CN112300274B - 新型冠状病毒特异性抗原肽的人源抗体、制备方法及用途 - Google Patents
新型冠状病毒特异性抗原肽的人源抗体、制备方法及用途 Download PDFInfo
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Abstract
本公开涉及一种新型冠状病毒特异性抗原肽的人源抗体、制备方法及用途。具体来说,本公开涉及一种抗SARS‑CoV‑2抗体或其抗原结合片段,以及其在疾病诊断、制备COVID‑19疫苗、制备预防、治疗COVID‑19的药物中的用途。
Description
技术领域
本公开涉及一种新型冠状病毒特异性抗原肽结合的人抗体及其用途。具体来说,本公开涉及一种单克隆抗体,所述抗体特异性结合SARS-CoV-2病毒的抗原肽S672-691,该抗原肽位于SARS-CoV-2病毒S蛋白的TMPRSS2位点,及其前述抗原肽在制备COVID-19疫苗、制备预防、治疗COVID-19的药物中的用途。
背景技术
新型冠状病毒(SARS-CoV-2)属于乙型冠状病毒属,线性单链RNA(ssRNA)病毒。其基因组全长约29903个核苷酸,共包含10个基因。自2020年1月10日,第一个SARS-CoV-2基因组序列数据被公布,此后陆续有多个从患者身上分离的新型冠状病毒的基因组序列发布。2020年1月22日,基因组科学数据中心正式发布2019新型冠状病毒资源库。经数据分析,2019新型冠状病毒(SARS-CoV-2)与2003年爆发的SARS病毒基因组序列相似度为80%,与2017年2月从国内的蝙蝠中采集到的Bat SARS-like coronavirus isolate bat-SL-CoVZC45基因组序列相似性最高,相似度为88%。截止到2020年1月30日,全球已有6家机构在“全球共享流感病毒数据库GISAID”上发布了13例新型冠状基因组序列。
新型冠状病毒(SARS-CoV-2)是一种包膜的正链RNA病毒,含有30kb的基因组和四种结构蛋白,即刺突蛋白(S)、包膜蛋白(E)、膜蛋白(M)和核衣壳蛋白(N)。S蛋白调节病毒对目标宿主细胞上受体的附着。E蛋白质的功能是组装病毒并充当离子通道;M蛋白与E蛋白一起在病毒组装中发挥作用,并参与新病毒颗粒的生物合成;N蛋白与病毒RNA形成核糖核蛋白复合物。新型冠状病毒的表面刺状糖蛋白(S蛋白)负责通过与宿主细胞表面受体(ACE2)的相互作用附着到宿主细胞上。S蛋白以同型三聚体的形式存在,每个单体含有1200多个氨基酸。在SARS-CoV-2的S蛋白中,一个含有306-575残基的小结构域被鉴定为受体结合域(RBD),其中被称为受体结合基序(RBM)的439-508残基直接介导了与ACE2的相互作用。冠状病毒进入细胞依赖于病毒突刺蛋白与细胞受体的结合以及宿主细胞蛋白酶对S蛋白的启动。阐明哪些细胞因子被2019-nCoV利用可能为病毒的传播和治疗靶点的发现提供新的思路。有研究表明2019-nCoV使用SARS-CoV受体ACE2进入,此外,TMPRSS2的定向表达挽救了E64d对2019-nCoV驱动的病毒进入细胞的抑制,这表明新冠病毒的S蛋白由TMPRSS2启动。经批准用于临床的TMPRSS2抑制剂被阻断进入,可能构成一种治疗方案以及抗病毒干预的潜在靶点。
噬菌体展示技术最初是由英国医学研究理事会(Medical Research Council)于1990年开发,是通过制备人源抗体文库(library),并以抗体片段(Fab,ScFv)的形式表达在噬菌体表面上,从而筛选特异性抗原的抗体克隆技术。已经提出了可以从单pot抗体文库系统中筛选出几乎所有与抗原发生特异性反应的重组人单克隆抗体可能性,因此,当使用噬菌体展示抗体技术时,能获得可应用于体内诊断或治疗的各种抗体片段(Fab或ScFv)。
发明内容
发明要解决的问题
本公开的一个目的在于,提供一种与新型冠状病毒特异性结合的单克隆抗体。
本公开的另一个目的在于,提供所述单克隆抗体的用途。
本公开的另一个目的在于,本公开还提供一种包含用于编码所述单克隆抗体的多核苷酸的表达载体。
本公开的另一个目的在于,本公开还提供一种转化成所述表达载体的转化体。
用于解决问题的方案
本公开中以COVID-19患者PBMC构建噬菌体人源抗体库,通过与特异性抗原肽相互结合,从而筛选特异性抗原的抗体克隆技术。
具体来说,本公开记载了以下技术方案。
(1).一种抗SARS-CoV-2抗体或其抗原结合片段,所述抗体特异性地结合所述SARS-CoV-2表位,其中,所述SARS-CoV-2表位包含如SEQ ID NO:23所示的序列。
(2).根据(1)所述的抗体或其抗原结合片段,其包含重链可变区,其中,编码所述重链可变区的序列包含如下所示序列中的一种或多种:
(i)如SEQ ID NO:11-13任一项所示的序列;
(ii)如SEQ ID NO:17-19任一项所示的序列。
(3).根据(1)-(2)任一项所述的抗体或其抗原结合片段,其包含轻链可变区,其中,编码所述轻链可变区的序列包含如下所示序列中的一种或多种:
(i)如SEQ ID NO:14-16任一项所示的序列;
(ii)如SEQ ID NO:20-22任一项所示的序列。
(4).根据(1)-(3)任一项所述的抗体或其抗原结合片段,其包含连接子,其中,编码所述连接子的序列包含如SEQ ID NO:3所示的序列。
(5).根据权利要求1所述的抗体或其抗原结合片段,其包含重链可变区(VH)和轻链可变区(VL),所述VH包含VH互补决定区(CDR)1、VH互补决定区(CDR)2和VH互补决定区(CDR)3,以及所述VL包含VLCDR1、VLCDR2和VLCDR3,其中,
所述VH由如下氨基酸编码:VHCDR1包含如SEQ ID NO:11或17所示的氨基酸序列,VHCDR2包含如SEQ ID NO:12或18所示的氨基酸序列,和VHCDR3包含如SEQ ID NO:13或19所示的氨基酸序列;并且
所述VL由如下氨基酸编码:VLCDR1包含如SEQ ID NO:14或20所示的氨基酸序列,VLCDR2包含如SEQ ID NO:15或21所示的氨基酸序列,和VLCDR3包含如SEQ ID NO:16或22所示的氨基酸序列。
(6).根据(5)所述的抗体或其抗原结合片段,其中,编码所述抗体或其抗原结合片段包含如下所示序列中的一种或多种:
(i)VH包含如SEQ ID NO:1所示的氨基酸序列,和VL包含如SEQ ID NO:5所示的氨基酸序列;
