CN109575140B - 靶向pd-1或pd-l1且靶向vegf家族的双靶向融合蛋白及其用途 - Google Patents

靶向pd-1或pd-l1且靶向vegf家族的双靶向融合蛋白及其用途 Download PDF

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CN109575140B
CN109575140B CN201710905683.9A CN201710905683A CN109575140B CN 109575140 B CN109575140 B CN 109575140B CN 201710905683 A CN201710905683 A CN 201710905683A CN 109575140 B CN109575140 B CN 109575140B
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胡品良
邹敬
洪伟东
何芸
白洁
宋凌云
杨文第
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Beijing Beyond Biotechnology Co ltd
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Abstract

本发明提供了靶向PD‑1或PD‑L1且靶向VEGF家族这两者的双靶向融合蛋白，所述双靶向融合蛋白包含(i)抗PD‑1抗体或者抗PD‑L1抗体和(ii)在所述抗PD‑1抗体或者抗PD‑L1抗体的两条重链中的每一重链的C端有效连接的一个VID。本发明还提供了编码所述双靶向融合蛋白的多核苷酸、包含所述多核苷酸的载体、包含所述多核苷酸或载体的宿主细胞、以及所述双靶向融合蛋白在个体中治疗、预防和/或诊断与PD‑1活性、PD‑L1活性和VEGF家族活性相关的疾病中的用途。

Description

靶向PD-1或PD-L1且靶向VEGF家族的双靶向融合蛋白及其 用途
技术领域
本发明总体上涉及医药生物技术领域。具体地，本发明涉及靶向程序性死亡蛋白-1(programmed death-1(PD-1))或程序性死亡蛋白配体1(programmed death-1ligand(PD-L1))、且靶向血管内皮细胞生长因子(Vascular Endothelial Cell Growth Factor(VEGF))家族这两者的双靶向融合蛋白、编码所述双靶向融合蛋白的多核苷酸、包含所述多核苷酸的载体、包含所述多核苷酸或载体的宿主细胞、以及所述双靶向融合蛋白在个体中治疗、预防和/或诊断与PD-1或PD-L1活性和VEGF家族活性相关的疾病中的用途。
背景技术
免疫检查点(immune checkpoint)是免疫系统中存在的一类抑制性信号分子，通过调节外周组织中免疫反应的持续性和强度避免组织损伤，并参与维持对于自身抗原的耐受(Pardoll DM.,The blockade of immune checkpoints in cancer immunotherapy.NatRev Cancer,2012,12(4):252-264)。研究发现，肿瘤细胞能够逃避体内免疫系统而失控增殖的原因之一是利用了免疫检查点的抑制性信号通路，由此抑制了T淋巴细胞活性，使得T淋巴细胞不能有效发挥对肿瘤的杀伤效应(Yao S,Zhu Y和Chen L.，Advances intargeting cell surface signaling molecules for immune modulation.Nat Rev DrugDiscov,2013,12(2):130-146)。
程序性死亡蛋白-1(PD-1)是一种重要的免疫检查点蛋白，目前也是肿瘤免疫治疗的一个重要靶标。PD-1在1992年首次被发现，对其基因的克隆和表达表明PD-1活化后能够诱导T细胞程序性死亡。在活化的T细胞、B细胞和髓样细胞上均发现存在PD-1蛋白。PD-1还诱导性地表达于巨噬细胞、树突状细胞以及单核细胞。在静息的淋巴细胞表面无PD-1表达。
PD-1是一种55kDa I型跨膜蛋白，其胞浆区含有一个免疫受体酪氨酸抑制基序，与CD28和CTLA-4具有同源性。已鉴定到PD-1的两种细胞表面糖蛋白配体，分别为程序性死亡蛋白配体1(PD-L1)和程序性死亡蛋白配体2(PD-L2)。已经在许多癌细胞上发现了PD-1的配体表达，包括人肺癌、卵巢癌、结肠癌和多种骨髓瘤。另外，在各类上皮癌、血液癌和其他恶性肿瘤细胞表面上高表达PD-1的配体。在肿瘤患者中，PD-1的配体如PD-L1的表达经常与癌的预后不良相关(Iwai Y等人，Involvement of PD-L1 on tumor cells in the escapefrom host immune system and tumor immunotherapy by PD-L1 blockade，PNAS，2002，99(19):12293-7)。
PD-1与PD-1的配体的结合对于调节T淋巴细胞活性和维持外周免疫耐受发挥重要作用。PD-1与PD-1的配体结合后可导致T细胞凋亡、免疫无应答、T细胞“耗竭”和分泌IL-10等。因此，PD-1发挥限制T细胞活化、抑制T细胞增殖和提高对抗原的耐受性的功能。活化的淋巴细胞表面PD-1的表达上调能够导致对获得性或者固有的免疫反应的抑制，由此导致了(包括T淋巴细胞在内的)肿瘤浸润性淋巴细胞虽然具有肿瘤抗原特异性，但由于肿瘤细胞上PD-1的配体与肿瘤浸润性淋巴细胞上的PD-1结合产生了抑制肿瘤浸润性淋巴细胞活化的信号，从而肿瘤细胞能够逃避免疫系统对肿瘤细胞的杀伤。
研究表明，这些表达PD-1的肿瘤浸润性淋巴细胞是功能障碍型淋巴细胞，所述淋巴细胞的生物学功能可以通过阻断PD-1与PD-1的配体结合的抗体恢复。目前，抑制PD-1与PD-1的配体结合的抗体主要包括抗PD-1单克隆抗体和抗PD-L1单克隆抗体，但也有针对PD-L2的产品。
当前，研究比较成熟的抗PD-1抗体有百时美施贵宝(BMS)公司的纳武单抗(Nivolumab)和默克(Merck)公司的派姆单抗(Pembrolizumab)。纳武单抗(商品名
Figure BDA0001423925870000021
)为完全人源化的IgG4抗体分子，派姆单抗(商品名
Figure BDA0001423925870000022
)为人源化IgG4抗体分子。所述抗PD-1单克隆抗体与T淋巴细胞上的PD-1结合后能够抑制PD-1与其配体PD-L1和PD-L2的结合，由此促进T淋巴细胞活化、增殖和产生免疫活化型细胞因子如IL-2，并解除PD-1对具有抗肿瘤活性的T淋巴细胞免疫监视的抑制。美国食品药品监督管理局目前批准的纳武单抗适应症包括：黑素瘤、非小细胞肺癌、肾癌、头颈部肿瘤等；派姆单抗的适应症包括：头颈部肿瘤、非小细胞肺癌、黑素瘤等。关于抗PD-L1抗体，罗氏(Roche)研发的atezolizumab、德国默克(Merck KGaA)和美国辉瑞(Pfizer)合作开发的avelumab、阿斯利康研发的durvalumab也显示了对肿瘤的治疗效果。
虽然抗PD-1抗体、抗PD-L1抗体对肿瘤具有治疗效果，但它们平均的治疗有效率仅为20％左右，肺癌的五年生存率仅16％。仍有相当一部分的肿瘤患者对使用抗PD-1抗体、抗PD-L1抗体的治疗无应答。因此，如何提高肿瘤治疗的有效性仍是目前肿瘤治疗领域迫切需要解决的一个难题。
另一方面，肿瘤血管形成(angiogenesis)也是肿瘤快速生长的一个重要原因(Ferrara N和Alitalo K，Clinical applications of angiogenic growth factors andtheir inhibitors，Nat Med.,1999；5(12):1359-64)。在肿瘤的表面和深处，到处都可以见到粗细不等的血管，生命的营养物质和氧气通过这些血管被运送至肿瘤组织。肿瘤血管形成是一个相当复杂的过程，受多种因子的正负调控。在这多种因子中，血管内皮细胞生长因子家族是一类作用最强的正性调控因子，发挥着刺激新生血管形成的功能。正常组织内血管内皮细胞生长因子和血管内皮细胞生长抑制因子同时存在，且保持相对平衡，这种平衡使得人体血管可以正常地生成和分化。但是，在肿瘤生长过程中，VEGF家族分子数量激增，与血管生成抑制因子之间的调节失衡，由此，极大地促进了血管内皮细胞的分裂增殖和迁移、提高了血管通透性、抑制肿瘤细胞凋亡，为肿瘤的生长和转移提供了良好的微环境。
VEGF家族包含六种密切相关的多肽，分别是高度保守的同源二聚体糖蛋白，有六个亚型：VEGF-A、-B、-C、-D、-E、和胎盘生长因子(placental growth factor(PLGF))，分子量从35至44kDa不等。VEGF-A(包括其剪接物如VEGF165)的表达与一些实体瘤的微血管密度具有相关性，并且组织中VEGF-A的浓度与乳腺癌、肺癌、前列腺癌和结肠癌等实体瘤的预后有关。每个VEGF家族成员的生物学活性通过细胞表面VEGF受体(VEGFR)家族中的一种或多种介导，所述VEGFR家族包括VEGFR1(也称为Flt-1)、VEGFR2(也称为KDR、Flk-1)、VEGFR3(也称为Flt-4)等，其中VEGFR1、VEGFR2与血管的生成关系密切，VEGF-C/D/VEGFR3则与淋巴管生成密切相关。VEGF家族的主要生物学功能包括：(1)选择性促进血管内皮细胞有丝分裂，刺激内皮细胞增殖并促进血管形成；(2)提高血管尤其是微小血管的通透性，使血浆大分子外渗沉积在血管外的基质中，为肿瘤细胞的生长和新生毛细血管网的建立提供营养；(3)促进肿瘤的增殖和转移，所述肿瘤的增殖和转移依赖VEGF家族使血管内皮细胞分泌胶原酶和纤溶酶原,借以降解血管基底膜，同时，肿瘤组织内部新形成的微血管基膜不完善,这种性质使肿瘤易于进入血循环；(4)其他作用：VEGF家族可诱导上皮细胞出现间隙及开窗现象，可活化上皮细胞的胞质小泡及细胞器；VEGF家族直接刺激内皮细胞释放蛋白水解酶，降解基质，释放更多的VEGF家族分子，加速肿瘤的发展，细胞外蛋白酶又可激活细胞外基质的结合性和VEGF家族的释放；VEGF家族通过增加血管通透性使血浆蛋白(包括纤维蛋白原)释放，形成纤维素网络，为肿瘤生长、发展和转移提供了良好的基质；(5)VEGF家族抑制机体的免疫反应,促进恶性肿瘤的浸润与转移(Lapeyre-Prost A等人，ImmunomodulatoryActivity of VEGF in Cancer，Int Rev Cell Mol Biol.2017；330:295-342)。
在VEGF家族中，胎盘生长因子(PLGF)的40％氨基酸序列和VEGF-A同源，参与病理状态下的新生血管和侧支血管的形成。PLGF的生物学功能是通过特异结合其受体VEGFR1/Flt-1来激活的。VEGFR1/Flt-1具有很强的生物学活性，结合其配体后可介导内皮细胞与基质细胞的作用，也影响内皮细胞的分化成熟。PLGF能促进早孕时滋养细胞增殖与分化，可诱导内皮细胞增殖、迁移，抗内皮细胞凋亡，并能增加血管的通透性，能增强低浓度VEGF的生物学活性，是参与多种肿瘤血管生成的重要的促血管生长因子。过度的PLGF表达导致肿瘤生长的增加和血管的存活。在原发肿瘤中观察到PLGF在所有血管丰富的肿瘤中表达，而血管少的肿瘤只部分表达PLGF。因此，PLGF可用来解释脑部肿瘤血管新生的机制，而通过抑制PLGF的生物学活性可以达到抑制肿瘤生长的目的。
临床研究显示利用单克隆抗体或可溶性VEGFR能够阻断VEGF家族与其受体的结合，阻碍VEGF家族信号通路的传导也是目前治疗肿瘤的方法之一。基因泰克(Genentech)公司研发的贝伐单抗(Bevacizumab，商品名Avastin)是一种重组的人鼠嵌合抗VEGF抗体，可通过阻断VEGF-A与VEGFR的结合，使VEGFR无法活化，由此发挥抗血管生成的作用。贝伐单抗目前用于一线治疗转移性结直肠癌，将来有可能用于转移性肺癌、乳癌、胰脏癌、肾脏癌等疾病的治疗。贝伐单抗也是开发较为成功的抗体药物之一。Sanofi-aventis公司和Regeneron公司研制的阿柏西普(aflibercept)是一种VEGF-Trap，其是将VEGFR1胞外第2个结构域和VEGFR2胞外第3个结构域与人IgG1恒定区融合获得的一种融合蛋白，能通过抑制血管生成而对一部分肿瘤患者发挥抗肿瘤作用。
仍有大部分的肿瘤患者对目前可用的抗PD-1抗体、抗PD-L1抗体、抗VEGF抗体或VEGF-Trap的单独治疗无反应。
鉴于免疫检查点蛋白PD-1、PD-L1在调节免疫应答中的重要性，以及VEGF家族在肿瘤微环境中抑制抗肿瘤免疫和促进肿瘤血管形成的作用，本领域仍需要治疗肿瘤的备选疗法。优选地，这类备选疗法能够既靶向免疫抑制性蛋白PD-1或PD-L1，又靶向具有免疫抑制作用和促血管生成作用的VEGF家族分子，从而导致免疫系统的激活和肿瘤血管的消退，从而对靶向PD-1或PD-L1的单一治疗无反应或者靶向VEGF家族的单一治疗无反应的患者显示功效。这类备选疗法的一个方案是共施用靶向PD-1或PD-L1和靶向VEGF家族分子的两种不同的生物制品。共施用需要注射两个独立的产品或单次注射两种不同蛋白的联合制剂。尽管两次注射允许给药量和给药时间的灵活性，但是它造成了患者不便依从和疼痛。另外，尽管联合制剂可能提供在给药量方面的某种灵活性，但它通常难以找到在溶液中允许两种蛋白的化学和物理稳定性的配制条件，原因在于两种蛋白的分子特征不同。另外，共施用和联合制剂两种不同药物的疗法可能增加患者和/或付款人的额外花费。因此，仍需要治疗肿瘤的备选疗法，并且优选地这类备选疗法包含靶向PD-1或PD-L1且靶向VEGF家族的双靶向融合蛋白。
本发明提供了靶向PD-1或PD-L1且靶向VEGF家族的双靶向新型融合蛋白，其能够抑制对PD-1途径或对PD-L1途径和对VEGF家族信号传导途径的激活，并用于在个体中治疗、预防和/或诊断与PD-1、PD-L1活性和VEGF家族活性相关的疾病。
发明概述
本发明公开了一种新型的靶向PD-1或PD-L1且靶向VEGF家族的双靶向融合蛋白、编码所述双靶向融合蛋白的多核苷酸、包含所述多核苷酸的载体、包含所述多核苷酸或载体的宿主细胞、以及所述双靶向融合蛋白在个体中治疗、预防和/或诊断与PD-1、PD-L1活性和VEGF家族活性相关的疾病中的用途。
因此，在一个方面，本发明提供了靶向PD-1或PD-L1且靶向VEGF家族的双靶向融合蛋白，所述双靶向融合蛋白抑制PD-1与其配体的结合或抑制PD-L1与其受体的结合、且抑制VEGF家族的信号传导途径，其包含(i)抗PD-1抗体或者抗PD-L1抗体；和(ii)与所述抗PD-1抗体或者抗PD-L1抗体有效连接的至少两个抑制VEGF家族的结构域(VEGFs familiyinhibiting domain，下文中缩写为VID)。
在一个实施方案中，本发明的双靶向融合蛋白包含(i)抗PD-1抗体或者抗PD-L1抗体；和(ii)在所述抗PD-1抗体或者抗PD-L1抗体的两条重链中的每一重链的C端有效连接的一个VID，任选地，所述(i)和所述(ii)通过肽接头有效连接，由此，两个相同或者不同的VID各自在它们的N端氨基酸处与所述抗PD-1抗体或者抗PD-L1抗体的重链之一的C端氨基酸融合，任选地通过肽接头融合，优选地，所述VID包含VEGF家族的受体的胞外结构域的一部分。
所述双靶向融合蛋白中包含的抗PD-1抗体可以是任何抗PD-1抗体，只要是能够抑制或减少PD-1与其配体结合的抗体即可，包括现有技术中已知的抗PD-1抗体和将来研发出的抗PD-1抗体。在一个实施方案中，所述抗PD-1抗体包含选自SEQ ID NO：1/2、3/4、5/6、7/8、9/10、11/12、13/14、15/16、17/18、19/20、21/22、23/24、和120/121的成对重链可变区序列/轻链可变区序列中所含的全部重链CDR与轻链CDR，优选地，所述抗PD-1抗体包含选自SEQ ID NO：1/2、3/4、5/6、7/8、9/10、11/12、13/14、15/16、17/18、19/20、21/22、23/24、和120/121的成对重链可变区序列/轻链可变区序列，或与所述成对重链可变区序列/轻链可变区序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多序列同一性的序列；更优选地，所述抗PD-1抗体包含选自纳武单抗、pidilizumab和派姆单抗的抗PD-1抗体的重链可变区和轻链可变区，特别地，所述抗PD-1抗体选自纳武单抗、pidilizumab和派姆单抗。
所述双靶向融合蛋白中包含的抗PD-L1抗体可以是任何抗PD-L1抗体，只要是能够抑制或减少PD-L1与其受体结合(例如与PD-1或CD80(B7-1)或与这两者结合)的抗体即可，包括现有技术中已知的抗PD-L1抗体和将来研发出的抗PD-L1抗体。在一个实施方案中，本发明融合蛋白中的抗PD-L1抗体包含选自SEQ ID NO：25/26、27/28和29/30的成对重链可变区序列/轻链可变区序列中所含的全部重链CDR与轻链CDR。优选地，所述抗PD-L1抗体包含选自SEQID NO：25/26、27/28和29/30的成对重链可变区序列/轻链可变区序列，或与所述成对重链可变区序列/轻链可变区序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多序列同一性的序列；更优选地，所述抗PD-L1抗体选自atezolizumab、avelumab和durvalumab。
在一个实施方案中，所述抗PD-1抗体或抗PD-L1抗体是IgG类抗体，特别地是IgG1亚类、IgG2亚类、IgG4亚类抗体。在一个优选的实施方案中，包含于本发明融合蛋白中的所述抗PD-1抗体或抗PD-L1抗体是IgG4亚类抗体，特别地是人IgG4亚类抗体。在一个实施方案中，所述IgG4亚类抗体在Fc区中第S228位置(EU编号)处包含氨基酸置换，特别是氨基酸置换S228P。SEQ ID NO:33中显示了示例性IgG1亚类抗PD-1抗体的重链恒定区氨基酸序列。SEQID NO:34中显示了示例性IgG2亚类抗PD-1抗体的重链恒定区氨基酸序列。SEQ ID NO:35中显示了示例性IgG4亚类抗PD-1抗体的重链恒定区氨基酸序列。
在一个实施方案中，所述抗PD-1抗体或抗PD-L1抗体包含完全抗体的可变区和恒定区。本发明的双靶向融合蛋白中的抗体轻链恒定区型别可以是κ型或λ型，优选地是κ型。SEQ ID NO:31中显示了示例性抗PD-1抗体的κ型轻链恒定区氨基酸序列。SEQ ID NO:32中显示了示例性抗PD-1抗体的λ型轻链恒定区氨基酸序列。
所述双靶向融合蛋白中包含的VID包含VEGF家族的受体的胞外结构域的一部分。在一个实施方案中，所述VID包含VEGFR1的免疫球蛋白(Ig)样结构域2(Domain 2，缩写为D2)和VEGFR2的Ig样结构域3(Domain 3，缩写为D3)。在一个具体实施方案中，所述VEGFR1-D2/VEGFR2-D3具有SEQ ID NO：63所示的氨基酸序列，或与SEQ ID NO：63的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多同一性的氨基酸序列。在一个实施方案中，所述VID包含VEGFR1-D2以及VEGFR2-D3和VEGFR2的Ig样结构域4(Domain 4，缩写为D4)。在一个具体实施方案中，所述VEGFR1-D2/VEGFR2-D3-D4具有SEQ IDNO：64所示的氨基酸序列，或与SEQ ID NO：64的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多同一性的氨基酸序列。在一个实施方案中，所述VID包含VEGFR1-D2。在一个具体实施方案中，所述VEGFR1-D2具有SEQ ID NO：65所示的氨基酸序列，或与SEQ ID NO：65的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多同一性的氨基酸序列。
在一个实施方案中，在所述抗PD-1抗体或抗PD-L1抗体的重链的C端连接所述VID的肽接头包含一个或多个氨基酸，优选地包含选自SEQ ID NO：36-62的肽接头。
在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:73的抗PD-1抗体轻链亚基和SEQ ID NO:75的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.3。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:77的抗PD-1抗体轻链亚基和SEQ ID NO:79的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.7。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:81的抗PD-1抗体轻链亚基和SEQ ID NO:83的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.4。在一个具体实施方案中，所述融合蛋白包含SEQID NO:85的抗PD-1抗体轻链亚基和SEQ ID NO:87的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.5。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:89的抗PD-1抗体轻链亚基和SEQ ID NO:91的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.6。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:93的抗PD-1抗体轻链亚基和SEQ ID NO:95的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.8。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:97的抗PD-1抗体轻链亚基和SEQ IDNO:99的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.9。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:101的抗PD-1抗体轻链亚基和SEQ ID NO:103的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.10。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:105的抗PD-1抗体轻链亚基和SEQ ID NO:107的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.11。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:109的抗PD-1抗体轻链亚基和SEQ ID NO:111的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.12。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:113的抗PD-1抗体轻链亚基和SEQ ID NO:115的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.13。在一个具体实施方案中，所述融合蛋白包含SEQ ID NO:117的抗PD-1抗体轻链亚基和SEQ ID NO:119的抗PD-1抗体重链-VID融合亚基，下文中称为融合蛋白BY24.14。
在一个具体实施方案中，所述融合蛋白包含(i)一个选自atezolizumab、avelumab和durvalumab的抗PD-L1抗体和(ii)在所述抗PD-L1抗体的两条重链中的每一重链的C端有效连接的一个VID分子。
在一个实施方案中，所述融合蛋白特异性地靶向PD-1或PD-L1和VEGF家族分子，抑制由PD-1或PD-L1和VEGF家族分子介导的信号传导。本发明的融合蛋白不仅在N端能高亲和性结合PD-1或PD-L1，而且在C端也能高亲和性地结合多种VEGF因子。本发明所设计的融合蛋白的结构充分保证了该融合蛋白与两类靶标结合的合适物理空间距离，这种结构的融合蛋白与PD-1或PD-L1和VEGF家族分子中的一种分子特异性结合后不影响该融合蛋白与PD-1或PD-L1和VEGF家族分子中另一种分子的特异性结合。
本发明还提供了编码本发明融合蛋白的多核苷酸、包含编码本发明融合蛋白的多核苷酸的载体，优选地表达载体，最优选地具有双表达盒的谷氨酰胺合成酶表达载体。在另一个方面，本发明提供了包含本发明多核苷酸或载体的宿主细胞。本发明也提供了一种用于产生本发明融合蛋白的方法，包括步骤(i)在适于表达本发明融合蛋白的条件下培养本发明的宿主细胞，和(ii)回收本发明的融合蛋白。
在一个方面，本发明提供了一种包含本发明融合蛋白的诊断试剂盒和药物组合物。进一步地，还提供了本发明的融合蛋白、诊断试剂盒或药物组合物的用途，用于治疗、预防和/或诊断与PD-1或PD-L1活性和VEGF家族活性相关的疾病，特别地用于治疗、预防和/或诊断癌性疾病(例如，实体瘤和软组织瘤)，最特别地用于治疗、预防和/或诊断黑素瘤、乳腺癌、结肠癌、食管癌、胃肠道间质肿瘤(GIST)、肾癌(例如，肾细胞癌)、肝癌、非小细胞肺癌(NSCLC)、卵巢癌、胰腺癌、前列腺癌、头颈部肿瘤、胃癌、血液学恶性病(例如，淋巴瘤)。
除非另外限定，否则本文中所用的全部技术与科学术语具有如本发明所属领域的普通技术人员通常理解的相同含义。本文所提及的全部出版物、专利申请、专利和其他参考文献通过引用的方式完整地并入。此外，本文中所述的材料、方法和例子仅是说明性的并且不意在是限制性的。本发明的其他特征、目的和优点将从本说明书及附图并且从后附的权利要求书中显而易见。
附图简述
结合以下附图一起阅读时，将更好地理解以下详细描述的本发明的优选实施方案。出于说明本发明的目的，图中显示了目前优选的实施方案。然而，应当理解本发明不限于图中所示实施方案的精确安排和手段。
图1：例示了本发明的靶向PD-1或PD-L1且靶向VEGF家族的双靶向融合蛋白的结构示意图。
图2：显示了实施例2中制备并纯化的本发明融合蛋白在还原剂(5mM 1,4-二硫苏糖醇)存在下通过SDS-PAGE并用考马斯蓝染色后的结果。图2A中的泳道1：蛋白分子量标准标志物；泳道2：融合蛋白BY24.3；泳道3：融合蛋白BY24.4；泳道4：融合蛋白BY24.5；泳道5：融合蛋白BY24.6；泳道6：融合蛋白BY24.8；泳道7：融合蛋白BY24.9；泳道8：融合蛋白BY24.10；泳道9：融合蛋白BY24.11；图2B中的泳道1：蛋白分子量标准标志物；泳道2：融合蛋白BY24.12；泳道3：融合蛋白BY24.13；泳道4：融合蛋白BY24.14；泳道5：抗体BY18.1；泳道6：蛋白301-8；泳道7：融合蛋白BY24.7。
图3：显示了本发明的融合蛋白BY24.3、抗体BY18.1和蛋白301-8对实验动物体重的影响。
图4：显示了将本发明的融合蛋白BY24.3与抗体BY18.1和蛋白301-8的体内抗肿瘤作用进行比较的示意图。
发明详述
本发明提供了阻断免疫检查点PD-1途径或PD-L1途径和VEGF家族信号传导途径的融合蛋白和药物组合物。本发明还提供了用于产生该融合蛋白的方法，以及该融合蛋白在个体中治疗、预防和/或诊断与PD-1或PD-L1活性和VEGF家族活性相关的疾病中的用途。
除非下文中另外定义，否则本说明书中的术语如本领域通常所用那样使用。
I.定义
术语“约”在与数字数值联合使用时意为涵盖具有比指定数字数值小5％的下限和比指定数字数值大5％的上限的范围内的数字数值。
如本文中所用，术语“包含”或“包括”意指包括所述的要素、整数或步骤，但是不排除任意其他要素、整数或步骤。
“PD-1途径”是指任何通过与PD-1结合而引发的细胞内信号传导途径，包括但不限于PD-1与PD-L1结合而引发的细胞内信号传导途径、或PD-1与PD-L2结合而引发的细胞内信号传导途径、或者PD-1与PD-L1和PD-L2这两者结合而引发的细胞内信号传导途径。
“PD-L1途径”是指任何通过与PD-L1结合而引发的细胞内信号传导途径，包括但不限于PD-L1与PD-1结合而引发的细胞内信号传导途径、或PD-L1与CD80(B7-1)结合而引发的细胞内信号传导途径、或者PD-L1与PD-1和CD80(B7-1)这两者结合而引发的细胞内信号传导途径。
如本文所用，术语“特异性结合”意指对抗原或目的分子的结合具有选择性并且可以与不想要的或非特异的相互作用区别。所述特异性结合可以通过酶联免疫吸附测定(ELISA)或本领域技术人员熟悉的其他技术，例如表面等离子体共振(SPR)技术(在BIAcore仪上分析)(Liljeblad等人,Analysis of agalacto-IgG in rheumatoid arthritisusing surface plasmon resonance，Glyco J.,2000，17,323-329)测量。
“亲和力”或“结合亲和力”指反映结合对子的成员之间相互作用的固有结合亲和力。分子X对其配偶物Y的亲和力可以通常由解离常数(KD)代表，解离常数是解离速率常数和缔合速率常数(分别是koff和kon)的比例。亲和力可以由本领域已知的常见方法测量。用于测量亲和力的一个具体方法是表面等离子体共振法(SPR)。
术语“抗体”在本文中以最广意义使用并且包括但不限于单克隆抗体、多克隆抗体、多特异性抗体(例如，双特异性抗体)、只要它们显示出所需的抗原结合活性即可。抗体可以是完整抗体分子，也可以是完整抗体分子的功能性片段，包括但不限于例如Fab、F(ab')2。抗体的恒定区可以经改变(例如经突变)以修饰抗体特性(例如，以增加或减少以下一个或多个特性：抗体糖基化、半胱氨酸残基数目、效应细胞功能或补体功能)。
术语“全抗体”、“全长抗体”、“完全抗体”和“完整抗体”在本文中可互换地用来指这样的抗体，所述抗体具有基本上与天然抗体结构相似的结构。
术语“抗体重链”指在抗体分子中存在的两种类型多肽链中的较大者，其在正常情况下决定抗体所属的类别。
术语“抗体轻链”指在抗体分子中存在的两种类型多肽链中的较小者。κ轻链和λ轻链指两个主要的抗体轻链同种型。
氨基酸序列的“同一性百分数(％)”是指将候选序列与本说明书中所示的具体氨基酸序列进行比对并且如有必要的话为达到最大序列同一性百分数而引入空位后，并且不考虑任何保守置换作为序列同一性的一部分时，候选序列中与本说明书中所示的具体氨基酸序列的氨基酸残基相同的氨基酸残基百分数。
术语“有效连接”意指指定的各组分处于一种允许它们以预期的方式起作用的关系。
“信号序列”是连接至蛋白质的N-端部分的氨基酸的序列，其促进蛋白质分泌至细胞外。细胞外蛋白质的成熟形式没有信号序列，其在分泌过程期间被切除。
术语“N端”指N端的最末氨基酸，术语“C端”指C端的最末氨基酸。
术语“融合”指将两个或多个组分由肽键直接连接或借助一个或多个肽接头有效连接。
如本文所用，术语“融合蛋白”指包含抗体轻链亚基和抗体重链-VID融合亚基的融合多肽分子，其中抗体轻链亚基是融合蛋白中存在的多肽链中的较小者，抗体重链-VID融合亚基是融合蛋白中存在的多肽链中的较大者。
术语“宿主细胞”指已经向其中引入外源多核苷酸的细胞，包括这类细胞的子代。宿主细胞包括“转化体”和“转化的细胞”，这包括原代转化的细胞和从其衍生的子代。宿主细胞是可以用来产生本发明融合蛋白的任何类型的细胞系统。宿主细胞包括培养的细胞，也包括转基因动物、转基因植物或培养的植物组织或动物组织内部的细胞。
术语“个体”或“受试者”可互换地使用，是指哺乳动物。哺乳动物包括但不限于驯化动物(例如，奶牛、绵羊、猫、犬和马)、灵长类(例如，人和非人灵长类如猴)、兔和啮齿类(例如，小鼠和大鼠)。特别地，个体是人。
术语“治疗”指意欲改变正在接受治疗的个体中疾病之天然过程的临床介入。想要的治疗效果包括但不限于防止疾病出现或复发、减轻症状、减小疾病的任何直接或间接病理学后果、防止转移、降低病情进展速率、改善或缓和疾病状态，以及缓解或改善预后。在一些实施方案中，本发明的融合蛋白用来延缓疾病发展或用来减慢疾病的进展。
术语“抗肿瘤作用”指可以通过多种手段展示的生物学效果，包括但不限于例如，肿瘤体积减少、肿瘤细胞数目减少、肿瘤细胞增殖减少或肿瘤细胞存活减少。术语“肿瘤”和“癌症”在本文中互换地使用，涵盖实体瘤和液体肿瘤。
II.融合蛋白
本发明提供了靶向PD-1或PD-L1且靶向VEGF家族的双靶向融合蛋白，其包含(i)抗PD-1抗体或者抗PD-L1抗体；和(ii)与所述抗PD-1抗体或者抗PD-L1抗体有效连接的至少两个VID，其中融合蛋白的这两种组分彼此通过肽键直接或经肽接头连接。另外，融合蛋白中的组分(i)抗PD-1抗体或抗PD-L1抗体的各条肽链可以例如通过二硫键连接。
在一些实施方案中，本发明的融合蛋白是由二硫键键合的两条抗体轻链亚基和两条抗体重链-VID融合亚基组成的异四聚体糖蛋白。从N端至C端，每条抗体重链-VID融合亚基具有一个抗体重链，随后是一个VID，其中抗体重链和VID由肽键直接连接或借助一个或多个肽接头连接。
本发明的融合蛋白阻断免疫检查点PD-1途径或PD-L1途径且抑制VEGF家族信号传导途径。该融合蛋白阻断的免疫检查点PD-1途径是PD-1与其配体结合所介导的信号传导途径。该融合蛋白阻断的PD-L1途径是PD-L1与其受体结合所介导的信号传导途径。该融合蛋白抑制的VEGF家族信号传导途径是由VEGF-A、-B、-C、-D、-E和PLGF与VEGF家族的受体(例如VEGFR1、VEGFR2和VEGFR3)结合所介导的信号传导途径。
在一些实施方案中，本发明的融合蛋白以10-8M或更小、例如以10-9M至10-12M的解离常数(KD)与PD-1或PD-L1结合；且以10-8M或更小、例如以10-9M至10-12M的解离常数(KD)与VEGF家族特异性结合。
－抗PD-1抗体或抗PD-L1抗体
本发明融合蛋白中包含的抗PD-1抗体或抗PD-L1抗体是由二硫键键合的两条轻链和两条重链组成的异四聚体糖蛋白。
在一个实施方案中，从N端至C端，每条抗PD-1抗体或抗PD-L1抗体的重链具有一个可变区(VH)，也称作可变重链域或重链可变结构域，随后是三个恒定结构域(CH1、CH2和CH3)，也称作重链恒定区。类似地，从N端至C端，每条抗PD-1抗体或抗PD-L1抗体的轻链具有一个可变区(VL)，也称作可变轻链域或轻链可变结构域，随后一个恒定轻链(CL)结构域，也称作轻链恒定区。抗PD-1抗体或抗PD-L1抗体基本上由借助抗PD-1抗体或抗PD-L1抗体的铰链区连接的两个Fab分子和一个Fc结构域组成。
本发明融合蛋白中包含的抗PD-1抗体或抗PD-L1抗体能够以高的亲和力，例如以10-8M或更小、优选地以10-9M至10-12M的KD，分别与PD-1或PD-L1特异性结合，并由此阻断PD-1与配体PD-L1/PD-L2结合所介导的信号传导途径或阻断PD-L1与受体PD-1/CD80(B7-1)结合所介导的信号传导途径。
本文在下表1A中提供了本发明融合蛋白中包含的抗PD-1抗体的成对重链可变区(VH)和轻链可变区(VL)的例子。另外，本文在下表1B中提供了本发明融合蛋白中包含的抗PD-L1抗体的成对重链可变区(VH)和轻链可变区(VL)的例子。在一些实施方案中，本发明融合蛋白中的抗PD-1抗体或抗PD-L1抗体分别包含与表1A或表1B中所示的氨基酸序列基本上同一的序列，例如，与表1A或表1B所示的成对重链可变区序列/轻链可变区序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多序列同一性的序列。
表1A.融合蛋白中的抗PD-1抗体的重链可变区和轻链可变区序列的例子
Figure BDA0001423925870000141
Figure BDA0001423925870000151
Figure BDA0001423925870000161
表1B.融合蛋白中的抗PD-L1抗体的重链可变区和轻链可变区序列的例子
Figure BDA0001423925870000162
在一个实施方案中，本发明融合蛋白中的抗PD-1抗体包含选自SEQ ID NO：1/2、3/4、5/6、7/8、9/10、11/12、13/14、15/16、17/18、19/20、21/22、23/24、和120/121的成对重链可变区序列/轻链可变区序列中所含的全部重链CDR与轻链CDR。在一个实施方案中，本发明融合蛋白中的抗PD-L1抗体包含选自SEQ ID NO：25/26、27/28和29/30的成对重链可变区序列/轻链可变区序列中所含的全部重链CDR与轻链CDR。用于鉴定重链可变区与轻链可变区的氨基酸序列中的CDR的方法及技术为本领域中已知的，且可用于鉴定本文公开的特定重链可变区及/或轻链可变区的氨基酸序列中的CDR。可用于鉴定CDR边界的示例性公知技术包括例如Kabat界定法、Chothia界定法以及AbM界定法。参见，例如Kabat,Sequences ofProteins of Immunological Interest,National Institutes of Health,Bethesda,Md.(1991)；Al-Lazikani等人，Standard conformations for the canonical structures ofimmunoglobulins.，J.Mol.Biol.273:927-948(1997)；以及Martin AC等人，Modelingantibody hypervariable loops:a combined algorithm，Proc.Natl.Acad.Sci.USA 86:9268-9272(1989)。
本发明融合蛋白中的抗PD-1抗体或抗PD-L1抗体可以基于其轻链恒定区的氨基酸序列而划分为κ型或λ型，优选为κ型。
本文在下表2中提供了本发明融合蛋白中的抗PD-1抗体轻链恒定区的氨基酸序列的例子。
表2.融合蛋白中的抗PD1抗体轻链恒定区序列的例子
Figure BDA0001423925870000171
本发明融合蛋白中的抗PD-1抗体或抗PD-L1抗体基于其重链恒定区的氨基酸序列优选地是IgG类抗体，特别地是IgG1亚类、IgG2亚类、IgG4亚类抗体，更特别地是IgG4亚类抗体。优选地，所述IgG4亚类抗PD-1抗体或抗PD-L1抗体在Fc区中第S228位置处包含防止发生臂交换(arm-exchange)的氨基酸置换，特别地是氨基酸置换S228P。
本文在下表3中提供了本发明融合蛋白中的抗PD-1抗体重链恒定区的氨基酸序列的例子。
表3.融合蛋白中的抗PD1抗体重链恒定区序列的例子
Figure BDA0001423925870000172
Figure BDA0001423925870000181
－抑制VEGF家族的结构域(VID)
本发明融合蛋白中的“抑制VEGF家族的结构域(VID)”包含VEGFR的胞外结构域的一部分。VEGFR受体是位于细胞表面的一种酪氨酸激酶受体，其胞外区由7个免疫球蛋白(Ig)样结构域组成。例如，人VEGFR1包含编号为1、2、3、4、5、6和7的七个Ig样结构域，Ig样结构域1在胞外结构域的N端，Ig样结构域7在胞外结构域的C端。除非本文另外指出，否则Ig样结构域从VEGFR蛋白的N端至C端顺序编号。在一些实施方案中，VID包含选自VEGFR1、VEGFR2和VEGFR3的一种或多种VEGFR的至少一个Ig样结构域。在一些方面，VID包含VEGFR的至少1、2、3、4、5、6个但不超过7个Ig样结构域。在另一方面，VID包含VEGFR的1至7、1至6、1至5、1至4、1至3或1至2个Ig样结构域。
本文还考虑了包含两种或多种VEGFR的至少一个Ig样结构域的VID。在一些实施方案中，VID包含来自两种或多种选自VEGFR1、VEGFR2和VEGFR3的VEGFR的至少一个Ig样结构域。本文考虑了包含每种VEGFR的七个Ig样结构域的任意组合的VID。例如，VID可以包含VEGFR1(例如人VEGFR1)的Ig样结构域2和VEGFR2(例如人VEGFR2)的Ig样结构域3。在另一实施方案中，VID可以包含VEGFR1(例如人VEGFR1)的Ig样结构域1-3、VEGFR1(例如人VEGFR1)的Ig样结构域2-3、VEGFR2(例如人VEGFR2)的Ig样结构域1-3、VEGFR1(例如人VEGFR1)的Ig样结构域2和VEGFR2(例如人VEGFR2)的Ig样结构域3-4，或VEGFR1(例如人VEGFR1)的Ig样结构域2和VEGFR3(例如人VEGFR3)的Ig样结构域3。这些Ig样结构域和其他可以用作VID的部分的Ig样结构域的更详细描述见美国专利号7531173；Yu DC等，Soluble vascularendothelial growth factor decoy receptor FP3 exerts potent antiangiogeniceffects，Mol.Ther.，2012，20(3)：938-947和Holash，J.等，VEGF-Trap:a VEGF blockerwith potent antitumor effects，PNAS，2002，99(17)：11393-11398，全部文献在此以其整体引入作为参考。在一些实施方案中，VID具有任一选自表4中SEQ ID NO：63－65所示的氨基酸序列或与SEQ ID NO：63－65所示的氨基酸序列具有至少90％、91％、92％、93％、94％、95％、96％、97％、98％、99％或更多同一性的氨基酸序列。
表4融合蛋白中的VID氨基酸序列的例子
Figure BDA0001423925870000191
本发明融合蛋白中包含的VID能够以高的亲和力，例如以10-8M或更小、优选地以10-9M至10-12M的KD，与VEGF家族特异性结合，并由此抑制VEGF家族与细胞表面VEGFR的结合和随后的信号传导。
－肽接头
本发明融合蛋白中在抗PD-1抗体或抗PD-L1抗体的重链C端和VID N端任选包含的“肽接头”是一个或多个氨基酸、一般约2－20个氨基酸的肽。本领域已知或本文中描述了肽接头。
在一些实施方案中，所述肽接头包含至少5个氨基酸，优选地包含选自AKTTPKLEEGEFSEAR(SEQ ID NO：36)；AKTTPKLEEGEFSEARV(SEQ ID NO：37)；AKTTPKLGG(SEQID NO：38)；SAKTTPKLGG(SEQ ID NO：39)；SAKTTP(SEQ ID NO：40)；RADAAP(SEQ ID NO：41)；RADAAPTVS(SEQ ID NO：42)；RADAAAAGGPGS(SEQ ID NO：43)；RADAAAA(SEQ ID NO：44)；SAKTTPKLEEGEFSEARV(SEQ ID NO：45)；ADAAP(SEQ ID NO：46)；DAAPTVSIFPP(SEQ ID NO：47)；TVAAP(SEQ ID NO：48)；TVAAPSVFIFPP(SEQ ID NO：49)；QPKAAP(SEQ ID NO：50)；QPKAAPSVTLFPP(SEQ ID NO：51)；AKTTPP(SEQ ID NO：52)；AKTTPPSVTPLAP(SEQ ID NO：53)；AKTTAP(SEQ ID NO：54)；AKTTAPSVYPLAP(SEQ ID NO：55)；ASTKGP(SEQ ID NO：56)；ASTKGPSVFPLAP(SEQ ID NO：57)；GGGGSGGGGSGGGGS(SEQ ID NO：58)；GENKVEYAPALMALS(SEQID NO：59)；GPAKELTPLKEAKVS(SEQ ID NO：60)；GHEAAAVMQVQYPAS(SEQ ID NO：61)；和GGGGSGGGGSGGGGSA(SEQ ID NO：62)的肽接头。
III.本发明的双靶向融合蛋白的生产和纯化
本发明的双靶向融合蛋白可以例如通过固态肽合成(例如Merrifield固相合成)或重组生产获得。为了重组生产，将编码所述双靶向融合蛋白的抗体轻链亚基的多核苷酸和/或编码所述双靶向融合蛋白的抗体重链-VID融合亚基的多核苷酸分离并插入一个或多个载体中以便进一步在宿主细胞中克隆和/或表达。使用常规方法，可以轻易地分离所述多核苷酸并将其测序。在一个实施方案中，提供了包含本发明的一种或多种多核苷酸的载体，优选地表达载体。
可以使用本领域技术人员熟知的方法来构建表达载体。表达载体包括但不限于病毒、质粒、粘粒、λ噬菌体或酵母人工染色体(YAC)。在一个优选的实施方案中，使用了具有双表达盒的谷氨酰胺合成酶高效表达载体。
一旦已经制备了用于表达的包含本发明的一种或多种多核苷酸的表达载体，则可以将表达载体转染或引入适宜的宿主细胞中。多种技术可以用来实现这个目的，例如，原生质体融合、磷酸钙沉淀、电穿孔、逆转录病毒的转导、病毒转染、基因枪、基于脂质体的转染或其他常规技术。
在一个实施方案中，提供了包含一种或多种本发明多核苷酸的宿主细胞。在一些实施方案中，提供了包含本发明表达载体的宿主细胞。如本文所用，术语“宿主细胞”指可以工程化以产生本发明的双靶向融合蛋白的任何种类的细胞系统。适于复制和支持本发明的双靶向融合蛋白表达的宿主细胞是本领域熟知的。根据需要，这类细胞可以用特定表达载体转染或转导，并且可以培育大量含有载体的细胞用于接种大规模发酵器以获得足够量的本发明双靶向融合蛋白用于临床应用。合适的宿主细胞包括原核微生物，如大肠杆菌，真核微生物如丝状真菌或酵母，或各种真核细胞，如中国仓鼠卵巢细胞(CHO)、昆虫细胞等。可以使用适于悬浮培养的哺乳动物细胞系。有用的哺乳动物宿主细胞系的例子包括SV40转化的猴肾CV1系(COS-7)；人胚肾系(293或293F细胞)、幼仓鼠肾细胞(BHK)、猴肾细胞(CV1)、非洲绿猴肾细胞(VERO-76)、人宫颈癌细胞(HELA)、犬肾细胞(MDCK)、布法罗大鼠肝脏细胞(BRL3A)、人肺细胞(W138)、人肝脏细胞(Hep G2)、CHO细胞、骨髓瘤细胞系如YO、NS0、P3X63和Sp2/0等。适于产生蛋白质的哺乳动物宿主细胞系的综述参见例如Yazaki和Wu，Methods inMolecular Biology，第248卷(B.K.C.Lo编著，Humana Press，Totowa，NJ)，第255-268页(2003)。
本领域已知在这些宿主细胞系统中表达外源基因的标准技术。在一个实施方案中，提供了产生本发明的双靶向融合蛋白的方法，其中所述方法包括在适于表达所述双靶向融合蛋白的条件下培养如本文中提供的宿主细胞，所述宿主细胞包含编码所述双靶向融合蛋白的多核苷酸，并且从宿主细胞(或宿主细胞培养基)回收所述双靶向融合蛋白。
如本文所述制备的双靶向融合蛋白可以通过已知的现有技术如高效液相色谱、离子交换层析、凝胶电泳、亲和层析、大小排阻层析等纯化。用来纯化特定蛋白质的实际条件还取决于如净电荷、疏水性、亲水性等因素，并且这些对本领域技术人员是显而易见的。
可以通过多种熟知分析方法中的任一种方法确定本发明的双靶向融合蛋白的纯度，所述熟知分析方法包括凝胶电泳、高效液相色谱等。可以通过本领域已知的多种测定法，鉴定、筛选或表征本文提供的双靶向融合蛋白的物理/化学特性和/或生物学活性。
IV.药物组合物和试剂盒
在另一个方面，本发明提供了组合物，例如，药物组合物，所述组合物包含与可药用载体配制在一起的本文所述的双靶向融合蛋白。如本文所用，“可药用载体”包括生理上相容的任何和全部溶剂、分散介质、等渗剂和吸收延迟剂等。本发明的药物组合物适于静脉内、肌内、皮下、肠胃外、直肠、脊髓或表皮施用(例如，通过注射或输注)。
本发明的组合物可以处于多种形式。这些形式例如包括液体、半固体和固体剂型，如液态溶液剂(例如，可注射用溶液剂和可输注溶液剂)、分散体剂或混悬剂、脂质体剂和栓剂。优选的形式取决于预期的施用模式和治疗用途。常见的优选组合物处于可注射用溶液剂或可输注溶液剂形式。优选的施用模式是肠胃外(例如，静脉内、皮下、腹腔(i.p.)、肌内)注射。在一个优选实施方案中，通过静脉内输注或注射施用双靶向融合蛋白。在另一个优选实施方案中，通过肌内、腹腔或皮下注射施用双靶向融合蛋白。
