CN115819596A - 一种靶向人ceacam5/6的抗体、制备方法和应用 - Google Patents

一种靶向人ceacam5/6的抗体、制备方法和应用 Download PDF

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CN115819596A
CN115819596A CN202111093726.0A CN202111093726A CN115819596A CN 115819596 A CN115819596 A CN 115819596A CN 202111093726 A CN202111093726 A CN 202111093726A CN 115819596 A CN115819596 A CN 115819596A
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徐伟
莫世甫
赵勇
顾莉蕴
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Youmai Biotechnology Lianyungang Co ltd
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    • A—HUMAN NECESSITIES
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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    • C07K16/46—Hybrid immunoglobulins

Abstract

本发明提供了一种靶向人癌胚抗原CEACAM5/6的单克隆抗体，和一种能同时靶向人癌胚抗原CEACAM5/6和人CD47的双特异性抗体、制备方法和应用。具体地，涉及一种双特异性抗体能同时(1)特异性结合人CD47蛋白并阻断其与SIRPα的结合，从而因此阻断CD47对表达SIRPα的巨噬细胞的抑制作用的抗人CD47抗体及其抗原结合片段；(2)特异性结合人癌胚蛋白CEACAM5/6。本发明还涉及所述双特异性抗体的制备方法和用途。

Description

一种靶向人CEACAM5/6的抗体、制备方法和应用
技术领域
本发明属于生物免疫技术领域。具体地，本发明提供了一种靶向人 CEACAM5/6的抗体、制备方法和应用。
背景技术
人癌胚抗原细胞粘附分子(CEACAM)基因家族早在上世纪60年代就被发 现，包括位于19号染色体(q13.1-13.3之间)的35个基因/伪基因，其中21个编 码蛋白质。CEACAM属于免疫球蛋白超家族黏附分子，其结构域高度糖基化， 通常包括1-2个免疫球蛋白可变区样结构域(N domain)和0-6个免疫球蛋白恒 定区样结构域。CEACAM蛋白位于细胞膜外，其中CEACAM1、CEACAM3和 CEACAM4通过疏水跨膜结构域与细胞膜连接；CEACAM5-8则通过糖基磷酰肌 醇连接到细胞膜上。这些细胞外域通常作为细胞(例如上皮、内皮、树突状和 白细胞)间的粘附分子。
CEACAM涉及多种细胞功能，以细胞间的黏附功能为基础，通过信号转导 调节细胞生长和分化，并在胰岛素稳态、血管生成及免疫调节中发挥重要作用。 CEACAM基因家族成员参与各种各样的病理生理作用，包括作为微生物病原体 的受体。它们在致癌中起重要作用，特别是在癌症检测、进展和转移中。癌胚 抗原细胞粘附分子5(CEACAM5，简称为CEA，又称为CD66e)是具有约180kDa 分子量的糖蛋白。CEACAM5含有经由糖基磷脂酰肌醇(GPI)锚与细胞膜连接 的7个结构域。7个结构域包括单一N端Ig可变域和与Ig恒定域同源的6个结构域 (A1-B1-A2-B2-A3-B3)。CEACAM5最初分类为仅在胎儿组织中表达的蛋白 质，现在已经在几种正常成年组织中被鉴定出来。CEACAM5的过量表达在许 多类型的癌症中观察到，例如，在结肠癌患者的血液中可以检测到CEACAM5， 并且进一步研究确定了它的过度表达与许多恶性肿瘤，通常是不良预后相关。 在前列腺癌和结直肠癌中，CEACAM5的过度表达被证明可以作为肿瘤生物标 志物。
此外，在多种恶性肿瘤，如乳房、胰腺、卵巢、结肠、肺和胃腺肿瘤中 CEACAM5/CEACAM6亦被发现呈过度表达，并且与肿瘤的侵袭性和转移有关。 在肝转移的起始过程中，CEACAM5与其受体CEAr结合，它们的相互作用导致 促炎细胞因子的活化和产生，主要是IL-1、IL-6、IL-10和TNF-α。总之，这些 细胞因子改变肝细胞和Kupffer细胞的微环境，以及它们与肝窦细胞的相互作用。 这些相互作用不仅影响肿瘤细胞或其他肝细胞，似乎也促进了脑脊液中CSC和 其他循环肿瘤细胞的活力。
综上，CEACAM5已成为用于靶向治疗的潜在有用的肿瘤相关抗原，而本 领域当前已有的靶向CEACAM5的药物仍有诸多不足。因此，本领域迫切需要 开发一种靶向CEACAM5的特异性抗体分子。
发明内容
本发明的目的在于提供一种靶向人CEACAM5/6的抗体、制备方法和应用。
在本发明的第一方面，提供了一种抗人CEACAM5/6抗体的重链可变区，所 述的重链可变区包括以下三个互补决定区：
SEQ ID NO.1所示的VH-CDR1，
SEQ ID NO.2所示的VH-CDR2，和
SEQ ID NO.3所示的VH-CDR3；其中，上述氨基酸序列中任意一种氨基酸序 列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留 对CEACAM5/6的结合亲和力的衍生序列。
在另一优选例中，所述重链可变区的氨基酸序列选自下组：SEQ ID NO.7、11 或13。
在本发明的第二方面，提供了一种抗人CEACAM5/6抗体的重链，所述的重链 具有如本发明的第一方面所述的重链可变区。
在另一优选例中，所述的抗体的重链还包括重链恒定区。
在另一优选例中，所述的重链恒定区为人源、鼠源、骆驼源或兔源的，较佳 地为人源的。
在另一优选例中，所述的重链恒定区选自人IgG1、IgG2、IgG3或IgG4的重链 恒定区，较佳地，为人IgG1的重链恒定区。
在另一优选例中，所述的抗体的重链的序列如SEQ ID NO.9、14或15所示。
在本发明的第三方面，提供了一种抗人CEACAM5/6抗体的轻链可变区，所述 的轻链可变区包括以下三个互补决定区CDR：
SEQ ID NO.4所示的VL-CDR1，
SEQ ID NO.5所示的VL-CDR2，和
SEQ ID NO.6所示的VL-CDR3；其中，上述氨基酸序列中任意一种氨基酸序列 还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留对 CEACAM5/6的结合亲和力的衍生序列。
在另一优选例中，所述轻链可变区的序列如SEQ ID NO.8或12所示。
在本发明的第四方面，提供了一种抗人CEACAM5/6抗体的轻链，所述的轻链 具有如本发明的第三方面所述的轻链可变区。
在另一优选例中，所述抗体的轻链还包括轻链恒定区。
在另一优选例中，所述的轻链恒定区为人源、鼠源、骆驼源或兔源的，较佳 地为人源的。
在另一优选例中，所述的轻链恒定区选自人κ或λ亚型的轻链恒定区。
在另一优选例中，所述的抗体的轻链如SEQ ID NO.10或16所示。
在本发明的第五方面，提供了一种抗人CEACAM5/6抗体，所述抗体具有：
(1)如本发明的第一方面所述的重链可变区；和/或
(2)如本发明的第三方面所述的轻链可变区；
或者，所述抗体具有：如本发明的第二方面所述的重链；和/或如本发明的第 四方面所述的轻链。
在另一优选例中，所述抗体为人源化抗体。
在另一优选例中，所述抗体特异性结合人CEACAM5和/或人CEACAM6。
在另一优选例中，所述的抗体为双链抗体、或单链抗体。
在另一优选例中，所述的抗体为单克隆抗体。
在另一优选例中，所述的抗体包括单特异性、双特异性、三特异性或多特异 性抗体。
在另一优选例中，所述的抗体的轻链包括所述的三个轻链CDR以及用于连接 轻链CDR的轻链框架区；和所述的抗体的重链包括所述的三个重链CDR以及用于 连接重链CDR的重链框架区。
在另一优选例中，所述的抗体的重链可变区如SEQ ID NO.7、11、13所示； 和/或所述的抗体的轻链可变区如SEQ ID NO.8、12所示。
在另一优选例中，所述的抗体的重链可变区如SEQ ID NO.7所示，并且所述 抗体的轻链可变区如SEQ ID NO.8所示。
在另一优选例中，所述的抗体的重链可变区如SEQ ID NO.11所示，并且所述 抗体的轻链可变区如SEQ ID NO.12所示。
在另一优选例中，所述的抗体的重链可变区如SEQ ID NO.13所示，并且所述 抗体的轻链可变区如SEQ ID NO.12所示。
在另一优选例中，所述的多特异性抗体，包括：
(1)如本发明的第五方面中所述的CEACAM5/6抗体或其抗原结合片段；
(2)结合其他靶点的抗体或其抗原结合片段。
在另一优选例中，所述其他靶点选自下组：CD47、CD73、CD47、CD3、CTLA4、 PD-1、PD-L1、CD28、CD40、OX40、LAG3、DR5、TIM-3、TIGIT、VEGF、VEGFR、 Ang2、CD39、CD73、GITR。
在本发明的第六方面，提供了一种多特异性抗原结合分子，包含：
第一抗原结合域D1；和
第二抗原结合域D2；
其中，D1特异性结合靶分子CEACAM5/6蛋白；
D2特异性结合靶分子CD47蛋白；
所述D1为特异性结合CEACAM5/6蛋白的抗体或其抗原结合片段；
所述D2为特异性结合CD47蛋白的抗体或其抗原结合片段；
其中，所述D1具有包含以下三个互补决定区CDR的重链可变区：
SEQ ID NO.1所示的VH-CDR1，
SEQ ID NO.2所示的VH-CDR2，和
SEQ ID NO.3所示的VH-CDR3；以及
包含以下三个互补决定区CDR的轻链可变区：
SEQ ID NO.4所示的VL-CDR1，
SEQ ID NO.5所示的VL-CDR2，和
SEQ ID NO.6所示的VL-CDR3；
其中，所述抗原结合片段的结构选自下组：(i)Fab片段；(ii)F(ab')2片段；(iii)Fd 片段；(iv)Fv片段；(v)单链Fv(scFv)分子；或(vi)dAb片段。
在另一优选例中，所述特异性结合CD47蛋白的抗体的重链包括：具有SEQ IDNO.17所示的重链可变区；和所述特异性结合CD47蛋白的抗体的轻链包括：具有 SEQ IDNO.18所示的轻链可变区。
在另一优选例中，所述抗人CD47抗体的重链可变区或轻链可变区各自包含1-6 个突变，较佳地，1-4个突变。
在另一优选例中，所述抗人CD47抗体的重链可变区的突变选自下组：
H-I37V、H-P45L、H-I48V、H-T52S、H-T52L、H-T52I、H-T52H、H-T52A、 H-T52G、H-T52N、H-T52Q、H-T64K、H-R94K、H-P101D、或其组合。
在另一优选例中，所述抗人CD47抗体的轻链可变区的突变选自下组：
L-L4M、L-I46L、L-V58I、L-T93N、或其组合。
在另一优选例中，所述特异性结合CD47蛋白的抗体的重链包括：具有SEQ IDNO.19所示的重链可变区；和所述特异性结合CD47蛋白的抗体的轻链包括：具有 SEQ IDNO.20所示的轻链可变区。
在另一优选例中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过 添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留对CD47的结合亲和力 的衍生序列。
在另一优选例中，所述的双特异性抗原结合分子是双特异性抗体。
在另一优选例中，所述的D1和/或D2为单链抗体(scFv)。
在另一优选例中，所述的双特异性抗原结合分子中，仅D2为单链抗体(scFv)。
在另一优选例中，所述的D1和D2通过接头连接，所述接头包括抗体恒定区。
在另一优选例中，所述的D1为抗人CEACAM5/6抗体。
在另一优选例中，所述的D2为抗CD47单链抗体(scFv)。
在另一优选例中，所述的抗CEACAM5/6人源化抗体包括：单链抗体、双链抗 体、单克隆抗体、嵌合抗体、鼠源抗体、人源化抗体、和双特异性抗体。
在另一优选例中，所述的抗人CEACAM5/6抗体包含以下三个互补决定区CDR 的重链可变区：
SEQ ID NO.1所示的VH-CDR1，
SEQ ID NO.2所示的VH-CDR2，和
SEQ ID NO.3所示的VH-CDR3；以及
包含以下三个互补决定区CDR的轻链可变区：
SEQ ID NO.4所示的VL-CDR1，
SEQ ID NO.5所示的VL-CDR2，和
SEQ ID NO.6所示的VL-CDR3。
在另一优选例中，所述的双特异性抗体包含选自表4所示的重链和/或轻链：
表4.
