋势几乎没有差别。
采用以下配方组成，重复实施上述实验步骤，进一步考察在抗氧化剂存在的情况下，pH值变化对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响。检测结果显示于图5A至图5F中。
表8液体制剂的配方组成
Figure BDA0003580083210000101
√：表示处方中含有所对应的物质；
20mM组氨酸盐为组氨酸和盐酸组氨酸组成的包含20mM组氨酸/组氨酸离子的缓冲体系
从图5A至图5F中可以看出，在pH为5.2～6.2范围内，SEC-HPLC和非还原CE-SDS纯度下降的趋势基本没有差别。pH为5.2和pH为5.5的制剂样品中CEX碱性峰增加的速度稍快。因此，pH为5.2～6.2范围内，包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性基本不会受到显著影响。
实施例4.缓冲体系对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响
除了使用醋酸盐缓冲体系代替组氨酸盐缓冲体系并将制剂的最终pH值调整为5.0-5.8以外，采用类似于实施例1中所述的制备方法并根据下表中所示的配方组成制备液体制剂H58T、A50T、A52T、A55T和A58T。
表9液体制剂的配方组成
Figure BDA0003580083210000111
√：表示处方中含有所对应的物质；
20mM组氨酸盐为组氨酸和盐酸组氨酸组成的包含20mM组氨酸/组氨酸离子的缓冲体系；
20mM醋酸盐：为冰醋酸和醋酸钠组成的包含20mM醋酸/醋酸根的缓冲体系。
采用类似于实施例1中所述的储存条件、采样时间点和检测项目对各个制剂样品的稳定性进行检测。检测结果显示于下表和图6A至图6F中。
表10缓冲体系对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的Tm值的影响
制剂编号 H58T A50T A52T A55T A58T
Tm值(℃) 60.9 61.1 61.2 60.9 60.4
从上表和图6A至图6F中可以看出，在醋酸盐和组氨酸盐缓冲液中，S2E3-SCFV-FCH+C34M-IGG1K抗体的Tm值基本没有差别。在37℃条件下放置8周后，在pH为5.0～5.8的范围内，S2E3-SCFV-FCH+C34M-IGG1K抗体在醋酸盐缓冲液中的SEC聚体增加的速度稍高于在pH5.8的组氨酸盐缓冲液中，然而两者的CEX-HPLC和非还原CE-SDS纯度下降趋势基本没有差别。
实施例5.聚山梨酯80对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响
取适量S2E3-SCFV-FCH+C34M-IGG1K抗体样品，超滤换液置换至缓冲盐buffer(20mM组氨酸/盐酸组氨酸，pH5.8)中，并用缓冲盐buffer稀释至2.5mg/ml，即得S2E3-SCFV-FCH+C34M-IGG1K抗体溶液。配制表11所示处方的5×制剂buffer母液(包含1倍浓度的缓冲盐、5倍浓度的海藻糖和乙二胺四乙酸二钠，pH约为5.8)。然后，取32ml S2E3-SCFV-FCH+C34M-IGG1K抗体溶液、8ml 5×制剂buffer母液，混匀。分别取5ml各加入一定体积的10％聚山梨酯80，分别配制成含0％、0.005％、0.01％、0.02％、0.03％、0.04％、0.05％、0.10％聚山梨酯80的制剂样品。
表11配方组成
Figure BDA0003580083210000112
Figure BDA0003580083210000121
表11中的“％”意指重量/体积百分数或％w/v。例如，0.05％聚山梨酯80意指制剂中聚山梨酯80的含量为0.5mg/mL。另外，将表11中含0.05％聚山梨酯80的制剂编号为H58TE(2)0.05％吐温80，以及将表11中不含聚山梨酯80的制剂编号为H58TE-ps(2)无吐温80。
将表11中的各个制剂样品进行振摇实验，以1500次/mim的速率振摇3天后，检测各个样品中的不溶性微粒数量(光阻法)和SEC-HPLC聚体的水平。振摇实验的结果显示于图7A-7E中。当制剂样品中不含聚山梨酯80时，振摇3天后，样品的不溶性微粒数量和SEC聚体的比例均明显升高，这表明包含S2E3-SCFV-FCH+C34M-IGG1K抗体的制剂容易受到振摇等机械作用力的影响而引起蛋白质聚集。在包含S2E3-SCFV-FCH+C34M-IGG1K抗体的制剂中加入0.005％～0.10％浓度的聚山梨酯80时，振摇3天后，样品的不溶性微粒数量均基本没有增加。含有0.005％～0.02％浓度的聚山梨酯80的制剂样品在振摇后SEC-HPLC聚体水平有所升高。制剂中的聚山梨酯80浓度升高至0.03％及以上时，制剂样品中的SEC-HPLC聚体比例基本不会升高。
采用类似于实施例1中所述的储存条件、采样时间点和检测项目对编号为H58TE(2)0.05％吐温80和H58TE-ps(2)无吐温80的制剂样品的稳定性进行检测，以考察在储存过程中聚山梨酯80可能产生的降解产物对制剂稳定性的影响。实验结果显示于图8A-8F中。在37℃条件下，含有0.5mg/ml聚山梨酯80的制剂样品与不含聚山梨酯80的制剂样品中SEC-HPLC、CEX-HPLC和非还原CE-SDS纯度下降的趋势基本没有差别，这表明包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性基本不受在储存过程中聚山梨酯80可能产生的降解产物的影响。
实施例6.蛋白质浓度对包含S2E3-SCFV-FCH+C34M-IGG1K抗体的药物制剂的稳定性的影响
取S2E3-SCFV-FCH+C34M-IGG1K抗体原液(含有20mM组氨酸/盐酸组氨酸、90mg/ml海藻糖、1mM乙二胺四乙酸二钠、0.5mg/ml聚山梨酯80，pH 5.8)适量，分别使用制剂buffer稀释至20mg/ml、1mg/ml、0.5mg/ml、0.2mg/ml，混匀，制备得到不同蛋白质浓度的制剂样品。
表12配方组成
Figure BDA0003580083210000122
Figure BDA0003580083210000131
将各个处方的样品置于25±2℃下储存以进行加速稳定性试验，并在第0天、1个月、2个月、3个月和6个月进行取样，并采用类似于实施例1中所述的检测条件，检测样品中SEC-HPLC、CEX-HPLC和非还原CE-SDS的水平。检测结果显示于下表和图9A至图9F中。另外，还将各个处方的样品置于5±3℃下储存以进行长期稳定性试验，并在第0天、3个月、6个月、9个月和12个月进行取样，并采用类似于实施例1中所述的检测条件，检测样品中SEC-HPLC、CEX-HPLC和非还原CE-SDS的水平。检测结果显示于图10A至图10F中。
从图9A至图10F中可以看出，在加速(25±2℃)条件下，S2E3-SCFV-FCH+C34M-IGG1K抗体浓度为0.2mg/ml和20mg/ml的制剂样品在前3个月时SEC聚体比例增加的速度较快，4种不同蛋白浓度的制剂样品的CEX-HPLC和非还原CE-SDS的纯度变化趋势一致。在5±3℃下，S2E3-SCFV-FCH+C34M-IGG1K抗体浓度为0.2mg/ml的制剂样品的SEC聚体比例较高，而S2E3-SCFV-FCH+C34M-IGG1K抗体浓度为0.5mg/ml、1.0mg/ml和20mg/ml的制剂样品中SEC聚体比例变化趋势基本一致。4种不同蛋白浓度的制剂样品的CEX-HPLC和非还原CE-SDS的纯度变化趋势一致。
由此可见，S2E3-SCFV-FCH+C34M-IGG1K抗体浓度为0.2mg/ml、0.5mg/ml、1.0mg/ml和20mg/ml的制剂样品在加速(25±2℃)和长期(5±3℃)条件下放置时，CEX-HPLC和非还原CE-SDS的纯度变化没有差别，而S2E3-SCFV-FCH+C34M-IGG1K抗体浓度为0.2mg/ml的制剂样品在长期(5±3℃)条件下聚体水平较高。
序列表
<110> 重庆智翔金泰生物制药股份有限公司
智翔（上海）医药科技有限公司
<120> 包含针对狂犬病病毒G蛋白的双特异性抗体的药物制剂及其制备方法
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<212> PRT
<213> 智人(Homo sapiens)
<400> 10
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
130 135 140
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 11
<211> 232
<212> PRT
<213> 小家鼠(Mus musculus)
<400> 11
Pro Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala
1 5 10 15
Pro Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile
20 25 30
Lys Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val
35 40 45
Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val
50 55 60
Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp
65 70 75 80
Tyr Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln
85 90 95
Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys Asp
100 105 110
Leu Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val
115 120 125
Arg Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr
130 135 140
Lys Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu
145 150 155 160
Asp Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr
165 170 175
