CN115043934B - 靶向新冠病毒的纳米抗体及其制备方法和应用 - Google Patents
靶向新冠病毒的纳米抗体及其制备方法和应用 Download PDFInfo
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Abstract
本发明实施例公开了一种靶向新冠病毒的纳米抗体及其制备方法和应用，所述纳米抗体包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，提供的纳米抗体对新冠病毒野生株、新冠病毒变异株阿尔法(Alpha)、新冠病毒贝塔(Beta)、新冠病毒德尔塔(Delta)、新冠病毒Omicron及其亚型BA.2的RBD结构域具有较高的亲和力，能够快速且特异性地中和不同的新冠病毒，有效抑制其活性；同时，提供的纳米抗体具有更长的CDR3序列，有利于纳米抗体进行灵活调节其构象，提高与病毒的结合作用，有利于广泛应用。
Description
本申请是靶向新冠病毒的纳米抗体及其制备方法和应用的分案申请，原申请的申请日为2022年5月23日，申请号为202210330386.7，发明创造名称为靶向新冠病毒的纳米抗体及其制备方法和应用。
技术领域
本发明涉及生物技术领域，尤其涉及一种靶向新冠病毒的纳米抗体及其制备方法和应用。
背景技术
新型冠状病毒肺炎(Corona Virus Disease 2019，COVID-19)可通过飞沫传播、接触呼吸道分泌物、气溶胶传播等途径传播，带来的后果十分严重。引起 COVID-19的急性呼吸系统综合症冠状病毒2(Severe acute respiratory syndrome coronavirus 2，SARS-CoV-2)主要表达4种结构蛋白，分别为刺突蛋白(spike，S)、包膜蛋白(envelope)、膜蛋白(membrane)和核衣壳蛋白(nucleocapsid)。其中S蛋白包括N端的S1亚基和C末端的S2亚基，S1亚基包含大约200个氨基酸的受体结合域(receptor binding domain，RBD)负责病毒与宿主受体结合，S2亚基负责病毒与宿主细胞膜融合。S蛋白在病毒附着、感染和传播中发挥至关重要的作用，通过与宿主细胞的血管紧张素转化酶2(angiotensin convertingenzyme2，ACE2) 结合进而感染细胞，是抗SARS-CoV-2药物研发的重要靶点。
中和抗体是对抗COVID-19的重要疗法。已有研究发现通过给重症 COVID-19患者输送康复者的血清能够显著改善临床结果，表明中和抗体疗法对新冠重症患者也有较好的缓解作用。其作用机理为中和抗体与新冠病毒表面的刺突糖蛋白结合进而影响SARS-CoV-2与宿主细胞的表面受体结合，从而使病毒无法感染靶细胞。目前已有多个中和抗体获批上市用于COVID-19的治疗，这些单抗靶点主要集中在S蛋白的RBD结构域。然而，面对不断突变的SARS-CoV-2，已有中和抗体很难靶向新的新冠变异株。最近的一项研究表明，omicron亚型BA.2 可逃避几乎所有的中和抗体治疗。因此，迫切需要开发出新的可以同时靶向不同新冠变异株特别是omicron及其亚型BA.2的中和抗体。
发明内容
本申请的目的在于提供一种靶向新冠病毒的纳米抗体及其制备方法和应用，旨在解决现有技术中缺乏新冠变异株Omicron及其亚型BA.2的中和抗体的问题。
为实现上述申请目的，本申请采用的技术方案如下：
第一方面，本申请公开了一种靶向新冠病毒的纳米抗体，纳米抗体包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，其中，纳米抗体NbN2的氨基酸序列如Seq.ID NO.1所示，纳米抗体NbN3的氨基酸序列如Seq.ID NO.2所示，纳米抗体NbN4 的氨基酸序列如Seq.ID NO.3所示，纳米抗体NbN5的氨基酸序列如Seq.ID NO.4 所示，纳米抗体NbN6的氨基酸序列如Seq.ID NO.5所示，纳米抗体NbN11的氨基酸序列如Seq.ID NO.6所示。
第二方面，本申请公开了一种靶向新冠病毒的纳米抗体的制备方法，包括如下步骤：
提供目的蛋白，将目的蛋白包被在免疫管上进行富集筛选，得到噬菌体文库；
将噬菌体文库的洗脱液进行二代测序，并根据测序结果合成原始计数占比前 10的纳米抗体的基因序列；
将纳米抗体的基因序列克隆至表达载体中得到重组质粒，将重组质粒转入宿主细胞中诱导表达并进行纯化，得到靶向新冠病毒的纳米抗体。
第三方面，本申请公开了靶向新冠病毒的纳米抗体在制备预防和/或治疗新型冠状病毒肺炎的药物中的应用。
第四方面，本申请公开了靶向新冠病毒的纳米抗体在制备检测新型冠状病毒的检测试剂或检测试剂盒中的应用。
本申请第一方面提供的新冠病毒Omicron及其亚型BA.2的纳米抗体，包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体 NbN6、纳米抗体NbN11中的至少一种，提供的纳米抗体对新冠病毒野生株、新冠病毒变异株阿尔法(Alpha)、新冠病毒贝塔(Beta)、新冠病毒德尔塔(Delta)、新冠病毒Omicron及其亚型BA.2的RBD结构域具有较高的亲和力，能够快速且特异性地中和不同的新冠病毒，有效抑制其活性；同时，提供的纳米抗体具有更长的CDR3序列，有利于纳米抗体进行灵活调节其构象，提高与病毒的结合作用，且制备成本较低，有利于广泛应用。
本申请第二方面提供的一种靶向新冠病毒Omicron及其亚型BA.2的纳米抗体的制备方法，该制备方法基于噬菌体展示筛选技术，以新冠病毒Omicron及其亚型BA.2的RBD蛋白为靶标构建了纳米抗体库，并结合二代测序、序列合成、克隆、表达、纯化，得靶向新冠病毒Omicron及其亚型BA.2的纳米抗体；该制备方法利用噬菌体在宿主菌中快速复制的生物特性，能够快速、高通量地筛选与特定变异株奥密克戎及其亚型BA.2RBD蛋白结合的纳米抗体，制备方法快速、简便，有利于大量筛选，提高了筛选效率。
本申请第三方面提供的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体在制备预防和/或治疗新型冠状病毒肺炎的药物中的应用，由于筛选得到的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，对新冠病毒野生株及各变异株的病毒具有较好的中和活性能力和抑制活性能力，适用于制备预防和/或治疗新型冠状病毒肺炎的相关药物。
本申请第四方面提供的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体在制备检测新型冠状病毒的检测试剂或检测试剂盒中的应用，提供的靶向新冠病毒 Omicron及其亚型BA.2的纳米抗体包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，对新冠病毒野生株及各变异株的病毒具有较高得结合能力，适用于制备检测新型冠状病毒的检测试剂或检测试剂盒。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
其中：
图1为Omicron RBD蛋白纳米抗体筛选示意图。
图2为Omicron RBD筛选第三轮噬菌体洗脱液二代测序top 10序列示意图。
图3为Omicron RBD纳米抗体表达纯化示意图。
图4为纳米抗体免疫印迹(Western Blot，WB)检测示意图。
图5为Omicron RBD纳米抗体ELISA验证示意图。
图6为SARS-CoV-2野生株(Wild Type，WT)RBD纳米抗体ELISA验证示意图。
图7为Delta RBD纳米抗体ELISA验证示意图。
图8为纳米抗体与阴性对照蛋白牛血清白蛋白(Bovine Serum Albumin，BSA)ELISA验证示意图。
图9为Biacore检测纳米抗体N2与WT RBD蛋白的亲和常数示意图。
图10为Biacore检测纳米抗体N2与Alpha RBD蛋白的亲和常数示意图。
图11为Biacore检测纳米抗体N2与Beta RBD蛋白的亲和常数示意图。
图12为Biacore检测纳米抗体N2与Delta RBD蛋白的亲和常数示意图。
图13为Biacore检测纳米抗体N2与Omicron RBD蛋白的亲和常数示意图。
图14为Biacore检测纳米抗体N2与Omicron BA.2RBD蛋白的亲和常数示意图。
图15为Biacore检测纳米抗体N3与WT RBD蛋白的亲和常数示意图。
图16为Biacore检测纳米抗体N3与Alpha RBD蛋白的亲和常数示意图。
图17为Biacore检测纳米抗体N3与Beta RBD蛋白的亲和常数示意图。
图18为Biacore检测纳米抗体N3与Delta RBD蛋白的亲和常数示意图。
图19为Biacore检测纳米抗体N3与Omicron RBD蛋白的亲和常数示意图。
图20为Biacore检测纳米抗体N3与Omicron BA.2RBD蛋白的亲和常数示意图。
图21为Biacore检测纳米抗体N4与WT RBD蛋白的亲和常数示意图。