Figure BDA0003268530520000051
Figure BDA0003268530520000061
在本发明的第七方面，提供了一种重组蛋白，所述的重组蛋白具有：
(i)如本发明第一方面所述的重链可变区、如本发明第二方面所述的重链、如 本发明第三方面所述的轻链可变区、如本发明第四方面所述的轻链、如本发明第五 方面所述的抗体或如本发明第六方面所述的多特异性抗原结合分子；以及
(ii)任选的协助表达和/或纯化的标签序列。
在另一优选例中，所述的标签序列包括6His标签。
在另一优选例中，所述的重组蛋白(或多肽)包括融合蛋白。
在另一优选例中，所述的重组蛋白为单体、二聚体、或多聚体。
在另一优选例中，所述的重组蛋白还包括靶向其他肿瘤靶点的蛋白或多肽药 物。
在另一优选例中，所述蛋白或多肽药物包含具有调节免疫系统功能的细胞因 子。
在另一优选例中，所述细胞因子包括野生型或突变型。
在另一优选例中，所述细胞因子选自下组：IL-1、IL-2、IL-7、IL8、IL-10、 IL-12、IL-15、IL-18、IL-21、GM-CSF、IFNα/β/γ，OX40-L、4-1BB-L、CTLA-4、 或其组合。
在本发明的第八方面，提供了一种CAR构建物，所述的CAR构建物的抗原结 合结构域的scFv区段特异性结合于人CEACAM5/6，并且所述scFv区段包含如本发 明的第一方面所述的重链可变区和如本发明的第三方面所述的轻链可变区。
在本发明的第九方面，提供了一种重组的免疫细胞，所述的免疫细胞表达外 源的如本发明的第八方面所述的CAR构建物。
在另一优选例中，所述的免疫细胞选自下组：NK细胞、T细胞。
在另一优选例中，所述的免疫细胞来自人或非人哺乳动物(如鼠)。
在本发明的第十方面，提供了一种抗体药物偶联物，所述的抗体药物偶联物 含有：
(a)抗体部分，所述抗体部分选自下组：如本发明第一方面所述的重链可变区、 如本发明第二方面所述的重链、如本发明第三方面所述的轻链可变区、如本发明第 四方面所述的轻链、如本发明第五方面所述的抗体、或如本发明的第六方面所述的 多特异性抗原结合分子、或其组合；和
(b)与所述抗体部分偶联的偶联部分，所述偶联部分选自下组：可检测标记物、 药物、毒素、细胞因子、酶、或其组合。
在另一优选例中，所述的抗体部分与所述的偶联部分通过化学键或接头进行 偶联。
在本发明的第十一方面，提供了一种活性成分的用途，所述活性成分选自下 组：如本发明第一方面所述的重链可变区、如本发明第二方面所述的重链、如本发 明第三方面所述的轻链可变区、如本发明第四方面所述的轻链、如本发明第五方面 所述的抗体、如本发明的第六方面所述的多特异性抗原结合分子、如本发明的第七 方面所述的重组蛋白、如本发明的第八方面所述的CAR构建物、如本发明的第九 方面所述的重组的免疫细胞、如本发明的第十方面所述的抗体药物偶联物、或其组 合，所述活性成分用于
(a)制备检测试剂或试剂盒；
(b)制备预防和/或治疗人CEACAM5/6相关疾病的药物或制剂；和/或
(c)制备预防和/或治疗人CEACAM5/6相关的癌症或肿瘤的药物或制剂。
在另一优选例中，所述癌症或肿瘤选自下组：结肠癌、直肠癌、淋巴瘤、胰 腺癌、肺癌、胃癌、肝细胞瘤、乳腺癌或甲状腺癌、或其组合。
在本发明的第十二方面，提供了一种药物组合物，所述的药物组合物含有：
(i)活性成分，所述活性成分选自下组：如本发明第一方面所述的重链可变区、 如本发明第二方面所述的重链、如本发明第三方面所述的轻链可变区、如本发明第 四方面所述的轻链、如本发明第五方面所述的抗体、如本发明第六方面所述的多特 异性抗原结合分子、如本发明第七方面所述的重组蛋白、如本发明第八方面所述的 CAR构建物、如本发明第九方面所述的重组的免疫细胞、如本发明第十方面所述 的抗体药物偶联物、或其组合；以及
(ii)药学上可接受的载体。
在另一优选例中，所述的药物组合物为液态制剂。
在另一优选例中，所述的药物组合物为注射剂。
在另一优选例中，所述的药物组合物用于治疗肿瘤。
在另一优选例中，所述肿瘤为高表达人CEACAM5/6的肿瘤。
在本发明的第十三方面，提供了一种多核苷酸，所述的多核苷酸编码选自下 组的多肽：
(1)如本发明第一方面所述的重链可变区、如本发明第二方面所述的重链、如 本发明第三方面所述的轻链可变区、如本发明第四方面所述的轻链、如本发明第五 方面所述的抗体；或
(2)如本发明第六方面所述的多特异性抗原结合分子；
(3)如本发明第七方面所述的重组蛋白；或
(4)如本发明第八方面所述的CAR构建物。
在本发明的第十四方面，提供了一种载体，所述的载体含有如本发明的第十 三方面所述的多核苷酸。
在另一优选例中，所述的载体包括：细菌质粒、噬菌体、酵母质粒、植物细 胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒、或其他载体。
在本发明的第十五方面，提供了一种遗传工程化的宿主细胞，所述的宿主细 胞含有如本发明的第十四方面所述的载体或基因组中整合有如本发明的第十三方 面所述的多核苷酸。
在另一优选例中，所述的宿主细胞为哺乳动物细胞，较佳地，为人、鼠、羊、 马、狗或猫的细胞，更佳地，为中国仓鼠卵巢细胞。
在本发明的第十六方面，提供了一种体外非诊断的检测(包括诊断性或非诊断性)样品中人CEACAM5/6蛋白的方法，所述方法包括步骤：
(1)在体外，将所述样品与如本发明的第五方面所述的抗体接触；
(2)检测是否形成抗原-抗体复合物，其中形成复合物就表示样品中存在CEACAM5/6蛋白。
在本发明的第十七方面，提供了一种治疗人CEACAM5/6相关疾病的方法，所 述方法包括：
给需要的对象施用如本发明的第五方面所述的抗体、所述抗体的多特异性抗 体、所述抗体的药物偶联物、或表达所述抗体的CAR-T细胞、或其组合。
在另一优选例中，所述人CEACAM5/6相关疾病为CEACAM5/6高表达的肿瘤， 优选为结直肠癌、胰腺癌、肺癌、胃癌、肝细胞瘤、乳腺癌和甲状腺癌。
在另一优选例中，所述人CEACAM5/6相关疾病为CEACAM5/6和CD47高表达 的肿瘤，优选为结直肠癌。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例) 中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方 案。限于篇幅，在此不再一一累述。
附图说明
图1显示了UM05-C9与人CEACAM5蛋白的ELISA。
图2显示了UM05-C9与CHO-CEACAM5细胞的结合。
图3显示了UM05-L9和双特异性抗体L34与食蟹猴(A)CEACAM5蛋白和 (B)CEACAM6蛋白的ELISA。
图4显示了UM05-C9的细胞内化结果。
图5显示了CD47-CEACAM5/6双特异性抗体的不同结构组合。其中，绿色 或橙色分别标志不同抗体的抗原结合片段(CD47抗体或CEACAM5/6抗体的 VH/VL)。
图6显示了UM11-L34分子对(A)CEACAM5和(B)CD47分子分别进行的亲和 力检测。
图7显示了UM11-L34分子对CEACAM5和CD47分子顺序进行的亲和力检 测。
具体实施方式
本发明人经过广泛而深入地研究，通过大量筛选，首次开发了一种活性优 异的抗CEACAM5的抗体，并且意外发现，所述的抗体竟然对CEACAM6也具有 优异的结合活性。此外，本发明还首次提供了一种高效、安全的抗人 CD47-CEACAM5/6的双特异性抗体、及其制备方法和用途。本发明的双特异性 抗体对CEACAM5/6和CD47高表达的肿瘤细胞有更好的靶向和特异性杀伤作用， 同时可以降低与红细胞和/或血小板的结合作用。具体地，本发明的抗CEACAM5/6和CD47双特异性抗体与CD47+-CEACAM5/6+肿瘤细胞结合选择性 更强，与红细胞和血小板表面CD47蛋白结合更弱，通过调节人免疫功能治疗多 种癌症。在此基础上完成了本发明。
术语
如本文所用，术语“抗体”指免疫球蛋白，是由两条相同的重链和两条相同 的轻链通过链间二硫键连接而成的四肽链结构。免疫球蛋白重链恒定区的氨基 酸组成和排列顺序不同，故其抗原性也不同。据此，可将免疫球蛋白分为五类， 或称为免疫球蛋白的不同种型，即IgM、IgD、IgG、IgA和IgE，对应于不同类 免疫球蛋白的重链恒定区分别称为α、δ、ε、γ、和μ。IgG代表免疫球蛋白中最 重要的一类，由于化学结构和生物功能差异，它又可以分为4个子类：IgG1、IgG2、 IgG3和IgG4。轻链通过恒定区的不同分为κ或λ链。不同类免疫球蛋白的亚单位 结构和三维构型是本领域人员所熟知的。
互补结合决定区(complement determine region，CDR)是决定抗体与抗原 特异性结合的关键序列结构。重链和轻链各包含了3个CDR，分别命名为HCDR1、 HCDR2和HCDR3，以及LCDR1、LCDR2和LCDR3。在一个给定的抗体的轻链 可变区或重链可变区氨基酸序列中，各CDR的精确氨基酸序列边界可以使用许 多公知的抗体CDR指派系统的任一种或其组合确定，所述指派系统包括例如： 基于抗体的三维结构和CDR环的拓扑学的Chothia(Chothia等人.(1989)Nature 342:877-883，Al-Lazikani等人,“Standard conformations for thecanonical structures of immunoglobulins”,Journal of Molecular Biology,273,927-948(1997))， 基于抗体序列可变性的Kabat(Kabat等人,Sequences of Proteins ofImmunological Interest,第4版,U.S.Department of Health and Human Services,National Institutes of Health(1987)),AbM(University of Bath)，Contact(University College London)，国际Immuno GeneTics database(IMGT)，以及基于loop结构位 置的Chothia定义。
例如，根据不同的CDR确定方案，每一个CDR的残基如下所述。
表1 CDR定义方案
Figure BDA0003268530520000101
注1：不同文献中的标注略有不同，特别是Chothia标注方案。
注2：除了contact标注方案使用Chothia或者Martin编号方案外，其他的标注 方案都能与各种编号方案兼容。
注3：Chothia CDR-H1环的末端在使用Kabat编号惯例时根据环的长度在 H32和H34之间变化。(这是因为Kabat编号方案将插入放置在H35A和H35B处) 如果H35A和H35B都不存在时，那么CDR-H1结束在32位；如果只有H35A存在 时，那么CDR-H1结束在33位；如果H35A和H35B同时存在，那么CDR-H1结束 在34位。
除非另有说明，否则在本发明中，术语“CDR”或“CDR序列”涵盖以上述任 一种方式确定的CDR序列。
除非另有说明，否则在本发明中，当提及抗体可变区中的残基位置(包括重 链可变区残基和轻链可变区残基)时，是指根据Kabat编号系统(Kabat等人， Sequences ofProteins of Immunological Interest,5th Ed.Public Health Service,NationalInstitutes of Health,Bethesda,Md.(1991))的编号位置。
在一个实施方案中，本发明抗体的CDR通过Kabat规则确定边界，或通过 IMGT规则确定，或通过其组合确定规则。
具有不同特异性(即，针对不同抗原的不同结合位点)的抗体具有不同的CDR。然而，尽管CDR在抗体与抗体之间是不同的，但是CDR内只有有限数量 的氨基酸位置直接参与抗原结合。使用Kabat、Chothia、AbM、Contact和IMGT 方法中的至少两种，可以确定最小重叠区域，从而提供用于抗原结合的“最小结 合单位”。最小结合单位可以是CDR的一个子部分。正如本领域技术人员熟知 的，通过抗体的结构和蛋白折叠，可以确定CDR序列其余部分的残基。因此， 本发明也考虑本文所给出的任何CDR的变体。例如，在一个CDR的变体中，最小结合单位的氨基酸残基可以保持不变，而根据Kabat或IMGT定义的其余CDR 残基可以被保守氨基酸残基替代。
如本文所用，术语“框架区”(FR)指插入CDR间的氨基酸序列，即指在单 一物种中不同的免疫球蛋白间相对保守的免疫球蛋白的轻链和重链可变区的 那些部分。免疫球蛋白的轻链和重链各具有四个FR，分别称为FR1-L、FR2-L、 FR3-L、FR4-L和FR1-H、FR2-H、FR3-H、FR4-H。相应地，轻链可变结构域可 因此称作(FR1-L)-(CDR1-L)-(FR2-L)-(CDR2-L)-(FR3-L)-(CDR3-L)-(FR4-L)且重 链可变结构域可因此表示为(FR1-H)-(CDR1-H)-(FR2-H)-(CDR2-H)-(FR3-H)- (CDR3-H)-(FR4-H)。优选地，本发明的FR是人抗体FR或其衍生物，所述人抗 体FR的衍生物与天然存在的人抗体FR基本相同，即序列同一性达到85％、90％、95％、96％、97％、98％或99％。
获知CDR的氨基酸序列，本领域的技术人员可轻易确定框架区FR1-L、 FR2-L、FR3-L、FR4-L和/或FR1-H、FR2-H、FR3-H、FR4-H。
如本文所用，术语″人框架区″是与天然存在的人抗体的框架区基本相同 的(约85％或更多，具体地90％、95％、97％、99％或100％)框架区。
如本文所用，术语“单克隆抗体”或“mAb”指针对特定抗原的具有单一 氨基酸组成的抗体分子，且不应理解为需要通过任何特定方法产生该抗体。单 克隆抗体可由B细胞或杂交瘤的单一克隆产生，但还可为重组的，即通过蛋白 工程产生。
如本文所用，术语“抗原”或“靶抗原”指能够由抗体或抗体样结合蛋白 所结合的分子或分子的部分。该术语进一步指能够用于动物以产生能够与该抗 原的表位结合的抗体的分子或分子的部分。靶抗原可具有一个或多个表位。对 于每种由抗体或由抗体样结合蛋白识别的靶抗原，抗体样结合蛋白能够与识别 靶抗原的完整抗体竞争。
如本文所用，术语“亲和力”理论上通过完整抗体和抗原间的平衡缔合来 定义。本发明双抗的亲和力可以通过KD值(解离常数)(或其它测定方式)进行评 估或测定，例如生物膜层干涉技术(Bio-layer interferometry BLI)，使用Biacore 8K测量确定。
抗CEACAM5/6抗体
CEACAM属于免疫球蛋白超家族黏附分子，其结构域高度糖基化，通常包 括1-2个免疫球蛋白可变区样结构域(N domain)和0-6个免疫球蛋白恒定区样 结构域。CEACAM蛋白位于细胞膜外，其中CEACAM1、CEACAM3和CEACAM4 通过疏水跨膜结构域与细胞膜连接；CEACAM5-8则通过糖基磷酰肌醇连接到 细胞膜上。这些细胞外域通常作为细胞(例如上皮、内皮、树突状和白细胞) 间的粘附分子。在多种恶性肿瘤，如乳房、胰腺、卵巢、结肠、肺和胃腺肿瘤 中CEACAM5/CEACAM6被发现呈过度表达，并且与肿瘤的侵袭性和转移有关。
如本文所用，“抗CEACAM5/6抗体”、“抗CEACAM5/CEACAM6抗体” 是指本发明中能够与CEACAM5分子和CEACAM6分子有较强特异性结合的抗 体，即所述抗体具有针对CEACAM5和CEACAM6分子的交叉反应性。
本发明提供了一种抗CEACAM5/6抗体(例如，UM05-L9)，其与CEACAM5 分子有强结合外，亦与CEACAM6分子有较强结合。
优选地，所述抗体包括重链和轻链，所述重链含有重链可变区(VH)氨基酸 序列，所述轻链含有轻链可变区(VL)氨基酸序列。
所述的重链可变区(VH)具有选自下组的互补决定区VH-CDR：
SEQ ID NO.1所示的VH-CDR1，
SEQ ID NO.2所示的VH-CDR2，和
SEQ ID NO.3所示的VH-CDR3；
所述的轻链可变区(VL)具有选自下组的互补决定区VL-CDR：
SEQ ID NO.4所示的VL-CDR1，
SEQ ID NO.5所示的VL-CDR2，和
SEQ ID NO.6所示的VL-CDR3；
其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺 失、修饰和/或取代至少一个氨基酸的，并能够保留CEACAM5和/或CEACAM6 结合亲和力的衍生序列。
在另一优选例中，所述经过添加、缺失、修饰和/或取代至少一个氨基酸序 列所形成的序列优选为同源性或序列相同性为至少80％，较佳地至少85％，更 佳地至少为90％，最佳地至少95％的氨基酸序列。
本发明上述内容中，所述添加、缺失、修饰和/或取代的氨基酸数量，优选 为不超过初始氨基酸序列总氨基酸数量的40％，更优选为不超过35％，更优选 为1-33％，更优选为5-30％，更优选为10-25％，更优选为15-20％。
本发明上述内容中，更优选地，所述添加、缺失、修饰和/或取代的氨基酸 数量，可以是1-7个，更优选为1-5个，更优选为1-3个，更优选为1-2个。
较佳地，本文所述抗体为抗体全长蛋白、抗原抗体结合域蛋白质片段、双 特异性抗体、多特异性抗体、单链抗体(single chain antibody fragment，scFv)、 单域抗体(single domain antibody，sdAb)和单区抗体(Signle-domain antibody)中 的一种或多种，以及上述抗体所制得的单克隆抗体或多克隆抗体。所述单克隆 抗体可以由多种途径和技术进行研制，包括杂交瘤技术、噬菌体展示技术、单 淋巴细胞基因克隆技术等，主流是通过杂交瘤技术从野生型或转基因小鼠制备 单克隆抗体。
所述的抗体全长蛋白为本领域常规的抗体全长蛋白，其包括重链可变区、 轻链可变区、重链恒定区和轻链恒定区。所述的蛋白质的重链可变区和轻链可 变区与人源重链恒定区和人源轻链恒定区构成全人源抗体全长蛋白。较佳地， 所述的抗体全长蛋白为IgG1、IgG2、IgG3或IgG4。所述人源轻链恒定区选自人 κ或λ亚型的轻链恒定区；进一步优选地，所述人源重链恒定区为人IgG1的重 链恒定区，所述人源轻链恒定区为κ链。
本发明中的抗体(抗CEACAM5/6的抗体)可以是全长蛋白(如IgG1,IgG2a, IgG2b或者IgG2c)，也可以是包含抗原抗体结合域或抗原结合片段的蛋白片段 (例如Fab，F(ab'),sdAb,scFv片段)。
如本文所用，抗原结合片段的非限制性例子包括：(i)Fab片段；(ii)F(ab')2片段；(iii)Fd片段；(iv)Fv片段；(v)单链Fv(scFv)分子；(vi)dAb片段；和(vii)由 模拟抗体高变区的氨基酸残基组成的最小识别单位(例如，独立的互补性决定区 (CDR)如CDR3肽)或受约束的FR3-CDR3-FR4肽。在另一优选例，本发明的 CEACAM5/6的抗体或其抗原结合片段选自下组：驼源单域抗体、scFv、scFv 二聚体、BsFv、dsFv、dsFv2、dsFv-dsFv'、Fv片段、Fab、Fab'、F(ab')2、ds双 功能抗体、纳米抗体、域抗体或双价域抗体。
本发明中的抗体(抗CEACAM5/6的抗体)可以是野生型蛋白，也可以是经过 特定突变已达到某种特定效果的突变型蛋白，例如利用突变消除抗体的效应子 功能。
本发明的抗体可以是双链或单链抗体，并且可以是选自动物源抗体、嵌合 抗体、人源化抗体，更优选为人源化抗体、人-动物嵌合抗体，更优选为全人源 化抗体。
本发明所述抗体衍生物可以是单链抗体、和/或抗体片段，如：Fab、Fab'、 (Fab')2或该领域内其他已知的抗体衍生物等，以及IgA、IgD、IgE、IgG以及IgM 抗体或其他亚型的抗体中的任意一种或几种。所述的单链抗体为本领域常规的 单链抗体，其包括重链可变区、轻链可变区和15～20个氨基酸的短肽。
其中，所述动物优选为哺乳动物，如鼠。
本发明抗体可以是靶向CEACAM5/6(例如人CEACAM5/6)的嵌合抗体、人 源化抗体、CDR嫁接和/或修饰的抗体。
在另一优选例中，所述抗体的重链可变区含有SEQ ID NO.7、11或13所示 的氨基酸序列。
在另一优选例中，所述抗体的轻链可变区含有SEQ ID NO.8或12所示的氨 基酸序列。
在另一优选例中，所述靶向CEACAM5/6的抗体的重链可变区(VH)氨基酸 序列，和/或轻链可变区(VL)氨基酸序列如下表2所示：
表2.抗体编号及VH、VL的序列编号(SEQ ID NO.)
抗体编号 VH序列编号 VL序列编号
UM05-L9 7 8
UM05-C9 11 12
UM05-C9’ 13 12
其中，UM05-L9是人-鼠嵌合抗体，UM05-C9是人源化抗体，UM05-C9’ 是另一个版本的人源化抗体，三者的VH-CDR以及VL-CDR相同。
CD47及其抗体
CD47是45～50kD的跨膜糖蛋白，属于免疫球蛋白(Ig)超家族的成员。 研究发现，肿瘤细胞上高表达的CD47与巨噬细胞上的配体SIRPα结合，使得 SIRPα酪氨酸磷酸化，从而发出抑制性调节信号，抑制巨噬细胞的吞噬作用。 对应的，阻断该通路可以解除巨噬细胞对肿瘤细胞吞噬的抑制作用，提高机体 对肿瘤细胞的免疫反应，为肿瘤的免疫治疗提供一条新途径。
在以CD47-SIRPα轴为靶标的肿瘤治疗中，最主要的机制是巨噬细胞的激 活，从而增强巨噬细胞对肿瘤细胞的吞噬作用。其次，阻断CD47可能进一步将 巨噬细胞募集到肿瘤组织中，以及向肿瘤组织募集额外的免疫细胞的细胞因子 和趋化因子，如单核细胞趋化蛋白3(MCP-3)，这些细胞因子的分泌有助于 CD47阻断疗法的功效。再次，靶向CD47-SIRPα的疗法也可能改变肿瘤中巨噬 细胞的极化状态。巨噬细胞按其表型和功能活性又可以分为M1(Ⅰ型)和M2 (Ⅱ型)两种，M1巨噬细胞能产生大量的促炎细胞因子，并介导抵抗细胞内寄生虫和抑制肿瘤生长；M2巨噬细胞则产生较少的促炎分子，参与组织损伤修复、 血管生成和促进肿瘤生长。研究发现，阻断CD47-SIRPα后可显著增加小鼠微 环境中的M1巨噬细胞肿，而小鼠M2巨噬细胞则没有显著的增加。最后，其它 免疫细胞也可以响应CD47阻断疗法。SIRPα在骨髓免疫细胞上高度表达，因 此它可能是骨髓谱系的关键调节剂。在小鼠中，CD47通过SIRPα+树突状细胞 来调节抗原摄取，使用同源免疫活性肿瘤模型发现，CD47阻断的治疗效果取决 于树突状细胞。通过刺激巨噬细胞或树突细胞的抗原呈递，靶向CD47-SIRPα轴的疗法可以促进肿瘤的适应性免疫应答。
CD47广泛表达于多种细胞，特别是在新生红细胞上高表达。因此靶向CD47 的治疗性抗体有可能会引起贫血。另一方面，体内游离抗人CD47抗体的浓度也 会由于红细胞对抗人CD47抗体的吸附作用而大大降低。
因此，开发新的治疗性抗人CD47抗体，使得其与肿瘤细胞表面CD47蛋白 的结合选择性更强，与红细胞表面CD47蛋白结合更弱，成为新型治疗性抗人 CD47抗体的目标之一。
本发明中优选的抗人CD47抗体包括中国专利202010240238.7中描述的抗 人CD47抗体或其抗原结合片段，其通过引用并入本文(如本文所用，以代号 UM03-C4表示，其重链可变区如SEQ ID NO.17；轻链可变区如SEQ ID NO.18 所示)。该CD47抗体的IgG4亚型在本专利中以代号UM03-C4表示。此外，在 该抗人CD47抗体基础上，还包括对其VH、VL区进行突变而得的衍生抗体人 CD47抗体(如本文所用，以代号UM03-C4’表示)。所涉及的突变抗人CD47抗体 包括如下突变或者下述突变的组合：
重链：H-I37V、H-P45L、H-I48V、H-R94K、H-T52R、H-T52S、H-T52L、 H-T52I、H-T52H、H-T52A、H-T52N、H-T52Q；
轻链：L-L4M、L-I46L、L-V58I、L-T93N。
优选地，将抗人CD47抗体或其抗原结合域与抗人CEACAM5/6的抗体或其 抗原结合域进行不同组合从而获得本发明中所述的双特异性抗体。
双特异性抗体
双特异性抗体(Bispecific Antibody，bsAb)是一种非天然抗体，它能同时靶 向两种不同的抗原或蛋白，阻断两种不同的信号通路，激发具有特异性的免疫 反应，其特异性和双功能性在肿瘤免疫治疗中的作用越来越重要，已成为当今 世界抗体工程治疗肿瘤方面的研究热点。研究表明，双特异性抗体在肿瘤免疫 治疗方面主要有介导免疫细胞对肿瘤的杀伤；结合双靶点，阻断双信号通路， 发挥独特的或重叠的功能，可以有效防止耐药性；具有强特异性、靶向性和降 低脱靶毒性；有效降低治疗成本等优势(摘自抗体圈)，因此采用双特异性抗体 药物可以降低肿瘤细胞逃逸几率，清除肿瘤细胞，提高疗效。
双特异性抗体可通过双杂交瘤细胞，化学偶联，重组基因等手段制备，其 中重组基因技术在结合位点以及产量等方面灵活性强。据不完全统计，目前已 有60多种双特异性抗体，根据其特点以及结构差异性，双特异性抗体结构主要 有含Fc片段的双特异性抗体(IgG-like双特异性抗体，具有Fc介导的效应功能)和 不含Fc片段的双特异性抗体(non-IgG-like双特异性抗体，通过抗原结合力发挥 作用，具有分子量小、免疫原性低等优势)两种结构。
本发明还包括同时结合抗人CEACAM5/6抗体和其他靶点的抗体。较佳地， 所述的其他靶点选自下组：CD47、CD73、CD47、CD3、CTLA4、PD-1、PD-L1、CD28、CD40、OX40、LAG3、DR5、TIM-3、TIGIT、VEGF、VEGFR、Ang2、 CD39、CD73、GITR、或其组合。
在另一优选例中，本发明的双特异性抗体中结合其他靶点的抗体为抗人 CD47抗体。
如本文所用，术语“双特异性抗体”、“双功能抗体”“本发明抗体”、 “本发明双抗”“双抗”、“双功能融合抗体”、“抗人CEACAM5/6和CD47 双特异性抗体”、“抗CEACAM5/6和CD47抗体”可互换使用，是指同时结合 CEACAM5/6和CD47的双特异性抗体。
如本文所用，术语“接头(linker)”是指插入免疫球蛋白结构域中为轻链 和重链的结构域提供足够的可动性以折叠成交换双重可变区免疫球蛋白的一 个或多个氨基酸残基。
如本文所用，所述接头为柔性接头，优选为肽接头。合适的肽接头实例包 括单甘氨酸(Gly)、或丝氨酸(Ser)残基，肽接头中氨基酸残基的标识和序列可随 着肽接头中需要实现的次级结构要素的类型而变化。
本发明中提供了一种基于前述抗人CEACAM5/6抗体和抗人CD47抗体或其 抗原结合片段而构建的双特异性抗体。这些双特异性抗体通过接头(linker)把 抗人CEACAM5/6抗体和抗人CD47抗体或其抗原结合片段连接起来，组成对称 或者不对称结构的多价抗体，达到可同时结合CEACAM5/6和CD47蛋白的效果。 这里可以选取使用的接头序列包括：
(G)n、(GGGGS)n、(GGGSG)n、(GGSGG)n、
GGGGSGSAGSAAGSGEFGGGGSGGG(SEQ ID NO.65)、
SSSSKAPPPSLPSPSRLPGPSDTPILPQ(SEQ ID NO.66)、
KESGSVSSEQLAQFRSLD(SEQ ID NO.67)、
EGKSSGSGSESKST(SEQ ID NO.68)、
GSAGSAAGSGEF(SEQ ID NO.69)等。优选地，所述接头为(GGGGS)n， 其中n为0-5之间的整数；或SSSSKAPPPSLPSPSRLPGPSDTPILPQ，或 GGGSSSSKAPPPSLPSPSRLPGPSDTPILPQGGG；更优选地，接头为 GGGSSSSKAPPPSLPSPSRLPGPSDTPILPQGGG。
在本发明的一个具体实施例中，优选的接头序列如SEQ ID NO.64中的 119-152位或SEQ ID NO.66所示。
本发明所描述的抗人CEACAM5/6抗体和抗人CD47抗体或其抗原结合片段 而构建的双特异性抗体可以还包含人源重链恒定区和/或人源轻链恒定区；更优 选地，所述人源重链恒定区选自人IgG1、IgG2或IgG4的重链恒定区，所述人源 轻链恒定区选自人κ或λ链的轻链恒定区；进一步优选地，所述人源重链恒定 区为人IgG1的重链恒定区，所述人源轻链恒定区为κ链；进一步优选地，所述 人源重链恒定区为人IgG1的重链恒定区可以携带本领域技术人员所通常知晓 得突变，以增强或减弱IgG1抗体恒定区介导的生物学功能，包括ADCC、ADCP (Antibody-dependent cellular phagocytosis)、CDC和FcRn结合活性等。这些突 变包括但不限于如文献(Conceptual Approaches to Modulating Antibody EffectorFunctions and Circulation Half-Life.Front.Immunol.,2019,10:1296)中所列的突变：
表3 IgG1重链恒定区突变及其功能
Figure BDA0003268530520000171
Figure BDA0003268530520000181
根据上述技术，本发明人设计了不同结构形式的抗人CEACAM5/6-CD47 双特异性抗体。优选地，组成的抗人CEACAM5/6-CD47双特异性抗体的序列如 表4所示。
表4.抗人CEACAM5/6-CD47双特异性抗体的序列
Figure BDA0003268530520000191
与现有技术相比，本发明的抗人CEACAM5/6-CD47抗体与 CEACAM5/6+-CD47+双阳性肿瘤细胞表面CD47蛋白的结合选择性更强，杀伤作 用更强，与红细胞和血小板表面CD47蛋白结合更弱，并且对红细胞没有凝集作 用。
在本发明中，本发明的双特异性抗体还包括其保守性变异体，指与本发明 双抗的氨基酸序列相比，有至多10个，较佳地至多8个，更佳地至多5个，最佳 地至多3个氨基酸被性质相似或相近的氨基酸所替换而形成多肽。这些保守性 变异多肽最好根据表A进行氨基酸替换而产生。
表A
最初的残基 代表性的取代 优选的取代
Ala(A) Val；Leu；Ile Val
Arg(R) Lys；Gln；Asn Lys
Asn(N) Gln；His；Lys；Arg Gln
Asp(D) Glu Glu
Cys(C) Ser Ser
Gln(Q) Asn Asn
Glu(E) Asp Asp
Gly(G) Pro；Ala Ala
His(H) Asn；Gln；Lys；Arg Arg
Ile(I) Leu；Val；Met；Ala；Phe Leu
Leu(L) Ile；Val；Met；Ala；Phe Ile
Lys(K) Arg；Gln；Asn Arg
Met(M) Leu；Phe；Ile Leu
Phe(F) Leu；Val；Ile；Ala；Tyr Leu
Pro(P) Ala Ala
Ser(S) Thr Thr
Thr(T) Ser Ser
Trp(W) Tyr；Phe Tyr
Tyr(Y) Trp；Phe；Thr；Ser Phe
Val(V) Ile；Leu；Met；Phe；Ala Leu
编码核酸和表达载体
本发明还提供了编码上述抗体或其片段或其融合蛋白的多核苷酸分子。本 发明的多核苷酸可以是DNA形式或RNA形式。DNA形式包括cDNA、基因组 DNA或人工合成的DNA。DNA可以是单链的或是双链的。DNA可以是编码链 或非编码链。编码本发明的成熟多肽的多核苷酸包括：只编码成熟多肽的编码 序列；成熟多肽的编码序列和各种附加编码序列；成熟多肽的编码序列(和任选 的附加编码序列)以及非编码序列。
术语“编码多肽的多核苷酸”可以是包括编码此多肽的多核苷酸，也可以 是还包括附加编码和/或非编码序列的多核苷酸。
本发明的核酸(以及核酸组合)可用于在合适的表达系统产生本发明的重组 抗体。
本发明还涉及与上述的序列杂交且两个序列之间具有至少50％，较佳地至 少70％，更佳地至少80％相同性的多核苷酸。本发明特别涉及在严格条件下与 本发明所述多核苷酸可杂交的多核苷酸。在本发明中，“严格条件”是指：(1) 在较低离子强度和较高温度下的杂交和洗脱，如0.2×SSC，0.1％SDS，60℃； 或(2)杂交时加有变性剂,如50％(v/v)甲酰胺，0.1％小牛血清/0.1％Ficoll，42℃ 等；或(3)仅在两条序列之间的相同性至少在90％以上，更好是95％以上时才发 生杂交。并且,可杂交的多核苷酸编码的多肽与成熟多肽有相同的生物学功能 和活性。
本发明的抗体的核苷酸全长序列或其片段通常可以用PCR扩增法、重组法 或人工合成的方法获得。一种可行的方法是用人工合成的方法来合成有关序列， 尤其是片段长度较短时。通常，通过先合成多个小片段，然后再进行连接可获 得序列很长的片段。此外，还可将重链的编码序列和表达标签(如6His)融合在 一起，形成融合蛋白。
一旦获得了有关的序列，就可以用重组法来大批量地获得有关序列。这通 常是将其克隆入载体，再转入细胞，然后通过常规方法从增殖后的宿主细胞中 分离得到有关序列。本发明所涉及的生物分子(核酸、蛋白等)包括以分离的形 式存在的生物分子。
目前，已经可以完全通过化学合成来得到编码本发明蛋白(或其片段，或其 衍生物)的DNA序列。然后可将该DNA序列引入本领域中已知的各种现有的 DNA分子(或如载体)和细胞中。此外，还可通过化学合成将突变引入本发明蛋 白序列中。
本发明还涉及包含上述的适当DNA序列以及适当启动子或者控制序列的 载体。这些载体可以用于转化适当的宿主细胞，以使其能够表达蛋白质。
宿主细胞可以是原核细胞，如细菌细胞；或是低等真核细胞，如酵母细胞； 或是高等真核细胞，如哺乳动物细胞。代表性例子有：大肠杆菌，链霉菌属； 鼠伤寒沙门氏菌的细菌细胞；真菌细胞如酵母；果蝇S2或Sf9的昆虫细胞；CHO、 COS7、293细胞的动物细胞等。
较佳地，本发明还提供了包含本发明的载体的宿主细胞，优选地，所述宿 主细胞是哺乳动物细胞，更优选为人、鼠、羊、马、狗或猫的细胞，进一步优 选为中国仓鼠卵巢细胞。
用重组DNA转化宿主细胞可用本领域技术人员熟知的常规技术进行。当宿 主为原核生物如大肠杆菌时，能吸收DNA的感受态细胞可在指数生长期后收获， 用CaCl2法处理，所用的步骤在本领域众所周知。另一种方法是使用MgCl2。如 果需要，转化也可用电穿孔的方法进行。当宿主是真核生物，可选用如下的DNA 转染方法：磷酸钙共沉淀法，常规机械方法如显微注射、电穿孔,脂质体包装 等。
获得的转化子可以用常规方法培养，表达本发明的基因所编码的多肽。根 据所用的宿主细胞，培养中所用的培养基可选自各种常规培养基。在适于宿主 细胞生长的条件下进行培养。当宿主细胞生长到适当的细胞密度后，用合适的 方法(如温度转换或化学诱导)诱导选择的启动子，将细胞再培养一段时间。
在早期培养条件中，双特异性抗体的表达量可达3.9g/L，纯度均在97％以上， 且在培养过程中可以很好地代谢乳酸。
在上面的方法中的重组多肽可在细胞内、或在细胞膜上表达、或分泌到细 胞外。如果需要，可利用其物理的、化学的和其它特性通过各种分离方法分离 和纯化重组的蛋白。这些方法是本领域技术人员所熟知的。这些方法的例子包 括但并不限于：常规的复性处理、用蛋白沉淀剂处理(盐析方法)、离心、渗透 破菌、超处理、超离心、分子筛层析(凝胶过滤)、吸附层析、离子交换层析、 高效液相层析(HPLC)和其它各种液相层析技术及这些方法的结合。