如本文所用的短语“肠胃外施用“和“肠胃外方式施用”意指除了肠施用和局部施用之外的施用模式，通常通过注射施用，并且包括但不限于静脉内、肌内、动脉内、皮内、腹腔、经气管、皮下注射和输注。
治疗性组合物一般应当是无菌的并且在制造和储存条件下稳定。可以将组合物配制为溶液、微乳液、分散体、脂质体或冻干形式。可以通过将活性化合物(即双靶向融合蛋白)以要求的量加入适宜的溶剂中，随后过滤消毒，制备无菌可注射溶液剂。通常，通过将所述活性化合物并入无菌溶媒中来制备分散体，所述无菌溶媒含有基础分散介质和其他成分。可以使用包衣剂如卵磷脂等。在分散体的情况下，可以通过使用表面活性剂来维持溶液剂的适宜流动性。可以通过在组合物中包含延迟吸收的物质例如单硬脂酸盐和明胶而引起可注射组合物的延长吸收。
在某些实施方案中，可以口服施用本发明的双靶向融合蛋白，例如随惰性稀释剂或可食用载体一起经口施用。本发明的双靶向融合蛋白也可以封闭在硬壳或软壳明胶胶囊中、压缩成片剂或直接掺入受试者的膳食中。对于口服治疗施用，所述化合物可以随赋形剂一起掺入并且以可摄取的片剂、颊用片剂、锭剂(troche)、胶囊剂、酏剂、混悬剂、糖浆剂、糯米纸囊剂(wafer)等形式使用。为了通过非肠胃外施用方法施用本发明的双靶向融合蛋白，可能需要将所述双靶向融合蛋白与防止其失活的材料包衣或随这种材料共施用。还可以用本领域已知的医疗装置施用治疗组合物。
本发明的药物组合物可以包含“治疗有效量”或“预防有效量”的本发明所述双靶向融合蛋白。“治疗有效量”指以需要的剂量并持续需要的时间段，有效实现所需治疗结果的量。可以根据多种因素如疾病状态、个体的年龄、性别和重量等变动治疗有效量。治疗有效量是任何有毒或有害作用不及治疗有益作用的量。相对于未治疗的受试者，“治疗有效量”优选地抑制可度量参数(例如肿瘤生长率)至少约20％、更优选地至少约40％、甚至更优选地至少约60％和仍更优选地至少约80％。可以在预示人肿瘤中的功效的动物模型系统中评价本发明的双靶向融合蛋白抑制可度量参数(例如，肿瘤体积)的能力。
“预防有效量”指以需要的剂量并持续需要的时间段，有效实现所需预防结果的量。通常，由于预防性剂量在受试者中在疾病较早阶段之前或在疾病较早阶段使用，故预防有效量小于治疗有效量。
包含本文所述双靶向融合蛋白的试剂盒也处于本发明的范围内。试剂盒可以包含一个或多个其他要素，例如包括：使用说明书；其他试剂，例如标记物或用于偶联的试剂；可药用载体；和用于施用至受试者的装置或其他材料。
V.双靶向融合蛋白的用途
本文公开的双靶向融合蛋白具有体外和体内诊断用途以及治疗性和预防性用途。例如，可以将这些分子施用至体外或离体的培养细胞或施用至受试者，例如，人类受试者，以治疗、预防和/或诊断多种与PD-1活性、PD-L1活性和VEGF家族活性相关的疾病，例如癌症。
在一个方面，本发明提供了体外或体内检测生物样品，例如血清、精液或尿或组织活检样品(例如，来自过度增生性或癌性病灶)中存在PD-1或PD-L1和VEGF家族分子的诊断方法。该诊断方法包括：(i)在允许相互作用发生的条件下使样品(和任选地，对照样品)与如本文所述的双靶向融合蛋白接触或向受试者施用所述双靶向融合蛋白和(ii)检测所述双靶向融合蛋白和样品(和任选地，对照样品)之间复合物的形成。复合物的形成表示存在PD-1或PD-L1和VEGF家族分子，并且可以显示本文所述治疗和/或预防的适用性或需求。
在一些实施方案中，在治疗之前，例如，在起始治疗之前或在治疗间隔后的某次治疗之前检测PD-1或PD-L1和VEGF家族分子。可以使用的检测方法包括免疫组织化学、免疫细胞化学、FACS、ELISA测定、PCR-技术(例如，RT-PCR)或体内成像技术。一般地，体内和体外检测方法中所用的双靶向融合蛋白直接或间接地用可检测物质标记以促进检测结合的或未结合的结合物。合适的可检测物质包括多种生物学活性酶、辅基、荧光物质、发光物质、顺磁(例如，核磁共振活性)物质和放射性物质。
在一些实施方案中，体内确定PD-1或PD-L1和VEGF家族分子的水平和/或分布，例如，以非侵入方式确定(例如，通过使用合适的成像技术(例如，正电子发射断层摄影术(PET)扫描)检测可检测物标记的本发明双靶向融合蛋白。在一个实施方案中，例如，通过检测用PET试剂(例如，18F-氟脱氧葡萄糖(FDG))以可检测方式标记的本发明双靶向融合蛋白，体内测定PD-1或PD-L1和VEGF家族分子的水平和/或分布。
在一个实施方案中，本发明提供了包含本文所述双靶向融合蛋白和使用说明书的诊断试剂盒。
在另一个方面，本发明涉及使用双靶向融合蛋白体内用来治疗或预防需要在受试者中增强免疫应答并减少血管形成的疾病，从而抑制或减少相关疾病如癌性肿瘤的生长或出现、转移或复发。可以单独使用双靶向融合蛋白以抑制癌性肿瘤的生长或者预防其出现。备选地，双靶向融合蛋白可以与其他癌症治疗剂/预防剂组合施用。当本发明的双靶向融合蛋白与一种或多种其他药物组合施用时，这种组合可以按任何顺序施用或者同时施用。
因此，在一个实施方案中，本发明提供一种抑制受试者中肿瘤细胞生长的方法，所述方法包括向受试者施用治疗有效量的本文所述的双靶向融合蛋白。在另一个实施方案中，本发明提供一种防止受试者中肿瘤细胞出现或者转移或者复发的方法，所述方法包括向受试者施用预防有效量的本文所述的双靶向融合蛋白。
在一些实施方案中，用双靶向融合蛋白治疗和/或预防的癌包括但不限于实体瘤、血液学癌(例如，白血病、淋巴瘤、骨髓瘤，例如，多发性骨髓瘤)及转移性病灶。在一个实施方案中，癌是实体瘤。实体瘤的例子包括恶性肿瘤，例如，多个器官系统的肉瘤和癌，如侵袭肺、乳房、卵巢、淋巴样、胃肠道的(例如，结肠)、肛门、生殖器和生殖泌尿道(例如，肾、膀胱上皮、膀胱细胞、前列腺)、咽、CNS(例如，脑、神经的或神经胶质细胞)、头和颈、皮肤(例如，黑素瘤)、鼻咽(例如，分化或未分化的转移性或局部复发性鼻咽癌)和胰的那些癌、以及腺癌，包括恶性肿瘤，如结肠癌、直肠癌、肾细胞癌、肝癌、非小细胞肺癌、小肠癌和食道癌。癌症可以处于早期、中期或晚期或是转移性癌。
在一些实施方案中，癌选自黑素瘤、乳腺癌、结肠癌、食管癌、胃肠道间质肿瘤(GIST)、肾癌(例如，肾细胞癌)、肝癌、非小细胞肺癌(NSCLC)、卵巢癌、胰腺癌、前列腺癌、头颈部肿瘤、胃癌、血液学恶性病(例如，淋巴瘤)。
描述以下实施例以辅助对本发明的理解。不意在且不应当以任何方式将实施例解释成限制本发明的保护范围。
实施例
实施例1、包含目的基因的谷氨酰胺合成酶高效表达载体的构建
(1)作为对照的抗PD1抗体BY18.1的编码核苷酸的合成及表达载体的构建
根据International Nonproprietary Name(INN)数据库中编号为9623的纳武单抗的氨基酸序列数据，优化为适合在中国仓鼠卵巢癌细胞(CHO)中表达的下述核苷酸序列，并委托上海捷瑞生物工程有限公司合成该核苷酸序列。所述核苷酸序列表达后产生的抗PD1抗体在本文中表示为抗体BY18.1。
抗PD1抗体BY18.1的轻链(BY18.1L)核苷酸序列(SEQ ID NO:66)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGAGATTGTGCTGACACAGTCCCCCGCTACTCTGAGCCTGAGCCCTGGCGAGAGGGCTACACTGTCTTGCAGAGCTTCTCAGTCCGTGTCTTCTTACCTCGCTTGGTATCAGCAGAAGCCCGGCCAGGCTCCAAGACTGCTGATCTATGACGCTTCTAACCGCGCTACAGGCATTCCTGCTAGGTTCAGCGGCAGCGGCTCTGGCACCGACTTCACACTCACAATTAGCTCTCTTGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTCTAGCAACTGGCCTAGAACATTCGGCCAGGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
抗PD1抗体BY18.1的轻链(BY18.1L)氨基酸序列(SEQ ID NO:67)：
METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
抗PD1抗体BY18.1的重链(BY18.1H)核苷酸序列(SEQ ID NO:68)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTGCAGCTCGTGGAGTCCGGCGGCGGCGTGGTGCAGCCCGGCAGATCCCTCAGACTGGACTGCAAGGCATCCGGCATTACATTCTCTAACTCTGGAATGCACTGGGTGAGACAGGCTCCTGGCAAGGGCCTGGAATGGGTGGCCGTGATTTGGTACGACGGCTCTAAGAGATACTACGCTGACTCCGTGAAGGGCCGGTTCACAATTAGCAGAGACAACTCCAAGAACACTCTGTTCCTCCAGATGAACAGCCTGAGAGCCGAGGACACCGCTGTGTACTACTGCGCCACCAACGACGACTACTGGGGCCAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCAAGTAA GTCGAC
抗PD1抗体BY18.1的重链(BY18.1H)氨基酸序列(SEQ ID NO:69)：
METDTLLLWVLLLWVPGSTGQVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK
其中带下划线部分“METDTLLLWVLLLWVPGSTG”为信号肽序列。
上海捷瑞生物工程有限公司合成了上述BY18.1L编码核苷酸序列和BY18.1H编码核苷酸序列。分别将BY18.1L编码核苷酸用XhoI-EcoRI双酶切，将具有双表达盒的谷氨酰胺合成酶高效表达载体(专利授权号：CN104195173B，获自北京比洋生物技术有限公司)用XhoI-EcoRI双酶切，再通过连接酶将经XhoI-EcoRI双酶切的BY18.1L编码核苷酸连接入经XhoI-EcoRI双酶切的具有双表达盒的谷氨酰胺合成酶高效表达载体，获得已导入了BY18.1L编码核苷酸的具有双表达盒的谷氨酰胺合成酶高效表达载体；然后，分别将BY18.1H编码核苷酸用XbaI-SalI双酶切，将已导入了BY18.1L编码核苷酸的具有双表达盒的谷氨酰胺合成酶高效表达载体用XbaI-SalI双酶切，再通过连接酶将经XbaI-SalI双酶切的BY18.1H编码核苷酸连接入经XbaI-SalI双酶切的已导入了BY18.1L编码核苷酸的具有双表达盒的谷氨酰胺合成酶高效表达载体，由此获得了已导入BY18.1L编码核苷酸和BY18.1H编码核苷酸的具有双表达盒的谷氨酰胺合成酶高效表达载体，经测序验证正确后表达，获得抗PD1抗体BY18.1。
备选地，也可以将BY18.1L编码核苷酸连接入已导入了BY18.1H编码核苷酸的具有双表达盒的谷氨酰胺合成酶高效表达载体，表达并获得抗体BY18.1。
(2)作为对照的蛋白301-8编码核苷酸的合成及表达载体构建
根据International Nonproprietary Name(INN)数据库中编号为8739的阿柏西普(aflibercept)的氨基酸序列数据，优化为适合在中国仓鼠卵巢癌细胞(CHO)中表达的下述核苷酸序列，并委托上海捷瑞生物工程有限公司合成该核苷酸序列。所述核苷酸序列表达后产生的蛋白产物在本文中表示为蛋白301-8。
蛋白301-8的编码核苷酸序列(SEQ ID NO:70)：
AAGCTTGCCACCATGGAGACCGACACCCTGCTGCTCTGGGTGCTGCTGCTCTGGGTGCCCGGCTCCACCGGATCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGGATAAGACCCATACATGTCCCCCATGCCCCGCTCCAGAACTGCTGGGCGGACCTTCCGTGTTTCTGTTCCCACCCAAACCAAAGGACACACTGATGATCAGCAGAACCCCTGAGGTGACTTGCGTGGTCGTGGACGTGAGCCATGAGGACCCCGAGGTGAAGTTCAACTGGTATGTGGATGGCGTGGAAGTGCATAATGCCAAGACAAAACCTAGGGAAGAGCAGTACAACAGCACCTACAGGGTGGTGAGCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAATACAAGTGCAAGGTGTCCAATAAGGCTCTGCCTGCACCTATCGAGAAGACCATCAGCAAAGCCAAGGGCCAACCCAGAGAGCCTCAAGTCTACACCCTGCCCCCAAGCAGGGATGAGCTGACCAAAAATCAAGTGAGCCTGACATGCCTGGTCAAAGGCTTCTACCCTAGCGACATCGCCGTGGAGTGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACTCCCCCCGTCCTGGATAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACAGTCGATAAAAGCAGGTGGCAGCAAGGCAATGTCTTTAGCTGTAGCGTGATGCACGAGGCCCTGCATAACCACTACACTCAAAAGTCCCTGTCCCTGAGCCCCGGA
蛋白301-8的氨基酸序列(SEQ ID NO:71)：
METDTLLLWVLLLWVPGSTGSDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
分别将蛋白301-8的编码核苷酸序列用XhoI-EcoRI双酶切，将具有双表达盒的谷氨酰胺合成酶高效表达载体(专利授权号：CN104195173B，获自北京比洋生物技术有限公司)用XhoI-EcoRI双酶切，再通过连接酶将经XhoI-EcoRI双酶切的蛋白301-8编码核苷酸连接入经XhoI-EcoRI双酶切的具有双表达盒的谷氨酰胺合成酶高效表达载体，获得已导入了蛋白301-8的编码核苷酸序列的具有双表达盒的谷氨酰胺合成酶高效表达载体。经测序验证正确后用于表达，所表达的蛋白命名为蛋白301-8。蛋白301-8的氨基酸序列与现有技术中公开的阿柏西普的氨基酸序列相同。
(3)PD-1和VEGF家族的双靶向融合蛋白编码核苷酸的合成及表达载体的构建
根据表1A中抗PD-1抗体的重链可变区和轻链可变区序列、表2中抗体的轻链恒定区序列、表3中抗体的重链恒定区序列、表4中的VID序列、以及SEQ ID NO：36－62的肽接头序列，优化为适合在中国仓鼠卵巢癌细胞(CHO)中表达的核苷酸序列，并委托上海捷瑞生物工程有限公司合成如下SEQ ID NO:72－118中偶数编号所示的多核苷酸序列。
融合蛋白BY24.3(κ,IgG4)的轻链亚基(BY24.3L)核苷酸序列(SEQ ID NO:72)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGAGATTGTGCTGACACAGTCCCCCGCTACTCTGAGCCTGAGCCCTGGCGAGAGGGCTACACTGTCTTGCAGAGCTTCTCAGTCCGTGTCTTCTTACCTCGCTTGGTATCAGCAGAAGCCCGGCCAGGCTCCAAGACTGCTGATCTATGACGCTTCTAACCGCGCTACAGGCATTCCTGCTAGGTTCAGCGGCAGCGGCTCTGGCACCGACTTCACACTCACAATTAGCTCTCTTGAACCTGAGGACTTCGCCGTGTACTACTGCCAGCAGTCTAGCAACTGGCCTAGAACATTCGGCCAGGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.3(κ,IgG4)的轻链亚基(BY24.3L)氨基酸序列(SEQ ID NO:73)：METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.3(κ,IgG4)的重链-VID融合亚基(BY24.3H)核苷酸序列(SEQ IDNO:74)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTGCAGCTCGTGGAGTCCGGCGGCGGCGTGGTGCAGCCCGGCAGATCCCTCAGACTGGACTGCAAGGCATCCGGCATTACATTCTCTAACTCTGGAATGCACTGGGTGAGACAGGCTCCTGGCAAGGGCCTGGAATGGGTGGCCGTGATTTGGTACGACGGCTCTAAGAGATACTACGCTGACTCCGTGAAGGGCCGGTTCACAATTAGCAGAGACAACTCCAAGAACACTCTGTTCCTCCAGATGAACAGCCTGAGAGCCGAGGACACCGCTGTGTACTACTGCGCCACCAACGACGACTACTGGGGCCAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.3(κ,IgG4)的重链-VID融合亚基(BY24.3H)氨基酸序列(SEQ IDNO:75)：
METDTLLLWVLLLWVPGSTGQVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.7(κ,IgG2)的轻链亚基(BY24.7L)核苷酸序列(SEQ ID NO:76)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGAGATCAAGCGGACCGTGGCCGCCCCATCCGTGTTCATTTTCCCACCTTCCGAGATTGTGCTGACACAGTCCCCCGCTACTCTGAGCCTGAGCCCTGGCGAGAGGGCTACACTGTCTTGCAGAGCTTCCAAGGGCGTGAGCACATCCGGCTACTCCTACCTCCACTGGTATCAGCAGAAGCCAGGCCAGGCCCCAAGACTGCTGATATACCTCGCTTCTTACTTAGAGTCTGGCGTGCCCGCTCGGTTCAGCGGCTCCGGCTCTGGCACCGACTTCACCCTGACAATTTCTAGCCTGGAGCCCGAGGACTTCGCCGTGTACTACTGCCAGCACTCTAGGGACCTGCCTCTCACATTCGGCGGCGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.7(κ,IgG2)的轻链亚基(BY24.7L)氨基酸序列(SEQ ID NO:77)：
METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.7(κ,IgG2)的重链-VID融合亚基(BY24.7H)核苷酸序列(SEQ IDNO:78)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTGCAGCTCGTGCAGTCTGGCGTGGAGGTGAAGAAGCCTGGCGCCTCTGTGAAGGTGTCTTGCAAGGCTTCCGGCTACACTTTCACTAACTACTACATGTACTGGGTGAGACAGGCTCCCGGCCAGGGCCTAGAGTGGATGGGCGGCATTAACCCTAGCAACGGCGGCACAAACTTCAACGAGAAGTTCAAGAACCGCGTGACCCTGACCACAGACTCTAGCACAACAACTGCTTACATGGAGCTGAAGTCTCTCCAGTTCGACGACACCGCTGTGTACTACTGCGCTCGGAGGGACTACAGATTCGACATGGGCTTCGACTACTGGGGCCAGGGCACCACTGTGACAGTGTCTACAGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCAACTTCGGCACCCAGACATACACATGCAACGTGGACCACAAGCCTTCTAACACAAAGGTGGACAAGACCGTGGAGCGGAAGTGCTGCGTGGAGTGCCCACCTTGCCCCGCTCCTCCTGTGGCCGGCCCTTCTGTGTTCCTGTTCCCACCTAAGCCAAAGGACACACTCATGATCAGCAGAACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGAGCCACGAGGACCCCGAGGTGCAGTTCAACTGGTATGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAGCCTAGAGAAGAACAGTTCAACAGCACATTCAGAGTGGTGTCCGTGCTCACCGTGGTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCAGCCCCTATCGAAAAAACAATCAGCAAGACCAAGGGCCAGCCTAGAGAGCCTCAGGTGTACACACTGCCTCCATCTCGGGAAGAAATGACAAAGAACCAGGTGTCCCTCACATGCCTCGTGAAGGGCTTCTACCCATCCGACATCGCTGTGGAGTGGGAGTCTAACGGCCAGCCCGAGAACAACTACAAGACCACCCCTCCTATGCTCGACTCCGACGGCTCTTTCTTCCTGTACTCTAAGCTGACCGTGGACAAGTCCAGATGGCAGCAGGGCAACGTGTTCTCTTGCAGCGTGATGCACGAGGCTCTCCACAACCACTACACCCAGAAGTCCCTGAGCCTGTCTCCAGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCTAAGTCGAC
融合蛋白BY24.7(κ,IgG2)的重链-VID融合亚基(BY24.7H)氨基酸序列(SEQ IDNO:79)：
METDTLLLWVLLLWVPGSTGQVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSTASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNT
融合蛋白BY24.4(κ,IgG4)的轻链亚基(BY24.4L)核苷酸序列(SEQ ID NO:80)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGAGATCGTGCTGACACAGAGTCCTAGTTCCCTGAGCGCATCCGTCGGCGATAGGGTGACTATCACTTGTAGCGCACGCAGTAGCGTGTCTTACATGCACTGGTTTCAGCAGAAGCCCGGCAAGGCACCCAAGCTGTGGATCTACCGGACCAGTAACCTCGCCTCTGGAGTGCCATCCAGGTTTAGTGGCTCCGGAAGTGGAACTTCTTACTGCCTCACAATTAATAGTCTCCAGCCCGAGGATTTTGCAACATACTACTGTCAGCAGCGGTCTAGCTTTCCCCTGACATTCGGCGGAGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.4(κ,IgG4)的轻链亚基(BY24.4L)氨基酸序列(SEQ ID NO:81)：METDTLLLWVLLLWVPGSTGEIVLTQSPSSLSASVGDRVTITCSARSSVSYMHWFQQKPGKAPKLWIYRTSNLASGVPSRFSGSGSGTSYCLTINSLQPEDFATYYCQQRSSFPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.4(κ,IgG4)的重链-VID融合亚基(BY24.4H)核苷酸序列(SEQ IDNO:82)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTGCAGCTGGTCCAGAGCGGAAGCGAGCTTAAGAAGCCTGGAGCATCTGTCAAGATTAGTTGTAAGGCAAGTGGCTACACATTTACCAACTACGGAATGAATTGGGTGCGCCAGGCACCCGGACAGGGCCTCCAGTGGATGGGATGGATCAATACCGATAGCGGCGAGTCTACATACGCTGAGGAGTTTAAGGGCCGGTTCGTGTTCAGTCTCGATACAAGCGTGAACACAGCTTACCTCCAAATCACTTCTCTGACTGCTGAGGACACCGGCATGTACTTTTGCGTCCGCGTCGGCTACGACGCACTCGATTACTGGGGACAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.4(κ,IgG4)的重链-VID融合亚基(BY24.4H)氨基酸序列(SEQ IDNO:83)：
METDTLLLWVLLLWVPGSTGQVQLVQSGSELKKPGASVKISCKASGYTFTNYGMNWVRQAPGQGLQWMGWINTDSGESTYAEEFKGRFVFSLDTSVNTAYLQITSLTAEDTGMYFCVRVGYDALDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.5(κ,IgG4)的轻链亚基(BY24.5L)核苷酸序列(SEQ ID NO:84)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGACATTCAGATGACCCAGTCTCCAAGCTCTCTGAGCGCTTCCGTGGGCGACCGCGTGACAATTACATGCCTCGCATCTCAGACCATTGGCACCTGGCTGACATGGTATCAGCAGAAGCCTGGCAAGGCCCCTAAGCTGCTGATTTACACCGCTACCTCCCTCGCCGACGGCGTGCCATCTAGGTTCTCTGGCTCCGGCTCCGGCACAGACTTCACACTCACTATTTCTTCCCTCCAGCCCGAGGACTTCGCCACATACTACTGCCAGCAGGTGTACTCTATCCCTTGGACTTTCGGCGGCGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.5(κ,IgG4)的轻链亚基(BY24.5L)氨基酸序列(SEQ ID NO:85)：METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASVGDRVTITCLASQTIGTWLTWYQQKPGKAPKLLIYTATSLADGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVYSIPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.5(κ,IgG4)的重链-VID融合亚基(BY24.5H)核苷酸序列(SEQ IDNO:86)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCGAGGTCCAGCTCGTCGAGAGTGGAGGCGGCCTCGTGCAGCCCGGCGGAAGCCTCAGACTGTCTTGTGCTGCATCTGGCTTCACTTTCTCTAGTTACATGATGAGTTGGGTGAGACAGGCACCAGGAAAGGGATTGGAGTGGGTCGCAACAATCAGTGGCGGAGGAGCAAACACATACTACCCCGATAGCGTCAAGGGACGGTTCACCATTAGTCGCGATAACGCTAAGAACTCCCTGTACCTTCAGATGAATAGTCTCCGCGCTGAGGATACCGCCGTGTACTACTGCGCACGGCAGCTCTACTACTTCGATTACTGGGGCCAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.5(κ,IgG4)的重链-VID融合亚基(BY24.5H)氨基酸序列(SEQ IDNO:87)：
METDTLLLWVLLLWVPGSTGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYMMSWVRQAPGKGLEWVATISGGGANTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARQLYYFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.6(κ,IgG2)的轻链亚基(BY24.6L)核苷酸序列(SEQ ID NO:88)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCAATATTCAGATGACCCAGAGTCCTAGTAGCCTGAGCGCATCCGTCGGCGACCGCGTGACCATTACATGTAAGGCCGGACAGAACGTGAATAATTACCTCGCTTGGTATCAGCAGAAGCCAGGCAAGGCTCCAAAGGTGCTCATCTTCAATGCTAACAGTCTCCAGACTGGCGTCCCTTCCCGGTTTAGTGGAAGTGGATCTGGCACCGATTTCACACTCACTATCAGTTCTTTGCAACCCGAGGATTTTGCCACATACTACTGTCAGCAGTACAATAGCTGGACAACTTTCGGCGGAGGAACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.6(κ,IgG2)的轻链亚基(BY24.6L)氨基酸序列(SEQ ID NO:89)：METDTLLLWVLLLWVPGSTGNIQMTQSPSSLSASVGDRVTITCKAGQNVNNYLAWYQQKPGKAPKVLIFNANSLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSWTTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.6(κ,IgG2)的重链-VID融合亚基(BY24.6H)核苷酸序列(SEQ IDNO:90)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTCACACTCAAGGAGTCCGGCCCAGCTCTCGTGAAGCCTACACAGACCCTCACTCTCACCTGTACATTCAGCGGATTTAGCCTGAGCACTTCTGGAACATGCGTGTCTTGGATTCGGCAGCCACCCGGAAAGGCACTCGAATGGCTCGCAACCATCTGTTGGGAGGACAGTAAGGGCTACAATCCATCTCTGAAGTCTAGGCTGACAATTAGTAAGGACACCTCCAAGAATCAGGCCGTGCTGACTATGACTAATATGGACCCCGTCGATACCGCCACATACTACTGCGCTAGACGCGAGGATAGTGGCTACTTTTGGTTTCCTTACTGGGGCCAGGGAACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.6(κ,IgG2)的重链-VID融合亚基(BY24.6H)氨基酸序列(SEQ IDNO:91)：
METDTLLLWVLLLWVPGSTGQVTLKESGPALVKPTQTLTLTCTFSGFSLSTSGTCVSWIRQPPGKALEWLATICWEDSKGYNPSLKSRLTISKDTSKNQAVLTMTNMDPVDTATYYCARREDSGYFWFPYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.8(κ,IgG4)的轻链亚基(BY24.8L)核苷酸序列(SEQ ID NO:92)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGACATCGTGATGACCCAGAGTCCTGATAGTCTGGCCGTGTCCCTCGGCGAGCGCGCAACAATCAATTGCAAGGCATCTCAGTCCGTTTCCAACGATGTCGCATGGTATCAGCAGAAGCCTGGACAGCCACCTAAGCTGCTCATTAACTACGCCTTCCACAGATTCACTGGCGTGCCCGATCGGTTTTCCGGAAGTGGATACGGAACCGACTTTACACTGACTATTAGTTCTCTACAAGCTGAGGACGTCGCTGTGTACTACTGTCACCAGGCTTACTCTAGCCCATACACATTTGGAGGCGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.8(κ,IgG4)的轻链亚基(BY24.8L)氨基酸序列(SEQ ID NO:93)：
METDTLLLWVLLLWVPGSTGDIVMTQSPDSLAVSLGERATINCKASQSVSNDVAWYQQKPGQPPKLLINYAFHRFTGVPDRFSGSGYGTDFTLTISSLQAEDVAVYYCHQAYSSPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.8(κ,IgG4)的重链-VID融合亚基(BY24.8H)核苷酸序列(SEQ IDNO:94)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTCCAGCTCCAGGAGAGCGGACCTGGCCTCGTGAAGCCTAGCGAGACTCTGTCTCTGACATGTACAGTGTCCGGCTTTAGCCTCACTTCTTACGGCGTGCACTGGATTCGCCAGCCACCCGGAAAGGGATTGGAATGGCTCGGCGTCATTTGGGCCGGAGGCAGCACTAACTACAACCCATCTCTCAAGTCTAGGCTCACAATCAGCAAGGATAATAGTAAGAGTCAGGTGTCCCTGAAGATGAGTTCCGTCACCGCTGCCGATACCGCTGTGTACTACTGCGCACGCGCATACGGCAATTACTGGTACATCGACGTGTGGGGACAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.8(κ,IgG4)的重链-VID融合亚基(BY24.8H)氨基酸序列(SEQ IDNO:95)：
METDTLLLWVLLLWVPGSTGQVQLQESGPGLVKPSETLSLTCTVSGFSLTSYGVHWIRQPPGKGLEWLGVIWAGGSTNYNPSLKSRLTISKDNSKSQVSLKMSSVTAADTAVYYCARAYGNYWYIDVWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.9(κ,IgG4)的轻链亚基(BY24.9L)核苷酸序列(SEQ ID NO:96)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGATATTCAGATGACACAGTCCCCAAGTTCCCTGAGCGCCTCCGTCGGAGACCGGGTCACCATTACTTGTCGCGCTTCTCAGAGCGTGAGTAATTACCTCGATTGGTATCAGCAGAAGCCAGGAAAGGCTCCTAAGCTGCTCATCTACGACGCATCCACCCGCGCAACAGGCGTGCCTAGCCGGTTTAGCGGATCTGGAAGTGGCACTGATTTCACACTCACAATCTCTAGTCTGCAACCCGAGGACTTTGCTACATACTACTGTCAGCAGAACATGCAGCTGCCACTGACATTCGGCCAGGGAACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.9(κ,IgG4)的轻链亚基(BY24.9L)氨基酸序列(SEQ ID NO:97)：
METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASVGDRVTITCRASQSVSNYLDWYQQKPGKAPKLLIYDASTRATGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQNMQLPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.9(κ,IgG4)的重链-VID融合亚基(BY24.9H)核苷酸序列(SEQ IDNO:98)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCGTGCAGCTCGTCGAGTCCGGCGGAGGCGTGGTCCAGCCAGGACGCAGTCTGCGCCTGGATTGTAAGGCAAGCGGCATCACCTTTAGTAACTACGGTATGCACTGGGTGAGACAGGCTCCCGGAAAGGGCCTAGAATGGGTGGCCGTCATTTGGTACGACTCTTCTAGGAAGTACTACGCCGATAGTGTCAAGGGACGGTTCACAATCTCTCGCGATAATAGCAAGAATACACTGTTTTTGCAAATGAACTCCCTCAGAGCTGAGGATACCGCTGTGTACTACTGCGCAACCAACAATGATTACTGGGGACAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.9(κ,IgG4)的重链-VID融合亚基(BY24.9H)氨基酸序列(SEQ IDNO:99)：
METDTLLLWVLLLWVPGSTGVQLVESGGGVVQPGRSLRLDCKASGITFSNYGMHWVRQAPGKGLEWVAVIWYDSSRKYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNNDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.10(κ,IgG4)的轻链亚基(BY24.10L)核苷酸序列(SEQ ID NO:100)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCCTCAGTTACGTCCTCACACAGCCTCCATCCGTGTCTGTGAGTCCCGGACAGACCGCAAGAATCACATGTAGCGGCGACGCACTGCCTAAGCAGTACGCTTACTGGTATCAGCAGAAGCCAGGACAGGCACCTGTGCTGGTGATCTACAAGGATAGCGAGCGCCCAAGTGGCATTCCCGAGAGATTTAGTGGCTCTTCTAGTGGAACAACCGTCACCCTGACTATTTCCGGCGTGCAGGCCGAGGATGAGGCCGATTACTACTGTCAGTCTGCTGACTCTAGCGGAACATACGTCGTGTTTGGAGGCGGAACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.10(κ,IgG4)的轻链亚基(BY24.10L)氨基酸序列(SEQ ID NO:101)：METDTLLLWVLLLWVPGSTGLSYVLTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVLVIYKDSERPSGIPERFSGSSSGTTVTLTISGVQAEDEADYYCQSADSSGTYVVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.10(κ,IgG4)的重链-VID融合亚基(BY24.10H)核苷酸序列(SEQ IDNO:102)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCGACCTCGTGCAGTCCGGCGCCGAGGTGAAGAAGCCCGGCGCATCCGTCAAGGTGTCTTGCAAGGCAAGTGGCTACACTTTCACCAGTTACGGAATCAGTTGGGTCAGACAGGCACCTGGCCAGGGCCTGGAGTGGATGGGCTGGATTAGCGCTTACAACGGAAACACCAATTACGCTCAGAAGCTCCAGGGTCGGGTGACTATGACAACCGACACATCTACCAGCACCGCATACATGGAGCTGCGTAGTCTGAGATCCGACGATACCGCCGTGTACTACTGTGCTCGCGGCAGAGGATACTCCTACGGAATTGATGCATTCGATATTTGGGGACAGGGAACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.10(κ,IgG4)的重链-VID融合亚基(BY24.10H)氨基酸序列(SEQ IDNO:103)：
METDTLLLWVLLLWVPGSTGDLVQSGAEVKKPGASVKVSCKASGYTFTSYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARGRGYSYGIDAFDIWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.11(κ,IgG4)的轻链亚基(BY24.11L)核苷酸序列(SEQ ID NO:104)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGATGTCGTCATGACCCAGTCCCCTCTGTCTCTGCCCGTCACACTGGGACAGCCCGCATCCATTAGTTGTAGGTCTAGCCAGAGCATTGTGCACAGTAACGGCAATACATACCTGGAGTGGTATCTTCAAAAGCCTGGCCAGTCTCCTCAGCTGCTGATCTACAAGGTGAGTAATCGCTTTAGCGGCGTGCCTGATAGATTCAGCGGAAGTGGCTCCGGAACCGACTTCACACTCAAGATTTCTCGCGTGGAGGCCGAGGACGTCGGCGTGTACTACTGTTTTCAGGGGAGCCACGTGCCACTCACCTTTGGACAGGGCACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.11(κ,IgG4)的轻链亚基(BY24.11L)氨基酸序列(SEQ ID NO:105)：
METDTLLLWVLLLWVPGSTGDVVMTQSPLSLPVTLGQPASISCRSSQSIVHSNGNTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.11(κ,IgG4)的重链-VID融合亚基(BY24.11H)核苷酸序列(SEQ IDNO:106)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGGCCAGCTCGTGCAGAGTGGCGCAGAGGTGAAGAAGCCCGGAGCATCCGTCAAGGTCAGTTGCAAGGCCTCTGGATACACCTTTACCGATTACGAGATGCACTGGGTGCGGCAGGCTCCTGGACAGGGACTCGAATGGATGGGCGTCATTGAGTCCGAGACCGGCGGAACAGCTTACAATCAGAAGTTTCAGGGACGGGTGACACTCACTGCCGATAAGTCTTCTAGCACCGCCTACATGGAACTTTCCTCTCTGCGCTCAGAGGATACCGCTGTGTACTACTGTACACGCGAGGGAATCACAACTGTCGCAACCACATACTACTGGTACTTCGACGTGTGGGGCCAGGGAACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.11(κ,IgG4)的重链-VID融合亚基(BY24.11H)氨基酸序列(SEQIDNO:107)：
METDTLLLWVLLLWVPGSTGQGQLVQSGAEVKKPGASVKVSCKASGYTFTDYEMHWVRQAPGQGLEWMGVIESETGGTAYNQKFQGRVTLTADKSSSTAYMELSSLRSEDTAVYYCTREGITTVATTYYWYFDVWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.12(κ,IgG4)的轻链亚基(BY24.12L)核苷酸序列(SEQ ID NO:108)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGATATTGTGCTGACCCAGAGTCCCGCATCTCTCGCCGTCAGTCCTGGACAGCGCGCTACTATCACTTGCAGGGCTTCTGAGAGCGTCGATAATTACGGCATTTCCTTTATGAACTGGTATCAGCAGAAGCCTGGCCAGCCTCCAAAGCTGCTCATCTACACCTCTAGTAACAAGGATACAGGCGTGCCCGCAAGATTTAGCGGCTCCGGAAGTGGCACCGACTTCACACTCACAATCAACCCTATGGAGGCCGAGGATACCGCCGTGTACTACTGTCAGCAGTCTAAGGAGGTGCCTTGGACATTCGGCGGCGGAACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.12(κ,IgG4)的轻链亚基(BY24.12L)氨基酸序列(SEQ ID NO:109)：
METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSPGQRATITCRASESVDNYGISFMNWYQQKPGQPPKLLIYTSSNKDTGVPARFSGSGSGTDFTLTINPMEAEDTAVYYCQQSKEVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.12(κ,IgG4)的重链-VID融合亚基(BY24.12H)核苷酸序列(SEQ IDNO:110)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCGAGGTCCAGCTCGTCGAGTCCGGCGGAGGCCTCGTGCAGCCCGGCGGATCTCTGAGACTCAGTTGTGCCGCTAGTGGCTTTACATTTTCTTCCTACGGCATGTCTTGGGTGAGACAGGCTCCTGGAAAGGGATTAGAGTGGGTCGCAACTATTAGTGGCGGAGGAAGCGACACATACTACGCCGATTCCGTCAAGGGACGGTTCACCATCAGTCGCGATAACTCTAAGAACACACTGTACCTACAGATGAATAGCCTGAGAGCAGAGGATACCGCTGTGTACTACTGCGCACGCCAGCTCAATTACGCATGGTTTGCTTACTGGGGCCAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.12(κ,IgG4)的重链-VID融合亚基(BY24.12H)氨基酸序列(SEQ IDNO:111)：
METDTLLLWVLLLWVPGSTGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMSWVRQAPGKGLEWVATISGGGSDTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARQLNYAWFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.13(κ,IgG4)的轻链亚基(BY24.13L)核苷酸序列(SEQ ID NO:112)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCGATATTCAGATGACCCAGAGTCCATCTAGCGTGTCTGCTTCTGTGGGCGATCGGGTGACAATCACTTGTCGCGCAAGTCAGGGAATTAGTAGTTGGCTCGCATGGTATCAGCAGAAGCCTGGCAAGGCACCTAAGCTCCTCATTAGCGCCGCGTCATCCCTGCAATCCGGCGTGCCATCTAGGTTTAGTGGTTCCGGAAGCGGAACCGACTTTACACTCACTATCAGTTCTCTCCAGCCCGAGGATTTCGCAACATACTACTGTCAGCAGGCCAACCACCTGCCTTTCACATTTGGAGGCGGCACATTCGGCGGCGGAACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.13(κ,IgG4)的轻链亚基(BY24.13L)氨基酸序列(SEQ ID NO:113)：
METDTLLLWVLLLWVPGSTGDIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLISAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANHLPFTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.13(κ,IgG4)的重链-VID融合亚基(BY24.13H)核苷酸序列(SEQ IDNO:114)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGGTGCAGCTCGTCCAGTCTGGCGCTGAGGTCAAGAAGCCTGGATCTAGCGTGAAGGTGTCTTGTAAGGCAAGTGGCGGAACCTTTTCTAGTTACGCTATTAGTTGGGTGCGGCAGGCTCCCGGCCAGGGCCTGGAGTGGATGGGACTCATCATTCCTATGTTCGATACCGCCGGCTACGCCCAGAAGTTTCAGGGACGGGTCGCAATCACCGTAGATGAGAGTACCTCCACAGCATACATGGAGCTGAGTAGTCTCAGATCCGAGGACACTGCCGTGTACTACTGTGCTCGCGCAGAGCACTCTAGCACCGGAACATTTGATTACTGGGGACAGGGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.13(κ,IgG4)的重链-VID融合亚基(BY24.13H)氨基酸序列(SEQ IDNO:115)：
METDTLLLWVLLLWVPGSTGQVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGLIIPMFDTAGYAQKFQGRVAITVDESTSTAYMELSSLRSEDTAVYYCARAEHSSTGTFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
融合蛋白BY24.14(κ,IgG4)的轻链亚基(BY24.14L)核苷酸序列(SEQ ID NO:116)：
CTCGAGGCCACCATGGAGACCGACACACTCCTCCTGTGGGTGCTGCTGCTGTGGGTGCCTGGCTCCACTGGCCAGAGCGCTCTCACTCAGCCTGCTTCCGTGTCTGGAAGTCCCGGCCAGAGTATCACTATTTCTTGTACAGGAACTTCCTCCGACGTCGGATTTTACAATTACGTCAGTTGGTATCAGCAGCACCCCGGAAAGGCACCTGAACTAATGATCTACGATGTGTCTAACCGCCCAAGCGGCGTGAGCGATAGGTTCAGTGGCAGTAAGAGTGGCAACACCGCATCCCTGACCATTAGTGGATTACAGGCCGAGGACGAGGCTGATTACTACTGTTCTAGCTACACAAATATCTCCACATGGGTCTTCGGCGGAGGAACATTCGGCGGCGGAACTAAGGTGGAGATTAAGAGAACCGTGGCCGCCCCCAGCGTGTTCATCTTCCCTCCCAGCGACGAGCAGCTGAAGTCTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACTCCAAGGACAGCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGACTGTCTAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCTAAGAATTC
融合蛋白BY24.14(κ,IgG4)的轻链亚基(BY24.14L)氨基酸序列(SEQ ID NO:117)：
METDTLLLWVLLLWVPGSTGQSALTQPASVSGSPGQSITISCTGTSSDVGFYNYVSWYQQHPGKAPELMIYDVSNRPSGVSDRFSGSKSGNTASLTISGLQAEDEADYYCSSYTNISTWVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
融合蛋白BY24.14(κ,IgG4)的重链-VID融合亚基(BY24.14H)核苷酸序列(SEQ IDNO:118)：
TCTAGAGCCACCATGGAGACCGACACCCTGCTGCTGTGGGTGCTGCTCCTGTGGGTGCCTGGCTCCACAGGCCAGCTCCAGCTTCAGGAGAGCGGACCCGGCCTGGTCAAGCCATCCGAGACTCTCACTCTGACATGCACCGTGAGTGCTGATTCTATCAGTTCCACAACTTACTACTGGGTGTGGATTAGGCAGCCTCCCGGAAAGGGATTAGAATGGATCGGCAGCATTTCTTACAGTGGCTCCACATACTACAATCCTAGTCTGAAGTCTCGCGTGACCGTGTCCGTGGATACATCTAAGAACCAGTTTAGCCTCAAGCTGAATAGCGTCGCCGCAACAGATACCGCTCTGTACTACTGCGCACGCCACCTCGGCTACAATGGACGCTATCTGCCCTTCGATTACTGGGGCCAGGGAAGCACCCTCGTGACAGTGTCTTCCGCCTCCACCAAGGGCCCTTCCGTGTTCCCTCTGGCCCCTTGCTCCCGCTCCACCTCCGAGTCCACCGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCTGAGCCTGTGACCGTGTCCTGGAACTCCGGCGCCCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTGACCGTGCCTTCCTCCTCCCTGGGCACCAAGACCTACACCTGCAACGTGGACCACAAGCCTTCCAACACCAAGGTGGACAAGCGCGTGGAGTCCAAGTACGGCCCTCCTTGCCCTCCTTGCCCTGCCCCTGAGTTCCTGGGCGGCCCTTCCGTGTTCCTGTTCCCTCCTAAGCCTAAGGACACCCTGATGATCTCCCGCACCCCTGAGGTGACCTGCGTGGTGGTGGACGTGTCCCAGGAGGACCCTGAGGTGCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGCGAGGAGCAGTTCAACTCCACCTACCGCGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCTCCATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCTCCTTCCCAGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCTTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCCGCCTGACCGTGGACAAGTCCCGCTGGCAGGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCTGGGCGGCGGAGGATCTGGCGGCGGAGGCAGTGGAGGCGGCGGAAGCGCTTCCGACACCGGCCGCCCTTTCGTGGAGATGTACTCCGAGATCCCTGAGATCATCCACATGACCGAGGGCCGCGAGCTGGTGATCCCTTGCCGCGTGACCTCCCCTAACATCACCGTGACCCTGAAGAAGTTCCCTCTGGACACCCTGATCCCTGACGGCAAGCGCATCATCTGGGACTCCCGCAAGGGCTTCATCATCTCCAACGCCACCTACAAGGAGATCGGCCTGCTGACCTGCGAGGCCACCGTGAACGGCCACCTGTACAAGACCAACTACCTGACCCACCGCCAGACCAACACCATCATCGACGTGGTGCTGTCCCCTTCCCACGGCATCGAGCTGTCCGTGGGCGAGAAGCTGGTGCTGAACTGCACCGCCCGCACCGAGCTGAACGTGGGCATCGACTTCAACTGGGAGTACCCTTCCTCCAAGCACCAGCACAAGAAGCTGGTGAACCGCGACCTGAAGACCCAGTCCGGCTCCGAGATGAAGAAGTTCCTGTCCACCCTGACCATCGACGGCGTGACCCGCTCCGACCAGGGCCTGTACACCTGCGCCGCCTCCTCCGGCCTGATGACCAAGAAGAACTCCACCTTCGTGCGCGTGCACGAGAAGTAAGTCGAC
融合蛋白BY24.14(κ,IgG4)的重链-VID融合亚基(BY24.14H)氨基酸序列(SEQ IDNO:119)：
METDTLLLWVLLLWVPGSTGQLQLQESGPGLVKPSETLTLTCTVSADSISSTTYYWVWIRQPPGKGLEWIGSISYSGSTYYNPSLKSRVTVSVDTSKNQFSLKLNSVAATDTALYYCARHLGYNGRYLPFDYWGQGSTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGSGGGGSGGGGSASDTGRPFVEMYSEIPEIIHMTEGRELVIPCRVTSPNITVTLKKFPLDTLIPDGKRIIWDSRKGFIISNATYKEIGLLTCEATVNGHLYKTNYLTHRQTNTIIDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEK
其中氨基酸序列“METDTLLLWVLLLWVPGSTG”为信号肽。
使用上述实施例1(1)相同的方法，分别通过XhoI-EcoRI双酶切将BY24.3L、BY24.4L、LBY24.5L、BY24.6L、BY24.7L、BY24.8L、BY24.9L、BY24.10L、BY24.11L、BY24.12L、BY24.13L和BY24.14L编码核苷酸连接至具有双表达盒的谷氨酰胺合成酶高效表达载体(专利授权号：CN104195173B，获自北京比洋生物技术有限公司)；再通过XbaI-SalI双酶切将BY24.3H、BY24.4H、LBY24.5H、BY24.6H、BY24.7H、BY24.8H、BY24.9H、BY24.10H、BY24.11H、BY24.12H、BY24.13H和BY24.14H编码核苷酸分别克隆至已连接了相应的融合蛋白轻链亚基编码核苷酸的具有双表达盒的谷氨酰胺合成酶高效表达载体；或者反之亦然。将重组载体测序验证正确后用于表达。所表达的双靶向融合蛋白分别命名为融合蛋白BY24.3、BY24.4、BY24.5、BY24.6、BY24.7、BY24.8、BY24.9、BY24.10、BY24.11、BY24.12、BY24.13和BY24.14。
实施例2、融合蛋白的表达和纯化
(1)融合蛋白的瞬时表达
将293F(购自Invitrogen公司，目录号：11625-019)细胞悬浮培养于无血清CD 293培养液(购自Invitrogen公司，目录号：11913-019)中。转染前离心细胞培养物，获得细胞沉淀，用新鲜的无血清CD 293培养液悬浮细胞，将细胞浓度调整为1×106个细胞/ml。将细胞悬浮液置于摇瓶中。以100ml细胞悬浮液为例，分别将实施例1制备的重组表达载体质粒DNA250ug和聚乙烯亚胺(polyethylenimine(PEI))(Sigma，目录号：408727)500ug加入1ml无血清CD 293培养液中混匀，室温静置8分钟后，将PEI/DNA混悬液逐滴加入放置有100ml细胞悬浮液的摇瓶中。轻轻混匀，置于5％CO2、37℃摇床培养(120转/分钟)。5天后收集培养上清。
根据同样的方法，瞬时表达产生作为对照的抗体BY18.1和作为对照的蛋白301-8。
(2)表达蛋白的纯化
用pH 7.4 PBS溶液平衡的HiTrap MabSelect SuRe 1ml柱(GE Healthcare LifeSciences产品,目录号：11-0034-93)纯化上述实施例2(1)收集的培养上清中存在的融合蛋白。简而言之，用pH 7.4的PBS溶液以10个柱床体积平衡HiTrap MabSelect SuRe 1ml柱，流速为0.5ml/分钟；将上述实施例2(1)收集的培养上清用0.45μm滤膜过滤后，载样至用pH7.4 PBS溶液平衡的HiTrap MabSelect SuRe 1ml柱；装载上清液后，将该柱首先用pH 7.4的PBS溶液以流速0.5ml/分钟洗涤5-10个柱床体积，并随后用100mM柠檬酸缓冲液(pH 4.0)以流速0.5ml/分钟洗脱。收集洗脱峰，目的蛋白存在于洗脱峰中。
在还原剂(5mM 1,4-二硫苏糖醇)存在下通过SDS-PAGE并用考马斯蓝染色，分析融合蛋白的纯度和分子量。结果如图2所示。分子量理论预测值与实际测定值见表5。因真核表达系统中存在对蛋白质的糖基化作用，故分子量实际测定值略高于理论预测值。
表5经纯化的表达蛋白的分子量大小
Figure BDA0001423925870000461
实施例3、使用ELISA方法检测本发明的融合蛋白与人PD-1和重组人VEGF-A的结合
将抗原PD-1(北京义翘神州生物技术有限公司产品，目录号：10377-H08H)和抗原VEGF165(北京义翘神州生物技术有限公司产品，目录号：11066-HNAH)稀释至0.5μg/ml和0.02μg/ml并分别包被96孔ELISA板(购自Corning公司，货号：42592)。将上述实施例2(2)纯化的双靶向融合蛋白稀释至5μg/ml，然后进行3倍系列稀释，共稀释9个梯度，对每个浓度梯度进行复孔检测。将稀释样品50μl分别加入经抗原PD-1或抗原VEGF165包被的96孔板中，37℃孵育2小时。洗涤3次后，加入辣根过氧化物酶标记的山羊抗人二级抗体(北京中衫金桥公司产品，产品号：ZDR-5301)，37℃孵育1小时。洗涤3次后，加入3,3',5,5'-四甲基联苯胺(TMB)底物显色液(北京康为世纪生物科技有限公司，产品号：CW0050)50μl/孔。10分钟后，加入2N的H2SO4终止显色。使用ELISA读数仪在450nm处测定每孔的吸光度OD值。对作为对照的抗体BY18.1、作为对照的蛋白301-8实施同样的ELISA操作。
ELISA结果显示，与作为对照的抗体BY18.1同样地，本发明的融合蛋白BY24.3、BY24.4、BY24.5、BY24.6、BY24.7、BY24.8、BY24.9、BY24.10、BY24.11、BY24.12、BY24.13、BY24.14能特异性地结合PD-1；与作为对照的蛋白301-8同样地，本发明的融合蛋白BY24.3、BY24.4、LBY24.5、BY24.6、BY24.7、BY24.8、BY24.9、BY24.10、BY24.11、BY24.12、BY24.13、BY24.14也能特异性地结合VEGF-A。
应用GraphPadPrism5软件，将各融合蛋白的蛋白质浓度对吸光度OD值作图，并且拟合数据以产生融合蛋白介导的特异性结合作用的半数最大有效浓度EC50值。结果如下表6所示。
表6本发明的融合蛋白对PD-1、VEGF-A的结合作用
Figure BDA0001423925870000471
Figure BDA0001423925870000481
根据表6的结果可见，本发明所构建的新型融合蛋白均能够以高亲和力结合PD-1，其中融合蛋白BY24.4、BY24.5、BY24.7、BY24.8、BY24.10、BY24.11、BY24.12、BY24.14以比抗体BY18.1大的、甚至融合蛋白BY24.8以比抗体BY18.1大约10倍的亲和力结合PD-1，融合蛋白BY24.3与抗体BY18.1对PD-1的亲和力基本一致；本发明所构建的新型融合蛋白也均能够以高亲和力结合VEGF-A，其中融合蛋白BY24.3显示与作为对照的蛋白301-8相似的高亲和力结合VEGF-A。
实施例4、使用Biacore T100测定本发明的融合蛋白的亲和力
在
Figure BDA0001423925870000482
T100仪器(GE Healthcare Biosciences AB，瑞典)上于25℃进行表面等离子体共振测量。
首先，通过酰胺偶联将抗IgG抗体(GE Healthcare Life Sciences，目录号：BR-1008-39)共价固定在CM5芯片上。使用60μl N-乙基-N'-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDC)和60μl N-羟基琥珀酰亚胺(NHS)活化CM5芯片，然后将5μl抗IgG抗体加95μl稀释缓冲液HBST(0.1M HEPES,1.5M NaCl，pH7.4，加0.005％吐温20)经0.2um滤膜过滤后，通过酰胺偶联将抗IgG抗体共价固定在CM5芯片上，产生约9000-14000共振单位(RU)的捕获系统。使用120μl乙醇胺封闭CM5芯片。
然后，将实施例2制备的本发明的融合蛋白、抗体BY18.1和蛋白301-8分别稀释为5μg/ml，以流速10μL/分钟注射该稀释液2分钟，将1600RU的实施例2制备的本发明的融合蛋白、抗体BY18.1和蛋白301-8通过各自的Fc区非共价地捕获到CM5芯片表面上。通过与EDC/NHS交联来稳定所得的复合物，以避免在测量和再生期间的基线漂移。
分别将结合抗原PD-1(北京义翘神州生物技术有限公司产品，目录号：10377-H08H)、VEGF165(北京义翘神州生物技术有限公司产品，目录号：11066-HNAH)、VEGF-B(Biovision产品，目录号：4642-20)和PLGF-1(Biovision产品，目录号：4739-25)配制为如下浓度梯度：7nM、22nm、66nM、200nM、600nM。通过以流速30μl/分钟注射每个浓度180秒，解离时间600秒，测量结合。通过用3M MgCl2溶液以流速10μL/分钟洗涤30秒使表面再生。使用BIA评价软件(BIAevaluation 4.1software，来自GE Healthcare Biosciences AB，瑞典)进行数据分析，获得下表7所示的亲和力数据。
表7各蛋白质与VEGF家族分子、PD-1的结合
Figure BDA0001423925870000491
注：ND：未检测
根据表7所示的数据可见，融合蛋白BY24.3以高的亲和力与VEGF-A、VEGF-B和PLGF-1结合，亲和力分别达到9.59×10-12M、1.23×10-9M和1.82×10-10M，且对PD-1的亲和力(KD)与作为对照的抗体BY18.1基本一致，融合蛋白BY24.7以比抗体BY18.1大的亲和力(KD)结合PD-1；融合蛋白BY24.3显示与作为对照的蛋白301-8相似的高亲和力结合VEGF-A；融合蛋白BY24.3也以高亲和力结合VEGF-B和PLGF-1。
由此表明了本发明的双靶向融合蛋白以与抗PD-1抗体类似的、或者更好的亲和力与PD-1结合，并具有极好的与多种VEGF结合的能力。
表7的结果也表明了：实施例4的通过表面等离子体共振(SPR)技术测定的亲和力结果与实施例3的通过ELISA测定的亲和力结果呈现高度一致性。
实施例5、本发明的融合蛋白在人源化B-hPD-1小鼠模型中对肿瘤生长的抑制作用
将0.1mL DMEM培养基中的5×105个MC38鼠结肠癌细胞(获自ATCC，美国)接种于体重约18g，6周龄雌性B-hPD-1人源化小鼠(获自北京百奥赛图基因生物技术有限公司，产品编号：B-CM-001)右侧前胁肋部皮下。肿瘤在所述小鼠体内长大。待肿瘤体积达到约108mm3时将荷瘤小鼠随机分组，每组6只，共4组，分别为：PBS溶剂对照组、蛋白301-8组(3.3mg/kg)、融合蛋白BY24.3组(6.4mg/kg)和抗体BY18.1组(5mg/kg)，各给药组剂量以抗体BY18.1组的剂量为标准，对蛋白301-8组、融合蛋白BY24.3组和抗体BY18.1组而言所施用的剂量在摩尔量上是等同的。将第一次给药的时间设定为第0天。所有组给药途径均为腹腔(i.p.)注射，每三天给药1次，连续给药6次，末次给药3天后结束实验。每周测量肿瘤体积及小鼠体重2次，记录小鼠体重和肿瘤体积。实验结束时，将动物安乐死，剥取肿瘤称重、拍照，计算肿瘤生长抑制率(Tumor Growth Inhibition％)和肿瘤重量抑制率(Inhibition Rate ofTumor Weight％)。计算TGI％使用的公式是：[1－(给药组肿瘤体积变化的均值/PBS溶剂对照组肿瘤体积变化的均值)]x100％，计算IRTW％使用的公式是：[1－(给药组肿瘤重量/PBS溶剂对照组肿瘤重量)]x100％。该实验在北京百奥赛图基因生物技术有限公司实施。
结果见图3和图4。本实验观察了本发明的融合蛋白BY24.3以及作为药物对照的抗体BY18.1、蛋白301-8对MC38小鼠结肠癌皮下移植瘤生长的抑制作用。
整个实验过程中，所有动物精神状态良好，无动物死亡。实验结束时(首次给药后的第21天)，各组动物体重平均约为19g。将本发明的融合蛋白BY24.3组以及作为药物对照的抗体BY18.1组、蛋白301-8组的动物与PBS溶剂对照组动物进行体重比较，没有显著性差异(P>0.05)，表明动物对本发明的融合蛋白BY24.3耐受良好(图3)。
实验结束时，PBS溶剂对照组平均肿瘤体积±标准误为1386±170mm3,而融合蛋白BY24.3和蛋白301-8组平均肿瘤体积±标准误分别为452±69、1023±256，TGI％分别为73.3％、28.1％，IRTW％分别为74.6％、25.7％。融合蛋白BY24.3与PBS溶剂对照组的肿瘤体积相比有明显差异(P<0.05)，表明融合蛋白BY24.3有显著的肿瘤抑制作用。蛋白301-8有一定的肿瘤抑制作用，但是与PBS溶剂对照组相比，未见显著性差异(P>0.05)。作为对照的抗体BY18.1平均肿瘤体积±标准误为739±128，TGI％为50.6％，IRTW％为46.0％，与PBS溶剂对照组的肿瘤体积相比有显著差异(P<0.05)。融合蛋白BY24.3显示极佳的肿瘤抑制作用，与作为药物对照的抗体BY18.1组、蛋白301-8组的肿瘤抑制作用相比较，具有显著性差异。本实验的抗肿瘤药效结果在对小鼠肿瘤称重的比较上也得到进一步证实。
表8在人源化B-hPD-1小鼠模型中对肿瘤生长的抑制作用比较
Figure BDA0001423925870000511
尽管已经出于说明本发明的目的显示了某些代表性实施方案和细节，但是本领域技术人员显而易见的是可以对它们进行多种变化和修改而不脱离主题发明的范围。在这个方面，本发明范围仅由以下权利要求限定。
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<400> 21
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
20 25 30
Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45
Gly Leu Ile Ile Pro Met Phe Asp Thr Ala Gly Tyr Ala Gln Lys Phe
50 55 60
Gln Gly Arg Val Ala Ile Thr Val Asp Glu Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ala Glu His Ser Ser Thr Gly Thr Phe Asp Tyr Trp Gly Gln
100 105 110
Gly
<210> 22
<211> 101
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的轻链可变区
<400> 22
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Ser Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn His Leu Pro Phe
85 90 95
Thr Phe Gly Gly Gly
100
<210> 23
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的重链可变区
<400> 23
Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Thr Leu Thr Cys Thr Val Ser Ala Asp Ser Ile Ser Ser Thr
20 25 30
Thr Tyr Tyr Trp Val Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
35 40 45
Trp Ile Gly Ser Ile Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser
50 55 60
Leu Lys Ser Arg Val Thr Val Ser Val Asp Thr Ser Lys Asn Gln Phe
65 70 75 80
Ser Leu Lys Leu Asn Ser Val Ala Ala Thr Asp Thr Ala Leu Tyr Tyr
85 90 95
Cys Ala Arg His Leu Gly Tyr Asn Gly Arg Tyr Leu Pro Phe Asp Tyr
100 105 110
Trp Gly Gln Gly
115
<210> 24
<211> 104
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的轻链可变区
<400> 24
Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Phe Tyr
20 25 30
Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Glu Leu
35 40 45
Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asp Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Asn Ile
85 90 95
Ser Thr Trp Val Phe Gly Gly Gly
100
<210> 25
<211> 111
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-L1抗体的重链可变区
<400> 25
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser
20 25 30
Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly
100 105 110
<210> 26
<211> 101
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-L1抗体的轻链可变区
<400> 26
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45
Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Tyr His Pro Ala
85 90 95
Thr Phe Gly Gln Gly
100
<210> 27
<211> 114
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-L1抗体的重链可变区
<400> 27
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr
20 25 30
Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr Trp Gly
100 105 110
Gln Gly
<210> 28
<211> 102
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-L1抗体的轻链可变区
<400> 28
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Leu Pro
85 90 95
Trp Thr Phe Gly Gln Gly
100
<210> 29
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-L1抗体的重链可变区
<400> 29
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln
100 105 110
Gly
<210> 30
<211> 104
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-L1抗体的轻链可变区
<400> 30
Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln
1 5 10 15
Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr
20 25 30
Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu
35 40 45
Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe
50 55 60
Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu
65 70 75 80
Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser
85 90 95
Ser Thr Arg Val Phe Gly Thr Gly
100
<210> 31
<211> 113
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的轻链恒定区序列，κ型
<400> 31
Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile
1 5 10 15
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val
20 25 30
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys
35 40 45
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu
50 55 60
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu
65 70 75 80
Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr
85 90 95
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu
100 105 110
Cys
<210> 32
<211> 112
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的轻链恒定区序列，λ型
<400> 32
Thr Lys Leu Thr Val Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr
1 5 10 15
Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu
20 25 30
Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp
35 40 45
Lys Ala Asp Ser Ser Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro
50 55 60
Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu
65 70 75 80
Thr Pro Glu Gln Trp Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr
85 90 95
His Glu Gly Ser Thr Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser
100 105 110
<210> 33
<211> 337
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的重链恒定区序列，IgG1型
<400> 33
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
1 5 10 15
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
20 25 30
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
35 40 45
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
50 55 60
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
65 70 75 80
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
85 90 95
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
100 105 110
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
115 120 125
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
130 135 140
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
145 150 155 160
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
165 170 175
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
180 185 190
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
195 200 205
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
210 215 220
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
225 230 235 240
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
245 250 255
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
260 265 270
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
275 280 285
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
290 295 300
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
305 310 315 320
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
325 330 335
Lys
<210> 34
<211> 333
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的重链恒定区序列，IgG2型
<400> 34
Thr Thr Val Thr Val Ser Thr Ala Ser Thr Lys Gly Pro Ser Val Phe
1 5 10 15
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
20 25 30
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
35 40 45
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
50 55 60
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
65 70 75 80
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro
85 90 95
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu
100 105 110
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu
115 120 125
Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
130 135 140
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln
145 150 155 160
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
165 170 175
Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu
180 185 190
Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
195 200 205
Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
210 215 220
Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
225 230 235 240
Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
245 250 255
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
260 265 270
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly
275 280 285
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
290 295 300
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
305 310 315 320
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
325 330
<210> 35
<211> 334
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的重链恒定区序列，IgG4型
<400> 35
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
1 5 10 15
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
20 25 30
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
35 40 45
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
50 55 60
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
65 70 75 80
Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro
85 90 95
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro
100 105 110
Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe
115 120 125
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
130 135 140
Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val
145 150 155 160
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
165 170 175
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val
180 185 190
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
195 200 205
Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser
210 215 220
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
225 230 235 240
Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
245 250 255
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
260 265 270
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
275 280 285
Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp
290 295 300
Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
305 310 315 320
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
325 330
<210> 36
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 36
Ala Lys Thr Thr Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Arg
1 5 10 15
<210> 37
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 37
Ala Lys Thr Thr Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala Arg
1 5 10 15
Val
<210> 38
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 38
Ala Lys Thr Thr Pro Lys Leu Gly Gly
1 5
<210> 39
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 39
Ser Ala Lys Thr Thr Pro Lys Leu Gly Gly
1 5 10
<210> 40
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 40
Ser Ala Lys Thr Thr Pro
1 5
<210> 41
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 41
Arg Ala Asp Ala Ala Pro
1 5
<210> 42
<211> 9
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 42
Arg Ala Asp Ala Ala Pro Thr Val Ser
1 5
<210> 43
<211> 12
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 43
Arg Ala Asp Ala Ala Ala Ala Gly Gly Pro Gly Ser
1 5 10
<210> 44
<211> 7
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 44
Arg Ala Asp Ala Ala Ala Ala