(ii)VH包含如SEQ ID NO:7所示的氨基酸序列,和VL包含如SEQ ID NO:9所示的氨基酸序列;
(iii)如SEQ ID NO:24或26所示的氨基酸序列。
(7).一种多核苷酸,其中,所述多核苷酸选自(i)-(iv)中的任一项:
(i)包含如SEQ ID NO:2,6,8,10任一序列或其任意组合所示的核苷酸序列;
(ii)包含如SEQ ID NO:2,6,8,10任一序列或其任意组合所示的核苷酸序列的反向互补序列的核苷酸序列;
(iii)在高严格性杂交条件或非常高严格性杂交条件下,能够与(i)-(ii)中的任一项所示的核苷酸序列杂交的序列的反向互补序列;
(iv)与(i)-(iii)中的任一项所示的核苷酸序列具有至少90%,可选至少95%,优选至少97%,更优选至少98%,最优选至少99%的序列同一性的序列。
(8).根据(7)所述的多核苷酸,其中,所述多核苷酸选自(v)-(viii)中的任一项:
(v)包含如SEQ ID NO:2和SEQ ID NO:6所示的核苷酸序列,或包含如SEQ ID NO:8和SEQ ID NO:10所示的核苷酸序列;;
(vi)包含如(v)所示的核苷酸序列的反向互补序列的核苷酸序列;
(vii)在高严格性杂交条件或非常高严格性杂交条件下,能够与(v)-(vi)中的任一项所示的核苷酸序列杂交的序列的反向互补序列;
(viii)与(v)-(vii)中的任一项所示的核苷酸序列具有至少90%,可选至少95%,优选至少97%,更优选至少98%,最优选至少99%的序列同一性的序列。
(9).根据(8)所述的多核苷酸,其中,所述多核苷酸选自(ix)-(xii)中的任一项:
(ix)包含如SEQ ID NO:25或27任一序列所示的核苷酸序列;
(x)包含如SEQ ID NO:25,27任一序列所示的序列的反向互补序列的核苷酸序列;
(xi)在高严格性杂交条件或非常高严格性杂交条件下,能够与(ix)-(x)中的任一项所示的核苷酸序列杂交的序列的反向互补序列;
(xii)与(ix)-(xi)中的任一项所示的核苷酸序列具有至少90%,可选至少95%,优选至少97%,更优选至少98%,最优选至少99%的序列同一性的序列。
(10).一种载体,其中,所述载体包含根据(7)-(9)任一项所述的多核苷酸。
(11).一种分离的宿主细胞,其中,所述宿主细胞包含如(10)所述的载体。
(12).一种制备稳定表达目标蛋白的宿主细胞的方法,其中,所述方法包含利用(10)所述的载体,转化初始宿主细胞的步骤;可选的,所述宿主细胞为中国仓鼠卵巢细胞。
(13).一种制备目标蛋白的方法,所述方法包含利用(11)所述的宿主细胞、或通过(12)所述的方法,制备所述目标蛋白。
(14).一种抗体或其抗原结合片段,其中,所述抗体或其抗原结合片段为(13)所述的方法制备。
(15).一种试剂盒,其中,所述试剂盒包含根据(1)-(6)任一项或(14)所述的抗体或其抗原结合片段。
(16).(15)所述的试剂盒在制备用于检测COVID-19的试剂盒中的应用。
(17).一种药物组合物或疫苗,其中,所述药物组合物或疫苗中含有根据(1)-(6)任一项或(14)所述的抗体或其抗原结合片段。
(18).(1)-(6)任一项或(14)所述的抗体或其抗原结合片段,或(17)所述的药物组合物或疫苗在制备用于治疗或预防COVID-19的药物的应用。
(19).一种治疗或预防COVID-19的方法,其中,将(1)-(6)任一项或(14)所述的抗体或其抗原结合片段,或(17)所述的药物组合物或疫苗给予所述动物。
发明的效果
在一个实施方式中,本公开提供了能够特异性结合新型冠状病毒的人源抗体,其能够用于新型冠状病毒的诊断、预防或治疗。
在另一个实施方式中,本公开提供了用于制备能够特异性结合新型冠状病毒的人源抗体的方法。
附图说明
图1示出了构建VL和VH-VL文库时PCR琼脂糖凝胶电泳图,文库包括新冠病人PBMC和正常人PBMC为对照组。
图2示出了噬菌体展示文库测序的质量报告图,包括新冠病人PBMC和正常人PBMC构建的文库。
图3示出了噬菌体文库的筛选结果。
图4示出了抗体不同稀释浓度下的ELISA结果。
图5示出了抗体中和实验结果。
具体实施方式
定义
在本公开的权利要求和/或说明书中,词语“一(a)”或“一(an)”或“一(the)”可以指“一个”,但也可以指“一个或多个”、“至少一个”以及“一个或多于一个”。
如在权利要求和说明书中所使用的,词语“包含”、“具有”、“包括”或“含有”是指包括在内的或开放式的,并不排除额外的、未引述的元件或方法步骤。与此同时,“包含”、“具有”、“包括”或“含有”也可以表示封闭式的,排除额外的、未引述的元件或方法步骤。
在整个申请文件中,术语“约”表示:一个值包括测定该值所使用的装置或方法的误差的标准偏差。
虽然所公开的内容支持术语“或”的定义仅为替代物以及“和/或”,但除非明确表示仅为替代物或替代物之间相互排斥外,权利要求中的术语“或”是指“和/或”。
如本公开所使用的,术语“SARS-CoV-2”,也被称为“2019-nCoV”,其含义为2019新型冠状病毒。
如本公开所使用的,术语“COVID-19”的含义为新型冠状病毒肺炎(Corona VirusDisease 2019),简称“新冠肺炎”,是指2019新型冠状病毒(SARS-CoV-2)感染导致的肺炎。
本公开中的“序列同一性”和“同一性百分比”指两个或更多个多核苷酸或多肽之间相同(即同一)的核苷酸或氨基酸的百分比。两个或更多个多核苷酸或多肽之间的序列同一性可通过以下方法测定:将多核苷酸或多肽的核苷酸或氨基酸序列对准且对经对准的多核苷酸或多肽中含有相同核苷酸或氨基酸残基的位置数目进行评分,且将其与经对准的多核苷酸或多肽中含有不同核苷酸或氨基酸残基的位置数目进行比较。多核苷酸可例如通过含有不同核苷酸(即取代或突变)或缺失核苷酸(即一个或两个多核苷酸中的核苷酸插入或核苷酸缺失)而在一个位置处不同。多肽可例如通过含有不同氨基酸(即取代或突变)或缺失氨基酸(即一个或两个多肽中的氨基酸插入或氨基酸缺失)而在一个位置处不同。序列同一性可通过用含有相同核苷酸或氨基酸残基的位置数目除以多核苷酸或多肽中氨基酸残基的总数来计算。举例而言,可通过用含有相同核苷酸或氨基酸残基的位置数目除以多核苷酸或多肽中核苷酸或氨基酸残基的总数且乘以100来计算同一性百分比。
本公开中的术语“噬菌体展示技术”,是将外源蛋白或多肽的DNA序列插入到噬菌体外壳蛋白结构基因的适当位置,使外源基因随外壳蛋白的表达而表达,同时,外源蛋白随噬菌体的重新组装而展示到噬菌体表面的生物技术。