本发明的双抗可以单独使用，也可与可检测标记物(为诊断目的)、治疗剂、 或任何以上这些物质的组合结合或偶联。
用于诊断目的可检测标记物包括但不限于：荧光或发光标记物、放射性标 记物、MRI(磁共振成像)或CT(电子计算机X射线断层扫描技术)造影剂、或能够 产生可检测产物的酶。
可与本发明抗体结合或偶联的治疗剂包括但不限于：1.放射性核素；2.生 物毒；3.细胞因子如IL-2等；4.金纳米颗粒/纳米棒；5.病毒颗粒；6.脂质体； 7.纳米磁粒；8.肿瘤治疗剂(例如，顺铂)或任何形式的抗肿瘤药物等。
药物组合物
本发明还提供了一种组合物。优选地，所述的组合物是药物组合物，它含 有本发明上述的抗人CEACAM5/6抗体或其活性片段或其融合蛋白，以及药学 上可接受的载体。通常，可将这些物质配制于无毒的、惰性的和药学上可接受 的水性载体介质中，其中pH通常约为5-8，较佳地pH约为6-8，尽管pH值可随被 配制物质的性质以及待治疗的病症而有所变化。配制好的药物组合物可以通过 常规途径进行给药，其中包括(但并不限于)：静脉注射、静脉滴注、皮下注射、 局部注射、肌肉注射、瘤内注射、腹腔内注射(如腹膜内)、颅内注射、或腔内 注射。
本发明的药物组合物可直接用于结合CEACAM5/6，因而可用于治疗肿瘤。 此外，还可同时使用其他治疗剂。
本发明的药物组合物含有安全有效量(如0.001-99wt％，较佳地0.01-90wt％， 更佳地0.1-80wt％)的本发明的抗人CEACAM5/6抗体以及药学上可接受的载体 或赋形剂。这类载体包括(但并不限于)：盐水、缓冲液、葡萄糖、水、甘油、 乙醇、及其组合。药物制剂应与给药方式相匹配。本发明的药物组合物可以被 制成针剂形式，例如用生理盐水或含有葡萄糖和其他辅剂的水溶液通过常规方 法进行制备。药物组合物如针剂、溶液宜在无菌条件下制造。活性成分的给药 量是治疗有效量，例如每天约10微克/千克体重-约50毫克/千克体重。此外，本 发明的抗人CEACAM5/6抗体还可与其他治疗剂一起使用。
在本发明中，可单独使用本发明的抗人CEACAM5/6抗体，通过调整给药 方案以获得最佳目的反应。例如，单次给药，或在一段时间内多次给药，或者 可以随治疗情况的紧急程度按比例减少或增加剂量。
使用药物组合物时，是将安全有效量的本发明的抗人CEACAM5/6抗体施 用于哺乳动物，其中该安全有效量通常至少约10微克/千克体重，而且在大多数 情况下不超过约50毫克/千克体重，较佳地该剂量是约10微克/千克体重-约10毫 克/千克体重。当然，具体剂量还应考虑给药途径、病人健康状况等因素，这些 都是熟练医师技能范围之内的。
应用
本发明还提供了本发明抗体、双特异性抗体、抗体偶联物ADC、重组蛋白、 和/或免疫细胞的用途，例如用于制备诊断制剂或制备药物。进一步地，本发明 还提供了包含本发明的抗体或其抗原结合片段在制备药物或用于细胞疗法的 细胞中的用途，所述药物或细胞用于预防和/或治疗通过增强免疫应答而受益的 病症。
较佳地，所述的药物是用于预防和/或治疗与CEACAM5/6(CEACAM5蛋 白或CEACAM6蛋白)表达或功能异常相关的疾病的药物。
在本发明的一个实施方案中，其中所述病症是癌症，优选地，所述病症是 CEACAM5或CEACAM6高表达的肿瘤，优选为结直肠癌。
在另一优选例中，所述人CEACAM5/6相关疾病为CEACAM5/6和CD47同时 高表达的肿瘤，优选为结直肠癌。
本发明的主要优点包括
(1)本发明的抗人CEACAM5/6抗体具有同时结合CEACAM5以及 CEACAM6的交叉反应性；
(2)本发明的抗人CEACAM5/6和CD47双特异性抗体对CEACAM5/6和 CD47高表达的肿瘤细胞有更好的靶向和特异性杀伤作用，同时可以降低与红细 胞和/或血小板的结合作用。
下面结合具体实施例，进一步陈述本发明。应理解，这些实施例仅用于说 明本发明而不用于限制本发明的范围。下列实施例中未注明详细条件的实验方 法，通常按照常规条件如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring Harbor LaboratoryPress,1989)中所述的条件，或按照制造厂商所建议的 条件。除非另外说明，否则百分比和份数按重量计算。
本发明的抗体序列如表5所示：
表5.本发明的抗体序列
Figure BDA0003268530520000241
Figure BDA0003268530520000251
Figure BDA0003268530520000261
Figure BDA0003268530520000271
Figure BDA0003268530520000281
Figure BDA0003268530520000291
Figure BDA0003268530520000301
Figure BDA0003268530520000311
Figure BDA0003268530520000321
Figure BDA0003268530520000331
Figure BDA0003268530520000341
Figure BDA0003268530520000351
实施例1：人源化CD47单克隆抗体的突变
人源化CD47单克隆抗体的母本序列引自中国专利202010240238.7，其重链可 变区序列如SEQ ID NO:17所示；其轻链可变区序列如SEQ ID NO:18所示。该人 源化CD47单克隆抗体的IgG4亚型在本专利中以UM03-C4代号表示。
UM03-C4重链VH：
QVQLVESGGGVVQPGGSLRLSCAASGFTFSDYGMAWIRQAPGKGPEWIAFITNLASSIYYADTVTGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARAGDYRSFPYWGQGTLVTVSA(SEQ ID NO.17)
UM03-C4轻链VL：
EIVLTQSPATLSLSPGERATLSCSASSSVNYVNWYQQKPGQAPRILIYGISNLASGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSTFPPYTFGQGTKLEIK (SEQ ID NO.18)
其中，CD47抗体的重链和轻链中以Kabat定义的CDR序列用下划线标出。
此外，在该抗人CD47抗体基础上，还包括对其VH、VL区进行突变而得的 衍生抗体人CD47抗体(如本文所用，以代号UM03-C4’表示，其重链可变区序列 如SEQ ID NO:19所示；其轻链可变区序列如SEQ ID NO:20所示)。所涉及的突 变抗人CD47抗体包括如下突变或者下述突变的组合：
重链：H-I37V、H-P45L、H-I48V、H-R94K、H-P101D、H-T64K、H-T52R、 H-T52S、H-T52L、H-T52I、H-T52H、H-T52A、H-T52G、H-T52N、H-T52Q；
轻链：L-L4M、L-I46L、L-V58I、L-T93N。
其中，M18的突变都是在FR区，M19、M20和M21是在M18基础上添加了 CDR的1个或两个突变。
部分突变对CD47单抗的活性影响如下表6所列：
表6 CD47单抗突变及其功能影响
Figure BDA0003268530520000361
注：位置编号均根据Kabat规则。
实施例2：人源化CEACAM5/6单克隆抗体的获得
2.1小鼠抗人CEACAM5/6单克隆抗体的获得
本发明人构建了过表达人CEACAM5蛋白的CHO细胞株，以之免疫小鼠。取 免疫小鼠的脾细胞与SP2/0-AG14细胞进行杂交瘤细胞融合，并取适量融合后的细 胞铺至96孔板。融合后第10天取各孔上清，以ELISA法检测杂交瘤细胞分泌的 小鼠抗体与人CEACAM5的结合活性(方法参见实施例3)，得到了一系列具有较 高活性的杂交瘤细胞。
选择其中活性优异的杂交瘤细胞，其中，出乎意料的是一株抗体居然对 CEACAM6也具有优异的结合活性。
经测序获得其分泌抗体对应的重链可变区cDNA序列和轻链可变区cDNA序 列，其编码的重链可变区氨基酸序列如SEQ ID NO.7所示；编码的轻链可变区氨基 酸序列如SEQID NO.8所示。将小鼠抗体的重链可变区和轻链可变区分别连接至人 IgG1重链的恒定区和κ链的恒定区，得到人-鼠嵌合抗体UM05-L9，其重链序列如 SEQ ID NO.9所示，轻链序列如SEQ ID NO.10所示。
2.2小鼠抗人CEACAM5/6抗体的人源化
对实施例2中得到的小鼠抗体UM05-L9的重链可变区和轻链可变区序列进行 分析，得到其重链互补决定区CDR如下表所示：
表7 小鼠CEACAM5/6单抗UM05-L9的CDR区界定
Kabat定义 IMGT定义
H-CDR1 DYAMH GYTFTDYA
H-CDR2 VISTYSGHTNYNQKFKG ISTYSGHT
H-CDR3 GSTTAHYYTMDF VRGSTTAHYYTMDF
L-CDR1 GASENIYGTLN ENIYGT
L-CDR2 GATNLAD GAT
L-CDR3 QNVLSIPYT QNVLSIPYT
同时参考Kabat定义和IMGT定义的CDR。搜索人germline抗体序列数据库 (IGMT和IgBlast)，分别获得与小鼠抗体重/轻链可变区同源性较高的人germline 抗体序列，并将其框架区与上述小鼠抗体的CDR进行组合，即所谓CDR grafting， 同时对框架区的部分氨基酸进行了回复突变，最终获得了人源化抗体UM05-C9和 UM05-C9’，其重链可变区序列分别如SEQ ID NO.11和SEQ ID NO.13所示；轻链 可变区序列则两者相同，如SEQ ID NO.12所示。
实施例3：抗体UM05-C9/UM05-L9与CEACAM家族蛋白的结合
3.1抗体与人CEACAM5蛋白的结合
将浓度为1μg/mL的人CEACAM5蛋白溶液以100μL/well包被96孔高亲和力 板，4℃，振荡过夜。第二天先以300μL PBST(Tween20：0.5‰)洗涤3次，之后 用100μL/well的5％BSA/PBS封闭1小时，室温振荡。300μL PBST洗涤3次。用 PBS配制抗体样品的梯度稀释溶液。以100μL/孔加入96孔板，室温振荡1小时。 300μL PBST洗涤3次。配制二抗羊抗人(goatanti-human)IgG HRP溶液，以100μL/ 孔加入96孔板，室温振荡30min。300μL PBST洗涤4次。加入100μL/孔TMB， 显色20min。加入100μL/孔0.6N H2SO4，终止显色，检测OD450nm。
经检测，结果如图1所示，人源化抗体UM05-C9与人CEACAM5结合的EC50 为0.399nM。
3.2抗体与CHO-CEACAM5细胞的结合
用FACS检测抗体与过表达人CEACAM5的CHO细胞的结合情况。简单来说， 首先收集细胞，PBS洗涤一遍后计数，稀释成2×106/ml细胞悬液；分别取10μl抗 体工作液加入100μl细胞悬液中，4℃避光孵育30min；PBS洗涤2次后，加入对 应的荧光标记二抗Goat-anti-Human IgG(H+L)(Invitrogen)，4℃避光孵育30min， PBS洗涤2次后，以400μl FACS buffer悬起，以流式细胞仪检测抗体与细胞的结 合情况。
检测结果如图2所示，UM05-C9与CHO-CEACAM5结合的EC50为636.9ng/mL。
3.3 UM05-L9的亲和力检测
抗原抗体结合的亲和力用SPR技术(Surface Plasmon Resonance)进行检测。 简单来说，将4μg/mL浓度的嵌合抗体UM05-L9与protein A sensor chip(GE,Cat# 29127556)孵育30s进行抗体捕获。在抗原结合阶段，以梯度稀释的人CEACAM5 蛋白为流动相与sensorchip上捕获的UM05-L9抗体进行120s的结合。在解离阶段， 以HBS-EP缓冲液持续洗脱360s。CEACAM5蛋白在sensor chip上与抗体结合的 情况以Biacore T20x0(GE Healthcare)进行定量检测。检测结果如表8所示， UM05-L9的亲和力为0.68nM。
表8.UM05-L9与人CEACAM5的亲和力和动力学数据
受体 配体 ka(1/Ms) kd(1/s) KD(M)
CEACAM5 UM05-L9 6.54×10<sup>4</sup> 4.45×10<sup>-4</sup> 6.80×10<sup>-9</sup>
3.4 UM05-L9抗体与人CEACAM分子家族其它分子的结合
进一步使用UM05-L9检测了其与CEACAM分子家族其成员的结合情况。使 用了SW620-CEACAM1、SW620-CEACAM3、CHO-CEACAM6、CHO-CEACAM7、 CHO-CEACAM8等稳转细胞株。检测方法如3.2中所述。结果如下表9所示。
结果显示，UM05-L9分子除与CEACAM5分子有强结合外，亦与CEACAM6 分子有较强结合。而与其它被检测的分子，包括CEACAM1、3、7、8在内，都基 本没有结合。由于CEACAM5和CEACAM6都是肿瘤细胞特异性高表达的分子， 这提示UM05-L9(推测也包括UM05-C9分子)具有较好的肿瘤靶向性。
表9.UM05-L9与人CEACAM分子家族其它分子的结合
Figure BDA0003268530520000381
Figure BDA0003268530520000391
3.5 UM05-L9与食蟹猴CEACAM5和CEACAM6分子的结合
进一步使用UM05-L9以ELISA法检测了其与食蟹猴CEACAM5和CEACAM6 蛋白的结合情况。检测方法参考3.1中所述。结果图3所示。
结果显示UM05-L9与食蟹猴的CEACAM5和CEACAM6分子都有较强结合， 其结合EC50分别为0.6784nM和0.4629nM。
本发明的CEACAM5/6抗体可以与食蟹猴CEACAM5/6蛋白结合的特性，为将 相关抗体进行药物开发提供了动物模型研究的便利。
实施例4:UM05-C9的肿瘤组织特异性
UM05-C9的肿瘤组织特异性分析由四川夏派森医药科技有限公司采用上海芯 超生物科技有限公司的多器官肿瘤组织芯片进行(Cat No：HOrgC120PG04，Lot No： XT19-008)。
对免疫组化所得的结果进行图像扫描及半定量分析显示，UM05-C9检测到结 直肠癌肿瘤组织中有强染色，在部分肺癌和胰腺癌中亦有较强染色，而在对应的癌 旁组织中都呈阴性或低表达(表10)。
表10 UM05-C9对多器官肿瘤组织芯片染色结果(部分)
Figure BDA0003268530520000392
Figure BDA0003268530520000401
实施例5：CEACAM5/6抗体UM05-C9的细胞内吞性质
实验步骤：A549用F12K+10％FBS+1％P/S培养基培养，按照5万/孔，200μl/ 孔接96孔板中。用SiteClickTM Antibody Azido Modification Kit(Invitrogen,Cat #S20026)和Click-iTTM pHrodoTM iFL Red sDIBO Alkyne for Antibody Labeling (Invitrogen,Cat#C20034)试剂盒按说明书标记抗体，并将标记好的抗体按照1： 20、1：40、1：80、1：160、1：320、1：640用完全培养基稀释，至浓度为45、 22.5、11.25、5.62、2.8、1.4μg/ml。将前一天接板的细胞去上清，然后将上述配制 的抗体按照100μl/孔，加入到细胞中，在孵箱中继续孵育16h。用胰酶消化细胞， 收集细胞，流式细胞法检测抗体内吞情况(Ex＝560nM，Em＝585nM)。抗体被细 胞内吞后会有荧光被激发，而未被内吞的抗体不会有荧光被激发。
结果如图4所示。表明CEACAM5/6抗体结合到A549细胞后会发生内化(internalization)。
实施例6：CD47-CEACAM5/6双特异性抗体的设计
将抗人CD47抗体或其抗原结合域与抗人CEACAM5/6的抗体或其抗原结合域 进行不同组合，可以组成双特异性抗体。进行了如图5所示的几种双抗结构的尝试。
由此得到的双抗组合如表11所示。
表11.CEACAM5/6-CD47双特异性抗体序列及类型
Figure BDA0003268530520000402
Figure BDA0003268530520000411
实施例7：双特异性抗体的制备
将编码抗体重链或轻链的cDNA序列连接在编码信号肽的序列之后，分别克 隆到哺乳动物细胞表达载体pcDNA3.4上。将重链表达质粒和轻链表达质粒按2： 1的摩尔比用Lipofectamine 2000转染试剂(Invitrogen)转染入HEK293细胞，并 在37℃、5％二氧化碳条件下培养7天。收集培养液上清，并用Protein A亲和层 析法提纯上清中的抗体。纯化后抗体的质量可以用SDS-PAGE电泳和分子筛层析 等经典的蛋白分析方法分析。纯化后的抗体经PBS溶液透析和冷冻干燥浓缩后， 保存于-20℃。
实施例8：双特异性抗体与CD47蛋白或CEACAM家族蛋白的结合
8.1双特异性抗体与CD47蛋白的结合
将浓度为1μg/mL的人CD47蛋白溶液以100μL/孔包被96孔高亲和力板，4℃， 振荡过夜。第二天先以300μL PBST(Tween20：0.5‰)洗涤3次，之后用100μL/ 孔的5％BSA/PBS封闭1小时，室温振荡。300μL PBST洗涤3次。用PBS配制抗 体样品的梯度稀释溶液。以100μL/孔加入96孔板，室温振荡1小时。300μL PBST 洗涤3次。配制二抗羊抗鼠(goat anti-mouse)IgG HRP或羊抗人(goat anti-human) IgG HRP溶液，以100μL/孔加入96孔板，室温振荡1小时。300μLPBST洗涤4 次。加入100μL/孔TMB，显色20min。加入100μL/孔0.6N H2SO4，终止显色， 检测OD450 nm。
本发明涉及的抗体与CD47蛋白结合代表性数据如表12所示。
8.2双特异性抗体与CEACAM5蛋白的结合
双特异性抗体与人CEACAM5蛋白结合的检测方法同实施例5.1所描述，差别 在于将包被蛋白改为1μg/mL的人CEACAM5蛋白溶液。数据如表12所示。
表12 双抗与靶点结合活性
Figure BDA0003268530520000421
Figure BDA0003268530520000431
实施例9：双特异性抗体与CD47+-CEACAM5+的人肿瘤细胞A549的结合
肺癌细胞A549(ATCC)是同时表达CD47和CEACAM5的人源肿瘤细胞。 本申请发明人在A549细胞上使用流式细胞术检测了抗体的结合。用PBS配制待检 测抗体的浓度梯度溶液，配制成终浓度的2×工作液。收集A549细胞，PBS洗涤 一遍后计数，稀释成4×106/ml细胞悬液；取50μL抗体工作液加入50μL细胞悬液 中，4℃避光孵育60min；PBS洗两遍后加入对应的荧光标记二抗(anti-huIgG 633)，4℃避光孵育30min，PBS洗涤两次后，以400μL FACSbuffer悬起，以流式细胞仪 检测抗体与细胞的结合情况。数据如表13所示。
表13 双抗体与人肿瘤细胞A549的结合EC50(nM)
Figure BDA0003268530520000441
Figure BDA0003268530520000451
*表示这些分子在检测浓度范围内，检测信号没有达到平台，无法准确计算 EC50。“NA”代表未做检测。
实施例10：双特异性抗体与人红细胞的结合及对人红细胞的凝集作用
本发明人检测了本发明所设计的部分双特异性抗体以及阳性对照抗体Hu5F9 (美国专利US 9017675B2及文献PLoS ONE 2015,10(9):e0137345)与人红细胞的 结合以及对人红细胞的凝集作用。首先从志愿者外周血中分离红细胞，用生理盐水 悬成1％浓度红细胞悬液。用PBS配制抗体的浓度梯度溶液，配制成终浓度的2× 工作液。取50μL抗体工作液加入50μL红细胞悬液中，4℃避光孵育60min；PBS 洗两遍后加入对应的荧光标记二抗，4℃避光孵育30min，PBS洗涤两次后，以400μL FACS buffer悬起，以流式细胞仪检测抗体与红细胞的结合情况。对于红细胞凝集 检测，红细胞用生理盐水悬成2％浓度悬液，同样取50μL抗体工作液加入50μL 红细胞悬液中，置于96孔圆底板，室温静置2h后观察拍照。数据如表14所示。
表14 双特异性抗体与人红细胞的结合和凝集作用
Figure BDA0003268530520000452
Figure BDA0003268530520000461
注1：“+”个数代表与人红细胞的结合强弱。“-”代表在最高浓度为100nM的 抗体浓度检测范围内，未检测到抗体与红细胞的明显结合。“NA”代表未做检测。
注2：“+”个数代表抗体引起人红细胞凝聚的能力。“-”代表在最高浓度为200 μg/mL的抗体浓度检测范围内，抗体未引起明显的红细胞凝集。“NA”代表未做 检测。
实施例11：双特异性抗体抑制A549细胞上CD47与SIRPα蛋白的结合
肺癌细胞A549(ATCC)是同时表达CD47和CEACAM5的人源肿瘤细胞。 本申请发明人在A549细胞上使用流式细胞术检测了抗体抑制A549细胞上CD47 与SIRPα蛋白的结合。用PBS配制待检测抗体的浓度梯度溶液，配制成终浓度的 2×工作液。收集A549细胞，PBS洗涤一遍后计数，稀释成4×106/ml细胞悬液；取 50μL抗体工作液加入50μL细胞悬液中，4℃避光孵育60min；PBS洗两遍后加入 human-SIRPa-mouse-Fc tag蛋白(SIA-H52A8)，4℃避光孵育30min；PBS洗两遍 后加入羊抗鼠IgG1-Fc特异性检测二抗(115-095-071)，4℃避光孵育30min；PBS 洗两遍后以400μL FACS buffer悬起，以流式细胞仪检测标记二抗的结合情况，得 到抗体抑制A549细胞上CD47与SIRPα蛋白的结合的活性，结果如表15所示。
表15 双特异性抗体阻断A549细胞上CD47-SIRPα结合的活性
抗体代号 抑制(以UM03-C4为参考)
UM03-C4 +++
L8 +++
L10 +++
L11 +
L12 +++
L13 ++
L14 +++
L15 +++
L16 +++
L17 ++
L18 +
L23 +
L25 +++
L26 +++
L27 ++
L28 ++
L29 +
L30 +
L31 +
L32 +
L33 +
L34 ++
L35 +
L36 +
L37 +
L38 +
L39 +
L40 +++
L41 +++
注：“+”个数代表抗体阻断A549细胞上CD47与SIRPα结合的能力。
实施例12：双特异性抗体的ADCC
本申请发明人在A549细胞和人红细胞上检测了部分抗体对两个细胞的ADCC 杀伤作用。收集A549细胞，培养基重悬计数，稀释成1.5×105/ml的细胞悬液，A549 细胞以50ul/孔提前一天铺板，次日弃去培养基将配置好的抗体梯度稀释液加入孔 板中，37℃细胞培养箱孵育1小时；PBS洗两遍后，加入效应细胞 Jurkat-NFAT-luc/CD16A，细胞计数稀释成3×106/ml的细胞悬液，以30ul/孔加入孔 板中，继续放入37℃细胞培养箱孵育4小时，后加入30ul/孔One-Glo，等待3min 后用酶标仪检测生物发光强度。对红细胞杀伤作用的检测，首先从志愿者外周血中 分离红细胞，以培养基洗2遍之后计数，配置成1×107/mL细胞悬液，取50ul/well 细胞悬液与2×的抗体梯度稀释液50ul混合后置于37℃细胞培养箱孵育1小时；PBS 洗两遍后，加入效应细胞Jurkat-NFAT-CD16A，细胞计数稀释成3×106/ml的细胞悬液，以30ul/孔加入孔板中，继续放入37℃细胞培养箱孵育4小时，后加入30ul/ 孔One-Glo，等待3min后用酶标仪检测生物发光强度。检测出结果如下表16所示。
表16 双特异性抗体对A549及RBC细胞的ADCC作用
Figure BDA0003268530520000481
Figure BDA0003268530520000491
注：“+”个数定性的代表抗体介导的对靶细胞的ADCC活性。
“NA”代表未检测。
实施例13双特异性抗体的ADCP效应
本申请发明人在A549细胞和人红细胞上检测了部分抗体对两个细胞的ADCP 作用。收集A549细胞，培养基重悬计数，稀释成1×105/ml的细胞悬液，A549细 胞以50ul/孔提前一天铺板，次日弃去培养基将配置好的抗体梯度稀释液加入孔板 中，37℃细胞培养箱孵育1小时；PBS洗两遍后，加入效应细胞 Jurkat-NFAT-luc/CD32A，细胞计数稀释成3×106/ml的细胞悬液，以30ul/孔加入孔 板中，继续放入37℃细胞培养箱孵育4小时，后加入30ul/孔One-Glo，等待3min 后用酶标仪检测生物发光强度。结果如表17所示。
表17 双特异性抗体对A549及RBC细胞的ADCP作用
Figure BDA0003268530520000492
注：“+”个数定性的代表抗体介导的对靶细胞的ADCP活性。
“NA”代表未检测。
实施例14：抗原抗体亲和力检测
用SPR技术(Surface Plasmon Resonance)分别检测了L34分子与人CEACAM5 和人CD47的亲和力。简单来说，将4μg/mL浓度的抗体溶液与protein A sensor chip (GE,Cat#29127556)孵育30s进行抗体捕获。在抗原结合阶段，分别以人 CEACAM5蛋白和人CD47蛋白为流动相与sensor chip上捕获的抗体进行120s的 结合。在解离阶段，以HBS-EP缓冲液持续洗脱360s。抗原在sensor chip上与抗 体结合的情况以Biacore 8K进行定量检测。结果如图6和表18所示。
表18 L34分子与人CEACAM5和人CD47蛋白的亲和力
受体 ka(1/Ms) kd(1/s) KD(M)
CD47 6.86×10<sup>5</sup> 4.51×10<sup>-2</sup> 6.57×10<sup>-8</sup>
CEACAM5 2.33×10<sup>4</sup> 1.73×10<sup>-4</sup> 7.43×10<sup>-9</sup>
上述亲和力检测结果显示，以L34分子为代表的双抗分子对人CD47靶点的亲 和力显著弱化，而保持了与CEACAM5的亲和力。
在上述亲和力检测的基础上，用SPR技术还检测了两个抗原与双抗结合的表 位竞争情况，方法同前所述。在本实验中，先将抗体溶液与protein A sensor chip 孵育30s进行抗体捕获。在抗原结合阶段，先后以CEACAM5蛋白和CD47蛋白 为流动相与sensor chip上捕获的抗体进行120s的结合。抗原在sensor chip上与抗 体结合的情况以Biacore 8K进行定量检测。结果如图7所示。
实施例15：小鼠肿瘤药效模型
A549细胞(ATCC，人非小细胞肺癌细胞)为CD47+-CEACAM+的肿瘤细胞， 按6×106细胞/只的量将A549细胞接种于雌性NOD-Scid小鼠皮下。待肿瘤长至约 70mm3后，静脉给予配制在生理盐水中的抗体溶液。每组6只小鼠，每3天给药一 次，共给药3周。
实施例16：食蟹猴的药物代谢和毒理
对食蟹猴(n＝2)静脉注射抗体溶液，剂量为20mg/kg。在给药前后0、0.25、 4、8、24、48、72、96、144、192、240和336小时采血，ELISA法检测血药浓度， 并做药时曲线图。同时计算药代动力学参数。
讨论
除了已知的CEACAM5被用作肿瘤标记，测量癌症患者血液中升高的 CEACAM5的免疫学测定法已在临床上用于癌症的预后和控制之外，更重要的 是，CEACAM5已成为用于靶向治疗的潜在有用的肿瘤相关抗原。已经报道的 使用CEACAM5靶向免疫治疗癌症主要有2种主要方法。一种方法使用抗 CEACAM5抗体引发免疫细胞的溶解活性，特别是通过抗体依赖性细胞毒性 (ADCC)或补体依赖性细胞毒性(CDC)，以消除表达CEACAM5的肿瘤细 胞；另一种方法是通过抗CEACAM5抗体或抗体片段与例如药物、毒素、放射 性核苷酸、免疫调节剂或细胞因子等效应分子缀合，特异性靶向表达CEACAM5 的肿瘤细胞，从而发挥效应分子的治疗作用。鉴于CEACAM5更多地过量表达 于一些诸如结肠直肠癌、胰腺癌、肺癌、胃癌、肝细胞瘤、乳腺癌和甲状腺癌 等实体肿瘤中，因此当前的研究集中在抗CEACAM5抗体的抗原识别能力。
总之，当前的研究表明，靶向CEACAM5的治疗方法将有助于抑制肿瘤的 转移过程。因此，本发明提供的对CEACAM5和CEACAM6具有亲和力的抗 CEACAM5/6抗体，具有重要的应用价值。此外，本发明中的抗人CEACAM5/6 和CD47双特异性抗体对CEACAM5/6和CD47高表达的肿瘤细胞具有更好的靶 向和特异性杀伤作用外，还可以降低与红细胞和/或血小板的结合作用。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献 被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后， 本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申 请所附权利要求书所限定的范围。
序列表
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1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 14
<211> 451
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 14
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 Asp Tyr
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Arg Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys
450
<210> 15
<211> 451
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 15
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys
450
<210> 16
<211> 214
<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 Gly Ala Ser Glu Asn Ile Tyr Gly Thr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 17
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 17
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala
115
<210> 18
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 18
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 19
<211> 118
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 19
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser
115
<210> 20
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 20
Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 21
<211> 584
<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 Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Arg Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Glu Ser Gly
130 135 140
Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala
145 150 155 160
Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala Trp Ile Arg Gln Ala
165 170 175
Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile Thr Asn Leu Ala Ser
180 185 190
Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg Phe Thr Ile Ser Arg
195 200 205
Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala
210 215 220
Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser
225 230 235 240
Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Ala Ser
245 250 255
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
260 265 270
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
275 280 285
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
290 295 300
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
305 310 315 320
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
325 330 335
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