图22为Biacore检测纳米抗体N4与Alpha RBD蛋白的亲和常数示意图。
图23为Biacore检测纳米抗体N4与Beta RBD蛋白的亲和常数示意图。
图24为Biacore检测纳米抗体N4与Delta RBD蛋白的亲和常数示意图。
图25为Biacore检测纳米抗体N4与Omicron RBD蛋白的亲和常数示意图。
图26为Biacore检测纳米抗体N4与Omicron BA.2RBD蛋白的亲和常数示意图。
图27为Biacore检测纳米抗体N5与WT RBD蛋白的亲和常数示意图。
图28为Biacore检测纳米抗体N5与Alpha RBD蛋白的亲和常数示意图。
图29为Biacore检测纳米抗体N5与Beta RBD蛋白的亲和常数示意图。
图30为Biacore检测纳米抗体N5与Delta RBD蛋白的亲和常示意图。
图31为Biacore检测纳米抗体N5与Omicron RBD蛋白的亲和常数示意图。
图32为Biacore检测纳米抗体N5与Omicron BA.2RBD蛋白的亲和常数示意图。
图33为Biacore检测纳米抗体N6与WT RBD蛋白的亲和常数示意图。
图34为Biacore检测纳米抗体N6与Alpha RBD蛋白的亲和常数示意图。
图35为Biacore检测纳米抗体N6与Beta RBD蛋白的亲和常数示意图。
图36为Biacore检测纳米抗体N6与Delta RBD蛋白的亲和常数示意图。
图37为Biacore检测纳米抗体N6与Omicron RBD蛋白的亲和常数示意图。
图38为Biacore检测纳米抗体N6与Omicron BA.2RBD蛋白的亲和常数示意图。
图39为Biacore检测纳米抗体N11与WT RBD蛋白的亲和常数示意图。
图40为Biacore检测纳米抗体N11与Alpha RBD蛋白的亲和常数示意图。
图41为Biacore检测纳米抗体N11与Beta RBD蛋白的亲和常数示意图。
图42为Biacore检测纳米抗体N11与Delta RBD蛋白的亲和常数示意图。
图43为Biacore检测纳米抗体N11与Omicron RBD蛋白的亲和常数示意图。
图44为Biacore检测纳米抗体N11与Omicron BA.2RBD蛋白的亲和常数示意图。
图45为纳米抗体与ACE2竞争性ELISA示意图。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
本申请实施例第一方面提供的一种靶向新冠病毒的纳米抗体，纳米抗体包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体 NbN6、纳米抗体NbN11中的至少一种，其中，纳米抗体NbN2的氨基酸序列如 Seq.ID NO.1所示，纳米抗体NbN3的氨基酸序列如Seq.ID NO.2所示，纳米抗体 NbN4的氨基酸序列如Seq.ID NO.3所示，纳米抗体NbN5的氨基酸序列如Seq.ID NO.4所示，纳米抗体NbN6的氨基酸序列如Seq.ID NO.5所示，纳米抗体NbN11 的氨基酸序列如Seq.ID NO.6所示。
本申请实施例第一方面提供的新冠病毒Omicron及其亚型BA.2的纳米抗体，包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，提供的纳米抗体对新冠病毒野生株、新冠病毒变异株阿尔法(Alpha)、新冠病毒贝塔(Beta)、新冠病毒德尔塔(Delta)、新冠病毒Omicron及其亚型BA.2的RBD结构域具有较高的亲和力，能够快速且特异性地中和不同的新冠病毒，有效抑制其活性；同时，提供的纳米抗体具有更长的CDR3序列，有利于纳米抗体进行灵活调节其构象，提高与病毒的结合作用，且制备成本较低，有利于广泛应用。
纳米抗体NbN2的氨基酸序列如Seq.ID NO.1所示，Seq.ID NO.1如下： MAVQLVESGGGLVQAGGSLRLSCAATGLTFTGYVMAWFRQAPGKNREFVAAV SRSGVVTRYADSVKGRFTVSRDNAKNTVYLQMNTLNPEDTALYYCAADWHP LTGIMTTDSTEYDYWGQGTQVTVSS。
纳米抗体NbN2的碱基序列如Seq.ID NO.44所示，Seq.ID NO.44如下： ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGATTGGTGCAGGCTGGGGG CTCTCTGAGACTCTCCTGTGCAGCCACTGGACTCACCTTCACTGGTTATGTA ATGGCCTGGTTCCGCCAGGCTCCAGGGAAGAACCGTGAGTTTGTAGCGGCC GTTAGCCGGAGTGGTGTTGTTACACGCTATGCAGACTCCGTGAAGGGCCGA TTCACCGTCTCCAGAGACAACGCCAAGAACACGGTGTATCTGCAAATGAAC ACCCTGAATCCTGAGGACACGGCCCTTTATTACTGTGCAGCAGATTGGCATCCACTAACTGGAATAATGACGACTGACTCTACAGAGTATGACTACTGGGGCCAGGGGACCCAGGTCACCGTCTCCTCA。
纳米抗体NbN3的氨基酸序列如Seq.ID NO.2所示，Seq.ID NO.2如下： MAVQLVESGGGLVQAGGSLRLSCAASGRTFSRYAMGWFRQAPGKERQFVAGI SGSGIVTNYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAIYYCAPLILARTV DMNHWGKGTQVTVSS。
纳米抗体NbN3的碱基序列如Seq.ID NO.45所示，Seq.ID NO.45如下： ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGATTGGTGCAGGCTGGGGG CTCTCTGAGACTCTCCTGTGCAGCCTCTGGACGCACCTTCAGTAGGTATGCC ATGGGCTGGTTCCGCCAGGCTCCAGGGAAGGAGCGCCAGTTTGTAGCAGGT ATTAGCGGGAGTGGTATAGTCACAAACTATGCAGACTCCGTGAAGGGCCGA TTCACCATCTCCAGAGACAACGCCAAGAACACGGTGTACCTGCAAATGAAC AGCCTGAAACCTGAGGACACGGCCATTTATTACTGTGCACCGTTGATCCTAGCACGAACTGTGGACATGAACCACTGGGGCAAAGGGACCCAGGTCACCGTC TCCTCA。
纳米抗体NbN4的氨基酸序列如Seq.ID NO.3所示，Seq.ID NO.3如下： MAVQLVESGGGVVQPGGSLRLSCTASGFGFSLYGMSWYRQAPGKERELVASIA SGGDSTNYQDSVKGRFSMSRDNAKSTVYLQMNSLKPEDTAVYYCVAERVSSF LKIRTAYWGQGTQVTVSS。
纳米抗体NbN4的碱基序列如Seq.ID NO.46所示，Seq.ID NO.46如下： ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGAGTGGTGCAGCCTGGGGG GTCTCTGAGACTCTCCTGTACAGCCTCTGGATTCGGCTTCAGTCTTTATGGC ATGAGCTGGTACCGTCAGGCTCCAGGGAAGGAGCGCGAGTTGGTCGCATCT ATTGCCAGTGGTGGTGATAGTACAAACTATCAAGACTCCGTGAAGGGCCGA TTCTCCATGTCCAGAGACAATGCCAAGAGCACGGTGTATCTGCAAATGAAC AGCCTGAAACCTGAGGACACGGCCGTGTATTACTGCGTGGCAGAAAGAGT CTCGTCGTTCTTAAAAATTCGTACCGCCTACTGGGGCCAGGGGACCCAGGT CACCGTCTCCTCA。
纳米抗体NbN5的氨基酸序列如Seq.ID NO.4所示，Seq.ID NO.4如下： MAVQLVESGGGLVQAGGSLRVSCEASGFLFNGDAYVLGWYRQVPGRQRELVA TITSSGDTSYADSVKGRFTISRDNAKNTIFLQMSGLKPEDTAVYYCYAERWNGL LQRWGQGTQVTVSS。
纳米抗体NbN5的碱基序列如Seq.ID NO.47所示，Seq.ID NO.47如下： ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAGGCTGGGGG GTCTCTGAGAGTGTCCTGTGAAGCCTCTGGATTCCTTTTCAATGGCGATGCC TATGTCTTGGGCTGGTACCGCCAGGTTCCAGGTAGGCAGCGTGAATTGGTC GCAACTATTACGAGTAGCGGTGACACAAGCTATGCGGACTCCGTGAAGGGC CGATTCACCATCTCCAGAGACAACGCCAAGAATACAATTTTTCTGCAAATG AGCGGCCTGAAACCCGAGGACACGGCCGTCTATTACTGCTATGCAGAGAGG TGGAATGGGCTCCTTCAACGCTGGGGCCAGGGGACCCAGGTCACCGTCTCC TCA。