Lys Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr
180 185 190
Ser Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr
195 200 205
Ser Cys Ser Val Val His Glu Gly Leu His Asn His His Thr Thr Lys
210 215 220
Ser Phe Ser Arg Thr Pro Gly Lys
225 230
<210> 12
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Asn Arg Tyr
20 25 30
Thr Val Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 13
<211> 119
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 13
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Ala 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
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Leu Tyr Asp Gly Ser Asp Lys Phe Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Val Ala Val Ala Gly Thr His Phe Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Leu Val Thr Val Ser Ser
115
<210> 14
<211> 330
<212> PRT
<213> 小家鼠(Mus musculus)
<400> 14
Ala Lys Thr Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly
1 5 10 15
Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr
20 25 30
Phe Pro Glu Pro Val Thr Leu Thr Trp Asn Ser Gly Ser Leu Ser Ser
35 40 45
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu
50 55 60
Ser Ser Ser Val Thr Val Thr Ser Ser Thr Trp Pro Ser Gln Ser Ile
65 70 75 80
Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys
85 90 95
Ile Glu Pro Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys
100 105 110
Pro Ala Pro Asn Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro
115 120 125
Lys Ile Lys Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys
130 135 140
Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp
145 150 155 160
Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg
165 170 175
Glu Asp Tyr Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln
180 185 190
His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn
195 200 205
Lys Asp Leu Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly
210 215 220
Ser Val Arg Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu
225 230 235 240
Met Thr Lys Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met
245 250 255
Pro Glu Asp Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu
260 265 270
Asn Tyr Lys Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe
275 280 285
Met Tyr Ser Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn
290 295 300
Ser Tyr Ser Cys Ser Val Val His Glu Gly Leu His Asn His His Thr
305 310 315 320
Thr Lys Ser Phe Ser Arg Thr Pro Gly Lys
325 330
<210> 15
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Gln Ser Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Gly Tyr
20 25 30
Asn Phe Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp
85 90 95
Tyr Thr Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 16
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 16
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 Asp
20 25 30
Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ala Ala Ser Ser 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 Leu Asn Ser Tyr Pro Pro
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 17
<211> 106
<212> PRT
<213> 小家鼠(Mus musculus)
<400> 17
Gly Gln Pro Lys Ser Ser Pro Ser Val Thr Leu Phe Pro Pro Ser Ser
1 5 10 15
Glu Glu Leu Glu Thr Asn Lys Ala Thr Leu Val Cys Thr Ile Thr Asp
20 25 30
Phe Tyr Pro Gly Val Val Thr Val Asp Trp Lys Val Asp Gly Thr Pro
35 40 45
Val Thr Gln Gly Met Glu Thr Thr Gln Pro Ser Lys Gln Ser Asn Asn
50 55 60
Lys Tyr Met Ala Ser Ser Tyr Leu Thr Leu Thr Ala Arg Ala Trp Glu
65 70 75 80
Arg His Ser Ser Tyr Ser Cys Gln Val Thr His Glu Gly His Thr Val
85 90 95
Glu Lys Ser Leu Ser Arg Ala Asp Cys Ser
100 105
<210> 18
<211> 107
<212> PRT
<213> 小家鼠(Mus musculus)
<400> 18
Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu
1 5 10 15
Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe
20 25 30
Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg
35 40 45
Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser
50 55 60
Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu
65 70 75 80
Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser
85 90 95
Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys
100 105
<210> 19
<211> 232
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr
130 135 140
Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 20
<211> 232
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
1 5 10 15
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
20 25 30
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
35 40 45
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
50 55 60
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
65 70 75 80