340 345 350
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
355 360 365
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
370 375 380
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
385 390 395 400
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
405 410 415
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
420 425 430
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
435 440 445
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
450 455 460
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
465 470 475 480
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
485 490 495
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
500 505 510
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
515 520 525
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
530 535 540
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
545 550 555 560
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
565 570 575
Ser Leu Ser Leu Ser Pro Gly Lys
580
<210> 22
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 22
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 Gly Ala Ser Glu Asn Ile Tyr Gly Thr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln
115 120 125
Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser
130 135 140
Cys Ser Ala Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys
145 150 155 160
Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala
165 170 175
Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
180 185 190
Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
195 200 205
Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr
210 215 220
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
225 230 235 240
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
245 250 255
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
260 265 270
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
275 280 285
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
290 295 300
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
305 310 315 320
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
325 330 335
<210> 23
<211> 589
<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 Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Arg Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
130 135 140
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg
145 150 155 160
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala
165 170 175
Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile
180 185 190
Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg
195 200 205
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
210 215 220
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala
225 230 235 240
Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr
245 250 255
Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
260 265 270
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
275 280 285
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
290 295 300
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
305 310 315 320
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
325 330 335
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
340 345 350
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
355 360 365
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
370 375 380
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
385 390 395 400
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
405 410 415
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
420 425 430
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
435 440 445
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
450 455 460
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
465 470 475 480
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
485 490 495
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
500 505 510
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
515 520 525
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
530 535 540
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
545 550 555 560
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
565 570 575
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
580 585
<210> 24
<211> 341
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 24
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 Gly Ala Ser Glu Asn Ile Tyr Gly Thr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu
115 120 125
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu
130 135 140
Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Asn Tyr Val Asn
145 150 155 160
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr Gly
165 170 175
Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly
180 185 190
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp
195 200 205
Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr Thr
210 215 220
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro
225 230 235 240
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
245 250 255
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
260 265 270
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
275 280 285
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
290 295 300
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
305 310 315 320
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
325 330 335
Asn Arg Gly Glu Cys
340
<210> 25
<211> 589
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 25
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
130 135 140
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg
145 150 155 160
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala
165 170 175
Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile
180 185 190
Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg
195 200 205
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
210 215 220
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala
225 230 235 240
Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr
245 250 255
Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
260 265 270
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
275 280 285
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
290 295 300
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
305 310 315 320
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
325 330 335
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
340 345 350
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
355 360 365
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
370 375 380
Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu
385 390 395 400
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
405 410 415
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
420 425 430
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
435 440 445
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
450 455 460
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
465 470 475 480
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
485 490 495
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
500 505 510
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
515 520 525
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
530 535 540
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
545 550 555 560
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
565 570 575
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
580 585
<210> 26
<211> 589
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 26
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
130 135 140
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg
145 150 155 160
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala
165 170 175
Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile
180 185 190
Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg
195 200 205
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
210 215 220
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala
225 230 235 240
Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr
245 250 255
Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
260 265 270
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
275 280 285
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
290 295 300
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
305 310 315 320
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
325 330 335
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
340 345 350
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
355 360 365
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
370 375 380
Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu
385 390 395 400
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
405 410 415
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
420 425 430
Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu
435 440 445
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
450 455 460
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
465 470 475 480
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
485 490 495
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
500 505 510
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
515 520 525
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
530 535 540
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
545 550 555 560
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
565 570 575
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
580 585
<210> 27
<211> 472
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 27
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln
130 135 140
Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser
145 150 155 160
Cys Ser Ala Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys
165 170 175
Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala
180 185 190
Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
195 200 205
Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr
210 215 220
Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr
225 230 235 240
Lys Leu Glu Ile Lys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
245 250 255
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
260 265 270
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
275 280 285
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
290 295 300
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
305 310 315 320
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
325 330 335
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
340 345 350
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
355 360 365
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr
370 375 380
Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser
385 390 395 400
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
405 410 415
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val
420 425 430
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
435 440 445
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
450 455 460
Ser Leu Ser Leu Ser Pro Gly Lys
465 470
<210> 28
<211> 451
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 28
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
385 390 395 400
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415
Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met
420 425 430
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445
Pro Gly Lys
450
<210> 29
<211> 589
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 29
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
130 135 140
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg
145 150 155 160
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala
165 170 175
Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile
180 185 190
Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg
195 200 205
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
210 215 220
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala
225 230 235 240
Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr
245 250 255
Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
260 265 270
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
275 280 285
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
290 295 300
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
305 310 315 320
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
325 330 335
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
340 345 350
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
355 360 365
Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu
370 375 380
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
385 390 395 400
Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln
405 410 415
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
420 425 430
Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu
435 440 445
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
450 455 460
Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys
465 470 475 480
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
485 490 495
Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
500 505 510
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
515 520 525
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
530 535 540
Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln
545 550 555 560
Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
565 570 575
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
580 585
<210> 30
<211> 716
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln
130 135 140
Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val
145 150 155 160
Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Thr Leu Asn Trp
165 170 175
Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile Tyr Gly Ala
180 185 190
Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly Ser Gly Ser
195 200 205
Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val
210 215 220
Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr Thr Phe Gly
225 230 235 240
Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly
245 250 255
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu
260 265 270
Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg Leu
275 280 285
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala Trp
290 295 300
Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile Thr
305 310 315 320
Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg Phe
325 330 335
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn
340 345 350
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Gly
355 360 365
Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr Val
370 375 380
Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser
385 390 395 400
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys
405 410 415
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
420 425 430
Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
435 440 445
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
450 455 460
Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
465 470 475 480
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro
485 490 495
Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
500 505 510