1 5
<210> 45
<211> 18
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 45
Ser Ala Lys Thr Thr Pro Lys Leu Glu Glu Gly Glu Phe Ser Glu Ala
1 5 10 15
Arg Val
<210> 46
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 46
Ala Asp Ala Ala Pro
1 5
<210> 47
<211> 11
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 47
Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro
1 5 10
<210> 48
<211> 5
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 48
Thr Val Ala Ala Pro
1 5
<210> 49
<211> 12
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 49
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
1 5 10
<210> 50
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 50
Gln Pro Lys Ala Ala Pro
1 5
<210> 51
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 51
Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro
1 5 10
<210> 52
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 52
Ala Lys Thr Thr Pro Pro
1 5
<210> 53
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 53
Ala Lys Thr Thr Pro Pro Ser Val Thr Pro Leu Ala Pro
1 5 10
<210> 54
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 54
Ala Lys Thr Thr Ala Pro
1 5
<210> 55
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 55
Ala Lys Thr Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro
1 5 10
<210> 56
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 56
Ala Ser Thr Lys Gly Pro
1 5
<210> 57
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 57
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
1 5 10
<210> 58
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 58
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 59
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 59
Gly Glu Asn Lys Val Glu Tyr Ala Pro Ala Leu Met Ala Leu Ser
1 5 10 15
<210> 60
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 60
Gly Pro Ala Lys Glu Leu Thr Pro Leu Lys Glu Ala Lys Val Ser
1 5 10 15
<210> 61
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 61
Gly His Glu Ala Ala Ala Val Met Gln Val Gln Tyr Pro Ala Ser
1 5 10 15
<210> 62
<211> 16
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种肽接头
<400> 62
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala
1 5 10 15
<210> 63
<211> 205
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种VID的氨基酸序列
<400> 63
Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu
1 5 10 15
Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val
20 25 30
Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr
35 40 45
Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe
50 55 60
Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu
65 70 75 80
Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg
85 90 95
Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile
100 105 110
Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr
115 120 125
Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys
130 135 140
His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly
145 150 155 160
Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr
165 170 175
Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met
180 185 190
Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
195 200 205
<210> 64
<211> 303
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种VID的氨基酸序列
<400> 64
Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu
1 5 10 15
Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val
20 25 30
Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr
35 40 45
Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe
50 55 60
Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu
65 70 75 80
Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg
85 90 95
Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile
100 105 110
Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr
115 120 125
Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys
130 135 140
His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly
145 150 155 160
Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr
165 170 175
Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met
180 185 190
Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys Pro Phe Val
195 200 205
Ala Phe Gly Ser Gly Met Glu Ser Leu Val Glu Ala Thr Val Gly Glu
210 215 220
Arg Val Arg Ile Pro Ala Lys Tyr Leu Gly Tyr Pro Pro Pro Glu Ile
225 230 235 240
Lys Trp Tyr Lys Asn Gly Ile Pro Leu Glu Ser Asn His Thr Ile Lys
245 250 255
Ala Gly His Val Leu Thr Ile Met Glu Val Ser Glu Arg Asp Thr Gly
260 265 270
Asn Tyr Thr Val Ile Leu Thr Asn Pro Ile Ser Lys Glu Lys Gln Ser
275 280 285
His Val Val Ser Leu Val Val Tyr Val Pro Pro Gly Pro Gly Asp
290 295 300
<210> 65
<211> 100
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种VID的氨基酸序列
<400> 65
Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu
1 5 10 15
Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val
20 25 30
Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr
35 40 45
Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe
50 55 60
Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu
65 70 75 80
Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg
85 90 95
Gln Thr Asn Thr
100
<210> 66
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> 抗PD1抗体BY18.1的轻链（BY18.1L）核苷酸序列
<400> 66
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgagattgt gctgacacag tcccccgcta ctctgagcct gagccctggc 120
gagagggcta cactgtcttg cagagcttct cagtccgtgt cttcttacct cgcttggtat 180
cagcagaagc ccggccaggc tccaagactg ctgatctatg acgcttctaa ccgcgctaca 240
ggcattcctg ctaggttcag cggcagcggc tctggcaccg acttcacact cacaattagc 300
tctcttgaac ctgaggactt cgccgtgtac tactgccagc agtctagcaa ctggcctaga 360
acattcggcc agggcactaa ggtggagatt aagagaaccg tggccgcccc cagcgtgttc 420
atcttccctc ccagcgacga gcagctgaag tctggcaccg ccagcgtggt gtgcctgctg 480
aacaacttct acccccgcga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 540
ggcaacagcc aggagagcgt gaccgagcag gactccaagg acagcaccta cagcctgagc 600
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 660
acccaccagg gactgtctag ccccgtgacc aagagcttca accggggcga gtgctaagaa 720
ttc 723
<210> 67
<211> 234
<212> PRT
<213> 人工序列
<220>
<223> 抗PD1抗体BY18.1的轻链（BY18.1L）氨基酸序列
<400> 67
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
50 55 60
Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser
100 105 110
Asn Trp Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 68
<211> 1401
<212> DNA
<213> 人工序列
<220>
<223> 抗PD1抗体BY18.1的重链（BY18.1H）核苷酸序列
<400> 68
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtgca gctcgtggag tccggcggcg gcgtggtgca gcccggcaga 120
tccctcagac tggactgcaa ggcatccggc attacattct ctaactctgg aatgcactgg 180
gtgagacagg ctcctggcaa gggcctggaa tgggtggccg tgatttggta cgacggctct 240
aagagatact acgctgactc cgtgaagggc cggttcacaa ttagcagaga caactccaag 300
aacactctgt tcctccagat gaacagcctg agagccgagg acaccgctgt gtactactgc 360
gccaccaacg acgactactg gggccagggc accctcgtga cagtgtcttc cgcctccacc 420
aagggccctt ccgtgttccc tctggcccct tgctcccgct ccacctccga gtccaccgcc 480
gccctgggct gcctggtgaa ggactacttc cctgagcctg tgaccgtgtc ctggaactcc 540
ggcgccctga cctccggcgt gcacaccttc cctgccgtgc tgcagtcctc cggcctgtac 600
tccctgtcct ccgtggtgac cgtgccttcc tcctccctgg gcaccaagac ctacacctgc 660
aacgtggacc acaagccttc caacaccaag gtggacaagc gcgtggagtc caagtacggc 720
cctccttgcc ctccttgccc tgcccctgag ttcctgggcg gcccttccgt gttcctgttc 780
cctcctaagc ctaaggacac cctgatgatc tcccgcaccc ctgaggtgac ctgcgtggtg 840
gtggacgtgt cccaggagga ccctgaggtg cagttcaact ggtacgtgga cggcgtggag 900
gtgcacaacg ccaagaccaa gcctcgcgag gagcagttca actccaccta ccgcgtggtg 960
tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 1020
tccaacaagg gcctgccttc ctccatcgag aagaccatct ccaaggccaa gggccagcct 1080
cgcgagcctc aggtgtacac cctgcctcct tcccaggagg agatgaccaa gaaccaggtg 1140
tccctgacct gcctggtgaa gggcttctac ccttccgaca tcgccgtgga gtgggagtcc 1200
aacggccagc ctgagaacaa ctacaagacc acccctcctg tgctggactc cgacggctcc 1260
ttcttcctgt actcccgcct gaccgtggac aagtcccgct ggcaggaggg caacgtgttc 1320
tcctgctccg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgtccctg 1380
tccctgggca agtaagtcga c 1401
<210> 69
<211> 460
<212> PRT
<213> 人工序列
<220>
<223> 抗PD1抗体BY18.1的重链（BY18.1H）氨基酸序列
<400> 69
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
20 25 30
Gln Pro Gly Arg Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr
35 40 45
Phe Ser Asn Ser Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly
50 55 60
Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr
65 70 75 80
Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
85 90 95
Asn Thr Leu Phe Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu
115 120 125
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
130 135 140
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
145 150 155 160
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
165 170 175
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
180 185 190
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
195 200 205
Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn
210 215 220
Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro
225 230 235 240
Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe
245 250 255
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
260 265 270
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
275 280 285
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
290 295 300
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
305 310 315 320
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
325 330 335
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
340 345 350
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
355 360 365
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
370 375 380
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
385 390 395 400
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
405 410 415
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
420 425 430
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
435 440 445
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys
450 455 460
<210> 70
<211> 1365
<212> DNA
<213> 人工序列
<220>
<223> 蛋白301-8的编码核苷酸序列
<400> 70
aagcttgcca ccatggagac cgacaccctg ctgctctggg tgctgctgct ctgggtgccc 60
ggctccaccg gatccgacac cggccgccct ttcgtggaga tgtactccga gatccctgag 120
atcatccaca tgaccgaggg ccgcgagctg gtgatccctt gccgcgtgac ctcccctaac 180
atcaccgtga ccctgaagaa gttccctctg gacaccctga tccctgacgg caagcgcatc 240
atctgggact cccgcaaggg cttcatcatc tccaacgcca cctacaagga gatcggcctg 300
ctgacctgcg aggccaccgt gaacggccac ctgtacaaga ccaactacct gacccaccgc 360
cagaccaaca ccatcatcga cgtggtgctg tccccttccc acggcatcga gctgtccgtg 420
ggcgagaagc tggtgctgaa ctgcaccgcc cgcaccgagc tgaacgtggg catcgacttc 480
aactgggagt acccttcctc caagcaccag cacaagaagc tggtgaaccg cgacctgaag 540
acccagtccg gctccgagat gaagaagttc ctgtccaccc tgaccatcga cggcgtgacc 600
cgctccgacc agggcctgta cacctgcgcc gcctcctccg gcctgatgac caagaagaac 660
tccaccttcg tgcgcgtgca cgagaaggat aagacccata catgtccccc atgccccgct 720
ccagaactgc tgggcggacc ttccgtgttt ctgttcccac ccaaaccaaa ggacacactg 780
atgatcagca gaacccctga ggtgacttgc gtggtcgtgg acgtgagcca tgaggacccc 840
gaggtgaagt tcaactggta tgtggatggc gtggaagtgc ataatgccaa gacaaaacct 900
agggaagagc agtacaacag cacctacagg gtggtgagcg tgctgaccgt gctgcaccag 960
gattggctga acggcaagga atacaagtgc aaggtgtcca ataaggctct gcctgcacct 1020
atcgagaaga ccatcagcaa agccaagggc caacccagag agcctcaagt ctacaccctg 1080
cccccaagca gggatgagct gaccaaaaat caagtgagcc tgacatgcct ggtcaaaggc 1140
ttctacccta gcgacatcgc cgtggagtgg gagagcaatg gccagcctga gaacaactac 1200
aagaccactc cccccgtcct ggatagcgac ggcagcttct tcctgtactc caaactgaca 1260
gtcgataaaa gcaggtggca gcaaggcaat gtctttagct gtagcgtgat gcacgaggcc 1320
ctgcataacc actacactca aaagtccctg tccctgagcc ccgga 1365
<210> 71
<211> 451
<212> PRT
<213> 人工序列
<220>
<223> 蛋白301-8的氨基酸序列
<400> 71
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser
20 25 30
Glu Ile Pro Glu Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile
35 40 45
Pro Cys Arg Val Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe
50 55 60
Pro Leu Asp Thr Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser
65 70 75 80
Arg Lys Gly Phe Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu
85 90 95
Leu Thr Cys Glu Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr
100 105 110
Leu Thr His Arg Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro
115 120 125
Ser His Gly Ile Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys
130 135 140
Thr Ala Arg Thr Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr
145 150 155 160
Pro Ser Ser Lys His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys
165 170 175
Thr Gln Ser Gly Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile
180 185 190
Asp Gly Val Thr Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser
195 200 205
Ser Gly Leu Met Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu
210 215 220
Lys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
225 230 235 240
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
245 250 255
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
260 265 270
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
275 280 285
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
290 295 300
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
305 310 315 320
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
325 330 335
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
340 345 350
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
355 360 365
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
370 375 380
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
385 390 395 400
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr
405 410 415
Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
420 425 430
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
435 440 445
Ser Pro Gly
450
<210> 72
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.3（κ,IgG4）的轻链亚基（BY24.3L）核苷酸序列
<400> 72
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgagattgt gctgacacag tcccccgcta ctctgagcct gagccctggc 120
gagagggcta cactgtcttg cagagcttct cagtccgtgt cttcttacct cgcttggtat 180
cagcagaagc ccggccaggc tccaagactg ctgatctatg acgcttctaa ccgcgctaca 240
ggcattcctg ctaggttcag cggcagcggc tctggcaccg acttcacact cacaattagc 300
tctcttgaac ctgaggactt cgccgtgtac tactgccagc agtctagcaa ctggcctaga 360
acattcggcc agggcactaa ggtggagatt aagagaaccg tggccgcccc cagcgtgttc 420
atcttccctc ccagcgacga gcagctgaag tctggcaccg ccagcgtggt gtgcctgctg 480
aacaacttct acccccgcga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 540
ggcaacagcc aggagagcgt gaccgagcag gactccaagg acagcaccta cagcctgagc 600
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 660
acccaccagg gactgtctag ccccgtgacc aagagcttca accggggcga gtgctaagaa 720
ttc 723
<210> 73
<211> 234
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.3（κ,IgG4）的轻链亚基（BY24.3L）氨基酸序列
<400> 73
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
50 55 60
Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser
100 105 110
Asn Trp Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 74
<211> 2058
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.3（κ,IgG4）的重链-VID融合亚基（BY24.3H）核苷酸序列
<400> 74
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtgca gctcgtggag tccggcggcg gcgtggtgca gcccggcaga 120
tccctcagac tggactgcaa ggcatccggc attacattct ctaactctgg aatgcactgg 180
gtgagacagg ctcctggcaa gggcctggaa tgggtggccg tgatttggta cgacggctct 240
aagagatact acgctgactc cgtgaagggc cggttcacaa ttagcagaga caactccaag 300
aacactctgt tcctccagat gaacagcctg agagccgagg acaccgctgt gtactactgc 360
gccaccaacg acgactactg gggccagggc accctcgtga cagtgtcttc cgcctccacc 420
aagggccctt ccgtgttccc tctggcccct tgctcccgct ccacctccga gtccaccgcc 480
gccctgggct gcctggtgaa ggactacttc cctgagcctg tgaccgtgtc ctggaactcc 540
ggcgccctga cctccggcgt gcacaccttc cctgccgtgc tgcagtcctc cggcctgtac 600
tccctgtcct ccgtggtgac cgtgccttcc tcctccctgg gcaccaagac ctacacctgc 660
aacgtggacc acaagccttc caacaccaag gtggacaagc gcgtggagtc caagtacggc 720
cctccttgcc ctccttgccc tgcccctgag ttcctgggcg gcccttccgt gttcctgttc 780
cctcctaagc ctaaggacac cctgatgatc tcccgcaccc ctgaggtgac ctgcgtggtg 840
gtggacgtgt cccaggagga ccctgaggtg cagttcaact ggtacgtgga cggcgtggag 900
gtgcacaacg ccaagaccaa gcctcgcgag gagcagttca actccaccta ccgcgtggtg 960
tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 1020
tccaacaagg gcctgccttc ctccatcgag aagaccatct ccaaggccaa gggccagcct 1080
cgcgagcctc aggtgtacac cctgcctcct tcccaggagg agatgaccaa gaaccaggtg 1140
tccctgacct gcctggtgaa gggcttctac ccttccgaca tcgccgtgga gtgggagtcc 1200
aacggccagc ctgagaacaa ctacaagacc acccctcctg tgctggactc cgacggctcc 1260
ttcttcctgt actcccgcct gaccgtggac aagtcccgct ggcaggaggg caacgtgttc 1320
tcctgctccg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgtccctg 1380
tccctgggcg gcggaggatc tggcggcgga ggcagtggag gcggcggaag cgcttccgac 1440
accggccgcc ctttcgtgga gatgtactcc gagatccctg agatcatcca catgaccgag 1500
ggccgcgagc tggtgatccc ttgccgcgtg acctccccta acatcaccgt gaccctgaag 1560
aagttccctc tggacaccct gatccctgac ggcaagcgca tcatctggga ctcccgcaag 1620
ggcttcatca tctccaacgc cacctacaag gagatcggcc tgctgacctg cgaggccacc 1680
gtgaacggcc acctgtacaa gaccaactac ctgacccacc gccagaccaa caccatcatc 1740
gacgtggtgc tgtccccttc ccacggcatc gagctgtccg tgggcgagaa gctggtgctg 1800
aactgcaccg cccgcaccga gctgaacgtg ggcatcgact tcaactggga gtacccttcc 1860
tccaagcacc agcacaagaa gctggtgaac cgcgacctga agacccagtc cggctccgag 1920
atgaagaagt tcctgtccac cctgaccatc gacggcgtga cccgctccga ccagggcctg 1980
tacacctgcg ccgcctcctc cggcctgatg accaagaaga actccacctt cgtgcgcgtg 2040
cacgagaagt aagtcgac 2058
<210> 75
<211> 679
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.3（κ,IgG4）的重链-VID融合亚基（BY24.3H）氨基酸序列
<400> 75
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
20 25 30
Gln Pro Gly Arg Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr
35 40 45
Phe Ser Asn Ser Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly
50 55 60
Leu Glu Trp Val Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr
65 70 75 80
Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
85 90 95
Asn Thr Leu Phe Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu
115 120 125
Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
130 135 140
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
145 150 155 160
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser
165 170 175
Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
180 185 190
Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
195 200 205
Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn
210 215 220
Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro
225 230 235 240
Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe
245 250 255
Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
260 265 270
Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe
275 280 285
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
290 295 300
Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
305 310 315 320
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
325 330 335
Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala
340 345 350
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln
355 360 365
Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
370 375 380
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
385 390 395 400
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
405 410 415
Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu
420 425 430
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
435 440 445
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Gly Gly Gly Ser Gly
450 455 460
Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser Asp Thr Gly Arg Pro
465 470 475 480
Phe Val Glu Met Tyr Ser Glu Ile Pro Glu Ile Ile His Met Thr Glu
485 490 495
Gly Arg Glu Leu Val Ile Pro Cys Arg Val Thr Ser Pro Asn Ile Thr
500 505 510
Val Thr Leu Lys Lys Phe Pro Leu Asp Thr Leu Ile Pro Asp Gly Lys
515 520 525
Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe Ile Ile Ser Asn Ala Thr
530 535 540
Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu Ala Thr Val Asn Gly His
545 550 555 560
Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg Gln Thr Asn Thr Ile Ile
565 570 575
Asp Val Val Leu Ser Pro Ser His Gly Ile Glu Leu Ser Val Gly Glu
580 585 590
Lys Leu Val Leu Asn Cys Thr Ala Arg Thr Glu Leu Asn Val Gly Ile
595 600 605
Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys His Gln His Lys Lys Leu
610 615 620
Val Asn Arg Asp Leu Lys Thr Gln Ser Gly Ser Glu Met Lys Lys Phe
625 630 635 640
Leu Ser Thr Leu Thr Ile Asp Gly Val Thr Arg Ser Asp Gln Gly Leu
645 650 655
Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met Thr Lys Lys Asn Ser Thr
660 665 670
Phe Val Arg Val His Glu Lys
675
<210> 76
<211> 786
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.7（κ,IgG2）的轻链亚基（BY24.7L）核苷酸序列
<400> 76
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgagatcaa gcggaccgtg gccgccccat ccgtgttcat tttcccacct 120