纳米抗体NbN6的氨基酸序列如Seq.ID NO.5所示，Seq.ID NO.5如下： MAVQLVESGGGLVQPGGSLRLSCAASGIIFRFRTISWYRQAPGKQRELVASISG GSSTSYADTVKGRFTISRDNAKNTVYLQMNSLKPEDTGVYYCAASHTMVGWI YGMDYWGKGTQVTVSS。
纳米抗体NbN6的碱基序列如Seq.ID NO.48所示，Seq.ID NO.48如下： ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAACCTGGGGG GTCTCTGAGACTCTCCTGTGCAGCCTCTGGAATCATCTTCAGATTCAGAACC ATATCCTGGTACCGCCAGGCTCCAGGGAAGCAGCGCGAGTTGGTCGCAAGT ATTAGTGGCGGCAGTAGCACGTCCTATGCAGACACTGTGAAGGGCCGATTC ACCATCTCCAGAGACAACGCCAAGAACACGGTGTATCTGCAAATGAACAG CCTGAAACCTGAGGACACAGGCGTTTATTACTGTGCAGCGTCACATACTATG GTTGGGTGGATCTACGGCATGGACTACTGGGGCAAAGGGACCCAGGTCACC GTCTCCTCA。
纳米抗体NbN11的氨基酸序列如Seq.ID NO.6所示，Seq.ID NO.6如下： MAVQLVESGGGLVQAGGSLRLSCAASGSFFSINTMGWYRQAPGKQRELVAQIT NAGNTNYADSVKGRFTISRDNAKNTIYLQMNSLKPEDTAVYYCNAGQLWGSY RSWGQGTQVTVSS。
纳米抗体NbN11的碱基序列如Seq.ID NO.49所示，Seq.ID NO.49如下： ATGGCGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAGGCTGGGGG GTCTCTGAGACTCTCCTGTGCAGCCTCTGGAAGCTTCTTCAGTATCAATACC ATGGGCTGGTACCGCCAGGCTCCAGGGAAGCAGCGCGAGTTGGTCGCACA AATTACTAATGCAGGTAACACAAACTATGCAGACTCCGTGAAGGGCCGGTT CACCATCTCCAGAGACAACGCCAAGAACACGATATATCTGCAAATGAACAG CCTGAAACCTGAGGACACGGCCGTCTATTACTGCAATGCAGGACAGTTGTG GGGTAGTTACCGCTCCTGGGGCCAGGGGACCCAGGTCACCGTCTCCTCA。
在一些实施例中，纳米抗体包括4个框架区FR1、FR2、FR3、FR4和3个互补决定区CDR1、CDR2、CDR3；各个区域氨基酸序列的不同会影响纳米抗体与病毒的结合能力。
纳米抗体NbN2中，FR1的氨基酸序列如SEQ ID NO.7所示，具体为：AVQLVESGGGLVQAGGSLRLSCAATGLTFT；FR2的氨基酸序列如SEQ ID NO.8 所示，具体为：WFRQAPGKNREFVA；FR3的氨基酸序列如SEQ ID NO.9所示，具体为：RFTVSRDNAKNTVYLQMNTLNPEDTALYYCAA；FR4的氨基酸序列如SEQ ID NO.10所示，具体为：WGQGTQVTVSS；CDR1的氨基酸序列如SEQ ID NO.11所示，具体为：GYVMA；CDR2的氨基酸序列如SEQ ID NO.12所示，具体为：AVSRSGVVTRYADSVKG；CDR3的氨基酸序列如SEQ ID NO.13所示，具体为：DWHPLTGIMTTDSTEYDY。
纳米抗体NbN3中，FR1的氨基酸序列如SEQ ID NO.14所示，具体为：AVQLVESGGGLVQAGGSLRLSCAASGRTFS；FR2的氨基酸序列如SEQ ID NO.15所示，具体为：WFRQAPGKERQFVA；FR3的氨基酸序列如SEQ ID NO.16 所示，具体为：RFTISRDNAKNTVYLQMNSLKPEDTA I YYCAP；FR4的氨基酸序列如SEQ ID NO.17所示，具体为WGKGTQVTVSS；CDR1的氨基酸序列如 SEQ ID NO.18所示，具体为：RYAMG；CDR2的氨基酸序列如SEQ ID NO.19 所示，具体为：GI SGSGI VTNYADSVKG；CDR3的氨基酸序列如SEQ ID NO.20所示，具体为：LILARTVDMNH。
纳米抗体NbN4中，FR1的氨基酸序列如SEQ ID NO.21所示，具体为：AVQLVESGGGVVQPGGSLRLSCTASGFGFS；FR2的氨基酸序列如SEQ ID NO.22所示，具体为：WYRQAPGKERELVA；FR3的氨基酸序列如SEQ ID NO.23 所示，具体为：RFSMSRDNAKSTVYLQMNSLKPEDTAVYYCVA；FR4的氨基酸序列如SEQ ID NO.10所示，具体为：WGQGTQVTVSS；CDR1的氨基酸序列如SEQ ID NO.24所示，具体为：LYGMS；CDR2的氨基酸序列如SEQ ID NO.25 所示，具体为：SI ASGGDSTNYQDSVKG；CDR3的氨基酸序列如SEQ ID NO.26 所示，具体为：ERVSSFL KI RTAY。
纳米抗体NbN5中，FR1的氨基酸序列如SEQ ID NO.27所示，具体为：AVQLVESGGGLVQAGGSLRVSCEASGFLFN；FR2的氨基酸序列如SEQ ID NO.28所示，具体为：WYRQVPGRQRELVA；FR3的氨基酸序列如SEQ ID NO.29 所示，具体为：RFTISRDNAKNTIFLQMSGLKPEDTAVYYCYA；FR4的氨基酸序列如SEQ ID NO.10所示，具体为：WGQGTQVTVSS；CDR1的氨基酸序列如 SEQ ID NO.30所示，具体为：GDAYVLG；CDR2的氨基酸序列如SEQ ID NO.31 所示，具体为：TITSSGDTSYADSVKG；CDR3的氨基酸序列如SEQ ID NO.32 所示，具体为：ERWNGLLQR。
纳米抗体NbN6中，FR1的氨基酸序列如SEQ ID NO.33所示，具体为：AVQLVESGGGLVQPGGSLRLSCAASG I IFR；FR2的氨基酸序列如SEQ ID NO.34所示，具体为：WYRQAPGKQRELVA；FR3的氨基酸序列如SEQ ID NO.35 所示，具体为：RFTISRDNAKNTVYLQMNSLKPEDTGVYYCAA；FR4的氨基酸序列如SEQ ID NO.17所示，具体为：WGKGTQVTVSS；CDR1的氨基酸序列如SEQ ID NO.36所示，具体为：FRT I S；CDR2的氨基酸序列如SEQ ID NO.37 所示，具体为：SI SGGSSTSYADTVKG；CDR3的氨基酸序列如SEQ ID NO.38 所示，具体为：SHTMVGWIYGMDY。
纳米抗体NbN11中，FR1的氨基酸序列如SEQ ID NO.39所示，具体为：AVQLVESGGGLVQAGGSLRLSCAASGSFFS；FR2的氨基酸序列如SEQ ID NO.34所示，具体为：WYRQAPGKQRELVA；FR3的氨基酸序列如SEQ ID NO.40 所示，具体为：RFTISRDNAKNTIYLQMNSLKPEDTAVYYCNA；FR4的氨基酸序列如SEQ ID NO.10所示，具体为：WGQGTQVTVSS；CDR1的氨基酸序列如 SEQ ID NO.41所示，具体为：INTMG；CDR2的氨基酸序列如SEQ ID NO.42所示，具体为：QITNAGNTNYADSVKG；CDR3的氨基酸序列如SEQ ID NO.43所示，具体为：GQLWGSYRS。
本申请实施例第二方面提供了一种靶向新冠病毒的纳米抗体的制备方法，包括如下步骤：
S01.提供目的蛋白，将目的蛋白包被在免疫管上进行富集筛选，得到噬菌体文库；
S02.将噬菌体文库的洗脱液进行二代测序，并根据测序结果合成原始计数前占比前10的纳米抗体的基因序列；
S03.将纳米抗体的基因序列克隆至表达载体中得到重组质粒，将重组质粒转入宿主细胞中诱导表达并进行纯化，得到靶向新冠病毒的纳米抗体。