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
515 520 525
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
530 535 540
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
545 550 555 560
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
565 570 575
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
580 585 590
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
595 600 605
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
610 615 620
Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly
625 630 635 640
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
645 650 655
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
660 665 670
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
675 680 685
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
690 695 700
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
705 710 715
<210> 31
<211> 475
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln
130 135 140
Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val
145 150 155 160
Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Thr Leu Asn Trp
165 170 175
Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile Tyr Gly Ala
180 185 190
Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly Ser Gly Ser
195 200 205
Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val
210 215 220
Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr Thr Phe Gly
225 230 235 240
Gly Gly Thr Lys Val Glu Ile Lys Asp Lys Thr His Thr Cys Pro Pro
245 250 255
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
260 265 270
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
275 280 285
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
290 295 300
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
305 310 315 320
Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
325 330 335
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
340 345 350
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
355 360 365
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp
370 375 380
Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe
385 390 395 400
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
405 410 415
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
420 425 430
Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly
435 440 445
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
450 455 460
Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
465 470 475
<210> 32
<211> 214
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
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> 33
<211> 594
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 33
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Ser
115 120 125
Ala Gly Ser Ala Ala Gly Ser Gly Glu Phe Gly Gly Gly Gly Ser Gly
130 135 140
Gly Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
145 150 155 160
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
165 170 175
Asp Tyr Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu
180 185 190
Trp Ile Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
195 200 205
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
210 215 220
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
225 230 235 240
Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
245 250 255
Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val
260 265 270
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
275 280 285
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
290 295 300
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
305 310 315 320
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
325 330 335
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
340 345 350
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
355 360 365
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
370 375 380
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
385 390 395 400
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
405 410 415
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
420 425 430
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
435 440 445
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
450 455 460
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
465 470 475 480
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
485 490 495
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
500 505 510
Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
515 520 525
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
530 535 540
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
545 550 555 560
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
565 570 575
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
580 585 590
Gly Lys
<210> 34
<211> 470
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 34
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro
130 135 140
Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Gly
145 150 155 160
Ala Ser Glu Asn Ile Tyr Gly Thr Leu Asn Trp Tyr Gln Gln Lys Pro
165 170 175
Gly Lys Ser Pro Lys Leu Leu Ile Tyr Gly Ala Thr Asn Leu Ala Asp
180 185 190
Ala Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr
195 200 205
Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys
210 215 220
Gln Asn Val Leu Ser Ile Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val
225 230 235 240
Glu Ile Lys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
245 250 255
Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
260 265 270
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
275 280 285
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
290 295 300
Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
305 310 315 320
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
325 330 335
Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
340 345 350
Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
355 360 365
Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn
370 375 380
Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile
385 390 395 400
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
405 410 415
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys
420 425 430
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys
435 440 445
Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
450 455 460
Ser Leu Ser Pro Gly Lys
465 470
<210> 35
<211> 346
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 35
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Thr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile
35 40 45
Tyr Gly Ala Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Val Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Ser Ala Gly Ser Ala Ala Gly Ser Gly Glu Phe Gly Gly Gly Gly
115 120 125
Ser Gly Gly Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
130 135 140
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser
145 150 155 160
Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
165 170 175
Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg
180 185 190
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
195 200 205
Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr
210 215 220
Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
225 230 235 240
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
245 250 255
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
260 265 270
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
275 280 285
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
290 295 300
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
305 310 315 320
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
325 330 335
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
340 345
<210> 36
<211> 584
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 36
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
130 135 140
Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr
145 150 155 160
Thr Phe Thr Asp Tyr Ala Met His Trp Val Lys Gln Ala Pro Gly Gln
165 170 175
Gly Leu Glu Trp Ile Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn
180 185 190
Tyr Asn Gln Lys Phe Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser
195 200 205
Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr
210 215 220
Ala Val Tyr Tyr Cys Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr
225 230 235 240
Met Asp Phe Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser
245 250 255
Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr
260 265 270
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
275 280 285
Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
290 295 300
His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
305 310 315 320
Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
325 330 335
Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val
340 345 350
Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
355 360 365
Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
370 375 380
Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
385 390 395 400
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
405 410 415
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
420 425 430
Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
435 440 445
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
450 455 460
Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
465 470 475 480
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
485 490 495
Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
500 505 510
Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
515 520 525
Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
530 535 540
Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
545 550 555 560
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
565 570 575
Ser Leu Ser Leu Ser Pro Gly Lys
580
<210> 37
<211> 336
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 37
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln
115 120 125
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr
130 135 140
Cys Gly Ala Ser Glu Asn Ile Tyr Gly Thr Leu Asn Trp Tyr Gln Gln
145 150 155 160
Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile Tyr Gly Ala Thr Asn Leu
165 170 175
Ala Asp Ala Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
180 185 190
Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr
195 200 205
Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr Thr Phe Gly Gly Gly Thr
210 215 220
Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe
225 230 235 240
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
245 250 255
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
260 265 270
Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
275 280 285
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser
290 295 300
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His
305 310 315 320
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
325 330 335
<210> 38
<211> 589
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 38
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln
130 135 140
Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys
145 150 155 160
Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala Met His Trp Val Lys
165 170 175
Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Ser Thr Tyr
180 185 190
Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys Gly Lys Ala Ile Met
195 200 205
Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu
210 215 220
Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val Arg Gly Ser Thr Thr
225 230 235 240
Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln Gly Thr Thr Val Thr
245 250 255
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
260 265 270
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
275 280 285
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
290 295 300
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
305 310 315 320
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
325 330 335
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
340 345 350
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
355 360 365
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
370 375 380
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
385 390 395 400
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
405 410 415
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
420 425 430
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
435 440 445
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
450 455 460
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
465 470 475 480
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
485 490 495
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
500 505 510
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
515 520 525
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
530 535 540
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
545 550 555 560
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
565 570 575
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
580 585
<210> 39
<211> 341
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 39
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp
115 120 125
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp
130 135 140
Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Thr Leu
145 150 155 160
Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile Tyr
165 170 175
Gly Ala Thr Asn Leu Ala Asp Ala Val Pro Ser Arg Phe Ser Gly Ser
180 185 190
Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu
195 200 205
Asp Val Ala Thr Tyr Tyr Cys Gln Asn Val Leu Ser Ile Pro Tyr Thr
210 215 220
Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro
225 230 235 240
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr
245 250 255
Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
260 265 270
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu
275 280 285
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
290 295 300
Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala
305 310 315 320
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe
325 330 335
Asn Arg Gly Glu Cys
340
<210> 40
<211> 594
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 40
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
130 135 140
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
145 150 155 160
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
165 170 175
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
180 185 190
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
195 200 205
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
210 215 220
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
225 230 235 240
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
245 250 255
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
260 265 270
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
275 280 285
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
290 295 300
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
305 310 315 320
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
325 330 335
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
340 345 350
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
355 360 365