本公开中的术语“抗体”,指免疫球蛋白或其片段或它们的衍生物,并且包括其包含的抗原结合位点的任何多肽,而不管其是否是在体外或体内产生。该术语包括,但不限于,多克隆、单克隆、单特异性的、多特异性的、非特异性的、人源化、单链的、嵌合的、合成的、重组的、杂合的、突变的、嫁接的抗体。术语“抗体”还包括抗体片段例如Fab、F(ab’)2、FV、scFv、Fd、dAb和其它保留抗原结合功能的抗体片段。通常情况下,这样的片段将包括抗原结合片段。
本公开中的术语“单链抗体”(scFv),是由抗体重链可变区和轻链可变区通过有限个氨基酸的短肽(也被称为连接子,linker)连接而成的抗体。
本公开中的术语“外周血单个核细胞”(PBMC)是外周血中具有单个核的细胞,包括淋巴细胞和单核细胞。
本公开中的术语“S蛋白672-691多肽/抗原肽”,在本公开中也被称为“S672-691”,指的是氨基酸序列为ASYQTQTNSPRRARSVASQS的多肽。前述多肽对于新型冠状病毒具有良好的准确性和特异性,能够用于SARS-CoV-2的检测;与此同时,前述多肽对于SARS-CoV-2具有良好的抑制作用,能够用于SARS-CoV-2所导致的疾病的治疗。
本公开中的术语“IMGT编号方案”,是Lefranc等人为免疫球蛋白超家族的所有蛋白质序列引入了新的标准化编号系统,包括来自抗体轻链和重链的可变结构域以及来自不同物种的T细胞受体链。所述IMGT编号方法基于种系V序列(germ-line V)比对连续计数残基。
在本公开所描述的技术方案中,除非特别说明,否则本公开中针对抗体所采用的抗体编号方案均为IMGT编号方案。
在一些技术方案中,本公开涉及用于限定两个多核苷酸互补程度的杂交条件严格性。可选的,前述多核苷酸可以选自DNA。本公开使用的“严格性”指杂交期间的温度和离子强度条件以及是否存在某些有机溶剂。严格性越高,靶核苷酸序列与经标记的多核苷酸序列之间的互补程度越高。“严格条件”指仅具有高频率互补碱基的核苷酸序列将杂交的温度和离子条件。本文使用的术语“在高严格性或非常高的严格性条件下杂交”描述了用于杂交和洗涤的条件。用于进行杂交反应的指导可见于Current Protocols in MolecμLarBiology,John Wiley和Sons,N.Y.(1989),6.3.1-6.3.6中。本公开提及的具体杂交条件如下:1)高严格性杂交条件:在约45℃的6X氯化钠/柠檬酸钠(SSC)中,然后于65℃下用0.2XSSC、0.1%SDS洗涤一次或更多次;2)非常高严格性杂交条件:65℃的0.5M磷酸钠,7%SDS,然后于65℃下用0.2X SSC、1%SDS洗涤一次或更多次。
研究表明新型冠状病毒是通过S-蛋白与人ACE2互作的分子机制,来感染人的呼吸道上皮细胞,此外,新型冠状病毒的S蛋白由TMPRSS2启动。S蛋白调节冠状病毒对目标宿主细胞上受体的附着,在结合到宿主细胞产生相互作用时具有重要作用。因此我们通过对新型冠状病毒表面S蛋白进行特异性抗原表位进行研究并进行筛选。
我们在之前的研究中对新冠病毒特异性抗原肽进行筛选,并且得到了对于新型冠状病毒具有良好的准确性和特异性,能够用于SARS-CoV-2的检测,以及对于SARS-CoV-2具有良好的抑制作用,能够用于SARS-CoV-2所导致的疾病的治疗的抗原肽。
根据抗原肽在S蛋白的氨基酸位点的编号,上述抗原肽的名称为S672-691(其氨基酸序列为:ASYQTQTNSPRRARSVASQS),并且进行了一系列验证实验,这条多肽在检测以及抗病毒反应中都具有良好的效果。并且通过同源性序列比对,发现S672-691多肽位于新冠病毒TMPRSS2位点,且与其他SARS,MERS,human冠状病毒在相同区域没有很高的同源性,说明这个位点极有可能能够特异性检测,抑制新型冠状病毒。
本公开将COVID-19患者的PBMC取出,构建成噬菌体展示文库,通过与S672-691抗原肽进行生物淘选,进而筛选到了能够特异性结合新型冠状病毒的人源抗体。
本公开中采用的分子生物学方法,均可以参见“最新分子生物学实验方法汇编(Current Protocols in Molecular Biology,Wiley出版)”,“分子克隆实验指南(Molecular Cloning:A Laboratory Manual,冷泉港实验室出版)”等公开出版物中记载的相应方法。
实施例
本公开的其他目的、特征和优点将从以下详细描述中变得明显。但是,应当理解的是,详细描述和具体实施例(虽然表示本公开的具体实施方式)仅为解释性目的而给出,因为在阅读该详细说明后,在本公开的精神和范围内所作出的各种改变和修饰,对于本领域技术人员来说将变得显而易见。
实施例中采用的所有试剂,除非另有强调,否则均可以通过商业途径购买获得。
实施例1人源ScFv噬菌体展示文库的构建方法
表1本实施例中使用的主要试剂
试剂 | 编号 | 制造商 |
Phanta Max超保真DNA聚合酶试剂盒 | P505-d1-AA | Vazyme |
2×T5 Super PCR Mix(Colony) | TSE005 | TSINGKE |
FastPure Gel DNA Extraction Mini Kit | DC301 | Vazyme |
FastPure Plasmid Mini Kit | PM0201-200 | TSINGKE |
TG1 electrocompetent cells | 60502 | Lucigen |
T4 DNA Ligase | NEB | M0202S |
FastDigest NcoI | FD0575 | Thermo Scientific |
FastDigest XhoI | FD0694 | Thermo Scientific |
FastDigest NheI | FD0973 | Thermo Scientific |
FastDigest NotI | FD0595 | Thermo Scientific |
FastDigest sfiI | FD1824 | Thermo Scientific |
pATA-scFv-2Vector | - | ProteoGenix |
M13KO7 helper phage | N0315S | NEB |
1.文库构建
1.1组装重链可变区(VH)和轻链可变区(VL)
表2 PCR反应条件和步骤