本申请实施例第二方面提供的一种靶向新冠病毒Omicron及其亚型BA.2的纳米抗体的制备方法，该制备方法基于噬菌体展示筛选技术，以新冠病毒Omicron 及其亚型BA.2的RBD蛋白为靶标构建了纳米抗体库，并结合二代测序、序列合成、克隆、表达、纯化，得靶向新冠病毒Omicron及其亚型BA.2的纳米抗体；该制备方法利用噬菌体在宿主菌中快速复制的生物特性，能够快速、高通量地筛选与特定变异株奥密克戎及其亚型BA.2RBD蛋白结合的纳米抗体，制备方法快速、简便，有利于大量筛选，提高了筛选效率。
步骤S01中，提供目的蛋白，将目的蛋白包被在免疫管上进行富集筛选，得到噬菌体文库。
在一些实施例中，提供的目的蛋白为新型冠状病毒变异株奥密克戎(Omicron) 的棘突蛋白RBD结构域，并且提供的目的蛋白的浓度为10～12μg/mL。在具体实施例中，将目的蛋白按10μg/mL的浓度包被在免疫管上。
在一些实施例中，富集筛选的步骤中，进行2～3轮富集筛选。经过多轮富集，确保得到的文库容量大，更有利于进行阳性克隆子的筛选。
步骤S02中，将噬菌体文库的洗脱液进行二代测序，并根据测序结果合成原始计数前占比前10的纳米抗体的基因序列。
步骤S03中，将纳米抗体的基因序列克隆至表达载体中得到重组质粒，将重组质粒转入宿主细胞中诱导表达并进行纯化，得到靶向新冠病毒的纳米抗体。
在一些实施例中，表达载体选自pColdII载体，将纳米抗体的基因序列克隆至pColdII载体中得到重组质粒；进一步的，在克隆过程中同时融合表达血凝素标签(hemagglutinin HA tag)用于后续检测。
在一些实施例中，宿主细胞选自大肠杆菌，将重组质粒转入大肠杆菌细胞中诱导表达并进行纯化。
在一些实施例中，将重组质粒转入大肠杆菌细胞中诱导表达并进行纯化，步骤如下：(1)使用0.2mM浓度的IPTG在16℃低温进行诱导表达，能够防止形成包涵体和蛋白降解；(2)根据预实验诱导条件进行大量诱导表达，高压破菌仪工作条件1000W进行破菌；(3)于转速为17000g，4℃离心30min，取上清与 Ni填料4℃共孵育1小时，再进行洗脱纯化；(4)Ni柱纯化后进行分子筛分离， AKATA参数设置0.5mL流速/分钟，每1mL收集一次，得到纳米抗体。
本申请实施例第三方面公开了靶向新冠病毒的纳米抗体在制备预防和/或治疗新型冠状病毒肺炎的药物中的应用。
本申请实施例第三方面提供的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体在制备预防和/或治疗新型冠状病毒肺炎的药物中的应用，由于筛选得到的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体包括纳米抗体NbN2、纳米抗体 NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，对新冠病毒野生株及各变异株的病毒具有较好的中和活性能力和抑制活性能力，适用于制备预防和/或治疗新型冠状病毒肺炎的相关药物。
本申请实施例第四方面公开了靶向新冠病毒的纳米抗体在制备检测新型冠状病毒的检测试剂或检测试剂盒中的应用。
本申请实施例第四方面提供的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体在制备检测新型冠状病毒的检测试剂或检测试剂盒中的应用，提供的靶向新冠病毒Omicron及其亚型BA.2的纳米抗体包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，对新冠病毒野生株及各变异株的病毒具有较高得结合能力，适用于制备检测新型冠状病毒的检测试剂或检测试剂盒。
下面结合具体实施例进行说明。
实施例1
(一)靶向新冠病毒Omicron及其亚型BA.2的纳米抗体及其制备方法
试验步骤：
(1)将目的蛋白Omicron RBD按10μg/mL浓度包被在免疫管上，进行3 轮富集筛选；(2)使用第三轮噬菌体洗脱液送公司进行二代测序确定序列信息； (3)根据测序信息设计并合成筛选到的纳米抗体；(4)将纳米抗体基因序列克隆入pColdII载体，同时融合表达血凝素标签(hemagglutinin HA tag)用于后续检测；(5)使用0.2mM浓度的IPTG在16℃低温进行诱导(6)根据预实验诱导条件进行大量诱导表达，高压破菌仪工作条件1000W进行破菌；(7)17000g， 4℃离心30min，取上清与Ni填料4℃共孵育1小时；(8)Ni柱纯化后进行分子筛分离，AKATA参数设置0.5mL流速/分钟，每1mL收集一次，得到靶向新冠病毒Omicron及其亚型BA.2的纳米抗体。
结果分析：
Omicron RBD纳米抗体的筛选鉴定
对Omicron RBD进行了噬菌体纳米抗体文库的筛选，三轮筛选之后，如图1 所示，文库富集将近3000倍，将第三轮噬菌体洗脱液送二代测序，如图2所示，选择含量前10的序列进行表达，分别为：NbN2、NbN3、NbN4、NbN5、NbN6、 NbN7、NbN8、NbN9、NbN10、NbN11。
进一步进行纯化，得到高纯度的10条纳米抗体，纯化得到的10个纳米抗体的蛋白结果如图3所示，大小约15kD左右。并用anti-HA WB检测证实蛋白的正确性，如图4所示，筛选得到的10个克隆子均表达正确。
(二)ELISA初步鉴定纳米抗体与SARS-CoV-2各变异株RBD结合
试验步骤：
将HA标签融合到纳米抗体基因编码序列里，表达具有HA标签的10个纳米抗体NbN2、NbN3、NbN4、NbN5、NbN6、NbN7、NbN8、NbN9、NbN10、 NbN11，别用ELISA板包被OmicronRBD蛋白，封闭，之后加入各种浓度的纳米抗体在室温孵育1小时，PBS漂洗3次，anti-HA抗体室温孵育1小时后，辣根过氧化物酶标记的抗HA抗体放大信号，TMB显色，同时做无关纳米抗体的对照和无关蛋白抗原的空白对照。
结果分析
将上步纯化表达验证正确的10个纳米抗体与SARS-CoV-2WT、Delta (B.1.617.2)、Omicron(B.1.1.529)RBD蛋白和阴性对照蛋白牛血清白蛋白(Bovine Serum Albumin，BSA)进行ELISA验证，其中，10个纳米抗体NbN2、NbN3、 NbN4、NbN5、NbN6、NbN7、NbN8、NbN9、NbN10、NbN11在图中对应的图例为：NGS2、NGS3、NGS4、NGS5、NGS6、NGS7、NGS8、NGS9、NGS10、NGS11。
与Omicron RBD蛋白结合的分析结果如图5所示，可以看出，其中，NbN2、 NbN3、NbN4、NbN5、NbN6、NbN11六条纳米抗体与Omicron RBD显示结合活性。
与SARS-CoV-2野生株(Wild Type，WT)RBD蛋白结合的分析结果如图6 所示，可以看出，其中，NbN2、NbN3、NbN4、NbN5、NbN6、NbN11六条纳米抗体与Omicron RBD显示结合活性。
与Delta RBD蛋白结合的分析结果如图7所示，可以看出，其中，NbN2、 NbN3、NbN4、NbN5、NbN6、NbN11六条纳米抗体与Omicron RBD显示结合活性。
与阴性对照蛋白牛血清白蛋白结合的分析结果如图8所示，可以看出，其中，10个纳米抗体与BSA蛋白的结合，发现均无结合活性。
因此，确认了6个纳米抗体(Nb N2、NbN3、NbN4、NbN5、NbN6、NbN11) 与SARS-CoV-2各变异株RBD具有结合活性，而与阴性对照蛋白BSA无结合(图 5-8)。
(三)六个阳性纳米抗体与SARS-CoV-2各变异株RBD蛋白的亲和力
试验步骤：
提供6个纳米抗体(Nb N2、NbN3、NbN4、NbN5、NbN6、NbN11)，基于分子互相作用分析平台Biacore，分析检测纳米抗体与SARS-CoV-2各变异株 (WT、Alpha、Beta、Delta、Omicron和Omicron BA.2)RBD蛋白的亲和力。
结果分析
6个阳性纳米抗体Nb N2、NbN3、NbN4、NbN5、NbN6、NbN11分别与 SARS-CoV-2各变异株(WT、Alpha、Beta、Delta、Omicron和Omicron BA.2) RBD蛋白的亲和力如表1所示，可以看出，亲和力均在纳摩尔水平，具有较高的亲和力。
表1
(四)5个阳性纳米抗体与表面等离子共振(Surface Plasmon Resonance， SPR)分析
试验步骤：
实验用来验证在体外表达纯化的纳米抗体与体外纯化的抗原蛋白直接相互作用，并计算二者的平衡常数。在Biacore S200上运行多循环动力学以研究纳米抗体和OmicronRBD蛋白的结合动力学。提供5个阳性纳米抗体Nb D2、NbE8、 NbF2、NbG1和NbG4，分别在运行温度25℃下，新冠冠野生株(WT)、变异株阿尔法(Alpha)、贝塔(Beta)、德尔塔(Delta)和奥密克戎(Omicron)及其亚型BA.2RBD蛋白被固定在CM5 Series S传感器芯片表面，倍比稀释的纳米抗体以30μL/分钟的速度流过芯片表面，持续120秒，然后是240秒的解离，制作动力学曲线，计算各相关参数。
结果分析
(1)Nb N2纳米抗体表面等离子共振(Surface Plasmon Resonance，SPR) 分析