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
370 375 380
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
385 390 395 400
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
405 410 415
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
420 425 430
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
435 440 445
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
450 455 460
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
465 470 475 480
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
485 490 495
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
500 505 510
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
515 520 525
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
530 535 540
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
545 550 555 560
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
565 570 575
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
580 585 590
Gly Lys
<210> 41
<211> 346
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 41
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
115 120 125
Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
130 135 140
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn
145 150 155 160
Ile Tyr Gly Thr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro
165 170 175
Lys Leu Leu Ile Tyr Gly Ala Thr Asn Leu Ala Asp Ala Val Pro Ser
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser
195 200 205
Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys Gln Asn Val Leu
210 215 220
Ser Ile Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
225 230 235 240
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
245 250 255
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
260 265 270
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
275 280 285
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
290 295 300
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
305 310 315 320
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
325 330 335
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
340 345
<210> 42
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 42
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu
500 505 510
Trp Ile Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 43
<211> 589
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 43
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln
130 135 140
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg
145 150 155 160
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala
165 170 175
Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile
180 185 190
Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg
195 200 205
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met
210 215 220
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala
225 230 235 240
Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr
245 250 255
Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
260 265 270
Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
275 280 285
Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
290 295 300
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
305 310 315 320
Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
325 330 335
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
340 345 350
Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
355 360 365
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
370 375 380
Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu
385 390 395 400
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
405 410 415
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
420 425 430
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
435 440 445
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
450 455 460
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
465 470 475 480
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
485 490 495
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
500 505 510
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
515 520 525
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
530 535 540
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
545 550 555 560
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
565 570 575
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
580 585
<210> 44
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 44
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
500 505 510
Trp Val Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Lys Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Leu Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Ile
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 45
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 45
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
500 505 510
Trp Val Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Leu Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Ile
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Asn Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 46
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 46
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
500 505 510
Trp Val Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Lys Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Leu Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Ile
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Asn Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 47
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 47
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
500 505 510
Trp Val Ala Phe Ile Arg Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Leu Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Ile
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 48
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 48
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
245 250 255
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
260 265 270
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
275 280 285
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
290 295 300
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
305 310 315 320
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
325 330 335
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
340 345 350
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
355 360 365
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
370 375 380
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
385 390 395 400
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
405 410 415
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
420 425 430
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
435 440 445
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
450 455 460
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
465 470 475 480
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
485 490 495
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
500 505 510
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
515 520 525
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
530 535 540
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
545 550 555 560
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
565 570 575
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
580 585 590
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
595 600 605
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
610 615 620
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
625 630 635 640
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
645 650 655
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
660 665 670
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
675 680 685
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
690 695 700
Gly Lys
705
<210> 49
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 49
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Glu
115 120 125
Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala Trp Ile Arg
145 150 155 160
Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile Ala Phe Ile Thr Asn Leu
165 170 175
Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg Phe Thr Ile
180 185 190
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Gly Asp Tyr
210 215 220
Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
245 250 255
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
260 265 270
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
275 280 285
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
290 295 300
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
305 310 315 320
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
325 330 335
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
340 345 350
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
355 360 365
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
370 375 380
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
385 390 395 400
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
405 410 415
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
420 425 430
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
435 440 445
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
450 455 460
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
465 470 475 480
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
485 490 495
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
500 505 510
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
515 520 525
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
530 535 540
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
545 550 555 560
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
565 570 575
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
580 585 590
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
595 600 605
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
610 615 620
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
625 630 635 640
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
645 650 655
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
660 665 670
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
675 680 685
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
690 695 700
Gly Lys
705
<210> 50
<211> 711
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 50
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Phe Ile Arg Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Leu Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Ile Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
245 250 255
Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val
260 265 270
Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr
275 280 285
Phe Thr Asp Tyr Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly
290 295 300
Leu Glu Trp Ile Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr
305 310 315 320
Asn Gln Lys Phe Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile
325 330 335
Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala
340 345 350
Val Tyr Tyr Cys Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met
355 360 365
Asp Phe Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr
370 375 380
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
385 390 395 400
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
405 410 415
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
420 425 430
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
435 440 445
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
450 455 460
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
465 470 475 480
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
485 490 495
Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
500 505 510
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
515 520 525
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
530 535 540
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
545 550 555 560
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
565 570 575
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
580 585 590
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
595 600 605
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
610 615 620
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
625 630 635 640
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
645 650 655
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
660 665 670
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
675 680 685
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
690 695 700
Leu Ser Leu Ser Pro Gly Lys
705 710
<210> 51
<211> 711
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 51
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Phe Ile Arg Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Leu Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Ile Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
245 250 255
Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val
260 265 270
Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr
275 280 285
Phe Thr Asp Tyr Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly
290 295 300
Leu Glu Trp Ile Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr
305 310 315 320
Asn Gln Lys Phe Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile
325 330 335
Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala
340 345 350
Val Tyr Tyr Cys Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met
355 360 365
Asp Phe Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr
370 375 380
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
385 390 395 400
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
405 410 415
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
420 425 430
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
435 440 445
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
450 455 460
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
465 470 475 480
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
485 490 495
Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
500 505 510
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
515 520 525
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
530 535 540
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
545 550 555 560
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
565 570 575
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
580 585 590
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
595 600 605
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
610 615 620
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
625 630 635 640
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
645 650 655
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
660 665 670
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
675 680 685
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
690 695 700
Leu Ser Leu Ser Pro Gly Lys
705 710
<210> 52
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 52
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu
500 505 510
Trp Ile Ala Phe Ile Arg Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 53
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 53