tccgagattg tgctgacaca gtcccccgct actctgagcc tgagccctgg cgagagggct 180
acactgtctt gcagagcttc caagggcgtg agcacatccg gctactccta cctccactgg 240
tatcagcaga agccaggcca ggccccaaga ctgctgatat acctcgcttc ttacttagag 300
tctggcgtgc ccgctcggtt cagcggctcc ggctctggca ccgacttcac cctgacaatt 360
tctagcctgg agcccgagga cttcgccgtg tactactgcc agcactctag ggacctgcct 420
ctcacattcg gcggcggcac taaggtggag attaagagaa ccgtggccgc ccccagcgtg 480
ttcatcttcc ctcccagcga cgagcagctg aagtctggca ccgccagcgt ggtgtgcctg 540
ctgaacaact tctacccccg cgaggccaag gtgcagtgga aggtggacaa cgccctgcag 600
agcggcaaca gccaggagag cgtgaccgag caggactcca aggacagcac ctacagcctg 660
agcagcaccc tgaccctgag caaggccgac tacgagaagc acaaggtgta cgcctgcgag 720
gtgacccacc agggactgtc tagccccgtg accaagagct tcaaccgggg cgagtgctaa 780
gaattc 786
<210> 77
<211> 238
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.7（κ,IgG2）的轻链亚基（BY24.7L）氨基酸序列
<400> 77
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser
20 25 30
Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly
35 40 45
Val Ser Thr Ser Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro
50 55 60
Gly Gln Ala Pro Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser
65 70 75 80
Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
100 105 110
Gln His Ser Arg Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val
115 120 125
Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
130 135 140
Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu
145 150 155 160
Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175
Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser
180 185 190
Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala
195 200 205
Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
210 215 220
Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 78
<211> 1761
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.7（κ,IgG2）的重链-VID融合亚基（BY24.7H）核苷酸序列
<400> 78
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtgca gctcgtgcag tctggcgtgg aggtgaagaa gcctggcgcc 120
tctgtgaagg tgtcttgcaa ggcttccggc tacactttca ctaactacta catgtactgg 180
gtgagacagg ctcccggcca gggcctagag tggatgggcg gcattaaccc tagcaacggc 240
ggcacaaact tcaacgagaa gttcaagaac cgcgtgaccc tgaccacaga ctctagcaca 300
acaactgctt acatggagct gaagtctctc cagttcgacg acaccgctgt gtactactgc 360
gctcggaggg actacagatt cgacatgggc ttcgactact ggggccaggg caccactgtg 420
acagtgtcta cagcctccac caagggccct tccgtgttcc ctctggcccc ttgctcccgc 480
tccacctccg agtccaccgc cgccctgggc tgcctggtga aggactactt ccctgagcct 540
gtgaccgtgt cctggaactc cggcgccctg acctccggcg tgcacacctt ccctgccgtg 600
ctgcagtcct ccggcctgta ctccctgtcc tccgtggtga ccgtgccttc ctccaacttc 660
ggcacccaga catacacatg caacgtggac cacaagcctt ctaacacaaa ggtggacaag 720
accgtggagc ggaagtgctg cgtggagtgc ccaccttgcc ccgctcctcc tgtggccggc 780
ccttctgtgt tcctgttccc acctaagcca aaggacacac tcatgatcag cagaacccct 840
gaggtgacct gcgtggtggt ggacgtgagc cacgaggacc ccgaggtgca gttcaactgg 900
tatgtggacg gcgtggaggt gcacaacgct aagaccaagc ctagagaaga acagttcaac 960
agcacattca gagtggtgtc cgtgctcacc gtggtgcacc aggactggct gaacggcaaa 1020
gagtacaagt gcaaggtgtc caacaagggc ctgccagccc ctatcgaaaa aacaatcagc 1080
aagaccaagg gccagcctag agagcctcag gtgtacacac tgcctccatc tcgggaagaa 1140
atgacaaaga accaggtgtc cctcacatgc ctcgtgaagg gcttctaccc atccgacatc 1200
gctgtggagt gggagtctaa cggccagccc gagaacaact acaagaccac ccctcctatg 1260
ctcgactccg acggctcttt cttcctgtac tctaagctga ccgtggacaa gtccagatgg 1320
cagcagggca acgtgttctc ttgcagcgtg atgcacgagg ctctccacaa ccactacacc 1380
cagaagtccc tgagcctgtc tccaggcggc ggaggatctg gcggcggagg cagtggaggc 1440
ggcggaagcg cttccgacac cggccgccct ttcgtggaga tgtactccga gatccctgag 1500
atcatccaca tgaccgaggg ccgcgagctg gtgatccctt gccgcgtgac ctcccctaac 1560
atcaccgtga ccctgaagaa gttccctctg gacaccctga tccctgacgg caagcgcatc 1620
atctgggact cccgcaaggg cttcatcatc tccaacgcca cctacaagga gatcggcctg 1680
ctgacctgcg aggccaccgt gaacggccac ctgtacaaga ccaactacct gacccaccgc 1740
cagaccaaca cctaagtcga c 1761
<210> 79
<211> 580
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.7（κ,IgG2）的重链-VID融合亚基（BY24.7H）氨基酸序列
<400> 79
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys
20 25 30
Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
35 40 45
Phe Thr Asn Tyr Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly
50 55 60
Leu Glu Trp Met Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe
65 70 75 80
Asn Glu Lys Phe Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr
85 90 95
Thr Thr Ala Tyr Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp
115 120 125
Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Thr Ala Ser Thr Lys
130 135 140
Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
145 150 155 160
Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
165 170 175
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
180 185 190
Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
195 200 205
Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn
210 215 220
Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg
225 230 235 240
Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly
245 250 255
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
260 265 270
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
275 280 285
Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
290 295 300
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg
305 310 315 320
Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys
325 330 335
Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu
340 345 350
Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
355 360 365
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
370 375 380
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
385 390 395 400
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met
405 410 415
Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
420 425 430
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
435 440 445
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
450 455 460
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala
465 470 475 480
Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu
485 490 495
Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val
500 505 510
Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr
515 520 525
Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe
530 535 540
Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu
545 550 555 560
Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg
565 570 575
Gln Thr Asn Thr
580
<210> 80
<211> 720
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.4（κ,IgG4）的轻链亚基（BY24.4L）核苷酸序列
<400> 80
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgagatcgt gctgacacag agtcctagtt ccctgagcgc atccgtcggc 120
gatagggtga ctatcacttg tagcgcacgc agtagcgtgt cttacatgca ctggtttcag 180
cagaagcccg gcaaggcacc caagctgtgg atctaccgga ccagtaacct cgcctctgga 240
gtgccatcca ggtttagtgg ctccggaagt ggaacttctt actgcctcac aattaatagt 300
ctccagcccg aggattttgc aacatactac tgtcagcagc ggtctagctt tcccctgaca 360
ttcggcggag gcactaaggt ggagattaag agaaccgtgg ccgcccccag cgtgttcatc 420
ttccctccca gcgacgagca gctgaagtct ggcaccgcca gcgtggtgtg cctgctgaac 480
aacttctacc cccgcgaggc caaggtgcag tggaaggtgg acaacgccct gcagagcggc 540
aacagccagg agagcgtgac cgagcaggac tccaaggaca gcacctacag cctgagcagc 600
accctgaccc tgagcaaggc cgactacgag aagcacaagg tgtacgcctg cgaggtgacc 660
caccagggac tgtctagccc cgtgaccaag agcttcaacc ggggcgagtg ctaagaattc 720
<210> 81
<211> 233
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.4（κ,IgG4）的轻链亚基（BY24.4L）氨基酸序列
<400> 81
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Glu Ile Val Leu Thr Gln Ser Pro Ser Ser Leu Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Ser Ala Arg Ser Ser
35 40 45
Val Ser Tyr Met His Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys
50 55 60
Leu Trp Ile Tyr Arg Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg
65 70 75 80
Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Cys Leu Thr Ile Asn Ser
85 90 95
Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Ser
100 105 110
Phe Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr
115 120 125
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
130 135 140
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
145 150 155 160
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
165 170 175
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
180 185 190
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
195 200 205
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
210 215 220
Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 82
<211> 2070
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.4（κ,IgG4）的重链-VID融合亚基（BY24.4H）核苷酸序列
<400> 82
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtgca gctggtccag agcggaagcg agcttaagaa gcctggagca 120
tctgtcaaga ttagttgtaa ggcaagtggc tacacattta ccaactacgg aatgaattgg 180
gtgcgccagg cacccggaca gggcctccag tggatgggat ggatcaatac cgatagcggc 240
gagtctacat acgctgagga gtttaagggc cggttcgtgt tcagtctcga tacaagcgtg 300
aacacagctt acctccaaat cacttctctg actgctgagg acaccggcat gtacttttgc 360
gtccgcgtcg gctacgacgc actcgattac tggggacagg gcaccctcgt gacagtgtct 420
tccgcctcca ccaagggccc ttccgtgttc cctctggccc cttgctcccg ctccacctcc 480
gagtccaccg ccgccctggg ctgcctggtg aaggactact tccctgagcc tgtgaccgtg 540
tcctggaact ccggcgccct gacctccggc gtgcacacct tccctgccgt gctgcagtcc 600
tccggcctgt actccctgtc ctccgtggtg accgtgcctt cctcctccct gggcaccaag 660
acctacacct gcaacgtgga ccacaagcct tccaacacca aggtggacaa gcgcgtggag 720
tccaagtacg gccctccttg ccctccttgc cctgcccctg agttcctggg cggcccttcc 780
gtgttcctgt tccctcctaa gcctaaggac accctgatga tctcccgcac ccctgaggtg 840
acctgcgtgg tggtggacgt gtcccaggag gaccctgagg tgcagttcaa ctggtacgtg 900
gacggcgtgg aggtgcacaa cgccaagacc aagcctcgcg aggagcagtt caactccacc 960
taccgcgtgg tgtccgtgct gaccgtgctg caccaggact ggctgaacgg caaggagtac 1020
aagtgcaagg tgtccaacaa gggcctgcct tcctccatcg agaagaccat ctccaaggcc 1080
aagggccagc ctcgcgagcc tcaggtgtac accctgcctc cttcccagga ggagatgacc 1140
aagaaccagg tgtccctgac ctgcctggtg aagggcttct acccttccga catcgccgtg 1200
gagtgggagt ccaacggcca gcctgagaac aactacaaga ccacccctcc tgtgctggac 1260
tccgacggct ccttcttcct gtactcccgc ctgaccgtgg acaagtcccg ctggcaggag 1320
ggcaacgtgt tctcctgctc cgtgatgcac gaggccctgc acaaccacta cacccagaag 1380
tccctgtccc tgtccctggg cggcggagga tctggcggcg gaggcagtgg aggcggcgga 1440
agcgcttccg acaccggccg ccctttcgtg gagatgtact ccgagatccc tgagatcatc 1500
cacatgaccg agggccgcga gctggtgatc ccttgccgcg tgacctcccc taacatcacc 1560
gtgaccctga agaagttccc tctggacacc ctgatccctg acggcaagcg catcatctgg 1620
gactcccgca agggcttcat catctccaac gccacctaca aggagatcgg cctgctgacc 1680
tgcgaggcca ccgtgaacgg ccacctgtac aagaccaact acctgaccca ccgccagacc 1740
aacaccatca tcgacgtggt gctgtcccct tcccacggca tcgagctgtc cgtgggcgag 1800
aagctggtgc tgaactgcac cgcccgcacc gagctgaacg tgggcatcga cttcaactgg 1860
gagtaccctt cctccaagca ccagcacaag aagctggtga accgcgacct gaagacccag 1920
tccggctccg agatgaagaa gttcctgtcc accctgacca tcgacggcgt gacccgctcc 1980
gaccagggcc tgtacacctg cgccgcctcc tccggcctga tgaccaagaa gaactccacc 2040
ttcgtgcgcg tgcacgagaa gtaagtcgac 2070
<210> 83
<211> 683
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.4（κ,IgG4）的重链-VID融合亚基（BY24.4H）氨基酸序列
<400> 83
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys
20 25 30
Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr
35 40 45
Phe Thr Asn Tyr Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly
50 55 60
Leu Gln Trp Met Gly Trp Ile Asn Thr Asp Ser Gly Glu Ser Thr Tyr
65 70 75 80
Ala Glu Glu Phe Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val
85 90 95
Asn Thr Ala Tyr Leu Gln Ile Thr Ser Leu Thr Ala Glu Asp Thr Gly
100 105 110
Met Tyr Phe Cys Val Arg Val Gly Tyr Asp Ala Leu Asp Tyr Trp Gly
115 120 125
Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
130 135 140
Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala
145 150 155 160
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
165 170 175
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
180 185 190
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val
195 200 205
Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His
210 215 220
Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly
225 230 235 240
Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser
245 250 255
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
260 265 270
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro
275 280 285
Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
290 295 300
Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val
305 310 315 320
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
325 330 335
Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr
340 345 350
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
355 360 365
Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys
370 375 380
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
385 390 395 400
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp
405 410 415
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser
420 425 430
Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala
435 440 445
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Gly
450 455 460
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser Asp
465 470 475 480
Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu Ile Ile
485 490 495
His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val Thr Ser
500 505 510
Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr Leu Ile
515 520 525
Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe Ile Ile
530 535 540
Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu Ala Thr
545 550 555 560
Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg Gln Thr
565 570 575
Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile Glu Leu
580 585 590
Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr Glu Leu
595 600 605
Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys His Gln
610 615 620
His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly Ser Glu
625 630 635 640
Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr Arg Ser
645 650 655
Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met Thr Lys
660 665 670
Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680
<210> 84
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.5（κ,IgG4）的轻链亚基（BY24.5L）核苷酸序列
<400> 84
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgacattca gatgacccag tctccaagct ctctgagcgc ttccgtgggc 120
gaccgcgtga caattacatg cctcgcatct cagaccattg gcacctggct gacatggtat 180
cagcagaagc ctggcaaggc ccctaagctg ctgatttaca ccgctacctc cctcgccgac 240
ggcgtgccat ctaggttctc tggctccggc tccggcacag acttcacact cactatttct 300
tccctccagc ccgaggactt cgccacatac tactgccagc aggtgtactc tatcccttgg 360
actttcggcg gcggcactaa ggtggagatt aagagaaccg tggccgcccc cagcgtgttc 420
atcttccctc ccagcgacga gcagctgaag tctggcaccg ccagcgtggt gtgcctgctg 480
aacaacttct acccccgcga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 540
ggcaacagcc aggagagcgt gaccgagcag gactccaagg acagcaccta cagcctgagc 600
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 660
acccaccagg gactgtctag ccccgtgacc aagagcttca accggggcga gtgctaagaa 720
ttc 723
<210> 85
<211> 234
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.5（κ,IgG4）的轻链亚基（BY24.5L）氨基酸序列
<400> 85
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Leu Ala Ser Gln Thr
35 40 45
Ile Gly Thr Trp Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
50 55 60
Lys Leu Leu Ile Tyr Thr Ala Thr Ser Leu Ala Asp Gly Val Pro Ser
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Val Tyr
100 105 110
Ser Ile Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 86
<211> 2067
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.5（κ,IgG4）的重链-VID融合亚基（BY24.5H）核苷酸序列
<400> 86
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gcgaggtcca gctcgtcgag agtggaggcg gcctcgtgca gcccggcgga 120
agcctcagac tgtcttgtgc tgcatctggc ttcactttct ctagttacat gatgagttgg 180
gtgagacagg caccaggaaa gggattggag tgggtcgcaa caatcagtgg cggaggagca 240
aacacatact accccgatag cgtcaaggga cggttcacca ttagtcgcga taacgctaag 300
aactccctgt accttcagat gaatagtctc cgcgctgagg ataccgccgt gtactactgc 360
gcacggcagc tctactactt cgattactgg ggccagggca ccctcgtgac agtgtcttcc 420
gcctccacca agggcccttc cgtgttccct ctggcccctt gctcccgctc cacctccgag 480
tccaccgccg ccctgggctg cctggtgaag gactacttcc ctgagcctgt gaccgtgtcc 540
tggaactccg gcgccctgac ctccggcgtg cacaccttcc ctgccgtgct gcagtcctcc 600
ggcctgtact ccctgtcctc cgtggtgacc gtgccttcct cctccctggg caccaagacc 660
tacacctgca acgtggacca caagccttcc aacaccaagg tggacaagcg cgtggagtcc 720
aagtacggcc ctccttgccc tccttgccct gcccctgagt tcctgggcgg cccttccgtg 780
ttcctgttcc ctcctaagcc taaggacacc ctgatgatct cccgcacccc tgaggtgacc 840
tgcgtggtgg tggacgtgtc ccaggaggac cctgaggtgc agttcaactg gtacgtggac 900
ggcgtggagg tgcacaacgc caagaccaag cctcgcgagg agcagttcaa ctccacctac 960
cgcgtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa ggagtacaag 1020
tgcaaggtgt ccaacaaggg cctgccttcc tccatcgaga agaccatctc caaggccaag 1080
ggccagcctc gcgagcctca ggtgtacacc ctgcctcctt cccaggagga gatgaccaag 1140
aaccaggtgt ccctgacctg cctggtgaag ggcttctacc cttccgacat cgccgtggag 1200
tgggagtcca acggccagcc tgagaacaac tacaagacca cccctcctgt gctggactcc 1260
gacggctcct tcttcctgta ctcccgcctg accgtggaca agtcccgctg gcaggagggc 1320
aacgtgttct cctgctccgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1380
ctgtccctgt ccctgggcgg cggaggatct ggcggcggag gcagtggagg cggcggaagc 1440
gcttccgaca ccggccgccc tttcgtggag atgtactccg agatccctga gatcatccac 1500
atgaccgagg gccgcgagct ggtgatccct tgccgcgtga cctcccctaa catcaccgtg 1560
accctgaaga agttccctct ggacaccctg atccctgacg gcaagcgcat catctgggac 1620
tcccgcaagg gcttcatcat ctccaacgcc acctacaagg agatcggcct gctgacctgc 1680
gaggccaccg tgaacggcca cctgtacaag accaactacc tgacccaccg ccagaccaac 1740
accatcatcg acgtggtgct gtccccttcc cacggcatcg agctgtccgt gggcgagaag 1800
ctggtgctga actgcaccgc ccgcaccgag ctgaacgtgg gcatcgactt caactgggag 1860
tacccttcct ccaagcacca gcacaagaag ctggtgaacc gcgacctgaa gacccagtcc 1920
ggctccgaga tgaagaagtt cctgtccacc ctgaccatcg acggcgtgac ccgctccgac 1980
cagggcctgt acacctgcgc cgcctcctcc ggcctgatga ccaagaagaa ctccaccttc 2040
gtgcgcgtgc acgagaagta agtcgac 2067
<210> 87
<211> 682
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.5（κ,IgG4）的重链-VID融合亚基（BY24.5H）氨基酸序列
<400> 87
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
20 25 30
Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
35 40 45
Phe Ser Ser Tyr Met Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
50 55 60
Leu Glu Trp Val Ala Thr Ile Ser Gly Gly Gly Ala Asn Thr Tyr Tyr
65 70 75 80
Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys
85 90 95
Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Arg Gln Leu Tyr Tyr Phe Asp Tyr Trp Gly Gln
115 120 125
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
130 135 140
Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala
145 150 155 160
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
165 170 175
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
180 185 190
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
195 200 205
Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys
210 215 220
Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro
225 230 235 240
Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val
245 250 255
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr
260 265 270
Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu
275 280 285
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
290 295 300
Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser
305 310 315 320
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys
325 330 335
Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile
340 345 350
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
355 360 365
Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
370 375 380
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn
385 390 395 400
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser
405 410 415
Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg
420 425 430
Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
435 440 445
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Gly Gly
450 455 460
Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser Asp Thr
465 470 475 480
Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu Ile Ile His
485 490 495
Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val Thr Ser Pro
500 505 510
Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr Leu Ile Pro
515 520 525
Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe Ile Ile Ser
530 535 540
Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu Ala Thr Val
545 550 555 560
Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg Gln Thr Asn
565 570 575
Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile Glu Leu Ser
580 585 590
Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr Glu Leu Asn