SPR结果显示Nb N2纳米抗体与SARS-CoV-2各变异株(WT、Alpha、Beta、 Delta、Omicron和Omicron BA.2)RBD蛋白均有结合，如图9所示，Nb N2与 WT RBD蛋白的亲和力为24.45nM；如图10所示，Nb N2与Alpha RBD蛋白的亲和力为43nM；如图11所示，Nb N2与BetaRBD蛋白的亲和力为28.87nM；如图12所示，Nb N2与Delta RBD蛋白的亲和力为97.75nM；如图13所示，Nb N2与Omicron RBD蛋白的亲和力为55.44nM；如图14所示，Nb N2与OmicronBA.2RBD蛋白的亲和力为8.33nM。
(2)Nb N3纳米抗体表面等离子共振(Surface Plasmon Resonance，SPR) 分析
SPR结果显示Nb N3纳米抗体与SARS-CoV-2各变异株(WT、Alpha、Beta、 Delta、Omicron和Omicron BA.2)RBD蛋白均有结合，如图15所示，Nb N3 与WT RBD蛋白的亲和力为12.09nM；如图16所示，Nb N3与Alpha RBD蛋白的亲和力为21.24nM；如图17所示，Nb N3与Beta RBD蛋白的亲和力为16.92nM；如图18所示，Nb N3与Delta RBD蛋白的亲和力为18.65nM；如图19所示，Nb N3与Omicron RBD蛋白的亲和力为4.79nM；如图20所示，Nb N3与Omicron BA.2RBD蛋白的亲和力为8.77nM。
(3)Nb N4纳米抗体表面等离子共振(Surface Plasmon Resonance，SPR) 分析
SPR结果显示Nb N4纳米抗体与SARS-CoV-2各变异株(WT、Alpha、Beta、 Delta、Omicron和Omicron BA.2)RBD蛋白均有结合，如图21所示，Nb N4 与WT RBD蛋白的亲和力为2.89pM；如图22所示，Nb N4与Alpha RBD蛋白的亲和力为73.09nM；如图23所示，Nb N4与Beta RBD蛋白的亲和力为46.4nM；如图24所示，Nb N4与Delta RBD蛋白的亲和力为23.42nM；如图25所示，Nb N4与Omicron RBD蛋白的亲和力为5.09nM；如图26所示，Nb N4与Omicron BA.2RBD蛋白的亲和力为38.93nM。
(4)Nb N5纳米抗体表面等离子共振(Surface Plasmon Resonance，SPR) 分析
SPR结果显示Nb N5纳米抗体与SARS-CoV-2各变异株(WT、Alpha、Beta、 Delta、Omicron和Omicron BA.2)RBD蛋白均有结合，如图27所示，Nb N5 与WT RBD蛋白的亲和力为0.21nM；如图28所示Nb N5与Alpha RBD蛋白的亲和力为80.48nM；如图29所示，Nb N5与BetaRBD蛋白的亲和力为37.22nM；如图30所示，Nb N5与Delta RBD蛋白的亲和力为24.46nM；如图31所示，Nb N5与Omicron RBD蛋白的亲和力为90.88nM；如图32所示，Nb N5与OmicronBA.2RBD蛋白的亲和力为20.38nM。
(5)Nb N6纳米抗体表面等离子共振(Surface Plasmon Resonance，SPR) 分析
SPR结果显示Nb N6纳米抗体与SARS-CoV-2各变异株(WT、Alpha、Beta、 Delta、Omicron和Omicron BA.2)RBD蛋白均有结合，如图33所示，Nb N6 与WT RBD蛋白的亲和力为38.65nM；如图34所示，Nb N6与Alpha RBD蛋白的亲和力为101.3nM；如图35所示，Nb N6与Beta RBD蛋白的亲和力为65.36nM；如图36所示，Nb N6与Delta RBD蛋白的亲和力为26.91nM；如图37所示，Nb N6与Omicron RBD蛋白的亲和力为78.77nM；如图38所示，Nb N6与Omicron BA.2RBD蛋白的亲和力为89.52nM。
(6)Nb N11纳米抗体表面等离子共振(Surface Plasmon Resonance，SPR) 分析
SPR结果显示Nb N11纳米抗体与SARS-CoV-2各变异株(WT、Alpha、Beta、 Delta、Omicron和Omicron BA.2)RBD蛋白均有结合，如图39所示，Nb N11 与WT RBD蛋白的亲和力为36.06nM；如图40所示，Nb N11与Alpha RBD蛋白的亲和力为26.18nM；如图41所示，Nb N11与Beta RBD蛋白的亲和力为19.17 nM；如图42所示，Nb N11与Delta RBD蛋白的亲和力为28.66nM；如图43所示，Nb N11与Omicron RBD蛋白的亲和力为47.39nM；如图44所示，Nb N11与Omicron BA.2RBD蛋白的亲和力为105nM。
(五)六个阳性纳米抗体与ACE2竞争性ELISA实验
试验步骤
根据SPR结果，选择亲和力较好的纳米抗体进行竞争性ELISA实验验证。将OmicronRBD蛋白以0.5μg/mL的浓度包被在ELISA板上4℃过夜。次日用PBS 清洗一次，用3％的BSA进行室温封闭2小时。根据ACE2的最低饱和浓度进行纳米抗体倍比稀释，即稀释液为含0.5μg/mL ACE2的0.1％PBST溶液，室温孵育 2.5小时。0.1％PBST漂洗。
结果分析
如图45所示，检测了这6个纳米抗体是否与ACE2竞争结合omicron RBD 蛋白，用含ACE2最低饱和浓度的0.1％PBST溶液进行纳米抗体稀释，与提前包被有omicron RBD蛋白的ELISA板室温孵育2.5小时。0.1％PBST漂洗3次，用 anti-ACE2抗体检测ACE2的含量，同时做无纳米抗体和无ACE2蛋白的空白对照。结果显示Nb N2可明显竞争性抑制ACE2与omicronRBD蛋白的结合，半抑制浓度(half maximum inhibitory concentration，IC50)为0.09μM。结果显示 Nb N2纳米抗体可明显竞争性抑制ACE2与omicron RBD蛋白的结合。
因此，本申请提供的新冠病毒Omicron及其亚型BA.2的纳米抗体，包括纳米抗体NbN2、纳米抗体NbN3、纳米抗体NbN4、纳米抗体NbN5、纳米抗体NbN6、纳米抗体NbN11中的至少一种，提供的纳米抗体对新冠病毒野生株、新冠病毒变异株阿尔法(Alpha)、新冠病毒贝塔(Beta)、新冠病毒德尔塔(Delta)、新冠病毒Omicron及其亚型BA.2的RBD结构域具有较高的亲和力，能够快速且特异性地中和不同的新冠病毒，有效抑制其活性；同时，提供的纳米抗体具有更长的CDR3序列，有利于纳米抗体进行灵活调节其构象，提高与病毒的结合作用，且制备成本较低，有利于广泛应用。
以上所揭露的仅为本发明较佳实施例而已，当然不能以此来限定本发明之权利范围，因此依本发明权利要求所作的等同变化，仍属本发明所涵盖的范围。
序列表
<110> 深圳市人民医院
<120> 靶向新冠病毒的纳米抗体及其制备方法和应用
<140> 2022106375212
<141> 2022-03-31
<160> 49
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Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys Ala Pro
20 25 30
<210> 17
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 17
Trp Gly Lys Gly Thr Gln Val Thr Val Ser Ser
1 5 10
<210> 18
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 18
Arg Tyr Ala Met Gly
1 5
<210> 19
<211> 17
<212> PRT
<213> Artificial Sequence
<400> 19
Gly Ile Ser Gly Ser Gly Ile Val Thr Asn Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 20
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 20
Leu Ile Leu Ala Arg Thr Val Asp Met Asn His
1 5 10
<210> 21
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 21
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Gly Phe Ser
20 25 30
<210> 22
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 22
Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ala
1 5 10
<210> 23
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 23
Arg Phe Ser Met Ser Arg Asp Asn Ala Lys Ser Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Val Ala
20 25 30
<210> 24
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 24
Leu Tyr Gly Met Ser
1 5
<210> 25
<211> 17
<212> PRT
<213> Artificial Sequence
<400> 25
Ser Ile Ala Ser Gly Gly Asp Ser Thr Asn Tyr Gln Asp Ser Val Lys
1 5 10 15
Gly
<210> 26
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 26
Glu Arg Val Ser Ser Phe Leu Lys Ile Arg Thr Ala Tyr
1 5 10
<210> 27
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 27
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Val Ser Cys Glu Ala Ser Gly Phe Leu Phe Asn
20 25 30
<210> 28
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 28
Trp Tyr Arg Gln Val Pro Gly Arg Gln Arg Glu Leu Val Ala
1 5 10
<210> 29
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 29
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Ile Phe Leu Gln
1 5 10 15
Met Ser Gly Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Tyr Ala
20 25 30
<210> 30
<211> 7
<212> PRT
<213> Artificial Sequence
<400> 30
Gly Asp Ala Tyr Val Leu Gly
1 5
<210> 31
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 31
Thr Ile Thr Ser Ser Gly Asp Thr Ser Tyr Ala Asp Ser Val Lys Gly
1 5 10 15
<210> 32
<211> 9
<212> PRT
<213> Artificial Sequence
<400> 32
Glu Arg Trp Asn Gly Leu Leu Gln Arg
1 5
<210> 33
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 33
Ala 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 Ile Ile Phe Arg
20 25 30
<210> 34
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 34
Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val Ala
1 5 10
<210> 35
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 35
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Val Tyr Tyr Cys Ala Ala
20 25 30
<210> 36
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 36
Phe Arg Thr Ile Ser
1 5
<210> 37
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 37
Ser Ile Ser Gly Gly Ser Ser Thr Ser Tyr Ala Asp Thr Val Lys Gly
1 5 10 15
<210> 38
<211> 13
<212> PRT
<213> Artificial Sequence
<400> 38
Ser His Thr Met Val Gly Trp Ile Tyr Gly Met Asp Tyr
1 5 10
<210> 39
<211> 30
<212> PRT
<213> Artificial Sequence
<400> 39
Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Phe Phe Ser
20 25 30
<210> 40
<211> 32
<212> PRT
<213> Artificial Sequence
<400> 40
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Ile Tyr Leu Gln
1 5 10 15
Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn Ala
20 25 30
<210> 41
<211> 5
<212> PRT
<213> Artificial Sequence
<400> 41
Ile Asn Thr Met Gly
1 5
<210> 42
<211> 16
<212> PRT
<213> Artificial Sequence
<400> 42
Gln Ile Thr Asn Ala Gly Asn Thr Asn Tyr Ala Asp Ser Val Lys Gly
1 5 10 15
<210> 43
<211> 9
<212> PRT
<213> Artificial Sequence
<400> 43
Gly Gln Leu Trp Gly Ser Tyr Arg Ser
1 5
<210> 44
<211> 384
<212> DNA
<213> Artificial Sequence
<400> 44
atggcggtgc agctggtgga gtctggggga ggattggtgc aggctggggg ctctctgaga 60
ctctcctgtg cagccactgg actcaccttc actggttatg taatggcctg gttccgccag 120
gctccaggga agaaccgtga gtttgtagcg gccgttagcc ggagtggtgt tgttacacgc 180
tatgcagact ccgtgaaggg ccgattcacc gtctccagag acaacgccaa gaacacggtg 240
tatctgcaaa tgaacaccct gaatcctgag gacacggccc tttattactg tgcagcagat 300
tggcatccac taactggaat aatgacgact gactctacag agtatgacta ctggggccag 360
gggacccagg tcaccgtctc ctca 384
<210> 45
<211> 363
<212> DNA
<213> Artificial Sequence
<400> 45
atggcggtgc agctggtgga gtctggggga ggattggtgc aggctggggg ctctctgaga 60
ctctcctgtg cagcctctgg acgcaccttc agtaggtatg ccatgggctg gttccgccag 120
gctccaggga aggagcgcca gtttgtagca ggtattagcg ggagtggtat agtcacaaac 180
tatgcagact ccgtgaaggg ccgattcacc atctccagag acaacgccaa gaacacggtg 240
tacctgcaaa tgaacagcct gaaacctgag gacacggcca tttattactg tgcaccgttg 300
atcctagcac gaactgtgga catgaaccac tggggcaaag ggacccaggt caccgtctcc 360