其中,变性,退火,延伸(1)这三个步骤,重复30次。
引物序列:
Forward(F):
5′L-VH 1:ACAGGTGCCCACTCCCAGGTGCAG(SEQ ID NO:28)
5′L-VH 3:AAGGTGTCCAGTGTGARGTGCAG(SEQ ID NO:29)
5′L-VH 4/6:CCCAGATGGGTCCTGTCCCAGGTGCAG(SEQ ID NO:30)
5′L-VH 5/7:CAAGGAGTCTGTTCCGAGGTGCAG(SEQ ID NO:31)
5′L Vκ1/2:ATGAGGSTCCCYGCTCAGCTGCTGG(SEQ ID NO:32)
5′L Vκ3:CTCTTCCTCCTGCTACTCTGGCTCCCAG(SEQ ID NO:33)
5′L Vκ4/5:ATTTCTCTGTTGCTCTGGATCTCTG(SEQ ID NO:34)
5′L Vλ1:GGTCCTGGGCCCAGTCTGTGCTG(SEQ ID NO:35)
5′L Vλ2:GGTCCTGGGCCCAGTCTGCCCTG(SEQ ID NO:36)
5′L Vλ3:GCTCTGTGACCTCCTATGAGCTG(SEQ ID NO:37)
5′L Vλ4/5:GGTCTCTCTCSCAGCYTGTGCTG(SEQ ID NO:38)
5′L Vλ6:GTTCTTGGGCCAATTTTATGCTG(SEQ ID NO:39)
5′L Vλ7:GGTCCAATTCYCAGGCTGTGGTG(SEQ ID NO:40)
5′L Vλ8/9/10:GAGTGGATTCTCAGACTGTGGTG(SEQ ID NO:41)Reverse(R):
3′Cκ:TGCTGTCCTTGCTGTCCTGCT(SEQ ID NO:42)
3′Cλ:CACCAGTGTGGCCTTGTTGGCTTG(SEQ ID NO:43)
PCR后琼脂糖凝胶电泳检测结果如图1所示。
1.2构建轻链可变区噬菌体展示文库
1.2.1准备pATA-scFv-2载体为文库克隆
1.2.2消化载体和PCR产物
表3消化载体和PCR产物的反应体系
pATA-scFv-2载体 | 重链/轻链可变区PCR产物 | |
载体或PCR产物 | 25μg | 10μg |
Fast Digest NheI | 5μl | 2μl |
Fast Digest NotI | 5μl | 2μl |
10×Fast Digest Buffer | 25μl | 10μl |
ddH<sub>2</sub>O | 添加至反应体系共250μL | 添加至反应体系共100μL |
1.2.3连接
表4连接反应体系
T4 DNA Ligase(Thermo) | 8μl |
10×T4 DNA Ligase buffer | 15μl |
Vector(NheI/NotI) | 1.5μg |
VL fragment(NheI/NotI) | 0.5μg |
H<sub>2</sub>O | 添加至反应体系共150μl |
16℃孵育15h,65℃加热灭活10min。
1.2.4电转
1)TG1感受态细胞的制备。
2)37℃预温2ml SOC培养基(Sigma,S1797)。将电穿孔试管(0.2厘米的间隙)放在冰上(每个转换反应一个试管)。
3)从-80℃冰箱中取出电活性细胞,放在湿冰上,直到它们完全融化(5-10分钟)。细胞解冻后,轻轻拍打使其混合。
4)仔细移液管50μL DNA混合物倒入冷电穿孔试管不引入泡沫。快速地用你的手腕向下轻挥小口杯,将细胞沉淀到井底。
5)电穿孔600Ω,10μF和2.5kV。在脉冲后10秒内,立即在每个试管中加入2mL预加热SOC培养基。在37摄氏度下,220转/分钟,搅拌1小时。
6)收集所有的电转化介质。连续稀释10μL产物在100μL SOC中、并且在含有Amp/葡萄糖的LB培养基中传播。在37℃下放置过夜。通过菌落数量的计数,乘以培养体积,再除以电镀体积来计算转化子的总数。
1.3构建VL-VH噬菌体展示文库
1.3.1消化载体和PCR产物
表5消化反应体系
1.3.2连接
表6连接反应体系
T4 DNA Ligase(Thermo) | 8μl |
10×T4 DNA Ligase buffer | 15μl |
VL-vector(sfiI/XhoI) | 1.5μg |
VH(sfiI/XhoI) | 0.45μg |
H<sub>2</sub>O | 添加至反应体系共150μl |
16℃孵育15h,65℃加热灭活10min。
1.3.3电转
步骤同1.2.4
1.3.4文库评估
1)菌落PCR
以构建好的文库为模板,进行PCR
表7 PCR反应条件
其中,变性,退火,延伸(1)这三个步骤,重复30次。
Template:此处为构建好的抗体库
引物序列:
Forward(F):TGCTCGGGGATCCGAATTCT(SEQ ID NO:44)
Reverse(R):TCGAGTGCGGCCGCAAGCTT(SEQ ID NO:45)
2)测序
挑选阳性克隆送到武汉擎科生物科技有限公司测序。
测序质控结果如图2所示。
实施例2与新冠病毒特异性抗原肽特异性结合的单克隆抗体的制备
表8本实施例中使用的主要试剂
2.1第一轮
2.1.1生物淘选
(1)将100μl 1μg/ml SA-magnetic磁珠放置在EP管中。
(2)用0.05%PBST洗SA-magnetic磁珠,洗三遍。
(3)用200μl生物素偶联的新冠病毒S蛋白672-691多肽(S672-691-biotin)(100μg/ml)重悬磁珠,室温孵育一小时。
(4)用0.05%PBST洗SA-magnetic磁珠,洗三遍。
(5)用PBS稀释噬菌体文库.再用噬菌体文库重悬SA-magnetic磁珠,室温孵育1.5个小时。
(6)用0.05%PBST洗SA-magnetic磁珠,洗五遍。
(7)用200μL的甘氨酸-盐酸洗脱粘结剂,再用Tris-HCl中和至pH 7.0。
2.1.2测定稀释噬菌体的滴度
(1)培养大肠杆菌TG1直到OD600=0.4-0.6。
(2)混合10μL稀释的洗出液稀释E.coli TG1至180μL。
(3)培养混合物在37℃30分钟,然后倒到2×YT-A(Amp 100μg/ml)培养基中。将培养基倒扣培养在37℃处过夜。
2.1.3噬菌体文库扩增
(1)将10μl E.coli TG1加到800μl of 2YT培养液中在37℃混合培养知道OD600=0.4-0.6.