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
85 90 95
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
100 105 110
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
115 120 125
Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
130 135 140
Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser
145 150 155 160
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
165 170 175
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val
180 185 190
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
195 200 205
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
210 215 220
Ser Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 21
<211> 254
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 21
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Arg Tyr
20 25 30
Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser
130 135 140
Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val
145 150 155 160
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Ala Tyr Asp Tyr
165 170 175
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
180 185 190
Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala
210 215 220
Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp Tyr Thr
225 230 235 240
Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
245 250
<210> 22
<211> 254
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Arg Tyr
20 25 30
Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser
130 135 140
Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val
145 150 155 160
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Asp Gly Tyr Asp Phe
165 170 175
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
180 185 190
Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala
210 215 220
Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp Tyr Thr
225 230 235 240
Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
245 250
<210> 23
<211> 254
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 23
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Gly Tyr
20 25 30
Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser
130 135 140
Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val
145 150 155 160
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Leu Gly Gly Tyr Asp Phe
165 170 175
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
180 185 190
Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala
210 215 220
Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp Tyr Thr
225 230 235 240
Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
245 250
<210> 24
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Arg Tyr
20 25 30
Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 25
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Gln Ser Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Ala Tyr
20 25 30
Asp Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp
85 90 95
Tyr Thr Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 26
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Arg Tyr
20 25 30
Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 27
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Gln Ser Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Asp Gly Tyr
20 25 30
Asp Phe Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp
85 90 95
Tyr Thr Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 28
<211> 127
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Gly Tyr
20 25 30
Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 29
<211> 112
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Gln Ser Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Leu Gly Gly Tyr
20 25 30
Asp Phe Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp
85 90 95
Tyr Thr Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 30
<211> 214
<212> PRT
<213> 智人(Homo sapiens)
<400> 30
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 Asn Ile Arg Asn Ala
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Asp Ala Ser Thr Arg 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 Asn Ser Glu Phe 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> 31
<211> 448
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
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 Gly Ser Tyr
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Thr Ile Ser Tyr Asp Gly Ser Ile Lys Asp Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Lys Gly Asp Arg Thr Gly Asn Leu Asp Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro
115 120 125
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly
130 135 140
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn
145 150 155 160
Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser
180 185 190
Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
195 200 205
Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr
210 215 220
His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
225 230 235 240
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro
260 265 270
Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
275 280 285
Lys Thr Lys Pro Arg Glu Glu Gln Tyr 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 Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp 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> 32
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Gly Tyr
20 25 30
Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser
130 135 140
Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val
145 150 155 160
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Leu Gly Gly Tyr Asp Phe
165 170 175
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
180 185 190
Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala
210 215 220
Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp Tyr Thr
225 230 235 240
Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala Ser
245 250 255
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
260 265 270
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
275 280 285
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
290 295 300
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
305 310 315 320
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
325 330 335
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
340 345 350
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
355 360 365
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
370 375 380
Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
385 390 395 400
Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser
405 410 415
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
420 425 430
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val
435 440 445
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
450 455 460
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
465 470 475 480
Ser Leu Ser Leu Ser Pro Gly Lys
485
<210> 33
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Arg Tyr
20 25 30
Thr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser
130 135 140
Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val
145 150 155 160
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Gly Ala Tyr Asp Tyr
165 170 175
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
180 185 190
Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala
210 215 220
Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp Tyr Thr
225 230 235 240
Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala Ser
245 250 255
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
260 265 270
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
275 280 285
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
290 295 300
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
305 310 315 320
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
325 330 335
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
340 345 350
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
355 360 365
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
370 375 380
Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
385 390 395 400
Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser
405 410 415
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
420 425 430
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val
435 440 445
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
450 455 460
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
465 470 475 480
Ser Leu Ser Leu Ser Pro Gly Lys
485
<210> 34
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Tyr Ser Arg Tyr
20 25 30
Ser Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Arg Phe
50 55 60
Gln Gly Arg Leu Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Glu Asn Leu Asp Asn Ser Gly Thr Tyr Tyr Tyr Phe Ser Gly
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser
130 135 140
Ala Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val
145 150 155 160
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Ile Asp Gly Tyr Asp Phe
165 170 175
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu Met Ile
180 185 190
Tyr Asp Ala Thr Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly
195 200 205
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala
210 215 220
Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Asp Tyr Thr
225 230 235 240