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Arg Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
245 250 255
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
260 265 270
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
275 280 285
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
290 295 300
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
305 310 315 320
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
325 330 335
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
340 345 350
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
355 360 365
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
370 375 380
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
385 390 395 400
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
405 410 415
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
420 425 430
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
435 440 445
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
450 455 460
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
465 470 475 480
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
485 490 495
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
500 505 510
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
515 520 525
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
530 535 540
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
545 550 555 560
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
565 570 575
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
580 585 590
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
595 600 605
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
610 615 620
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
625 630 635 640
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
645 650 655
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
660 665 670
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
675 680 685
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
690 695 700
Gly Lys
705
<210> 54
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 54
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
450 455 460
Gly Ser Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
465 470 475 480
Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
485 490 495
Asp Tyr Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu
500 505 510
Trp Ile Ala Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp
515 520 525
Thr Val Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
530 535 540
Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
545 550 555 560
Tyr Cys Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln
565 570 575
Gly Thr Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly
580 585 590
Gly Ser Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala
595 600 605
Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala
610 615 620
Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln
625 630 635 640
Ala Pro Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val
645 650 655
Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
660 665 670
Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
675 680 685
Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu
690 695 700
Ile Lys
705
<210> 55
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 55
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Gly Pro Glu Trp Ile
35 40 45
Ala Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
245 250 255
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
260 265 270
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
275 280 285
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
290 295 300
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
305 310 315 320
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
325 330 335
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
340 345 350
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
355 360 365
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
370 375 380
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
385 390 395 400
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
405 410 415
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
420 425 430
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
435 440 445
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
450 455 460
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
465 470 475 480
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
485 490 495
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
500 505 510
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
515 520 525
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
530 535 540
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
545 550 555 560
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
565 570 575
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
580 585 590
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
595 600 605
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
610 615 620
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
625 630 635 640
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
645 650 655
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
660 665 670
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
675 680 685
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
690 695 700
Gly Lys
705
<210> 56
<211> 706
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 56
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile
35 40 45
Ala Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln
245 250 255
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
260 265 270
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
275 280 285
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
290 295 300
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
305 310 315 320
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
325 330 335
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
340 345 350
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
355 360 365
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
370 375 380
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
385 390 395 400
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
405 410 415
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
420 425 430
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
435 440 445
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
450 455 460
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
465 470 475 480
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
485 490 495
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
500 505 510
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
515 520 525
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
530 535 540
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
545 550 555 560
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
565 570 575
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
580 585 590
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
595 600 605
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
610 615 620
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
625 630 635 640
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
645 650 655
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
660 665 670
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
675 680 685
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
690 695 700
Gly Lys
705
<210> 57
<211> 719
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 57
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile
35 40 45
Ala Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
115 120 125
Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu
130 135 140
Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser
145 150 155 160
Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
165 170 175
Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala
180 185 190
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
195 200 205
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser
210 215 220
Thr Phe Pro Pro Tyr Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys
225 230 235 240
Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser Pro Ser Arg
245 250 255
Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln Val Gln Leu
260 265 270
Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser Val Lys Ile
275 280 285
Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala Met His Trp
290 295 300
Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Ser
305 310 315 320
Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys Gly Lys Ala
325 330 335
Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser
340 345 350
Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val Arg Gly Ser
355 360 365
Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln Gly Thr Thr
370 375 380
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
385 390 395 400
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
405 410 415
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
420 425 430
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
435 440 445
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
450 455 460
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
465 470 475 480
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
485 490 495
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
500 505 510
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
515 520 525
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
530 535 540
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
545 550 555 560
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
565 570 575
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
580 585 590
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
595 600 605
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
610 615 620
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
625 630 635 640
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
645 650 655
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
660 665 670
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
675 680 685
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
690 695 700
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
705 710 715
<210> 58
<211> 719
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 58
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 Ser Ala Ser Ser Ser Val Asn Tyr Val
20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr
35 40 45
Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser
50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu
65 70 75 80
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr
85 90 95
Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser
100 105 110
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Glu
115 120 125
Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys
130 135 140
Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala Trp Ile Arg
145 150 155 160
Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile Ala Phe Ile Asn Asn Leu
165 170 175
Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg Phe Thr Ile
180 185 190
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu
195 200 205
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Gly Asp Tyr
210 215 220
Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala
225 230 235 240
Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser Pro Ser Arg
245 250 255
Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln Val Gln Leu
260 265 270
Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser Val Lys Ile
275 280 285
Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala Met His Trp
290 295 300
Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Ser
305 310 315 320
Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys Gly Lys Ala
325 330 335
Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser
340 345 350
Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val Arg Gly Ser
355 360 365
Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln Gly Thr Thr
370 375 380
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
385 390 395 400
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
405 410 415
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
420 425 430
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
435 440 445
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
450 455 460
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn
465 470 475 480
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His
485 490 495
Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
500 505 510
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
515 520 525
Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
530 535 540
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
545 550 555 560
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
565 570 575
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
580 585 590
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
595 600 605
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
610 615 620
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
625 630 635 640
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
645 650 655
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
660 665 670
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
675 680 685
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
690 695 700
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
705 710 715
<210> 59
<211> 719
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 59
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser
450 455 460
Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln
465 470 475 480
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser
485 490 495
Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly
500 505 510
Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile Ala
515 520 525
Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr
530 535 540
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu
545 550 555 560
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
565 570 575
Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu
580 585 590
Val Thr Val Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
595 600 605
Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
610 615 620
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser
625 630 635 640
Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
645 650 655
Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg
660 665 670
Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
675 680 685
Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr
690 695 700
Phe Pro Pro Tyr Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys
705 710 715
<210> 60
<211> 719
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 60
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser
450 455 460
Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln Glu
465 470 475 480
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu
485 490 495
Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Asn Tyr Val Asn
500 505 510