(2)将培养至对数期的TG1转入10ml 2YT-G(终浓度2%葡萄糖)培养液,在摇床上37℃培养至OD600=0.4-0.6。
(3)加入洗脱后的产物,37℃孵育30分钟,37℃摇床摇30分钟。
(4)加入30ml 2YT-AG培养液(终浓度0.1%Amp,2%葡萄糖),37℃摇床培养1小时。
(5)加入M13KO7(M13KO7:TG1=20:1),37℃孵育30分钟,37℃摇床30分钟。
(6)菌液在5000rpm离心10分钟。用40ml2YT-AK重悬,30℃摇床孵育过夜。
(7)8000rpm离心10分钟,取出上清,用1ml PBS重悬,12000rpm离心5分钟,将上清转移至新的1.5ml离心管。
2.1.4扩增后的噬菌体文库滴度测定
步骤同2.1.2
2.2第二轮
2.2.1生物淘选
(1)用1ml 50μg牛血清白蛋白偶联的新冠病毒S蛋白672-691多肽(S672-691-BSA)覆盖EP管表面,4℃孵育过夜。
(2)用0.05%PBST洗三次。
(3)用5%牛奶装满EP管,30℃封闭两个小时。
(4)用0.05%PBST洗三次。
(5)用1%milk-PBS稀释噬菌体文库。将噬菌体文库加入到EP管中。37℃孵育两个小时。
(6)用0.05%PBST洗三次,用PBS洗两次。
(7)用1ml甘氨酸-盐酸洗脱粘结剂,再用Tris-HCl中和至pH 7.0。
表9生物淘选的结果
轮数 | 淘选前(噬斑形成单位) | 淘选后(噬斑形成单位) |
1 | 5×10<sup>11</sup> | 2.5×10<sup>6</sup> |
2 | 1.7×10<sup>12</sup> | 1.5×10<sup>5</sup> |
3 | 3×10<sup>12</sup> | 1.8×10<sup>8</sup> |
4 | 1.4×10<sup>12</sup> | 1.6×10<sup>9</sup> |
2.2.2噬菌体文库使用
(1)用5ml 5%牛奶/PBS覆盖EP管,4℃孵育过夜。
(2)用0.05%PBST冲洗3次。
(3)将洗脱噬菌体转入EP管中。在32℃下孵育1小时。
(4)将步骤7(1.2.1)中的噬菌体转入EP管中,32℃孵育1h,将噬菌体转入EP管。
2.2.3检测稀释噬菌体的滴度
步骤同2.1.2
2.2.4扩增噬菌体文库
步骤同2.1.3.
2.2.4测定扩增后的噬菌体滴度
步骤同2.1.2
2.3第三轮和第四轮
第三轮和第一轮相同,第四轮和第二轮相同。
表10每轮的选择条件
筛选结果如图3所示,接下来进行验证。
2.4多克隆噬菌体ELISA
(1)用抗原肽覆盖板子表面(实验组:4μg/ml 672-691-BSA;对照组1:10%BSA-PBS;对照组2:0μg/mL抗原肽)4℃孵育过夜。
(2)用0.05%PBST洗三次。
(3)用300μL 5%milk封闭,30℃封闭两个小时。
(4)用0.05%PBST洗三次。
(5)加入100μL稀释的噬菌体文库到每个孔中,如下表,37℃孵育两个小时。
(6)用0.05%PBST洗三次。
(7)用封闭液稀释抗-M13-HRP抗体(1:5000).每个孔中加100μL,37℃孵育1小时。
(8)用0.05%PBST洗三次。
(9)每孔加100μL TMB,室温孵育,每孔再加100μL 2M HCl终止反应。
(10)450nm-630nm读板。
表11多克隆噬菌体ELISA的结果
Ag:包被4μg/ml 672-691-BSA
Nc1:包被10%BSA-PBS
Nc2:包被PBS
2.5单克隆噬菌体ELISA(R3&R4)
(1)在板子上挑选192个克隆,检测洗脱液的滴度。在37℃条件下,摇床250rpm,直到OD600=0.4-0.6。(2)用M13KO7感染培养产物(MOI=20:1)在37℃孵育30分钟,然后37℃摇床30分钟.离心得到相同体积的沉积和上清2×YT-AK(Amp 100μg/ml,Kan100μg/ml)。30℃培养过夜。
(3)离心,上清液可用于酶联免疫吸附试验。
(4)用抗原肽覆盖板子(实验组:4μg/mL 672-691-BSA;对照组:10%BSA-PBS)4℃过夜培养,洗三次。
(5)用300μL 5%牛奶封闭液封闭,30℃2小时。洗三次。
(6)每孔加100μL噬菌体文库,在32℃培养2小时,洗三次。
(7)用封闭液稀释抗-M13-HRP抗体(1:5000)。然后每孔加100μL,37℃孵育1小时。洗三次。
(8)每孔加100μL TMB室温培养,然后每孔加100μL 2M HCl。
(9)450nm-620nm读板。然后对高特异性克隆进行测序。
我们得到的高特异性抗体的序列如下。
VH CDR
VL CDR
表12.单克隆噬菌体ELISA结果实验组(R3P1)
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
A | 0.01 | 0.01 | 0.01 | 0.02 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 |
B | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.53 | 0.01 | 0.01 | 0.01 |
C | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.02 | 0.01 |
D | 0.02 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.02 | 0.03 | 0.02 |
E | 0.03 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.02 | 0.01 | 0.02 |
F | 0.28 | 0.01 | 0.01 | 0.01 | 0.03 | 0.01 | 0.03 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 |
G | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.02 | 0.01 | 0.01 | 0.01 | 0.02 | 0.01 | 0.02 |
H | 0.02 | 0.02 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.02 | 0.02 | 0.02 | 0.02 |
表13.单克隆噬菌体ELISA结果实验组(R4P1)
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
A | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.02 |
B | 0.01 | 0.01 | 0.01 | 0.02 | 0.01 | 0.01 | 2.33 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 |
C | 0.01 | 0.61 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 2.32 | 0.01 | 0.01 | 1.34 | 0.01 |
D | 0.01 | 0.01 | 0.01 | 0.01 | 1.16 | 0.01 | 0.01 | 0.02 | 0.02 | 0.01 | 0.01 | 0.01 |
E | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 |
F | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.03 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 |
G | 0.01 | 0.01 | 2.39 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 |
H | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 0.01 | 2.01 | 0.01 | 0.01 |
2.6阳性克隆的ELISA验证
2.6.1噬菌体文库ELISA
(1)加50μL阳性克隆到2ml 2YT-AG的培养基产物中,直到OD600=0.4-0.6。
(2)用M13KO7感染培养产物(MOI=20:1)在37℃孵育30分钟,然后37℃摇床30分钟.离心得到相同体积的沉积和上清2×YT-AK(Amp 100μg/ml,Kan100μg/ml)。30℃培养过夜。
(3)离心,上清液可用于酶联免疫吸附试验。
(4)用抗原肽覆盖板子(实验组1:4μg/mL 672-691;实验组2:4μg/mL 672-691-BSA;对照组1:10%BSA-PBS;对照组2:4μg/mL SA;对照组3:0μg/mL antigen)4℃过夜培养,洗三次。
(5)用300μL 5%牛奶封闭液封闭,30℃2小时。洗三次。
(6)每孔加100μL噬菌体文库,在37℃培养1小时,洗三次。
(7)用封闭液稀释抗-M13-HRP抗体(1:5000)。然后每孔加100μL,37℃孵育1小时。洗三次。
(8)每孔加100μL TMB室温反应,然后每孔加100μL 2M HCl。
(9)450nm-620nm读板。
表14噬菌体ELISA验证结果
2.6.2蛋白质ELISA
(1)加500μL阳性克隆到5ml of 2YT-AG培养基中培养至OD600=0.8-1.
(2)离心4000rpm 10min,用5ml 2YT-AI(1mM IPTG)培养液重悬沉淀,30℃过夜培养;
(3)离心7000rpm 15min,弃掉上清。
(4)用1ml of pre-chilled TES buffer(lysozyme is now added)重悬细胞,冰上放置30分钟。
(5)离心7000rpm 15min,上清转移至新的EP管中。
(6)抗原肽覆盖板子(实验组1:4μg/mL 672-691;实验组2:4μg/mL 672-691-BSA;对照组1:10%BSA-PBS;对照组2:4μg/mL SA;对照组3:0μg/mL antigen)4℃过夜培养。
(7)用0.05%PBST洗三次。
(8)用300μL 5%milk-PBS封闭。
(9)用0.05%PBST洗三次。
(10)加100μL稀释的蛋白至每个孔中,37℃孵育1hours.