Pro Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala Ser
245 250 255
Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
260 265 270
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
275 280 285
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
290 295 300
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
305 310 315 320
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
325 330 335
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
340 345 350
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
355 360 365
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
370 375 380
Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
385 390 395 400
Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser
405 410 415
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
420 425 430
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val
435 440 445
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
450 455 460
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
465 470 475 480
Ser Leu Ser Leu Ser Pro Gly Lys
485
<210> 35
<211> 440
<212> PRT
<213> 狂犬病病毒(Rabies virus)
<400> 35
Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro Trp Ser Pro
1 5 10 15
Ile Asp Ile His His Leu Ser Cys Pro Asn Asn Leu Val Val Glu Asp
20 25 30
Glu Gly Cys Thr Asn Leu Ser Glu Phe Ser Tyr Met Glu Leu Lys Val
35 40 45
Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys Thr Gly Val
50 55 60
Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr Val Thr Thr
65 70 75 80
Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala Cys Arg Ala
85 90 95
Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu Glu Ser Leu
100 105 110
His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val Arg Thr Thr
115 120 125
Lys Glu Ser Leu Ile Ile Ile Ser Pro Ser Val Thr Asp Leu Asp Pro
130 135 140
Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly Lys Cys Ser
145 150 155 160
Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His Asp Tyr Thr
165 170 175
Ile Trp Met Pro Glu Asn Pro Arg Pro Arg Thr Pro Cys Asp Ile Phe
180 185 190
Thr Asn Ser Arg Gly Lys Arg Ala Ser Lys Gly Asn Lys Thr Cys Gly
195 200 205
Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly Ala Cys Arg
210 215 220
Leu Lys Leu Cys Gly Val Leu Gly Leu Arg Leu Met Asp Gly Thr Trp
225 230 235 240
Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro Pro Asp Gln
245 250 255
Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu His Leu Val
260 265 270
Val Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp Ala Leu Glu
275 280 285
Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu Ser His Leu
290 295 300
Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile Phe Asn Lys
305 310 315 320
Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Arg Thr Trp Asn
325 330 335
Glu Ile Ile Pro Ser Lys Gly Cys Leu Lys Val Gly Gly Arg Cys His
340 345 350
Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu Gly Pro Asp
355 360 365
Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu Gln Gln His
370 375 380
Met Glu Leu Leu Lys Ser Ser Val Ile Pro Leu Met His Pro Leu Ala
385 390 395 400
Asp Pro Ser Thr Val Phe Lys Glu Gly Asp Glu Ala Glu Asp Phe Val
405 410 415
Glu Val His Leu Pro Asp Val Tyr Lys Gln Ile Ser Gly Val Asp Leu
420 425 430
Gly Leu Pro Asn Trp Gly Lys Tyr
435 440
<210> 36
<211> 440
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro Trp Ser Pro
1 5 10 15
Ile Asp Ile His His Leu Ser Cys Pro Asn Asn Leu Val Val Glu Asp
20 25 30
Glu Gly Cys Thr Asn Leu Ser Glu Phe Ser Tyr Met Glu Leu Lys Val
35 40 45
Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys Thr Gly Val
50 55 60
Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr Val Thr Thr
65 70 75 80
Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala Cys Arg Ala
85 90 95
Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu Glu Ser Leu
100 105 110
His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val Arg Thr Thr
115 120 125
Lys Glu Ser Leu Ile Ile Ile Ser Pro Ser Val Thr Asp Leu Asp Pro
130 135 140
Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly Lys Cys Ser
145 150 155 160
Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His Asp Tyr Thr
165 170 175
Ile Trp Met Pro Glu Asn Pro Arg Pro Arg Thr Pro Cys Asp Ile Phe
180 185 190
Thr Asn Ser Arg Gly Lys Arg Ala Ser Lys Gly Asn Lys Thr Cys Gly
195 200 205
Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly Ala Cys Arg
210 215 220
Leu Lys Leu Cys Glu Val Leu Gly Leu Arg Leu Met Asp Gly Thr Trp
225 230 235 240
Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro Pro Asp Gln
245 250 255
Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu His Leu Val