tca 363
<210> 46
<211> 369
<212> DNA
<213> Artificial Sequence
<400> 46
atggcggtgc agctggtgga gtctggggga ggagtggtgc agcctggggg gtctctgaga 60
ctctcctgta cagcctctgg attcggcttc agtctttatg gcatgagctg gtaccgtcag 120
gctccaggga aggagcgcga gttggtcgca tctattgcca gtggtggtga tagtacaaac 180
tatcaagact ccgtgaaggg ccgattctcc atgtccagag acaatgccaa gagcacggtg 240
tatctgcaaa tgaacagcct gaaacctgag gacacggccg tgtattactg cgtggcagaa 300
agagtctcgt cgttcttaaa aattcgtacc gcctactggg gccaggggac ccaggtcacc 360
gtctcctca 369
<210> 47
<211> 360
<212> DNA
<213> Artificial Sequence
<400> 47
atggcggtgc agctggtgga gtctggggga ggcttggtgc aggctggggg gtctctgaga 60
gtgtcctgtg aagcctctgg attccttttc aatggcgatg cctatgtctt gggctggtac 120
cgccaggttc caggtaggca gcgtgaattg gtcgcaacta ttacgagtag cggtgacaca 180
agctatgcgg actccgtgaa gggccgattc accatctcca gagacaacgc caagaataca 240
atttttctgc aaatgagcgg cctgaaaccc gaggacacgg ccgtctatta ctgctatgca 300
gagaggtgga atgggctcct tcaacgctgg ggccagggga cccaggtcac cgtctcctca 360
<210> 48
<211> 366
<212> DNA
<213> Artificial Sequence
<400> 48
atggcggtgc agctggtgga gtctggggga ggcttggtgc aacctggggg gtctctgaga 60
ctctcctgtg cagcctctgg aatcatcttc agattcagaa ccatatcctg gtaccgccag 120
gctccaggga agcagcgcga gttggtcgca agtattagtg gcggcagtag cacgtcctat 180
gcagacactg tgaagggccg attcaccatc tccagagaca acgccaagaa cacggtgtat 240
ctgcaaatga acagcctgaa acctgaggac acaggcgttt attactgtgc agcgtcacat 300
actatggttg ggtggatcta cggcatggac tactggggca aagggaccca ggtcaccgtc 360
tcctca 366
<210> 49
<211> 354
<212> DNA
<213> Artificial Sequence
<400> 49
atggcggtgc agctggtgga gtctggggga ggcttggtgc aggctggggg gtctctgaga 60
ctctcctgtg cagcctctgg aagcttcttc agtatcaata ccatgggctg gtaccgccag 120
gctccaggga agcagcgcga gttggtcgca caaattacta atgcaggtaa cacaaactat 180
gcagactccg tgaagggccg gttcaccatc tccagagaca acgccaagaa cacgatatat 240
ctgcaaatga acagcctgaa acctgaggac acggccgtct attactgcaa tgcaggacag 300
ttgtggggta gttaccgctc ctggggccag gggacccagg tcaccgtctc ctca 354
Claims (5)
1.一种靶向新冠病毒的纳米抗体，其特征在于，所述纳米抗体为纳米抗体NbN6，其中，所述纳米抗体NbN6的氨基酸序列如 SEQ.ID NO.5所示。
2.根据权利要求1所述的靶向新冠病毒的纳米抗体，其特征在于，所述的纳米抗包括4个框架区FR1、FR2、FR3、FR4和3个互补决定区CDR1、CDR2、CDR3；
所述纳米抗体NbN6中，FR1的氨基酸序列如SEQ ID NO.33所示，FR2的氨基酸序列如SEQID NO.34所示，FR3的氨基酸序列如SEQ ID NO.35所示，FR4的氨基酸序列如SEQ ID NO.17所示，CDR1的氨基酸序列如SEQ ID NO.36所示，CDR2的氨基酸序列如SEQ ID NO.37所示，CDR3的氨基酸序列如SEQ ID NO.38所示。
3.根据权利要求1所述的靶向新冠病毒的纳米抗体，其特征在于，所述纳米抗体NbN6的碱基序列如SEQ.ID NO.48所示。
4.权利要求1~3任一所述的靶向新冠病毒的纳米抗体在制备预防和/或治疗新型冠状病毒肺炎的药物中的应用。
5.权利要求1~3任一所述的靶向新冠病毒的纳米抗体在制备检测新型冠状病毒的检测试剂或检测试剂盒中的应用。
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| CN114773464B (zh) * | 2022-06-20 | 2022-08-26 | 北京市疾病预防控制中心 | 一种针对新冠病毒omicron株S蛋白的单域抗体VHH-2及编码序列和应用 |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6018032A (en) * | 1995-09-11 | 2000-01-25 | Kyowa Hakko Kogyo Co., Ltd. | Antibody against human interleukin-5-receptor α chain |
| CN111647076A (zh) * | 2020-04-27 | 2020-09-11 | 南京医科大学 | 抗新型冠状病毒SARS-Cov-2的中和性单域抗体及其应用 |
| CN113754739A (zh) * | 2020-06-03 | 2021-12-07 | 中国人民解放军军事科学院军事医学研究院 | 一种冠状病毒s蛋白rbd糖蛋白的制备方法及其应用 |
Family Cites Families (30)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2007331672A1 (en) * | 2006-12-15 | 2008-06-19 | Ablynx N.V. | Amino acid sequences that modulate the interaction between cells of the immune system |
| GB201115214D0 (en) * | 2011-09-02 | 2011-10-19 | Health Prot Agency | Influenza virus antibody compositions |
| CA2910632A1 (en) * | 2013-04-29 | 2014-12-04 | Agrosavfe N.V. | Compositions comprising polypeptides that bind glucosylceramide of a fungal pest |
| WO2015156673A1 (en) * | 2014-04-10 | 2015-10-15 | Stichting Vu-Vumc | IMMUNOGLOBULINS BINDING HUMAN Vγ9Vδ2 T CELL RECEPTORS |
| CN111434692B (zh) * | 2019-01-15 | 2021-12-31 | 浙江道尔生物科技有限公司 | 抗cld18a2纳米抗体及其应用 |
| CN111116752B (zh) * | 2019-12-24 | 2021-09-03 | 北京纽安博生物技术有限公司 | 结合免疫球蛋白的单域抗体、抗禽流感单域抗体、双功能抗体及其应用 |
| WO2021156490A2 (en) * | 2020-02-06 | 2021-08-12 | Vib Vzw | Corona virus binders |