(11)用0.05%PBST洗三次。
(12)用封闭液稀释抗-cmyc-HRP抗体(1:1000),每孔加100μL,37℃孵育1hour.
(13)用0.05%PBST洗五次。
(14)每孔加100μL TMB室温培养,然后每孔加100μL 2M HCl。
(15)读板450nm-630nm。
表15.蛋白质ELISA验证结果
2.7抗体序列的测序
筛选得到的噬菌体阳性克隆,进行全序列测序,得到相应的抗体重链轻链,以及全序列如下:
R4P1-C2抗体重链氨基酸序列为如SEQ ID NO:1所示的序列;
R4P1-C2抗体重链核苷酸序列为如SEQ ID NO:2所示的序列;
R4P1-C2抗体轻链氨基酸序列为如SEQ ID NO:5所示的序列;
R4P1-C2抗体轻链核苷酸序列为如SEQ ID NO:6所示的序列;
R3P1-B9抗体重链氨基酸序列为如SEQ ID NO:7所示的序列;
R3P1-B9抗体重链核苷酸序列为如SEQ ID NO:8所示的序列;
R3P1-B9抗体轻链氨基酸序列为如SEQ ID NO:9所示的序列;
R3P1-B9抗体轻链核苷酸序列为如SEQ ID NO:10所示的序列;
R4P1-C2抗体氨基酸序列为如SEQ ID NO:24所示的序列;
R4P1-C2抗体核苷酸序列为如SEQ ID NO:25所示的序列;
R3P1-B9抗体氨基酸序列为如SEQ ID NO:26所示的序列;
R3P1-B9抗体核苷酸序列为如SEQ ID NO:27所示的序列。
实施例3 ELISA检测抗体不同稀释浓度条件下的OD值
酶联免疫吸附反应ELISA实验步骤:
1.将MES粉末(SIGMA,Lot#SLBZ3485)用ddH2O配制成0.1M、pH=6.0的MES buffer;将EDC(C8H17N3,Thermo Scientific,Lot#TB257918)用ddH2O稀释成10mg/ml。
2.将多肽用0.1M的MES buffer稀释成4μg/ml。
3.在微孔板设阴性对照,每孔中加入10μl EDC溶液和50μl多肽溶液;其余孔中加入10μl EDC溶液和50μl MES buffer。轻轻震动混匀。用不干胶条封板后于4℃过夜或室温放置两个小时以上。
4.去掉不干胶条,吸去孔内液体,每孔加300μl ddH2O,静置2分钟,弃去液体,拍干板子。再重复以上步骤2次。
5.用1X PBST配制1%BSA的封闭液(10X PBST:Solarbio,Cat#P1033-500;BSA:Solarbio,Cat#A8020),每孔加入200μl封闭液,室温封闭1小时。
6.弃去孔内液体,拍干板子。将待测血清用封闭液按1:500稀释,每孔加入100μl稀释血清,室温反应1小时。
7.弃去孔内液体,每孔加300μl 1X PBST,静置2分钟,弃去液体,拍干板子。再重复以上步骤2次。
8.将HRP标记的Goat Anti-Human IgG(Cwbio,Cat#CW0169S)用封闭液按1:5000稀释,每孔100μl,室温反应40分钟。
9.重复操作步骤7,用1XPBST洗板5次。
10.每孔加入TMB-ELISA显色液(Thermo Scientific,Lot#TK2666052)100μl,避光反应5-15分钟。
11.每孔加入2M H2SO4溶液50μl终止反应。
12.将酶标仪设定波长450nm测量各孔OD值,应在终止反应后30分钟内读值。
分别用S672-691多肽以及偶联BSA的S672-691多肽包被板子,检测不同浓度的单克隆抗体,ELISA检测结果如图4所示。
实施例4中和实验
4.1实验前准备
4.1.1平衡试剂
将保存在2-8℃的试剂(胰酶,DMEM完全培养基)取出,至室温平衡,30分钟以上
4.1.2操作人员
由经过培训的实验操作人员进行操作,实验操作前,在清洁区内更衣(穿好一次性无菌衣,换好工作鞋,戴好口罩,帽子,一次性医用乳胶手套)方可进入实验区内,进行实验操作。
4.2实验操作
4.2.1将待检测的血清(或血浆)于56℃水浴灭活30min,6000g离心3min,将上清转移至1.5ml离心管中待用。
4.2.2取96孔板,于第2列(细胞对照CC,见表2)加入DMEM完全培养基(1%双抗,25mM HEPES,10%FBS)150μl/孔,于第3~11列(第3列为病毒对照组VV,第4~11列为样品孔)加入DMEM完全培养基100μl/孔,于B4~B11孔中再加入DMEM完全培养基42.5μl/孔。
4.2.3于B4和B5孔加入血浆样品1(7.5μl)……以此类推,于B10和B11孔加入血浆样品4(7.5μl)。
4.2.4将多道移液器调至50μl,对B4~B11孔中液体轻柔的反复吹吸6~8次充分混匀,然后转移50μl液体至对应的C4~C11中吸弃50μl液体,加样顺序和加样方式参照表16。
表16加样顺序和加样方式
4.2.5用DMEM完全培养基将假病毒稀释至2*104TCID50/ml(按提供的稀释倍数稀释),于第3~11列每孔加50μl,使每孔含假病毒的量为1*103/孔。
4.2.6将上述96孔板置于细胞培养箱中(37℃,5%CO2)孵育1小时。
4.2.7当孵育时间至半小时,取出培养箱中事先准备好的Huh-7细胞(汇合率达80%~90%),以T75培养瓶为例,吸弃瓶中的培养基,加入5ml PBS缓冲液清洗细胞,倾去PBS后,加入3ml 0.25%胰酶-EDTA,使其浸没细胞消化1分钟,倾去胰酶,置于细胞培养箱中消化5分钟,轻轻拍打培养瓶侧壁使细胞脱落,加入10ml培养基中和胰酶,吹打几次后转移至离心管中,210g离心5分钟,倾去上清,用10ml DMEM完全培养基重悬细胞,细胞计数,用DMEM完全培养基将细胞稀释至5*105个/ml。
4.2.8孵育至1小时,向96孔板中每孔加100μl细胞,使每孔细胞为5*104个。
4.2.9将96孔板前后左右轻轻晃动,使细胞在孔中分散均匀,将96孔板放入细胞培养箱中,37℃,5%CO2培养20~28小时。
4.2.10 20~28小时后从细胞培养箱中取出96孔板,用多道移液器从每个上样孔中吸弃150μl上清,然后加入100μl荧光素酶检测试剂,室温避光反应2min。
4.2.11反应结束后,用多道移液器将反应孔中的液体反复吹吸6~8次,使细胞充分裂解,从每孔中吸出150μl液体,加入对应96孔化学发光检测板中,置于化学发光检测仪中读取发光值。
4.2.12计算中和抑制率:抑制率=[1-(样品组的发光强度均值-空白对照CC均值)/(阴性组的发光强度VC均值-空白对照CC均值)]*100%。
4.2.13根据中和抑制率结果,利用Reed-Muench法计算IC50。
中和实验结果如图5所示。
实验结果表明,图5中所涉及的两个多肽均具有良好的中和效果。
本公开并不旨在限于具体公开的实施方案的范围,提供所述实施方案例如来说明本公开的各方面。从本文的描述和教导,对所述组合物和方法的各种修改将变得明显。可以在不脱离本公开的真正范围和精神的情况下实践这类变化,并且这类变化旨在落入本公开的范围内。
序列表编号
本公开中,未在说明书中明确标识的序列所对应的含义如下:
SEQ ID NO:3示出了R4P1-C2抗体和R3P1-B9抗体的连接肽氨基酸序列;
SEQ ID NO:4示出了R4P1-C2抗体和R3P1-B9抗体的连接肽核苷酸序列;
SEQ ID NO:11-13依次示出了R4P1-C2抗体的重链的CDR1、CDR2、CDR3的氨基酸序列;
SEQ ID NO:14-16依次示出了R4P1-C2抗体的轻链的CDR1、CDR2、CDR3的氨基酸序列;
SEQ ID NO:17-19依次示出了R3P1-B9抗体的重链的CDR1、CDR2、CDR3的氨基酸序列;
SEQ ID NO:20-22依次示出了R3P1-B9抗体的轻链的CDR1、CDR2、CDR3的氨基酸序列;
SEQ ID NO:23示出了R4P1-C2抗体和R3P1-B9抗体所对应的抗原肽的氨基酸序列。
序列表
<110> 苏州方科生物科技有限公司
苏州系统医学研究所
<120> 新型冠状病毒特异性抗原肽的人源抗体、制备方法及用途
<160> 45
<170> SIPOSequenceListing 1.0
<210> 1
<211> 121
<212> PRT
<213> Artificial Sequence
<400> 1
Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Asn Ser Asp
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Thr Pro Glu Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Val Tyr His Ser Gly Ile Thr Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ser Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Asn Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Leu Gly Ser Thr Ser Ser Asp Gly Met Asp Val Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser
115 120
<210> 2
<211> 363
<212> DNA
<213> Artificial Sequence
<400> 2
cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60
acctgcgctg tctctggtgg ctccatcaac agtgataact ggtggagttg ggtccgccag 120
accccagaga aggggctgga gtggattgga gaagtctatc atagtggaat caccaactac 180