260 265 270
Val Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp Ala Leu Glu
275 280 285
Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu Ser His Leu
290 295 300
Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile Phe Asn Lys
305 310 315 320
Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Arg Thr Trp Asn
325 330 335
Glu Ile Ile Pro Ser Lys Gly Cys Leu Lys Val Gly Gly Arg Cys His
340 345 350
Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu Gly Pro Asp
355 360 365
Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu Gln Gln His
370 375 380
Met Glu Leu Leu Lys Ser Ser Val Ile Pro Leu Met His Pro Leu Ala
385 390 395 400
Asp Pro Ser Thr Val Phe Lys Glu Gly Asp Glu Ala Glu Asp Phe Val
405 410 415
Glu Val His Leu Pro Asp Val Tyr Lys Gln Ile Ser Gly Val Asp Leu
420 425 430
Gly Leu Pro Asn Trp Gly Lys Tyr
435 440
<210> 37
<211> 440
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro Trp Ser Pro
1 5 10 15
Ile Asp Ile His His Leu Ser Cys Pro Asn Asn Leu Val Val Glu Asp
20 25 30
Glu Gly Cys Thr Asn Leu Ser Glu Phe Ser Tyr Met Glu Leu Lys Val
35 40 45
Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys Thr Gly Val
50 55 60
Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr Val Thr Thr
65 70 75 80
Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala Cys Arg Ala
85 90 95
Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu Glu Ser Leu
100 105 110
His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val Arg Thr Thr
115 120 125
Lys Glu Ser Leu Ile Ile Ile Ser Pro Ser Val Thr Asp Leu Asp Pro
130 135 140
Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly Lys Cys Ser
145 150 155 160
Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His Asp Tyr Thr
165 170 175
Ile Trp Met Pro Glu Asn Pro Arg Pro Arg Thr Pro Cys Asp Ile Phe
180 185 190
Thr Asn Ser Arg Gly Lys Arg Ala Ser Lys Gly Asn Lys Thr Cys Gly
195 200 205
Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly Ala Cys Arg
210 215 220
Leu Lys Leu Cys Gly Val Leu Gly Leu Arg Leu Met Asp Gly Thr Trp
225 230 235 240
Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro Pro Asp Gln
245 250 255
Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu His Leu Val
260 265 270
Val Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp Ala Leu Glu
275 280 285
Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu Ser His Leu
290 295 300
Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile Phe Asn Lys
305 310 315 320
Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Arg Thr Trp Asn
325 330 335
Glu Thr Ile Pro Ser Lys Gly Cys Leu Lys Val Gly Gly Arg Cys His
340 345 350
Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu Gly Pro Asp
355 360 365
Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu Gln Gln His
370 375 380
Met Glu Leu Leu Lys Ser Ser Val Ile Pro Leu Met His Pro Leu Ala
385 390 395 400
Asp Pro Ser Thr Val Phe Lys Glu Gly Asp Glu Ala Glu Asp Phe Val
405 410 415
Glu Val His Leu Pro Asp Val Tyr Lys Gln Ile Ser Gly Val Asp Leu
420 425 430
Gly Leu Pro Asn Trp Gly Lys Tyr
435 440
<210> 38
<211> 32
<212> PRT
<213> 智人(Homo sapiens)
<400> 38
Val Ser Arg Leu Glu Glu Glu Met Arg Lys Leu Gln Ala Thr Val Gln
1 5 10 15
Glu Leu Gln Lys Arg Leu Asp Arg Leu Glu Glu Thr Val Gln Ala Lys
20 25 30
<210> 39
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro Trp Ser Pro
1 5 10 15
Ile Asp Ile His His Leu Ser Cys Pro Asn Asn Leu Val Val Glu Asp
20 25 30
Glu Gly Cys Thr Asn Leu Ser Glu Phe Ser Tyr Met Glu Leu Lys Val
35 40 45
Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys Thr Gly Val
50 55 60
Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr Val Thr Thr
65 70 75 80
Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala Cys Arg Ala
85 90 95
Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu Glu Ser Leu
100 105 110
His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val Arg Thr Thr
115 120 125
Lys Glu Ser Leu Ile Ile Ile Ser Pro Ser Val Thr Asp Leu Asp Pro
130 135 140
Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly Lys Cys Ser
145 150 155 160
Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His Asp Tyr Thr
165 170 175
Ile Trp Met Pro Glu Asn Pro Arg Pro Arg Thr Pro Cys Asp Ile Phe
180 185 190
Thr Asn Ser Arg Gly Lys Arg Ala Ser Lys Gly Asn Lys Thr Cys Gly
195 200 205
Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly Ala Cys Arg
210 215 220
Leu Lys Leu Cys Gly Val Leu Gly Leu Arg Leu Met Asp Gly Thr Trp
225 230 235 240
Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro Pro Asp Gln
245 250 255
Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu His Leu Val
260 265 270