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile Tyr Gly
515 520 525
Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly
530 535 540
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp
545 550 555 560
Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro Tyr Thr
565 570 575
Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly
580 585 590
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Glu Ser
595 600 605
Gly Gly Gly Val Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala
610 615 620
Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met Ala Trp Ile Arg Gln
625 630 635 640
Ala Pro Gly Lys Cys Pro Glu Trp Ile Ala Phe Ile Asn Asn Leu Ala
645 650 655
Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr Gly Arg Phe Thr Ile Ser
660 665 670
Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg
675 680 685
Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ala Gly Asp Tyr Arg
690 695 700
Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala
705 710 715
<210> 61
<211> 732
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 61
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile
35 40 45
Ala Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser
115 120 125
Leu Pro Ser Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu
130 135 140
Pro Gln Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser
145 150 155 160
Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Asn
165 170 175
Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu
180 185 190
Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser
195 200 205
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu
210 215 220
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro
225 230 235 240
Pro Tyr Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys Ser Ser Ser
245 250 255
Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser Pro Ser Arg Leu Pro Gly
260 265 270
Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln Val Gln Leu Val Gln Ser
275 280 285
Gly Ala Glu Val Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys
290 295 300
Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala Met His Trp Val Lys Gln
305 310 315 320
Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Ser Thr Tyr Ser
325 330 335
Gly His Thr Asn Tyr Asn Gln Lys Phe Lys Gly Lys Ala Ile Met Thr
340 345 350
Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg
355 360 365
Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val Arg Gly Ser Thr Thr Ala
370 375 380
His Tyr Tyr Thr Met Asp Phe Trp Gly Gln Gly Thr Thr Val Thr Val
385 390 395 400
Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser
405 410 415
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys
420 425 430
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
435 440 445
Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
450 455 460
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
465 470 475 480
Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
485 490 495
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro
500 505 510
Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
515 520 525
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
530 535 540
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
545 550 555 560
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
565 570 575
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
580 585 590
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
595 600 605
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
610 615 620
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
625 630 635 640
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
645 650 655
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
660 665 670
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
675 680 685
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
690 695 700
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
705 710 715 720
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
725 730
<210> 62
<211> 732
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 62
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Ala Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
35 40 45
Gly Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Val Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly
100 105 110
Gln Gly Thr Thr 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 Glu Glu Met 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 Lys Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser
450 455 460
Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln
465 470 475 480
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly Ser
485 490 495
Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly
500 505 510
Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile Ala
515 520 525
Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val Thr
530 535 540
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu
545 550 555 560
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
565 570 575
Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr Leu
580 585 590
Val Thr Val Ser Ala Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu
595 600 605
Pro Ser Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro
610 615 620
Gln Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro
625 630 635 640
Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Asn Tyr
645 650 655
Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu Ile
660 665 670
Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly
675 680 685
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
690 695 700
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro Pro
705 710 715 720
Tyr Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys
725 730
<210> 63
<211> 816
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 63
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile
35 40 45
Ala Phe Ile Thr Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser
115 120 125
Leu Pro Ser Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu
130 135 140
Pro Gln Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser
145 150 155 160
Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser Ala Ser Ser Ser Val Asn
165 170 175
Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ile Leu
180 185 190
Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser
195 200 205
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu
210 215 220
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Thr Phe Pro
225 230 235 240
Pro Tyr Thr Phe Gly Cys Gly Thr Lys Leu Glu Ile Lys Ser Ser Ser
245 250 255
Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser Pro Ser Arg Leu Pro Gly
260 265 270
Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln Val Gln Leu Val Gln Ser
275 280 285
Gly Ala Glu Val Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys
290 295 300
Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala Met His Trp Val Lys Gln
305 310 315 320
Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Val Ile Ser Thr Tyr Ser
325 330 335
Gly His Thr Asn Tyr Asn Gln Lys Phe Lys Gly Lys Ala Ile Met Thr
340 345 350
Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg
355 360 365
Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val Arg Gly Ser Thr Thr Ala
370 375 380
His Tyr Tyr Thr Met Asp Phe Trp Gly Gln Gly Thr Thr Val Thr Val
385 390 395 400
Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser
405 410 415
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys
420 425 430
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
435 440 445
Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
450 455 460
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
465 470 475 480
Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
485 490 495
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro
500 505 510
Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
515 520 525
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
530 535 540
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
545 550 555 560
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
565 570 575
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
580 585 590
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
595 600 605
Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
610 615 620
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
625 630 635 640
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
645 650 655
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
660 665 670
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
675 680 685
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
690 695 700
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
705 710 715 720
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Ser Leu Thr Cys
725 730 735
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
740 745 750
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
755 760 765
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
770 775 780
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
785 790 795 800
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
805 810 815
<210> 64
<211> 738
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 64
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30
Gly Met Ala Trp Ile Arg Gln Ala Pro Gly Lys Cys Pro Glu Trp Ile
35 40 45
Ala Phe Ile Asn Asn Leu Ala Ser Ser Ile Tyr Tyr Ala Asp Thr Val
50 55 60
Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Gly Asp Tyr Arg Ser Phe Pro Tyr Trp Gly Gln Gly Thr
100 105 110
Leu Val Thr Val Ser Ala Gly Gly Gly Ser Ser Ser Ser Lys Ala Pro
115 120 125
Pro Pro Ser Leu Pro Ser Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr
130 135 140
Pro Ile Leu Pro Gln Gly Gly Gly Glu Ile Val Leu Thr Gln Ser Pro
145 150 155 160
Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Ser
165 170 175
Ala Ser Ser Ser Val Asn Tyr Val Asn Trp Tyr Gln Gln Lys Pro Gly
180 185 190
Gln Ala Pro Arg Ile Leu Ile Tyr Gly Ile Ser Asn Leu Ala Ser Gly
195 200 205
Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
210 215 220
Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln
225 230 235 240
Gln Arg Ser Thr Phe Pro Pro Tyr Thr Phe Gly Cys Gly Thr Lys Leu
245 250 255
Glu Ile Lys Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser
260 265 270
Pro Ser Arg Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln Gln
275 280 285
Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala Ser
290 295 300
Val Lys Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asp Tyr Ala
305 310 315 320
Met His Trp Val Lys Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly
325 330 335
Val Ile Ser Thr Tyr Ser Gly His Thr Asn Tyr Asn Gln Lys Phe Lys
340 345 350
Gly Lys Ala Ile Met Thr Arg Asp Lys Ser Ile Ser Thr Ala Tyr Met
355 360 365
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Val
370 375 380
Arg Gly Ser Thr Thr Ala His Tyr Tyr Thr Met Asp Phe Trp Gly Gln
385 390 395 400
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
405 410 415
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
420 425 430
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
435 440 445
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
450 455 460
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
465 470 475 480
Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
485 490 495
Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
500 505 510
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
515 520 525
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
530 535 540
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
545 550 555 560
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
565 570 575
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
580 585 590
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
595 600 605
Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
610 615 620
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
625 630 635 640
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
645 650 655
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
660 665 670
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
675 680 685
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
690 695 700
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
705 710 715 720
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
725 730 735
Gly Lys
<210> 65
<211> 25
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 65
Gly Gly Gly Gly Ser Gly Ser Ala Gly Ser Ala Ala Gly Ser Gly Glu
1 5 10 15
Phe Gly Gly Gly Gly Ser Gly Gly Gly
20 25
<210> 66
<211> 28
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 66
Ser Ser Ser Ser Lys Ala Pro Pro Pro Ser Leu Pro Ser Pro Ser Arg
1 5 10 15
Leu Pro Gly Pro Ser Asp Thr Pro Ile Leu Pro Gln
20 25
<210> 67
<211> 18
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 67
Lys Glu Ser Gly Ser Val Ser Ser Glu Gln Leu Ala Gln Phe Arg Ser
1 5 10 15
Leu Asp
<210> 68
<211> 14
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 68
Glu Gly Lys Ser Ser Gly Ser Gly Ser Glu Ser Lys Ser Thr
1 5 10
<210> 69
<211> 12
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 69
Gly Ser Ala Gly Ser Ala Ala Gly Ser Gly Glu Phe
1 5 10

Claims (10)

1.一种抗人CEACAM5/6抗体的重链可变区，其特征在于，所述的重链可变区包括以下三个互补决定区：
SEQ ID NO.1所示的VH-CDR1，
SEQ ID NO.2所示的VH-CDR2，和
SEQ ID NO.3所示的VH-CDR3；其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留对CEACAM5/6的结合亲和力的衍生序列。
2.一种抗人CEACAM5/6抗体的重链，其特征在于，所述的重链具有如权利要求1所述的重链可变区。
3.一种抗人CEACAM5/6抗体的轻链可变区，其特征在于，所述的轻链可变区包括以下三个互补决定区CDR：
SEQ ID NO.4所示的VL-CDR1，
SEQ ID NO.5所示的VL-CDR2，和
SEQ ID NO.6所示的VL-CDR3；其中，上述氨基酸序列中任意一种氨基酸序列还包括任选地经过添加、缺失、修饰和/或取代至少一个氨基酸的，并能够保留对CEACAM5/6的结合亲和力的衍生序列。
4.一种抗人CEACAM5/6抗体的轻链，其特征在于，所述的轻链具有如权利要求3所述的轻链可变区。
5.一种抗人CEACAM5/6抗体，其特征在于，所述抗体具有：
(1)如权利要求1所述的重链可变区；和/或
(2)如权利要求3所述的轻链可变区；
或者，所述抗体具有：如权利要求2所述的重链；和/或如权利要求4所述的轻链。
6.一种多特异性抗原结合分子，其特征在于，包含：
第一抗原结合域D1；和
第二抗原结合域D2；
其中，D1特异性结合靶分子CEACAM5/6蛋白；
D2特异性结合靶分子CD47蛋白；
所述D1为特异性结合CEACAM5/6蛋白的抗体或其抗原结合片段；
所述D2为特异性结合CD47蛋白的抗体或其抗原结合片段；
其中，所述D1具有包含以下三个互补决定区CDR的重链可变区：
SEQ ID NO.1所示的VH-CDR1，
SEQ ID NO.2所示的VH-CDR2，和
SEQ ID NO.3所示的VH-CDR3；以及
包含以下三个互补决定区CDR的轻链可变区：
SEQ ID NO.4所示的VL-CDR1，
SEQ ID NO.5所示的VL-CDR2，和
SEQ ID NO.6所示的VL-CDR3；
其中，所述抗原结合片段的结构选自下组：(i)Fab片段；(ii)F(ab')2片段；(iii)Fd片段；(iv)Fv片段；(v)单链Fv(scFv)分子；或(vi)dAb片段。
7.一种重组蛋白，其特征在于，所述的重组蛋白具有：
(i)如权利要求1所述的重链可变区、如权利要求2所述的重链、如权利要求3所述的轻链可变区、如权利要求4所述的轻链、如权利要求5所述的抗体或如权利要求6所述的多特异性抗原结合分子；以及
(ii)任选的协助表达和/或纯化的标签序列。
8.一种CAR构建物，其特征在于，所述的CAR构建物的抗原结合结构域的scFv区段特异性结合于人CEACAM5/6，并且所述scFv区段包含如权利要求1所述的重链可变区和如权利要求3所述的轻链可变区。
9.一种重组的免疫细胞，其特征在于，所述的免疫细胞表达外源的如权利要求8所述的CAR构建物。
10.一种抗体药物偶联物，其特征在于，所述的抗体药物偶联物含有：
(a)抗体部分，所述抗体部分选自下组：如权利要求1所述的重链可变区、如权利要求2所述的重链、如权利要求3所述的轻链可变区、如权利要求4所述的轻链、如权利要求5所述的抗体、或如权利要求6所述的多特异性抗原结合分子、或其组合；和
(b)与所述抗体部分偶联的偶联部分，所述偶联部分选自下组：可检测标记物、药物、毒素、细胞因子、酶、或其组合。
CN202111093726.0A 2021-09-17 2021-09-17 一种靶向人ceacam5/6的抗体、制备方法和应用 Pending CN115819596A (zh)

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US7232888B2 (en) * 2002-07-01 2007-06-19 Massachusetts Institute Of Technology Antibodies against tumor surface antigens
CN101607985B (zh) * 2008-12-24 2013-03-27 中国科学院生物物理研究所 抗人cea的单克隆抗体，包含其的组合物，及其用途
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