aacccgtccc tcaagagtcg agtctccata tctgtagaca agtccaagaa ccagttctcc 240
ctgaacctga attctgtgac cgccgcggac acggccgtgt attactgtgc gagagatctc 300
gggagcacct cgtctgacgg tatggacgtc tggggccaag ggaccacggt caccgtctcg 360
agt 363
<210> 3
<211> 21
<212> PRT
<213> Artificial Sequence
<400> 3
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
1 5 10 15
Gly Gly Gly Ala Ser
20
<210> 4
<211> 63
<212> DNA
<213> Artificial Sequence
<400> 4
ggtggtggcg gttctggtgg tggtggtagc ggtggcggtg gtagtggcgg tggcggtgct 60
agc 63
<210> 5
<211> 108
<212> PRT
<213> Artificial Sequence
<400> 5
Gln Ala Gly Leu Thr Gln Pro Pro Ser Leu Ser Val Ser Pro Gly Gln
1 5 10 15
Thr Ala Arg Ile Thr Cys Ser Gly Asn Ala Leu Pro Lys Gln Tyr Ala
20 25 30
Tyr Trp Tyr Arg Gln Arg Pro Gly Gln Ala Pro Glu Leu Val Met Ser
35 40 45
Arg Asp Val Glu Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser
50 55 60
Ser Ser Gly Thr Thr Val Thr Leu Thr Ile Ser Gly Val Gln Ala Glu
65 70 75 80
Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Ala Asp Asn Ser Gly Thr Phe
85 90 95
Val Val Phe Gly Gly Gly Thr Gln Leu Ile Ile Leu
100 105
<210> 6
<211> 324
<212> DNA
<213> Artificial Sequence
<400> 6
caggcagggc tgactcagcc accctcgctg tcagtgtccc caggacagac ggccaggatc 60
acttgctctg gaaatgcgtt gccaaaacaa tatgcttatt ggtatcgcca gaggccaggc 120
caggcccctg aattggtgat gtctagagac gttgagaggc cctcagggat ccctgagcga 180
ttctctggct ccagctcagg gacaacagtc accttgacca ttagtggagt ccaggcagaa 240
gacgaggctg actattattg tcaatcagca gacaacagtg ggacttttgt ggttttcggc 300
ggaggaaccc agctgatcat ttta 324
<210> 7
<211> 127
<212> PRT
<213> Artificial Sequence
<400> 7
Gln Leu Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Arg Gly Phe Gly Glu Phe Gly Phe Val Pro Gly Ala Ser
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 125
<210> 8
<211> 381
<212> DNA
<213> Artificial Sequence
<400> 8
cagctgcagc tgcaggagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180
gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gagagatcgc 300
gggttcgggg agtttggttt tgtccccggc gcgagctggt tcgacccctg gggccaggga 360
accctggtca ccgtctcgag t 381
<210> 9
<211> 110
<212> PRT
<213> Artificial Sequence
<400> 9
Gln Pro Val Leu Thr Gln Pro Pro Ser Val Ser Ala Thr Pro Gly Gln
1 5 10 15
Thr Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Ser
20 25 30
Pro Val Asn Trp Tyr Gln Gln Leu Pro Gly Ala Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Thr Asn Asp Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Ala Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Val Trp His Asp Ser Leu
85 90 95
Asn Thr Tyr Val Phe Gly Thr Gly Thr Glu Leu Thr Val Leu
100 105 110
<210> 10
<211> 330
<212> DNA
<213> Artificial Sequence
<400> 10
cagcctgtgc tgactcagcc accctcagtg tctgccaccc ccgggcagac ggtcaccatc 60
tcttgttctg gaagcagctc caacatcgga agtagtcctg ttaactggta ccagcagctc 120
ccaggagcgg cccccaaact cctcatctat actaatgata agaggccctc aggggtccct 180
gaccgattct ccggctccaa gtctgccacc tcagcctccc tggccatcag tgggctccag 240
tctgaggatg aggcagatta ttactgtgca gtatggcatg acagcctgaa tacttatgtc 300
ttcggaactg ggacggagct gaccgtccta 330
<210> 11
<211> 9
<212> PRT
<213> Artificial Sequence
<400> 11
Gly Gly Ser Ile Asn Ser Asp Asn Trp
1 5
<210> 12
<211> 7
<212> PRT
<213> Artificial Sequence
<400> 12
Val Tyr His Ser Gly Ile Thr
1 5
<210> 13
<211> 14
<212> PRT
<213> Artificial Sequence
<400> 13
Ala Arg Asp Leu Gly Ser Thr Ser Ser Asp Gly Met Asp Val
1 5 10
<210> 14
<211> 6
<212> PRT
<213> Artificial Sequence
<400> 14
Ala Leu Pro Lys Gln Tyr
1 5
<210> 15
<211> 3
<212> PRT
<213> Artificial Sequence
<400> 15
Arg Asp Val
1
<210> 16
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 16
Gln Ser Ala Asp Asn Ser Gly Thr Phe Val Val
1 5 10
<210> 17
<211> 8
<212> PRT
<213> Artificial Sequence
<400> 17
Gly Phe Thr Phe Ser Ser Tyr Ala
1 5
<210> 18
<211> 8
<212> PRT
<213> Artificial Sequence
<400> 18
Ile Ser Tyr Asp Gly Ser Asn Lys
1 5
<210> 19
<211> 20
<212> PRT
<213> Artificial Sequence
<400> 19
Ala Arg Asp Arg Gly Phe Gly Glu Phe Gly Phe Val Pro Gly Ala Ser
1 5 10 15
Trp Phe Asp Pro
20
<210> 20
<211> 8
<212> PRT
<213> Artificial Sequence
<400> 20
Ser Ser Asn Ile Gly Ser Ser Pro
1 5
<210> 21
<211> 3
<212> PRT
<213> Artificial Sequence
<400> 21
Thr Asn Asp
1
<210> 22
<211> 11
<212> PRT
<213> Artificial Sequence
<400> 22
Ala Val Trp His Asp Ser Leu Asn Thr Tyr Val