Val Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp Ala Leu Glu
275 280 285
Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu Ser His Leu
290 295 300
Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile Phe Asn Lys
305 310 315 320
Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Arg Thr Trp Asn
325 330 335
Glu Ile Ile Pro Ser Lys Gly Cys Leu Lys Val Gly Gly Arg Cys His
340 345 350
Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu Gly Pro Asp
355 360 365
Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu Gln Gln His
370 375 380
Met Glu Leu Leu Lys Ser Ser Val Ile Pro Leu Met His Pro Leu Ala
385 390 395 400
Asp Pro Ser Thr Val Phe Lys Glu Gly Asp Glu Ala Glu Asp Phe Val
405 410 415
Glu Val His Leu Pro Asp Val Tyr Lys Gln Ile Ser Gly Val Asp Leu
420 425 430
Gly Leu Pro Asn Trp Gly Lys Tyr Gly Gly Gly Gly Ser Val Ser Arg
435 440 445
Leu Glu Glu Glu Met Arg Lys Leu Gln Ala Thr Val Gln Glu Leu Gln
450 455 460
Lys Arg Leu Asp Arg Leu Glu Glu Thr Val Gln Ala Lys Ala Ser Gly
465 470 475 480
Ala Ala His His His His His His
485
<210> 40
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro Trp Ser Pro
1 5 10 15
Ile Asp Ile His His Leu Ser Cys Pro Asn Asn Leu Val Val Glu Asp
20 25 30
Glu Gly Cys Thr Asn Leu Ser Glu Phe Ser Tyr Met Glu Leu Lys Val
35 40 45
Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys Thr Gly Val
50 55 60
Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr Val Thr Thr
65 70 75 80
Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala Cys Arg Ala
85 90 95
Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu Glu Ser Leu
100 105 110
His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val Arg Thr Thr
115 120 125
Lys Glu Ser Leu Ile Ile Ile Ser Pro Ser Val Thr Asp Leu Asp Pro
130 135 140
Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly Lys Cys Ser
145 150 155 160
Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His Asp Tyr Thr
165 170 175
Ile Trp Met Pro Glu Asn Pro Arg Pro Arg Thr Pro Cys Asp Ile Phe
180 185 190
Thr Asn Ser Arg Gly Lys Arg Ala Ser Lys Gly Asn Lys Thr Cys Gly
195 200 205
Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly Ala Cys Arg
210 215 220
Leu Lys Leu Cys Glu Val Leu Gly Leu Arg Leu Met Asp Gly Thr Trp
225 230 235 240
Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro Pro Asp Gln
245 250 255
Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu His Leu Val
260 265 270
Val Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp Ala Leu Glu
275 280 285
Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu Ser His Leu
290 295 300
Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile Phe Asn Lys
305 310 315 320
Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Arg Thr Trp Asn
325 330 335
Glu Ile Ile Pro Ser Lys Gly Cys Leu Lys Val Gly Gly Arg Cys His
340 345 350
Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu Gly Pro Asp
355 360 365
Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu Gln Gln His
370 375 380
Met Glu Leu Leu Lys Ser Ser Val Ile Pro Leu Met His Pro Leu Ala
385 390 395 400
Asp Pro Ser Thr Val Phe Lys Glu Gly Asp Glu Ala Glu Asp Phe Val
405 410 415
Glu Val His Leu Pro Asp Val Tyr Lys Gln Ile Ser Gly Val Asp Leu
420 425 430
Gly Leu Pro Asn Trp Gly Lys Tyr Gly Gly Gly Gly Ser Val Ser Arg
435 440 445
Leu Glu Glu Glu Met Arg Lys Leu Gln Ala Thr Val Gln Glu Leu Gln
450 455 460
Lys Arg Leu Asp Arg Leu Glu Glu Thr Val Gln Ala Lys Ala Ser Gly
465 470 475 480
Ala Ala His His His His His His
485
<210> 41
<211> 488
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro Trp Ser Pro
1 5 10 15
Ile Asp Ile His His Leu Ser Cys Pro Asn Asn Leu Val Val Glu Asp
20 25 30
Glu Gly Cys Thr Asn Leu Ser Glu Phe Ser Tyr Met Glu Leu Lys Val
35 40 45
Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys Thr Gly Val
50 55 60
Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr Val Thr Thr
65 70 75 80
Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala Cys Arg Ala
85 90 95
Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu Glu Ser Leu
100 105 110
His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val Arg Thr Thr
115 120 125
Lys Glu Ser Leu Ile Ile Ile Ser Pro Ser Val Thr Asp Leu Asp Pro
130 135 140
Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly Lys Cys Ser
145 150 155 160
Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His Asp Tyr Thr
165 170 175
Ile Trp Met Pro Glu Asn Pro Arg Pro Arg Thr Pro Cys Asp Ile Phe
180 185 190
Thr Asn Ser Arg Gly Lys Arg Ala Ser Lys Gly Asn Lys Thr Cys Gly
195 200 205
Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly Ala Cys Arg
210 215 220