595 600 605
Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys His Gln His
610 615 620
Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly Ser Glu Met
625 630 635 640
Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr Arg Ser Asp
645 650 655
Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met Thr Lys Lys
660 665 670
Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680
<210> 88
<211> 720
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.6（κ,IgG2）的轻链亚基（BY24.6L）核苷酸序列
<400> 88
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcaatattca gatgacccag agtcctagta gcctgagcgc atccgtcggc 120
gaccgcgtga ccattacatg taaggccgga cagaacgtga ataattacct cgcttggtat 180
cagcagaagc caggcaaggc tccaaaggtg ctcatcttca atgctaacag tctccagact 240
ggcgtccctt cccggtttag tggaagtgga tctggcaccg atttcacact cactatcagt 300
tctttgcaac ccgaggattt tgccacatac tactgtcagc agtacaatag ctggacaact 360
ttcggcggag gaactaaggt ggagattaag agaaccgtgg ccgcccccag cgtgttcatc 420
ttccctccca gcgacgagca gctgaagtct ggcaccgcca gcgtggtgtg cctgctgaac 480
aacttctacc cccgcgaggc caaggtgcag tggaaggtgg acaacgccct gcagagcggc 540
aacagccagg agagcgtgac cgagcaggac tccaaggaca gcacctacag cctgagcagc 600
accctgaccc tgagcaaggc cgactacgag aagcacaagg tgtacgcctg cgaggtgacc 660
caccagggac tgtctagccc cgtgaccaag agcttcaacc ggggcgagtg ctaagaattc 720
<210> 89
<211> 233
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.6（κ,IgG2）的轻链亚基（BY24.6L）氨基酸序列
<400> 89
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asn Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Gly Gln Asn
35 40 45
Val Asn Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
50 55 60
Lys Val Leu Ile Phe Asn Ala Asn Ser Leu Gln Thr Gly Val Pro Ser
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn
100 105 110
Ser Trp Thr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr
115 120 125
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
130 135 140
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
145 150 155 160
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
165 170 175
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
180 185 190
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
195 200 205
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
210 215 220
Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 90
<211> 2082
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.6（κ,IgG2）的重链-VID融合亚基（BY24.6H）核苷酸序列
<400> 90
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtcac actcaaggag tccggcccag ctctcgtgaa gcctacacag 120
accctcactc tcacctgtac attcagcgga tttagcctga gcacttctgg aacatgcgtg 180
tcttggattc ggcagccacc cggaaaggca ctcgaatggc tcgcaaccat ctgttgggag 240
gacagtaagg gctacaatcc atctctgaag tctaggctga caattagtaa ggacacctcc 300
aagaatcagg ccgtgctgac tatgactaat atggaccccg tcgataccgc cacatactac 360
tgcgctagac gcgaggatag tggctacttt tggtttcctt actggggcca gggaaccctc 420
gtgacagtgt cttccgcctc caccaagggc ccttccgtgt tccctctggc cccttgctcc 480
cgctccacct ccgagtccac cgccgccctg ggctgcctgg tgaaggacta cttccctgag 540
cctgtgaccg tgtcctggaa ctccggcgcc ctgacctccg gcgtgcacac cttccctgcc 600
gtgctgcagt cctccggcct gtactccctg tcctccgtgg tgaccgtgcc ttcctcctcc 660
ctgggcacca agacctacac ctgcaacgtg gaccacaagc cttccaacac caaggtggac 720
aagcgcgtgg agtccaagta cggccctcct tgccctcctt gccctgcccc tgagttcctg 780
ggcggccctt ccgtgttcct gttccctcct aagcctaagg acaccctgat gatctcccgc 840
acccctgagg tgacctgcgt ggtggtggac gtgtcccagg aggaccctga ggtgcagttc 900
aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcctcg cgaggagcag 960
ttcaactcca cctaccgcgt ggtgtccgtg ctgaccgtgc tgcaccagga ctggctgaac 1020
ggcaaggagt acaagtgcaa ggtgtccaac aagggcctgc cttcctccat cgagaagacc 1080
atctccaagg ccaagggcca gcctcgcgag cctcaggtgt acaccctgcc tccttcccag 1140
gaggagatga ccaagaacca ggtgtccctg acctgcctgg tgaagggctt ctacccttcc 1200
gacatcgccg tggagtggga gtccaacggc cagcctgaga acaactacaa gaccacccct 1260
cctgtgctgg actccgacgg ctccttcttc ctgtactccc gcctgaccgt ggacaagtcc 1320
cgctggcagg agggcaacgt gttctcctgc tccgtgatgc acgaggccct gcacaaccac 1380
tacacccaga agtccctgtc cctgtccctg ggcggcggag gatctggcgg cggaggcagt 1440
ggaggcggcg gaagcgcttc cgacaccggc cgccctttcg tggagatgta ctccgagatc 1500
cctgagatca tccacatgac cgagggccgc gagctggtga tcccttgccg cgtgacctcc 1560
cctaacatca ccgtgaccct gaagaagttc cctctggaca ccctgatccc tgacggcaag 1620
cgcatcatct gggactcccg caagggcttc atcatctcca acgccaccta caaggagatc 1680
ggcctgctga cctgcgaggc caccgtgaac ggccacctgt acaagaccaa ctacctgacc 1740
caccgccaga ccaacaccat catcgacgtg gtgctgtccc cttcccacgg catcgagctg 1800
tccgtgggcg agaagctggt gctgaactgc accgcccgca ccgagctgaa cgtgggcatc 1860
gacttcaact gggagtaccc ttcctccaag caccagcaca agaagctggt gaaccgcgac 1920
ctgaagaccc agtccggctc cgagatgaag aagttcctgt ccaccctgac catcgacggc 1980
gtgacccgct ccgaccaggg cctgtacacc tgcgccgcct cctccggcct gatgaccaag 2040
aagaactcca ccttcgtgcg cgtgcacgag aagtaagtcg ac 2082
<210> 91
<211> 687
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.6（κ,IgG2）的重链-VID融合亚基（BY24.6H）氨基酸序列
<400> 91
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Thr Leu Lys Glu Ser Gly Pro Ala Leu Val
20 25 30
Lys Pro Thr Gln Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser
35 40 45
Leu Ser Thr Ser Gly Thr Cys Val Ser Trp Ile Arg Gln Pro Pro Gly
50 55 60
Lys Ala Leu Glu Trp Leu Ala Thr Ile Cys Trp Glu Asp Ser Lys Gly
65 70 75 80
Tyr Asn Pro Ser Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser
85 90 95
Lys Asn Gln Ala Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr
100 105 110
Ala Thr Tyr Tyr Cys Ala Arg Arg Glu Asp Ser Gly Tyr Phe Trp Phe
115 120 125
Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
130 135 140
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser
145 150 155 160
Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
165 170 175
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
180 185 190
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
195 200 205
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys
210 215 220
Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu
225 230 235 240
Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu
245 250 255
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
260 265 270
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
275 280 285
Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
290 295 300
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr
305 310 315 320
Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn
325 330 335
Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser
340 345 350
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
355 360 365
Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val
370 375 380
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
385 390 395 400
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
405 410 415
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr
420 425 430
Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val
435 440 445
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
450 455 460
Ser Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
465 470 475 480
Ser Ala Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile
485 490 495
Pro Glu Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys
500 505 510
Arg Val Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu
515 520 525
Asp Thr Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys
530 535 540
Gly Phe Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr
545 550 555 560
Cys Glu Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr
565 570 575
His Arg Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His
580 585 590
Gly Ile Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala
595 600 605
Arg Thr Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser
610 615 620
Ser Lys His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln
625 630 635 640
Ser Gly Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly
645 650 655
Val Thr Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly
660 665 670
Leu Met Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680 685
<210> 92
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.8（κ,IgG4）的轻链亚基（BY24.8L）核苷酸序列
<400> 92
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgacatcgt gatgacccag agtcctgata gtctggccgt gtccctcggc 120
gagcgcgcaa caatcaattg caaggcatct cagtccgttt ccaacgatgt cgcatggtat 180
cagcagaagc ctggacagcc acctaagctg ctcattaact acgccttcca cagattcact 240
ggcgtgcccg atcggttttc cggaagtgga tacggaaccg actttacact gactattagt 300
tctctacaag ctgaggacgt cgctgtgtac tactgtcacc aggcttactc tagcccatac 360
acatttggag gcggcactaa ggtggagatt aagagaaccg tggccgcccc cagcgtgttc 420
atcttccctc ccagcgacga gcagctgaag tctggcaccg ccagcgtggt gtgcctgctg 480
aacaacttct acccccgcga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 540
ggcaacagcc aggagagcgt gaccgagcag gactccaagg acagcaccta cagcctgagc 600
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 660
acccaccagg gactgtctag ccccgtgacc aagagcttca accggggcga gtgctaagaa 720
ttc 723
<210> 93
<211> 234
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.8（κ,IgG4）的轻链亚基（BY24.8L）氨基酸序列
<400> 93
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala
20 25 30
Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Ser
35 40 45
Val Ser Asn Asp Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
50 55 60
Lys Leu Leu Ile Asn Tyr Ala Phe His Arg Phe Thr Gly Val Pro Asp
65 70 75 80
Arg Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys His Gln Ala Tyr
100 105 110
Ser Ser Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 94
<211> 2073
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.8（κ,IgG4）的重链-VID融合亚基（BY24.8H）核苷酸序列
<400> 94
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtcca gctccaggag agcggacctg gcctcgtgaa gcctagcgag 120
actctgtctc tgacatgtac agtgtccggc tttagcctca cttcttacgg cgtgcactgg 180
attcgccagc cacccggaaa gggattggaa tggctcggcg tcatttgggc cggaggcagc 240
actaactaca acccatctct caagtctagg ctcacaatca gcaaggataa tagtaagagt 300
caggtgtccc tgaagatgag ttccgtcacc gctgccgata ccgctgtgta ctactgcgca 360
cgcgcatacg gcaattactg gtacatcgac gtgtggggac agggcaccct cgtgacagtg 420
tcttccgcct ccaccaaggg cccttccgtg ttccctctgg ccccttgctc ccgctccacc 480
tccgagtcca ccgccgccct gggctgcctg gtgaaggact acttccctga gcctgtgacc 540
gtgtcctgga actccggcgc cctgacctcc ggcgtgcaca ccttccctgc cgtgctgcag 600
tcctccggcc tgtactccct gtcctccgtg gtgaccgtgc cttcctcctc cctgggcacc 660
aagacctaca cctgcaacgt ggaccacaag ccttccaaca ccaaggtgga caagcgcgtg 720
gagtccaagt acggccctcc ttgccctcct tgccctgccc ctgagttcct gggcggccct 780
tccgtgttcc tgttccctcc taagcctaag gacaccctga tgatctcccg cacccctgag 840
gtgacctgcg tggtggtgga cgtgtcccag gaggaccctg aggtgcagtt caactggtac 900
gtggacggcg tggaggtgca caacgccaag accaagcctc gcgaggagca gttcaactcc 960
acctaccgcg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaggag 1020
tacaagtgca aggtgtccaa caagggcctg ccttcctcca tcgagaagac catctccaag 1080
gccaagggcc agcctcgcga gcctcaggtg tacaccctgc ctccttccca ggaggagatg 1140
accaagaacc aggtgtccct gacctgcctg gtgaagggct tctacccttc cgacatcgcc 1200
gtggagtggg agtccaacgg ccagcctgag aacaactaca agaccacccc tcctgtgctg 1260
gactccgacg gctccttctt cctgtactcc cgcctgaccg tggacaagtc ccgctggcag 1320
gagggcaacg tgttctcctg ctccgtgatg cacgaggccc tgcacaacca ctacacccag 1380
aagtccctgt ccctgtccct gggcggcgga ggatctggcg gcggaggcag tggaggcggc 1440
ggaagcgctt ccgacaccgg ccgccctttc gtggagatgt actccgagat ccctgagatc 1500
atccacatga ccgagggccg cgagctggtg atcccttgcc gcgtgacctc ccctaacatc 1560
accgtgaccc tgaagaagtt ccctctggac accctgatcc ctgacggcaa gcgcatcatc 1620
tgggactccc gcaagggctt catcatctcc aacgccacct acaaggagat cggcctgctg 1680
acctgcgagg ccaccgtgaa cggccacctg tacaagacca actacctgac ccaccgccag 1740
accaacacca tcatcgacgt ggtgctgtcc ccttcccacg gcatcgagct gtccgtgggc 1800
gagaagctgg tgctgaactg caccgcccgc accgagctga acgtgggcat cgacttcaac 1860
tgggagtacc cttcctccaa gcaccagcac aagaagctgg tgaaccgcga cctgaagacc 1920
cagtccggct ccgagatgaa gaagttcctg tccaccctga ccatcgacgg cgtgacccgc 1980
tccgaccagg gcctgtacac ctgcgccgcc tcctccggcc tgatgaccaa gaagaactcc 2040
accttcgtgc gcgtgcacga gaagtaagtc gac 2073
<210> 95
<211> 684
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.8（κ,IgG4）的重链-VID融合亚基（BY24.8H）氨基酸序列
<400> 95
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val
20 25 30
Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Phe Ser
35 40 45
Leu Thr Ser Tyr Gly Val His Trp Ile Arg Gln Pro Pro Gly Lys Gly
50 55 60
Leu Glu Trp Leu Gly Val Ile Trp Ala Gly Gly Ser Thr Asn Tyr Asn
65 70 75 80
Pro Ser Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Ser
85 90 95
Gln Val Ser Leu Lys Met Ser Ser Val Thr Ala Ala Asp Thr Ala Val
100 105 110
Tyr Tyr Cys Ala Arg Ala Tyr Gly Asn Tyr Trp Tyr Ile Asp Val Trp
115 120 125
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
130 135 140
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr
145 150 155 160
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
165 170 175
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
180 185 190
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
195 200 205
Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp
210 215 220
His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr
225 230 235 240
Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro
245 250 255
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
260 265 270
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
275 280 285
Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
290 295 300
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val
305 310 315 320
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
325 330 335
Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys
340 345 350
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
355 360 365
Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
370 375 380
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
385 390 395 400
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
405 410 415
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys
420 425 430
Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu
435 440 445
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
450 455 460
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser
465 470 475 480
Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu Ile
485 490 495
Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val Thr
500 505 510
Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr Leu
515 520 525
Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe Ile
530 535 540
Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu Ala
545 550 555 560
Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg Gln
565 570 575
Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile Glu
580 585 590
Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr Glu
595 600 605
Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys His
610 615 620
Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly Ser
625 630 635 640
Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr Arg
645 650 655
Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met Thr
660 665 670
Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680
<210> 96
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.9（κ,IgG4）的轻链亚基（BY24.9L）核苷酸序列
<400> 96
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgatattca gatgacacag tccccaagtt ccctgagcgc ctccgtcgga 120
gaccgggtca ccattacttg tcgcgcttct cagagcgtga gtaattacct cgattggtat 180
cagcagaagc caggaaaggc tcctaagctg ctcatctacg acgcatccac ccgcgcaaca 240
ggcgtgccta gccggtttag cggatctgga agtggcactg atttcacact cacaatctct 300
agtctgcaac ccgaggactt tgctacatac tactgtcagc agaacatgca gctgccactg 360
acattcggcc agggaactaa ggtggagatt aagagaaccg tggccgcccc cagcgtgttc 420
atcttccctc ccagcgacga gcagctgaag tctggcaccg ccagcgtggt gtgcctgctg 480
aacaacttct acccccgcga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 540
ggcaacagcc aggagagcgt gaccgagcag gactccaagg acagcaccta cagcctgagc 600
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 660
acccaccagg gactgtctag ccccgtgacc aagagcttca accggggcga gtgctaagaa 720
ttc 723
<210> 97
<211> 234
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.9（κ,IgG4）的轻链亚基（BY24.9L）氨基酸序列
<400> 97
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser
35 40 45
Val Ser Asn Tyr Leu Asp Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
50 55 60
Lys Leu Leu Ile Tyr Asp Ala Ser Thr Arg Ala Thr Gly Val Pro Ser
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Asn Met
100 105 110
Gln Leu Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 98
<211> 2055
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.9（κ,IgG4）的重链-VID融合亚基（BY24.9H）核苷酸序列
<400> 98
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gcgtgcagct cgtcgagtcc ggcggaggcg tggtccagcc aggacgcagt 120
ctgcgcctgg attgtaaggc aagcggcatc acctttagta actacggtat gcactgggtg 180
agacaggctc ccggaaaggg cctagaatgg gtggccgtca tttggtacga ctcttctagg 240
aagtactacg ccgatagtgt caagggacgg ttcacaatct ctcgcgataa tagcaagaat 300
acactgtttt tgcaaatgaa ctccctcaga gctgaggata ccgctgtgta ctactgcgca 360
accaacaatg attactgggg acagggcacc ctcgtgacag tgtcttccgc ctccaccaag 420
ggcccttccg tgttccctct ggccccttgc tcccgctcca cctccgagtc caccgccgcc 480
ctgggctgcc tggtgaagga ctacttccct gagcctgtga ccgtgtcctg gaactccggc 540
gccctgacct ccggcgtgca caccttccct gccgtgctgc agtcctccgg cctgtactcc 600
ctgtcctccg tggtgaccgt gccttcctcc tccctgggca ccaagaccta cacctgcaac 660
gtggaccaca agccttccaa caccaaggtg gacaagcgcg tggagtccaa gtacggccct 720
ccttgccctc cttgccctgc ccctgagttc ctgggcggcc cttccgtgtt cctgttccct 780
cctaagccta aggacaccct gatgatctcc cgcacccctg aggtgacctg cgtggtggtg 840
gacgtgtccc aggaggaccc tgaggtgcag ttcaactggt acgtggacgg cgtggaggtg 900
cacaacgcca agaccaagcc tcgcgaggag cagttcaact ccacctaccg cgtggtgtcc 960
gtgctgaccg tgctgcacca ggactggctg aacggcaagg agtacaagtg caaggtgtcc 1020
aacaagggcc tgccttcctc catcgagaag accatctcca aggccaaggg ccagcctcgc 1080
gagcctcagg tgtacaccct gcctccttcc caggaggaga tgaccaagaa ccaggtgtcc 1140
ctgacctgcc tggtgaaggg cttctaccct tccgacatcg ccgtggagtg ggagtccaac 1200
ggccagcctg agaacaacta caagaccacc cctcctgtgc tggactccga cggctccttc 1260
ttcctgtact cccgcctgac cgtggacaag tcccgctggc aggagggcaa cgtgttctcc 1320
tgctccgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct gtccctgtcc 1380
ctgggcggcg gaggatctgg cggcggaggc agtggaggcg gcggaagcgc ttccgacacc 1440
ggccgccctt tcgtggagat gtactccgag atccctgaga tcatccacat gaccgagggc 1500
cgcgagctgg tgatcccttg ccgcgtgacc tcccctaaca tcaccgtgac cctgaagaag 1560
ttccctctgg acaccctgat ccctgacggc aagcgcatca tctgggactc ccgcaagggc 1620
ttcatcatct ccaacgccac ctacaaggag atcggcctgc tgacctgcga ggccaccgtg 1680
aacggccacc tgtacaagac caactacctg acccaccgcc agaccaacac catcatcgac 1740
gtggtgctgt ccccttccca cggcatcgag ctgtccgtgg gcgagaagct ggtgctgaac 1800
tgcaccgccc gcaccgagct gaacgtgggc atcgacttca actgggagta cccttcctcc 1860
aagcaccagc acaagaagct ggtgaaccgc gacctgaaga cccagtccgg ctccgagatg 1920
aagaagttcc tgtccaccct gaccatcgac ggcgtgaccc gctccgacca gggcctgtac 1980
acctgcgccg cctcctccgg cctgatgacc aagaagaact ccaccttcgt gcgcgtgcac 2040
gagaagtaag tcgac 2055
<210> 99
<211> 678
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.9（κ,IgG4）的重链-VID融合亚基（BY24.9H）氨基酸序列
<400> 99
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
20 25 30
Pro Gly Arg Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe
35 40 45
Ser Asn Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
50 55 60
Glu Trp Val Ala Val Ile Trp Tyr Asp Ser Ser Arg Lys Tyr Tyr Ala
65 70 75 80
Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
85 90 95
Thr Leu Phe Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
100 105 110
Tyr Tyr Cys Ala Thr Asn Asn Asp Tyr Trp Gly Gln Gly Thr Leu Val
115 120 125
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
130 135 140
Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu
145 150 155 160
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
165 170 175
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
180 185 190
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
195 200 205
Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr
210 215 220
Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro
225 230 235 240
Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro
245 250 255
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
260 265 270
Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn
275 280 285
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg
290 295 300
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
305 310 315 320
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
325 330 335
Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys
340 345 350
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu
355 360 365
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
370 375 380
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
385 390 395 400
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
405 410 415
Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly
420 425 430
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
435 440 445
Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Gly Gly Gly Ser Gly Gly
450 455 460
Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser Asp Thr Gly Arg Pro Phe
465 470 475 480
Val Glu Met Tyr Ser Glu Ile Pro Glu Ile Ile His Met Thr Glu Gly
485 490 495
Arg Glu Leu Val Ile Pro Cys Arg Val Thr Ser Pro Asn Ile Thr Val
500 505 510
Thr Leu Lys Lys Phe Pro Leu Asp Thr Leu Ile Pro Asp Gly Lys Arg
515 520 525
Ile Ile Trp Asp Ser Arg Lys Gly Phe Ile Ile Ser Asn Ala Thr Tyr
530 535 540
Lys Glu Ile Gly Leu Leu Thr Cys Glu Ala Thr Val Asn Gly His Leu
545 550 555 560
Tyr Lys Thr Asn Tyr Leu Thr His Arg Gln Thr Asn Thr Ile Ile Asp
565 570 575
Val Val Leu Ser Pro Ser His Gly Ile Glu Leu Ser Val Gly Glu Lys
580 585 590
Leu Val Leu Asn Cys Thr Ala Arg Thr Glu Leu Asn Val Gly Ile Asp
595 600 605
Phe Asn Trp Glu Tyr Pro Ser Ser Lys His Gln His Lys Lys Leu Val
610 615 620
Asn Arg Asp Leu Lys Thr Gln Ser Gly Ser Glu Met Lys Lys Phe Leu
625 630 635 640
Ser Thr Leu Thr Ile Asp Gly Val Thr Arg Ser Asp Gln Gly Leu Tyr
645 650 655
Thr Cys Ala Ala Ser Ser Gly Leu Met Thr Lys Lys Asn Ser Thr Phe
660 665 670
Val Arg Val His Glu Lys
675
<210> 100
<211> 729
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.10（κ,IgG4）的轻链亚基（BY24.10L）核苷酸序列
<400> 100
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcctcagtta cgtcctcaca cagcctccat ccgtgtctgt gagtcccgga 120
cagaccgcaa gaatcacatg tagcggcgac gcactgccta agcagtacgc ttactggtat 180
cagcagaagc caggacaggc acctgtgctg gtgatctaca aggatagcga gcgcccaagt 240
ggcattcccg agagatttag tggctcttct agtggaacaa ccgtcaccct gactatttcc 300
ggcgtgcagg ccgaggatga ggccgattac tactgtcagt ctgctgactc tagcggaaca 360
tacgtcgtgt ttggaggcgg aactaaggtg gagattaaga gaaccgtggc cgcccccagc 420
gtgttcatct tccctcccag cgacgagcag ctgaagtctg gcaccgccag cgtggtgtgc 480
ctgctgaaca acttctaccc ccgcgaggcc aaggtgcagt ggaaggtgga caacgccctg 540
cagagcggca acagccagga gagcgtgacc gagcaggact ccaaggacag cacctacagc 600
ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcacaaggt gtacgcctgc 660
gaggtgaccc accagggact gtctagcccc gtgaccaaga gcttcaaccg gggcgagtgc 720
taagaattc 729
<210> 101
<211> 236
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.10（κ,IgG4）的轻链亚基（BY24.10L）氨基酸序列
<400> 101
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Leu Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser
20 25 30
Val Ser Pro Gly Gln Thr Ala Arg Ile Thr Cys Ser Gly Asp Ala Leu
35 40 45
Pro Lys Gln Tyr Ala Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
50 55 60
Val Leu Val Ile Tyr Lys Asp Ser Glu Arg Pro Ser Gly Ile Pro Glu
65 70 75 80
Arg Phe Ser Gly Ser Ser Ser Gly Thr Thr Val Thr Leu Thr Ile Ser
85 90 95
Gly Val Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Ala Asp
100 105 110
Ser Ser Gly Thr Tyr Val Val Phe Gly Gly Gly Thr Lys Val Glu Ile
115 120 125
Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
130 135 140
Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn
145 150 155 160
Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu
165 170 175
Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
180 185 190
Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
195 200 205
Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser
210 215 220
Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 102
<211> 2079
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.10（κ,IgG4）的重链-VID融合亚基（BY24.10H）核苷酸序列
<400> 102
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gcgacctcgt gcagtccggc gccgaggtga agaagcccgg cgcatccgtc 120
aaggtgtctt gcaaggcaag tggctacact ttcaccagtt acggaatcag ttgggtcaga 180
caggcacctg gccagggcct ggagtggatg ggctggatta gcgcttacaa cggaaacacc 240
aattacgctc agaagctcca gggtcgggtg actatgacaa ccgacacatc taccagcacc 300
gcatacatgg agctgcgtag tctgagatcc gacgataccg ccgtgtacta ctgtgctcgc 360
ggcagaggat actcctacgg aattgatgca ttcgatattt ggggacaggg aaccctcgtg 420
acagtgtctt ccgcctccac caagggccct tccgtgttcc ctctggcccc ttgctcccgc 480
tccacctccg agtccaccgc cgccctgggc tgcctggtga aggactactt ccctgagcct 540
gtgaccgtgt cctggaactc cggcgccctg acctccggcg tgcacacctt ccctgccgtg 600
ctgcagtcct ccggcctgta ctccctgtcc tccgtggtga ccgtgccttc ctcctccctg 660
ggcaccaaga cctacacctg caacgtggac cacaagcctt ccaacaccaa ggtggacaag 720
cgcgtggagt ccaagtacgg ccctccttgc cctccttgcc ctgcccctga gttcctgggc 780
ggcccttccg tgttcctgtt ccctcctaag cctaaggaca ccctgatgat ctcccgcacc 840
cctgaggtga cctgcgtggt ggtggacgtg tcccaggagg accctgaggt gcagttcaac 900
tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcctcgcga ggagcagttc 960
aactccacct accgcgtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 1020
aaggagtaca agtgcaaggt gtccaacaag ggcctgcctt cctccatcga gaagaccatc 1080
tccaaggcca agggccagcc tcgcgagcct caggtgtaca ccctgcctcc ttcccaggag 1140
gagatgacca agaaccaggt gtccctgacc tgcctggtga agggcttcta cccttccgac 1200
atcgccgtgg agtgggagtc caacggccag cctgagaaca actacaagac cacccctcct 1260
gtgctggact ccgacggctc cttcttcctg tactcccgcc tgaccgtgga caagtcccgc 1320
tggcaggagg gcaacgtgtt ctcctgctcc gtgatgcacg aggccctgca caaccactac 1380
acccagaagt ccctgtccct gtccctgggc ggcggaggat ctggcggcgg aggcagtgga 1440
ggcggcggaa gcgcttccga caccggccgc cctttcgtgg agatgtactc cgagatccct 1500
gagatcatcc acatgaccga gggccgcgag ctggtgatcc cttgccgcgt gacctcccct 1560
aacatcaccg tgaccctgaa gaagttccct ctggacaccc tgatccctga cggcaagcgc 1620
atcatctggg actcccgcaa gggcttcatc atctccaacg ccacctacaa ggagatcggc 1680
ctgctgacct gcgaggccac cgtgaacggc cacctgtaca agaccaacta cctgacccac 1740
cgccagacca acaccatcat cgacgtggtg ctgtcccctt cccacggcat cgagctgtcc 1800
gtgggcgaga agctggtgct gaactgcacc gcccgcaccg agctgaacgt gggcatcgac 1860
ttcaactggg agtacccttc ctccaagcac cagcacaaga agctggtgaa ccgcgacctg 1920
aagacccagt ccggctccga gatgaagaag ttcctgtcca ccctgaccat cgacggcgtg 1980
acccgctccg accagggcct gtacacctgc gccgcctcct ccggcctgat gaccaagaag 2040
aactccacct tcgtgcgcgt gcacgagaag taagtcgac 2079
<210> 103
<211> 686
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.10（κ,IgG4）的重链-VID融合亚基（BY24.10H）氨基酸序列
<400> 103
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro
20 25 30
Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr
35 40 45
Ser Tyr Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
50 55 60
Trp Met Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln
65 70 75 80
Lys Leu Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr
85 90 95
Ala Tyr Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr
100 105 110
Tyr Cys Ala Arg Gly Arg Gly Tyr Ser Tyr Gly Ile Asp Ala Phe Asp
115 120 125
Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys
130 135 140
Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
145 150 155 160
Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
165 170 175
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
180 185 190
Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
195 200 205
Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn
210 215 220
Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser
225 230 235 240
Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly
245 250 255
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
260 265 270
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln
275 280 285
Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
290 295 300
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr
305 310 315 320
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
325 330 335
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile
340 345 350
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
355 360 365
Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser
370 375 380
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
385 390 395 400
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
405 410 415
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val
420 425 430
Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met
435 440 445
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
450 455 460
Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
465 470 475 480
Ala Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro
485 490 495
Glu Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg
500 505 510
Val Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp
515 520 525
Thr Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly
530 535 540
Phe Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys
545 550 555 560
Glu Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His
565 570 575
Arg Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly
580 585 590
Ile Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg
595 600 605
Thr Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser
610 615 620
Lys His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser
625 630 635 640
Gly Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val
645 650 655
Thr Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu
660 665 670
Met Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680 685
<210> 104
<211> 738
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.11（κ,IgG4）的轻链亚基（BY24.11L）核苷酸序列
<400> 104
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgatgtcgt catgacccag tcccctctgt ctctgcccgt cacactggga 120
cagcccgcat ccattagttg taggtctagc cagagcattg tgcacagtaa cggcaataca 180
tacctggagt ggtatcttca aaagcctggc cagtctcctc agctgctgat ctacaaggtg 240
agtaatcgct ttagcggcgt gcctgataga ttcagcggaa gtggctccgg aaccgacttc 300
acactcaaga tttctcgcgt ggaggccgag gacgtcggcg tgtactactg ttttcagggg 360
agccacgtgc cactcacctt tggacagggc actaaggtgg agattaagag aaccgtggcc 420
gcccccagcg tgttcatctt ccctcccagc gacgagcagc tgaagtctgg caccgccagc 480
gtggtgtgcc tgctgaacaa cttctacccc cgcgaggcca aggtgcagtg gaaggtggac 540
aacgccctgc agagcggcaa cagccaggag agcgtgaccg agcaggactc caaggacagc 600
acctacagcc tgagcagcac cctgaccctg agcaaggccg actacgagaa gcacaaggtg 660
tacgcctgcg aggtgaccca ccagggactg tctagccccg tgaccaagag cttcaaccgg 720
ggcgagtgct aagaattc 738
<210> 105
<211> 239
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.11（κ,IgG4）的轻链亚基（BY24.11L）氨基酸序列
<400> 105
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro
20 25 30
Val Thr Leu Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
35 40 45
Ile Val His Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys
50 55 60
Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe
65 70 75 80
Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
85 90 95
Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr
100 105 110
Cys Phe Gln Gly Ser His Val Pro Leu Thr Phe Gly Gln Gly Thr Lys
115 120 125
Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro
130 135 140
Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu
145 150 155 160
Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
165 170 175
Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp
180 185 190
Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys
195 200 205
Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln
210 215 220
Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 106
<211> 2094
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.11（κ,IgG4）的重链-VID融合亚基（BY24.11H）核苷酸序列
<400> 106
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccagggcca gctcgtgcag agtggcgcag aggtgaagaa gcccggagca 120
tccgtcaagg tcagttgcaa ggcctctgga tacaccttta ccgattacga gatgcactgg 180
gtgcggcagg ctcctggaca gggactcgaa tggatgggcg tcattgagtc cgagaccggc 240
ggaacagctt acaatcagaa gtttcaggga cgggtgacac tcactgccga taagtcttct 300
agcaccgcct acatggaact ttcctctctg cgctcagagg ataccgctgt gtactactgt 360
acacgcgagg gaatcacaac tgtcgcaacc acatactact ggtacttcga cgtgtggggc 420
cagggaaccc tcgtgacagt gtcttccgcc tccaccaagg gcccttccgt gttccctctg 480
gccccttgct cccgctccac ctccgagtcc accgccgccc tgggctgcct ggtgaaggac 540
tacttccctg agcctgtgac cgtgtcctgg aactccggcg ccctgacctc cggcgtgcac 600
accttccctg ccgtgctgca gtcctccggc ctgtactccc tgtcctccgt ggtgaccgtg 660
ccttcctcct ccctgggcac caagacctac acctgcaacg tggaccacaa gccttccaac 720
accaaggtgg acaagcgcgt ggagtccaag tacggccctc cttgccctcc ttgccctgcc 780
cctgagttcc tgggcggccc ttccgtgttc ctgttccctc ctaagcctaa ggacaccctg 840
atgatctccc gcacccctga ggtgacctgc gtggtggtgg acgtgtccca ggaggaccct 900
gaggtgcagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa gaccaagcct 960
cgcgaggagc agttcaactc cacctaccgc gtggtgtccg tgctgaccgt gctgcaccag 1020
gactggctga acggcaagga gtacaagtgc aaggtgtcca acaagggcct gccttcctcc 1080
atcgagaaga ccatctccaa ggccaagggc cagcctcgcg agcctcaggt gtacaccctg 1140
cctccttccc aggaggagat gaccaagaac caggtgtccc tgacctgcct ggtgaagggc 1200
ttctaccctt ccgacatcgc cgtggagtgg gagtccaacg gccagcctga gaacaactac 1260
aagaccaccc ctcctgtgct ggactccgac ggctccttct tcctgtactc ccgcctgacc 1320
gtggacaagt cccgctggca ggagggcaac gtgttctcct gctccgtgat gcacgaggcc 1380
ctgcacaacc actacaccca gaagtccctg tccctgtccc tgggcggcgg aggatctggc 1440
ggcggaggca gtggaggcgg cggaagcgct tccgacaccg gccgcccttt cgtggagatg 1500
tactccgaga tccctgagat catccacatg accgagggcc gcgagctggt gatcccttgc 1560
cgcgtgacct cccctaacat caccgtgacc ctgaagaagt tccctctgga caccctgatc 1620
cctgacggca agcgcatcat ctgggactcc cgcaagggct tcatcatctc caacgccacc 1680
tacaaggaga tcggcctgct gacctgcgag gccaccgtga acggccacct gtacaagacc 1740
aactacctga cccaccgcca gaccaacacc atcatcgacg tggtgctgtc cccttcccac 1800
ggcatcgagc tgtccgtggg cgagaagctg gtgctgaact gcaccgcccg caccgagctg 1860
aacgtgggca tcgacttcaa ctgggagtac ccttcctcca agcaccagca caagaagctg 1920
gtgaaccgcg acctgaagac ccagtccggc tccgagatga agaagttcct gtccaccctg 1980
accatcgacg gcgtgacccg ctccgaccag ggcctgtaca cctgcgccgc ctcctccggc 2040
ctgatgacca agaagaactc caccttcgtg cgcgtgcacg agaagtaagt cgac 2094
<210> 107
<211> 691
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.11（κ,IgG4）的重链-VID融合亚基（BY24.11H）氨基酸序列
<400> 107
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Gly Gln Leu Val Gln Ser Gly Ala Glu Val Lys
20 25 30
Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
35 40 45
Phe Thr Asp Tyr Glu Met His Trp Val Arg Gln Ala Pro Gly Gln Gly
50 55 60
Leu Glu Trp Met Gly Val Ile Glu Ser Glu Thr Gly Gly Thr Ala Tyr
65 70 75 80
Asn Gln Lys Phe Gln Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Ser
85 90 95
Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Thr Arg Glu Gly Ile Thr Thr Val Ala Thr Thr Tyr
115 120 125
Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser
130 135 140
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
145 150 155 160
Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
165 170 175
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
180 185 190
Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
195 200 205
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys
210 215 220
Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp
225 230 235 240
Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala
245 250 255
Pro Glu Phe 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 Gln Glu Asp Pro Glu Val Gln 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
Phe 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 Gly
340 345 350
Leu Pro Ser Ser 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 Gln Glu Glu Met Thr
370 375 380
Lys Asn Gln Val Ser Leu Thr Cys Leu 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 Tyr
420 425 430
Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu 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 Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
465 470 475 480
Gly Gly Gly Gly Ser Ala Ser Asp Thr Gly Arg Pro Phe Val Glu Met
485 490 495
Tyr Ser Glu Ile Pro Glu Ile Ile His Met Thr Glu Gly Arg Glu Leu
500 505 510
Val Ile Pro Cys Arg Val Thr Ser Pro Asn Ile Thr Val Thr Leu Lys
515 520 525
Lys Phe Pro Leu Asp Thr Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp
530 535 540
Asp Ser Arg Lys Gly Phe Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile
545 550 555 560
Gly Leu Leu Thr Cys Glu Ala Thr Val Asn Gly His Leu Tyr Lys Thr
565 570 575
Asn Tyr Leu Thr His Arg Gln Thr Asn Thr Ile Ile Asp Val Val Leu
580 585 590
Ser Pro Ser His Gly Ile Glu Leu Ser Val Gly Glu Lys Leu Val Leu
595 600 605
Asn Cys Thr Ala Arg Thr Glu Leu Asn Val Gly Ile Asp Phe Asn Trp
610 615 620
Glu Tyr Pro Ser Ser Lys His Gln His Lys Lys Leu Val Asn Arg Asp
625 630 635 640
Leu Lys Thr Gln Ser Gly Ser Glu Met Lys Lys Phe Leu Ser Thr Leu
645 650 655
Thr Ile Asp Gly Val Thr Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala
660 665 670
Ala Ser Ser Gly Leu Met Thr Lys Lys Asn Ser Thr Phe Val Arg Val
675 680 685
His Glu Lys
690
<210> 108
<211> 735
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.12（κ,IgG4）的轻链亚基（BY24.12L）核苷酸序列
<400> 108
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgatattgt gctgacccag agtcccgcat ctctcgccgt cagtcctgga 120
cagcgcgcta ctatcacttg cagggcttct gagagcgtcg ataattacgg catttccttt 180
atgaactggt atcagcagaa gcctggccag cctccaaagc tgctcatcta cacctctagt 240
aacaaggata caggcgtgcc cgcaagattt agcggctccg gaagtggcac cgacttcaca 300
ctcacaatca accctatgga ggccgaggat accgccgtgt actactgtca gcagtctaag 360
gaggtgcctt ggacattcgg cggcggaact aaggtggaga ttaagagaac cgtggccgcc 420
cccagcgtgt tcatcttccc tcccagcgac gagcagctga agtctggcac cgccagcgtg 480
gtgtgcctgc tgaacaactt ctacccccgc gaggccaagg tgcagtggaa ggtggacaac 540
gccctgcaga gcggcaacag ccaggagagc gtgaccgagc aggactccaa ggacagcacc 600
tacagcctga gcagcaccct gaccctgagc aaggccgact acgagaagca caaggtgtac 660
gcctgcgagg tgacccacca gggactgtct agccccgtga ccaagagctt caaccggggc 720
gagtgctaag aattc 735
<210> 109
<211> 238
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.12（κ,IgG4）的轻链亚基（BY24.12L）氨基酸序列
<400> 109
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala
20 25 30
Val Ser Pro Gly Gln Arg Ala Thr Ile Thr Cys Arg Ala Ser Glu Ser
35 40 45
Val Asp Asn Tyr Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro
50 55 60
Gly Gln Pro Pro Lys Leu Leu Ile Tyr Thr Ser Ser Asn Lys Asp Thr
65 70 75 80
Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
85 90 95
Leu Thr Ile Asn Pro Met Glu Ala Glu Asp Thr Ala Val Tyr Tyr Cys
100 105 110
Gln Gln Ser Lys Glu Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Val
115 120 125
Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
130 135 140
Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu
145 150 155 160
Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn
165 170 175
Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser
180 185 190
Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala
195 200 205
Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly
210 215 220
Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 110
<211> 2073
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.12（κ,IgG4）的重链-VID融合亚基（BY24.12H）核苷酸序列
<400> 110
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gcgaggtcca gctcgtcgag tccggcggag gcctcgtgca gcccggcgga 120
tctctgagac tcagttgtgc cgctagtggc tttacatttt cttcctacgg catgtcttgg 180
gtgagacagg ctcctggaaa gggattagag tgggtcgcaa ctattagtgg cggaggaagc 240
gacacatact acgccgattc cgtcaaggga cggttcacca tcagtcgcga taactctaag 300
aacacactgt acctacagat gaatagcctg agagcagagg ataccgctgt gtactactgc 360
gcacgccagc tcaattacgc atggtttgct tactggggcc agggcaccct cgtgacagtg 420
tcttccgcct ccaccaaggg cccttccgtg ttccctctgg ccccttgctc ccgctccacc 480
tccgagtcca ccgccgccct gggctgcctg gtgaaggact acttccctga gcctgtgacc 540
gtgtcctgga actccggcgc cctgacctcc ggcgtgcaca ccttccctgc cgtgctgcag 600
tcctccggcc tgtactccct gtcctccgtg gtgaccgtgc cttcctcctc cctgggcacc 660
aagacctaca cctgcaacgt ggaccacaag ccttccaaca ccaaggtgga caagcgcgtg 720
gagtccaagt acggccctcc ttgccctcct tgccctgccc ctgagttcct gggcggccct 780
tccgtgttcc tgttccctcc taagcctaag gacaccctga tgatctcccg cacccctgag 840
gtgacctgcg tggtggtgga cgtgtcccag gaggaccctg aggtgcagtt caactggtac 900
gtggacggcg tggaggtgca caacgccaag accaagcctc gcgaggagca gttcaactcc 960
acctaccgcg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaggag 1020
tacaagtgca aggtgtccaa caagggcctg ccttcctcca tcgagaagac catctccaag 1080
gccaagggcc agcctcgcga gcctcaggtg tacaccctgc ctccttccca ggaggagatg 1140
accaagaacc aggtgtccct gacctgcctg gtgaagggct tctacccttc cgacatcgcc 1200
gtggagtggg agtccaacgg ccagcctgag aacaactaca agaccacccc tcctgtgctg 1260
gactccgacg gctccttctt cctgtactcc cgcctgaccg tggacaagtc ccgctggcag 1320
gagggcaacg tgttctcctg ctccgtgatg cacgaggccc tgcacaacca ctacacccag 1380
aagtccctgt ccctgtccct gggcggcgga ggatctggcg gcggaggcag tggaggcggc 1440
ggaagcgctt ccgacaccgg ccgccctttc gtggagatgt actccgagat ccctgagatc 1500
atccacatga ccgagggccg cgagctggtg atcccttgcc gcgtgacctc ccctaacatc 1560
accgtgaccc tgaagaagtt ccctctggac accctgatcc ctgacggcaa gcgcatcatc 1620
tgggactccc gcaagggctt catcatctcc aacgccacct acaaggagat cggcctgctg 1680
acctgcgagg ccaccgtgaa cggccacctg tacaagacca actacctgac ccaccgccag 1740
accaacacca tcatcgacgt ggtgctgtcc ccttcccacg gcatcgagct gtccgtgggc 1800
gagaagctgg tgctgaactg caccgcccgc accgagctga acgtgggcat cgacttcaac 1860
tgggagtacc cttcctccaa gcaccagcac aagaagctgg tgaaccgcga cctgaagacc 1920