| BR112022016550A2 (pt) * | 2020-02-21 | 2022-11-16 | Harpoon Therapeutics Inc | Proteínas de ligação a flt3 e métodos de uso |
| CN111592595B (zh) * | 2020-04-27 | 2021-02-19 | 南京医科大学 | 抗新型冠状病毒SARS-Cov-2的中和性抗体及其应用 |
| WO2021224606A1 (en) * | 2020-05-04 | 2021-11-11 | The Rosalind Franklin Institute | Single domain antibodies binding to sars-cov-2 spike protein |
| US20220074938A1 (en) * | 2020-05-07 | 2022-03-10 | Senseutics Limited | Method of detecting pathogens and/or antigens in samples |
| CN115917061A (zh) * | 2020-05-15 | 2023-04-04 | 智利南方大学 | 用于生成纳米抗体的快速单步梯度法 |
| WO2022015573A2 (en) * | 2020-07-13 | 2022-01-20 | President And Fellows Of Harvard College | Sars-cov-2 antigen-binding proteins and uses thereof |
| WO2022015668A1 (en) * | 2020-07-15 | 2022-01-20 | Regents Of The University Of Minnesota | SARS-CoV-2 NANOBODIES AND METHODS OF USE THEREOF |
| WO2022011717A1 (zh) * | 2020-07-17 | 2022-01-20 | 上海洛启生物医药技术有限公司 | 针对新型冠状病毒的纳米抗体及其应用 |
| KR20220012810A (ko) * | 2020-07-23 | 2022-02-04 | (주)셀트리온 | 사스-코로나바이러스-2 s 단백질의 에피토프에 결합하는 사스-코로나바이러스-2 중화 결합 분자 |
| US20230235027A1 (en) * | 2020-07-23 | 2023-07-27 | The Usa, As Represented By The Secretary, Dept. Of Health And Human Services | Nanobodies directed to coronavirus spike protein receptor binding domain and uses thereof |
| EP3944877A1 (en) * | 2020-07-29 | 2022-02-02 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Nanobodies sars-cov2 |
| TW202221024A (zh) * | 2020-08-07 | 2022-06-01 | 美商索倫多醫療公司 | 結合sars-cov-2 s蛋白之中和抗體 |
| JP2023539109A (ja) * | 2020-08-19 | 2023-09-13 | ユニバーシティ オブ ピッツバーグ －オブ ザ コモンウェルス システム オブ ハイヤー エデュケイション | コロナウイルスナノボディならびにその使用及び同定の方法 |
| WO2022040603A2 (en) * | 2020-08-21 | 2022-02-24 | The Rockefeller University | Single-domain antibodies that bind sars-cov-2 |
| CN112010967B (zh) * | 2020-09-07 | 2021-03-23 | 康维众和(中山)生物药业有限公司 | 新型冠状病毒SARS-Cov-2中和毒性的纳米抗体及其制备方法与应用 |
| EP3970798A1 (en) * | 2020-09-18 | 2022-03-23 | NMI Naturwissenschaftliches und Medizinisches Institut an der Universität Tübingen | Sars-cov-2-nanobodies |
| CN112062840A (zh) * | 2020-09-22 | 2020-12-11 | 石河子大学 | 一种基于新型冠状病毒s蛋白的纳米抗体及其应用 |
| CN112062839A (zh) * | 2020-09-22 | 2020-12-11 | 石河子大学 | 一种基于新型冠状病毒s蛋白s1亚基的纳米抗体及其应用 |
| CN112094342B (zh) * | 2020-09-25 | 2022-05-13 | 中国科学技术大学 | 与SARS-CoV-2 RBD结合的羊驼源纳米抗体 |
| CN113563463B (zh) * | 2021-06-11 | 2022-05-17 | 中国医学科学院病原生物学研究所 | 一种抗新型冠状病毒SARS-CoV-2的中和纳米抗体及其应用 |
| CN114031685B (zh) * | 2022-01-10 | 2022-03-25 | 中国人民解放军军事科学院军事医学研究院 | 一种全人源抗新冠病毒中和抗体zw2g10及应用 |
| CN114106166B (zh) * | 2022-01-29 | 2022-05-06 | 中国疾病预防控制中心病毒病预防控制所 | 人源抗新冠病毒中和性抗体d2及其应用 |
| CN114478758B (zh) * | 2022-04-01 | 2022-06-21 | 深圳市人民医院 | 靶向SARS-CoV-2的纳米抗体及制备方法和应用 |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6018032A (en) * | 1995-09-11 | 2000-01-25 | Kyowa Hakko Kogyo Co., Ltd. | Antibody against human interleukin-5-receptor α chain |
| CN111647076A (zh) * | 2020-04-27 | 2020-09-11 | 南京医科大学 | 抗新型冠状病毒SARS-Cov-2的中和性单域抗体及其应用 |
| CN113754739A (zh) * | 2020-06-03 | 2021-12-07 | 中国人民解放军军事科学院军事医学研究院 | 一种冠状病毒s蛋白rbd糖蛋白的制备方法及其应用 |
Non-Patent Citations (2)
| Title |
|---|
| The role of single‑domain antibodies (or nanobodies) in SARS‑CoV‑2 neutralization;Arghavan Zebardast等;《Molecular Biology Reports 》;第49卷;第647-656页 * |
| 新冠病毒刺突蛋白B细胞线性表位发掘;简春利;《重庆理工大学学报(自然科学)》;第35卷(第1期);第230-235页 * |
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| Publication number | Publication date |
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| CN115043934A (zh) | 2022-09-13 |
| CN115043937B (zh) | 2023-06-02 |
| CN115043936B (zh) | 2023-06-27 |
| CN115043936A (zh) | 2022-09-13 |
| CN115043937A (zh) | 2022-09-13 |
| CN114478757B (zh) | 2022-07-05 |
| CN115043935A (zh) | 2022-09-13 |
| CN114478757A (zh) | 2022-05-13 |
| CN115043933B (zh) | 2023-08-08 |
| CN115043933A (zh) | 2022-09-13 |
| CN115043935B (zh) | 2023-06-27 |
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