Leu Lys Leu Cys Gly Val Leu Gly Leu Arg Leu Met Asp Gly Thr Trp
225 230 235 240
Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro Pro Asp Gln
245 250 255
Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu His Leu Val
260 265 270
Val Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp Ala Leu Glu
275 280 285
Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu Ser His Leu
290 295 300
Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile Phe Asn Lys
305 310 315 320
Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Arg Thr Trp Asn
325 330 335
Glu Thr Ile Pro Ser Lys Gly Cys Leu Lys Val Gly Gly Arg Cys His
340 345 350
Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu Gly Pro Asp
355 360 365
Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu Gln Gln His
370 375 380
Met Glu Leu Leu Lys Ser Ser Val Ile Pro Leu Met His Pro Leu Ala
385 390 395 400
Asp Pro Ser Thr Val Phe Lys Glu Gly Asp Glu Ala Glu Asp Phe Val
405 410 415
Glu Val His Leu Pro Asp Val Tyr Lys Gln Ile Ser Gly Val Asp Leu
420 425 430
Gly Leu Pro Asn Trp Gly Lys Tyr Gly Gly Gly Gly Ser Val Ser Arg
435 440 445
Leu Glu Glu Glu Met Arg Lys Leu Gln Ala Thr Val Gln Glu Leu Gln
450 455 460
Lys Arg Leu Asp Arg Leu Glu Glu Thr Val Gln Ala Lys Ala Ser Gly
465 470 475 480
Ala Ala His His His His His His
485

Claims (10)

1.药物组合物，其包含约0.2mg/ml～约20mg/ml的针对狂犬病病毒G蛋白的双特异性抗体、选自海藻糖和甘露醇的渗透压调节剂、金属离子螯合剂型抗氧化剂、非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.0-约6.2，其中所述双特异性抗体包含结合狂犬病病毒G蛋白的表位I的抗原结合片段和结合狂犬病病毒G蛋白的表位III的抗原结合片段，并且所述双特异性抗体具有中和狂犬病病毒的活性。
2.如权利要求1所述的药物组合物，其还包含缓冲盐，例如约10mM-约50mM的缓冲盐。
3.如权利要求2所述的药物组合物，其中所述药物组合物包含约0.5mg/ml～约2.0mg/ml的所述针对狂犬病病毒G蛋白的双特异性抗体、约10mM-约50mM的缓冲盐、约100mM-约400mM的选自海藻糖和甘露醇的渗透压调节剂、约0.2mM-约5mM的金属离子螯合剂型抗氧化剂(例如，乙二胺四乙酸盐)、约0.05mg/ml-约1.0mg/ml的非离子型表面活性剂和余量的水，其中所述药物组合物的pH值为约5.2-约6.2。
4.如权利要求2-3中任一项所述的药物组合物，其中所述缓冲盐是磷酸/磷酸盐缓冲对、组氨酸/组氨酸的无机盐缓冲对、醋酸/醋酸盐缓冲对、柠檬酸/柠檬酸盐缓冲对，优选组氨酸/组氨酸的无机盐缓冲对或醋酸/醋酸盐缓冲对，更优选组氨酸/盐酸组氨酸缓冲对或醋酸/醋酸钠缓冲对，还更优选组氨酸/盐酸组氨酸缓冲对。
5.如权利要求2-4中任一项所述的药物组合物，其中所述缓冲盐的浓度为约15mM-约30mM、约15mM-约25mM、约15mM-约20mM、约20mM-约25mM或约20mM。
6.如权利要求1-5中任一项所述的药物组合物，其中所述渗透压调节剂的浓度为约200mM-约300mM、约200mM-约250mM、约230mM-约260mM或约250mM-约300mM。
7.如权利要求1-6中任一项所述的药物组合物，其中所述金属离子螯合剂型抗氧化剂(例如，乙二胺四乙酸盐)的浓度为约0.5mM-约2.0mM、约0.5mM-约1.5mM、约0.5mM-约1.0mM、约1.0mM-约1.5mM或约1.0mM。
8.如权利要求1-7中任一项所述的药物组合物，其中所述非离子型表面活性剂为聚山梨酯型表面活性剂，例如聚山梨酯20和聚山梨酯80，其含量为约0.1mg/ml-约1.0mg/ml、约0.2mg/ml-约0.5mg/ml、约0.3mg/ml-约1.0mg/ml、约0.5mg/ml-约1.0mg/ml、约0.3mg/ml、约0.34mg/ml、约0.4mg/ml、约0.5mg/ml、约0.6mg/ml、约0.7mg/ml、约0.8mg/ml、约0.9mg/ml或约1.0mg/ml。
9.如权利要求1-8中任一项所述的药物组合物，其中所述药物组合物的pH值为约5.0-约6.0、约5.2-约6.2、约5.5-约6.0、约5.5-约5.8或约5.8。
10.权利要求1-9中任一项所述的药物组合物在制备用于预防或治疗狂犬病的药物中的用途。
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116327925A (zh) * 2022-09-27 2023-06-27 兴盟生物医药(苏州)有限公司 一种抗狂犬病毒人源抗体的组合制剂及应用
WO2023186173A1 (zh) * 2022-04-02 2023-10-05 重庆智翔金泰生物制药股份有限公司 包含针对狂犬病病毒g蛋白的双特异性抗体的药物制剂及其制备方法

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103505729A (zh) * 2013-05-24 2014-01-15 华北制药集团新药研究开发有限责任公司 一种稳定的狂犬病毒人源抗体组合制剂
CN110317267A (zh) * 2018-08-09 2019-10-11 北京智仁美博生物科技有限公司 针对狂犬病病毒的双特异性抗体及其用途
CN110623921A (zh) * 2019-08-15 2019-12-31 北京东方百泰生物科技有限公司 一种抗cd3和抗cd19的双特异性抗体注射制剂
CN110913904A (zh) * 2017-05-05 2020-03-24 美国安进公司 用于改善的储存和施用的包含双特异性抗体构建体的药物组合物
WO2021143826A1 (zh) * 2020-01-17 2021-07-22 信达生物制药(苏州)有限公司 重组抗程序性死亡受体1和抗分化抗原簇137双特异性抗体制剂及其用途
CN114146174A (zh) * 2020-07-24 2022-03-08 信达生物制药(苏州)有限公司 抗pd-l1/ox40双特异性抗体制剂及其制备方法和用途

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2668947C (en) * 2006-12-05 2017-02-07 Crucell Holland B.V. Liquid anti-rabies antibody formulations
JP2011072248A (ja) * 2009-09-30 2011-04-14 Evec Inc 狂犬病ウイルスに対するヒト抗体ならびにその組成物
CN107709360B (zh) * 2015-06-10 2021-10-19 赛特瑞恩股份有限公司 狂犬病毒g蛋白表位及与其特异性结合的中和狂犬病毒的结合分子
CN114748618B (zh) * 2022-04-02 2024-07-19 重庆智翔金泰生物制药股份有限公司 包含针对狂犬病病毒g蛋白的双特异性抗体的药物制剂及其制备方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103505729A (zh) * 2013-05-24 2014-01-15 华北制药集团新药研究开发有限责任公司 一种稳定的狂犬病毒人源抗体组合制剂
CN110913904A (zh) * 2017-05-05 2020-03-24 美国安进公司 用于改善的储存和施用的包含双特异性抗体构建体的药物组合物
CN110317267A (zh) * 2018-08-09 2019-10-11 北京智仁美博生物科技有限公司 针对狂犬病病毒的双特异性抗体及其用途
CN110623921A (zh) * 2019-08-15 2019-12-31 北京东方百泰生物科技有限公司 一种抗cd3和抗cd19的双特异性抗体注射制剂
WO2021143826A1 (zh) * 2020-01-17 2021-07-22 信达生物制药(苏州)有限公司 重组抗程序性死亡受体1和抗分化抗原簇137双特异性抗体制剂及其用途
CN114146174A (zh) * 2020-07-24 2022-03-08 信达生物制药(苏州)有限公司 抗pd-l1/ox40双特异性抗体制剂及其制备方法和用途

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023186173A1 (zh) * 2022-04-02 2023-10-05 重庆智翔金泰生物制药股份有限公司 包含针对狂犬病病毒g蛋白的双特异性抗体的药物制剂及其制备方法
CN116327925A (zh) * 2022-09-27 2023-06-27 兴盟生物医药(苏州)有限公司 一种抗狂犬病毒人源抗体的组合制剂及应用

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