cagtccggct ccgagatgaa gaagttcctg tccaccctga ccatcgacgg cgtgacccgc 1980
tccgaccagg gcctgtacac ctgcgccgcc tcctccggcc tgatgaccaa gaagaactcc 2040
accttcgtgc gcgtgcacga gaagtaagtc gac 2073
<210> 111
<211> 684
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.12（κ,IgG4）的重链-VID融合亚基（BY24.12H）氨基酸序列
<400> 111
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
20 25 30
Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
35 40 45
Phe Ser Ser Tyr Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
50 55 60
Leu Glu Trp Val Ala Thr Ile Ser Gly Gly Gly Ser Asp Thr Tyr Tyr
65 70 75 80
Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
85 90 95
Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Arg Gln Leu Asn Tyr Ala Trp Phe Ala Tyr Trp
115 120 125
Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
130 135 140
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr
145 150 155 160
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr
165 170 175
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
180 185 190
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr
195 200 205
Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp
210 215 220
His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr
225 230 235 240
Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro
245 250 255
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
260 265 270
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp
275 280 285
Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
290 295 300
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val
305 310 315 320
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
325 330 335
Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys
340 345 350
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
355 360 365
Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
370 375 380
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
385 390 395 400
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
405 410 415
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys
420 425 430
Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu
435 440 445
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly
450 455 460
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser
465 470 475 480
Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro Glu Ile
485 490 495
Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg Val Thr
500 505 510
Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp Thr Leu
515 520 525
Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly Phe Ile
530 535 540
Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys Glu Ala
545 550 555 560
Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His Arg Gln
565 570 575
Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly Ile Glu
580 585 590
Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg Thr Glu
595 600 605
Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser Lys His
610 615 620
Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser Gly Ser
625 630 635 640
Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val Thr Arg
645 650 655
Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu Met Thr
660 665 670
Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680
<210> 112
<211> 738
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.13（κ,IgG4）的轻链亚基（BY24.13L）核苷酸序列
<400> 112
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gcgatattca gatgacccag agtccatcta gcgtgtctgc ttctgtgggc 120
gatcgggtga caatcacttg tcgcgcaagt cagggaatta gtagttggct cgcatggtat 180
cagcagaagc ctggcaaggc acctaagctc ctcattagcg ccgcgtcatc cctgcaatcc 240
ggcgtgccat ctaggtttag tggttccgga agcggaaccg actttacact cactatcagt 300
tctctccagc ccgaggattt cgcaacatac tactgtcagc aggccaacca cctgcctttc 360
acatttggag gcggcacatt cggcggcgga actaaggtgg agattaagag aaccgtggcc 420
gcccccagcg tgttcatctt ccctcccagc gacgagcagc tgaagtctgg caccgccagc 480
gtggtgtgcc tgctgaacaa cttctacccc cgcgaggcca aggtgcagtg gaaggtggac 540
aacgccctgc agagcggcaa cagccaggag agcgtgaccg agcaggactc caaggacagc 600
acctacagcc tgagcagcac cctgaccctg agcaaggccg actacgagaa gcacaaggtg 660
tacgcctgcg aggtgaccca ccagggactg tctagccccg tgaccaagag cttcaaccgg 720
ggcgagtgct aagaattc 738
<210> 113
<211> 234
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.13（κ,IgG4）的轻链亚基（BY24.13L）氨基酸序列
<400> 113
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly
35 40 45
Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
50 55 60
Lys Leu Leu Ile Ser Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser
65 70 75 80
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn
100 105 110
His Leu Pro Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 114
<211> 2079
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.13（κ,IgG4）的重链-VID融合亚基（BY24.13H）核苷酸序列
<400> 114
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccaggtgca gctcgtccag tctggcgctg aggtcaagaa gcctggatct 120
agcgtgaagg tgtcttgtaa ggcaagtggc ggaacctttt ctagttacgc tattagttgg 180
gtgcggcagg ctcccggcca gggcctggag tggatgggac tcatcattcc tatgttcgat 240
accgccggct acgcccagaa gtttcaggga cgggtcgcaa tcaccgtaga tgagagtacc 300
tccacagcat acatggagct gagtagtctc agatccgagg acactgccgt gtactactgt 360
gctcgcgcag agcactctag caccggaaca tttgattact ggggacaggg caccctcgtg 420
acagtgtctt ccgcctccac caagggccct tccgtgttcc ctctggcccc ttgctcccgc 480
tccacctccg agtccaccgc cgccctgggc tgcctggtga aggactactt ccctgagcct 540
gtgaccgtgt cctggaactc cggcgccctg acctccggcg tgcacacctt ccctgccgtg 600
ctgcagtcct ccggcctgta ctccctgtcc tccgtggtga ccgtgccttc ctcctccctg 660
ggcaccaaga cctacacctg caacgtggac cacaagcctt ccaacaccaa ggtggacaag 720
cgcgtggagt ccaagtacgg ccctccttgc cctccttgcc ctgcccctga gttcctgggc 780
ggcccttccg tgttcctgtt ccctcctaag cctaaggaca ccctgatgat ctcccgcacc 840
cctgaggtga cctgcgtggt ggtggacgtg tcccaggagg accctgaggt gcagttcaac 900
tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcctcgcga ggagcagttc 960
aactccacct accgcgtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 1020
aaggagtaca agtgcaaggt gtccaacaag ggcctgcctt cctccatcga gaagaccatc 1080
tccaaggcca agggccagcc tcgcgagcct caggtgtaca ccctgcctcc ttcccaggag 1140
gagatgacca agaaccaggt gtccctgacc tgcctggtga agggcttcta cccttccgac 1200
atcgccgtgg agtgggagtc caacggccag cctgagaaca actacaagac cacccctcct 1260
gtgctggact ccgacggctc cttcttcctg tactcccgcc tgaccgtgga caagtcccgc 1320
tggcaggagg gcaacgtgtt ctcctgctcc gtgatgcacg aggccctgca caaccactac 1380
acccagaagt ccctgtccct gtccctgggc ggcggaggat ctggcggcgg aggcagtgga 1440
ggcggcggaa gcgcttccga caccggccgc cctttcgtgg agatgtactc cgagatccct 1500
gagatcatcc acatgaccga gggccgcgag ctggtgatcc cttgccgcgt gacctcccct 1560
aacatcaccg tgaccctgaa gaagttccct ctggacaccc tgatccctga cggcaagcgc 1620
atcatctggg actcccgcaa gggcttcatc atctccaacg ccacctacaa ggagatcggc 1680
ctgctgacct gcgaggccac cgtgaacggc cacctgtaca agaccaacta cctgacccac 1740
cgccagacca acaccatcat cgacgtggtg ctgtcccctt cccacggcat cgagctgtcc 1800
gtgggcgaga agctggtgct gaactgcacc gcccgcaccg agctgaacgt gggcatcgac 1860
ttcaactggg agtacccttc ctccaagcac cagcacaaga agctggtgaa ccgcgacctg 1920
aagacccagt ccggctccga gatgaagaag ttcctgtcca ccctgaccat cgacggcgtg 1980
acccgctccg accagggcct gtacacctgc gccgcctcct ccggcctgat gaccaagaag 2040
aactccacct tcgtgcgcgt gcacgagaag taagtcgac 2079
<210> 115
<211> 686
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.13（κ,IgG4）的重链-VID融合亚基（BY24.13H）氨基酸序列
<400> 115
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
20 25 30
Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr
35 40 45
Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly
50 55 60
Leu Glu Trp Met Gly Leu Ile Ile Pro Met Phe Asp Thr Ala Gly Tyr
65 70 75 80
Ala Gln Lys Phe Gln Gly Arg Val Ala Ile Thr Val Asp Glu Ser Thr
85 90 95
Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
100 105 110
Val Tyr Tyr Cys Ala Arg Ala Glu His Ser Ser Thr Gly Thr Phe Asp
115 120 125
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys
130 135 140
Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
145 150 155 160
Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
165 170 175
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
180 185 190
Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
195 200 205
Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn
210 215 220
Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser
225 230 235 240
Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly
245 250 255
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
260 265 270
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln
275 280 285
Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
290 295 300
His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr
305 310 315 320
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
325 330 335
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile
340 345 350
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
355 360 365
Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser
370 375 380
Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
385 390 395 400
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
405 410 415
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val
420 425 430
Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met
435 440 445
His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
450 455 460
Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
465 470 475 480
Ala Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr Ser Glu Ile Pro
485 490 495
Glu Ile Ile His Met Thr Glu Gly Arg Glu Leu Val Ile Pro Cys Arg
500 505 510
Val Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys Phe Pro Leu Asp
515 520 525
Thr Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp Ser Arg Lys Gly
530 535 540
Phe Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly Leu Leu Thr Cys
545 550 555 560
Glu Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn Tyr Leu Thr His
565 570 575
Arg Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser Pro Ser His Gly
580 585 590
Ile Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn Cys Thr Ala Arg
595 600 605
Thr Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu Tyr Pro Ser Ser
610 615 620
Lys His Gln His Lys Lys Leu Val Asn Arg Asp Leu Lys Thr Gln Ser
625 630 635 640
Gly Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr Ile Asp Gly Val
645 650 655
Thr Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala Ser Ser Gly Leu
660 665 670
Met Thr Lys Lys Asn Ser Thr Phe Val Arg Val His Glu Lys
675 680 685
<210> 116
<211> 747
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.14（κ,IgG4）的轻链亚基（BY24.14L）核苷酸序列
<400> 116
ctcgaggcca ccatggagac cgacacactc ctcctgtggg tgctgctgct gtgggtgcct 60
ggctccactg gccagagcgc tctcactcag cctgcttccg tgtctggaag tcccggccag 120
agtatcacta tttcttgtac aggaacttcc tccgacgtcg gattttacaa ttacgtcagt 180
tggtatcagc agcaccccgg aaaggcacct gaactaatga tctacgatgt gtctaaccgc 240
ccaagcggcg tgagcgatag gttcagtggc agtaagagtg gcaacaccgc atccctgacc 300
attagtggat tacaggccga ggacgaggct gattactact gttctagcta cacaaatatc 360
tccacatggg tcttcggcgg aggaacattc ggcggcggaa ctaaggtgga gattaagaga 420
accgtggccg cccccagcgt gttcatcttc cctcccagcg acgagcagct gaagtctggc 480
accgccagcg tggtgtgcct gctgaacaac ttctaccccc gcgaggccaa ggtgcagtgg 540
aaggtggaca acgccctgca gagcggcaac agccaggaga gcgtgaccga gcaggactcc 600
aaggacagca cctacagcct gagcagcacc ctgaccctga gcaaggccga ctacgagaag 660
cacaaggtgt acgcctgcga ggtgacccac cagggactgt ctagccccgt gaccaagagc 720
ttcaaccggg gcgagtgcta agaattc 747
<210> 117
<211> 237
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.14（κ,IgG4）的轻链亚基（BY24.14L）氨基酸序列
<400> 117
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly
20 25 30
Ser Pro Gly Gln Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp
35 40 45
Val Gly Phe Tyr Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys
50 55 60
Ala Pro Glu Leu Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val
65 70 75 80
Ser Asp Arg Phe Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr
85 90 95
Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser
100 105 110
Tyr Thr Asn Ile Ser Thr Trp Val Phe Gly Gly Gly Thr Lys Val Glu
115 120 125
Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser
130 135 140
Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn
145 150 155 160
Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala
165 170 175
Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys
180 185 190
Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp
195 200 205
Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu
210 215 220
Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230 235
<210> 118
<211> 2091
<212> DNA
<213> 人工序列
<220>
<223> 融合蛋白BY24.14（κ,IgG4）的重链-VID融合亚基（BY24.14H）核苷酸序列
<400> 118
tctagagcca ccatggagac cgacaccctg ctgctgtggg tgctgctcct gtgggtgcct 60
ggctccacag gccagctcca gcttcaggag agcggacccg gcctggtcaa gccatccgag 120
actctcactc tgacatgcac cgtgagtgct gattctatca gttccacaac ttactactgg 180
gtgtggatta ggcagcctcc cggaaaggga ttagaatgga tcggcagcat ttcttacagt 240
ggctccacat actacaatcc tagtctgaag tctcgcgtga ccgtgtccgt ggatacatct 300
aagaaccagt ttagcctcaa gctgaatagc gtcgccgcaa cagataccgc tctgtactac 360
tgcgcacgcc acctcggcta caatggacgc tatctgccct tcgattactg gggccaggga 420
agcaccctcg tgacagtgtc ttccgcctcc accaagggcc cttccgtgtt ccctctggcc 480
ccttgctccc gctccacctc cgagtccacc gccgccctgg gctgcctggt gaaggactac 540
ttccctgagc ctgtgaccgt gtcctggaac tccggcgccc tgacctccgg cgtgcacacc 600
ttccctgccg tgctgcagtc ctccggcctg tactccctgt cctccgtggt gaccgtgcct 660
tcctcctccc tgggcaccaa gacctacacc tgcaacgtgg accacaagcc ttccaacacc 720
aaggtggaca agcgcgtgga gtccaagtac ggccctcctt gccctccttg ccctgcccct 780
gagttcctgg gcggcccttc cgtgttcctg ttccctccta agcctaagga caccctgatg 840
atctcccgca cccctgaggt gacctgcgtg gtggtggacg tgtcccagga ggaccctgag 900
gtgcagttca actggtacgt ggacggcgtg gaggtgcaca acgccaagac caagcctcgc 960
gaggagcagt tcaactccac ctaccgcgtg gtgtccgtgc tgaccgtgct gcaccaggac 1020
tggctgaacg gcaaggagta caagtgcaag gtgtccaaca agggcctgcc ttcctccatc 1080
gagaagacca tctccaaggc caagggccag cctcgcgagc ctcaggtgta caccctgcct 1140
ccttcccagg aggagatgac caagaaccag gtgtccctga cctgcctggt gaagggcttc 1200
tacccttccg acatcgccgt ggagtgggag tccaacggcc agcctgagaa caactacaag 1260
accacccctc ctgtgctgga ctccgacggc tccttcttcc tgtactcccg cctgaccgtg 1320
gacaagtccc gctggcagga gggcaacgtg ttctcctgct ccgtgatgca cgaggccctg 1380
cacaaccact acacccagaa gtccctgtcc ctgtccctgg gcggcggagg atctggcggc 1440
ggaggcagtg gaggcggcgg aagcgcttcc gacaccggcc gccctttcgt ggagatgtac 1500
tccgagatcc ctgagatcat ccacatgacc gagggccgcg agctggtgat cccttgccgc 1560
gtgacctccc ctaacatcac cgtgaccctg aagaagttcc ctctggacac cctgatccct 1620
gacggcaagc gcatcatctg ggactcccgc aagggcttca tcatctccaa cgccacctac 1680
aaggagatcg gcctgctgac ctgcgaggcc accgtgaacg gccacctgta caagaccaac 1740
tacctgaccc accgccagac caacaccatc atcgacgtgg tgctgtcccc ttcccacggc 1800
atcgagctgt ccgtgggcga gaagctggtg ctgaactgca ccgcccgcac cgagctgaac 1860
gtgggcatcg acttcaactg ggagtaccct tcctccaagc accagcacaa gaagctggtg 1920
aaccgcgacc tgaagaccca gtccggctcc gagatgaaga agttcctgtc caccctgacc 1980
atcgacggcg tgacccgctc cgaccagggc ctgtacacct gcgccgcctc ctccggcctg 2040
atgaccaaga agaactccac cttcgtgcgc gtgcacgaga agtaagtcga c 2091
<210> 119
<211> 690
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白BY24.14（κ,IgG4）的重链-VID融合亚基（BY24.14H）氨基酸序列
<400> 119
Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro
1 5 10 15
Gly Ser Thr Gly Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val
20 25 30
Lys Pro Ser Glu Thr Leu Thr Leu Thr Cys Thr Val Ser Ala Asp Ser
35 40 45
Ile Ser Ser Thr Thr Tyr Tyr Trp Val Trp Ile Arg Gln Pro Pro Gly
50 55 60
Lys Gly Leu Glu Trp Ile Gly Ser Ile Ser Tyr Ser Gly Ser Thr Tyr
65 70 75 80
Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Val Ser Val Asp Thr Ser
85 90 95
Lys Asn Gln Phe Ser Leu Lys Leu Asn Ser Val Ala Ala Thr Asp Thr
100 105 110
Ala Leu Tyr Tyr Cys Ala Arg His Leu Gly Tyr Asn Gly Arg Tyr Leu
115 120 125
Pro Phe Asp Tyr Trp Gly Gln Gly Ser Thr Leu Val Thr Val Ser Ser
130 135 140
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
145 150 155 160
Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
165 170 175
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
180 185 190
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
195 200 205
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr
210 215 220
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
225 230 235 240
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro
245 250 255
Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
260 265 270
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
275 280 285
Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
290 295 300
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
305 310 315 320
Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
325 330 335
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu
340 345 350
Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
355 360 365
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys
370 375 380
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
385 390 395 400
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
405 410 415
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
420 425 430
Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
435 440 445
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
450 455 460
Leu Ser Leu Ser Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
465 470 475 480
Gly Gly Gly Ser Ala Ser Asp Thr Gly Arg Pro Phe Val Glu Met Tyr
485 490 495
Ser Glu Ile Pro Glu Ile Ile His Met Thr Glu Gly Arg Glu Leu Val
500 505 510
Ile Pro Cys Arg Val Thr Ser Pro Asn Ile Thr Val Thr Leu Lys Lys
515 520 525
Phe Pro Leu Asp Thr Leu Ile Pro Asp Gly Lys Arg Ile Ile Trp Asp
530 535 540
Ser Arg Lys Gly Phe Ile Ile Ser Asn Ala Thr Tyr Lys Glu Ile Gly
545 550 555 560
Leu Leu Thr Cys Glu Ala Thr Val Asn Gly His Leu Tyr Lys Thr Asn
565 570 575
Tyr Leu Thr His Arg Gln Thr Asn Thr Ile Ile Asp Val Val Leu Ser
580 585 590
Pro Ser His Gly Ile Glu Leu Ser Val Gly Glu Lys Leu Val Leu Asn
595 600 605
Cys Thr Ala Arg Thr Glu Leu Asn Val Gly Ile Asp Phe Asn Trp Glu
610 615 620
Tyr Pro Ser Ser Lys His Gln His Lys Lys Leu Val Asn Arg Asp Leu
625 630 635 640
Lys Thr Gln Ser Gly Ser Glu Met Lys Lys Phe Leu Ser Thr Leu Thr
645 650 655
Ile Asp Gly Val Thr Arg Ser Asp Gln Gly Leu Tyr Thr Cys Ala Ala
660 665 670
Ser Ser Gly Leu Met Thr Lys Lys Asn Ser Thr Phe Val Arg Val His
675 680 685
Glu Lys
690
<210> 120
<211> 106
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的重链可变区
<400> 120
Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Thr Cys Lys Ala Ser Gly Leu Thr Phe Ser Ser Ser
20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Thr Asn Asn Asp Tyr Trp Gly Gln Gly
100 105
<210> 121
<211> 101
<212> PRT
<213> 人工序列
<220>
<223> 融合蛋白中的一种抗PD-1抗体的轻链可变区
<400> 121
Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr
20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45
Tyr Thr Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
65 70 75 80
Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ser Asn Trp Pro Arg
85 90 95
Thr Phe Gly Gln Gly
100

Claims (25)

1.靶向PD-1且靶向VEGF家族的双靶向融合蛋白，所述双靶向融合蛋白抑制PD-1与其配体的结合、且抑制VEGF家族的信号传导途径，其包含
(i)抗PD-1抗体；和
(ii)在所述抗PD-1抗体的两条重链中的每一重链的C端有效连接的一个抑制VEGF家族的结构域，其中所述抑制VEGF家族的结构域也缩写为VID，所述VID是人VEGFR1的免疫球蛋白样结构域2和人VEGFR2的免疫球蛋白样结构域3，为SEQ ID NO：63所示的氨基酸序列；由此，两个相同的VID各自在它们的N端氨基酸处与所述抗PD-1抗体的重链之一的C端氨基酸通过肽接头有效连接，其中所述肽接头是选自SEQ ID NO：36－62的肽接头；
其中所述抗PD-1抗体包含SEQ ID NO：1/2的成对重链可变区序列/轻链可变区序列中所含的全部重链CDR与轻链CDR。
2.根据权利要求1所述的双靶向融合蛋白，其中所述抗PD-1抗体是IgG类抗体。
3.根据权利要求2所述的双靶向融合蛋白，其中所述抗PD-1抗体是IgG1亚类、IgG2亚类或IgG4亚类抗体。
4.根据权利要求3所述的双靶向融合蛋白，其中所述抗PD-1抗体是IgG4亚类抗体，且在Fc区中第S228位置处具有氨基酸置换。
5.根据权利要求4所述的双靶向融合蛋白，其中所述抗PD-1抗体是IgG4亚类抗体，且在Fc区中第S228位置处具有氨基酸置换S228P。
6.根据权利要求1所述的双靶向融合蛋白，其中所述抗PD-1抗体的轻链型别为κ型或λ型。
7.根据权利要求1所述的双靶向融合蛋白，其中所述抗PD-1抗体包含SEQ ID NO：1/2的成对重链可变区序列/轻链可变区序列。
8.根据权利要求7所述的双靶向融合蛋白，其中所述抗PD-1抗体是纳武单抗。
9.根据权利要求1所述的双靶向融合蛋白，其是
由SEQ ID NO: 73的抗PD-1抗体轻链亚基和SEQ ID NO: 75的抗PD-1抗体重链-VID融合亚基组成的双靶向融合蛋白。
10.多核苷酸，其编码权利要求1－9中任一项所述的双靶向融合蛋白。
11.载体，所述载体包含权利要求10所述的多核苷酸。
12.根据权利要求11所述的载体，其是表达载体。
13.根据权利要求11所述的载体，其是具有双表达盒的谷氨酰胺合成酶表达载体。
14.宿主细胞，其包含权利要求10所述的多核苷酸或权利要求11－13中任一项所述的载体。
15.用于产生权利要求1－9中任一项所述的双靶向融合蛋白的方法，所述方法包括步骤(i)在适于表达所述双靶向融合蛋白的条件下培养权利要求14所述的宿主细胞，和(ii)回收所述双靶向融合蛋白。
16.药物组合物，其包含权利要求1－9中任一项所述的双靶向融合蛋白和可药用载体。
17.权利要求1－9中任一项所述的双靶向融合蛋白和权利要求16所述的药物组合物用于制备在个体中治疗或预防与PD-1活性和VEGF家族活性相关的癌性疾病的药物中的用途，其中所述癌性疾病是黑素瘤、乳腺癌、结肠癌、食管癌、胃肠道间质肿瘤、肾癌、肝癌、非小细胞肺癌、卵巢癌、胰腺癌、前列腺癌、头颈部肿瘤、胃癌、血液学恶性病，所述个体是哺乳动物。
18.权利要求17所述的用途，其中所述个体是人。
19.权利要求18所述的用途，其中所述肾癌是肾细胞癌，所述血液学恶性病是淋巴瘤。
20.诊断试剂盒，其包含权利要求1－9中任一项所述的融合蛋白和任选地标记物或用于偶联的试剂。
21.根据权利要求20所述的诊断试剂盒，其包含用正电子发射断层摄影术可检测的标记物标记的权利要求1－9中任一项所述的融合蛋白。
22.根据权利要求21所述的诊断试剂盒，其中所述标记物是18F-氟脱氧葡萄糖。
23.权利要求20－22中任一项所述的诊断试剂盒用于制备在个体中诊断与PD-1活性相关的癌性疾病的产品中的用途，其中所述癌性疾病是黑素瘤、乳腺癌、结肠癌、食管癌、胃肠道间质肿瘤、肾癌、肝癌、非小细胞肺癌、卵巢癌、胰腺癌、前列腺癌、头颈部肿瘤、胃癌、血液学恶性病，所述个体是哺乳动物。
24.权利要求23所述的用途，其中所述个体是人。
25.权利要求24所述的用途，其中所述肾癌是肾细胞癌，所述血液学恶性病是淋巴瘤。
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