1 5 10
<210> 23
<211> 20
<212> PRT
<213> SARS-COV-2
<400> 23
Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val
1 5 10 15
Ala Ser Gln Ser
20
<210> 24
<211> 250
<212> PRT
<213> Artificial Sequence
<400> 24
Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
1 5 10 15
Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Asn Ser Asp
20 25 30
Asn Trp Trp Ser Trp Val Arg Gln Thr Pro Glu Lys Gly Leu Glu Trp
35 40 45
Ile Gly Glu Val Tyr His Ser Gly Ile Thr Asn Tyr Asn Pro Ser Leu
50 55 60
Lys Ser Arg Val Ser Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser
65 70 75 80
Leu Asn Leu Asn Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Leu Gly Ser Thr Ser Ser Asp Gly Met Asp Val Trp Gly
100 105 110
Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Ser Gln Ala
130 135 140
Gly Leu Thr Gln Pro Pro Ser Leu Ser Val Ser Pro Gly Gln Thr Ala
145 150 155 160
Arg Ile Thr Cys Ser Gly Asn Ala Leu Pro Lys Gln Tyr Ala Tyr Trp
165 170 175
Tyr Arg Gln Arg Pro Gly Gln Ala Pro Glu Leu Val Met Ser Arg Asp
180 185 190
Val Glu Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser Ser Ser
195 200 205
Gly Thr Thr Val Thr Leu Thr Ile Ser Gly Val Gln Ala Glu Asp Glu
210 215 220
Ala Asp Tyr Tyr Cys Gln Ser Ala Asp Asn Ser Gly Thr Phe Val Val
225 230 235 240
Phe Gly Gly Gly Thr Gln Leu Ile Ile Leu
245 250
<210> 25
<211> 750
<212> DNA
<213> Artificial Sequence
<400> 25
cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60
acctgcgctg tctctggtgg ctccatcaac agtgataact ggtggagttg ggtccgccag 120
accccagaga aggggctgga gtggattgga gaagtctatc atagtggaat caccaactac 180
aacccgtccc tcaagagtcg agtctccata tctgtagaca agtccaagaa ccagttctcc 240
ctgaacctga attctgtgac cgccgcggac acggccgtgt attactgtgc gagagatctc 300
gggagcacct cgtctgacgg tatggacgtc tggggccaag ggaccacggt caccgtctcg 360
agtggtggtg gcggttctgg tggtggtggt agcggtggcg gtggtagtgg cggtggcggt 420
gctagccagg cagggctgac tcagccaccc tcgctgtcag tgtccccagg acagacggcc 480
aggatcactt gctctggaaa tgcgttgcca aaacaatatg cttattggta tcgccagagg 540
ccaggccagg cccctgaatt ggtgatgtct agagacgttg agaggccctc agggatccct 600
gagcgattct ctggctccag ctcagggaca acagtcacct tgaccattag tggagtccag 660
gcagaagacg aggctgacta ttattgtcaa tcagcagaca acagtgggac ttttgtggtt 720
ttcggcggag gaacccagct gatcatttta 750
<210> 26
<211> 258
<212> PRT
<213> Artificial Sequence
<400> 26
Gln Leu Gln Leu Gln Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Asp Arg Gly Phe Gly Glu Phe Gly Phe Val Pro Gly Ala Ser
100 105 110
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly
115 120 125
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
130 135 140
Gly Gly Ala Ser Gln Pro Val Leu Thr Gln Pro Pro Ser Val Ser Ala
145 150 155 160
Thr Pro Gly Gln Thr Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn
165 170 175
Ile Gly Ser Ser Pro Val Asn Trp Tyr Gln Gln Leu Pro Gly Ala Ala
180 185 190
Pro Lys Leu Leu Ile Tyr Thr Asn Asp Lys Arg Pro Ser Gly Val Pro
195 200 205
Asp Arg Phe Ser Gly Ser Lys Ser Ala Thr Ser Ala Ser Leu Ala Ile
210 215 220
Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Val Trp
225 230 235 240
His Asp Ser Leu Asn Thr Tyr Val Phe Gly Thr Gly Thr Glu Leu Thr
245 250 255
Val Leu
<210> 27
<211> 774
<212> DNA
<213> Artificial Sequence
<400> 27
cagctgcagc tgcaggagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt caccttcagt agctatgcta tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactac 180
gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agctgaggac acggctgtgt attactgtgc gagagatcgc 300
gggttcgggg agtttggttt tgtccccggc gcgagctggt tcgacccctg gggccaggga 360
accctggtca ccgtctcgag tggtggtggc ggttctggtg gtggtggtag cggtggcggt 420
ggtagtggcg gtggcggtgc tagccagcct gtgctgactc agccaccctc agtgtctgcc 480
acccccgggc agacggtcac catctcttgt tctggaagca gctccaacat cggaagtagt 540
cctgttaact ggtaccagca gctcccagga gcggccccca aactcctcat ctatactaat 600
gataagaggc cctcaggggt ccctgaccga ttctccggct ccaagtctgc cacctcagcc 660
tccctggcca tcagtgggct ccagtctgag gatgaggcag attattactg tgcagtatgg 720
catgacagcc tgaatactta tgtcttcgga actgggacgg agctgaccgt ccta 774
<210> 28
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 28
acaggtgccc actcccaggt gcag 24
<210> 29
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 29
aaggtgtcca gtgtgargtg cag 23
<210> 30
<211> 27
<212> DNA
<213> Artificial Sequence
<400> 30
cccagatggg tcctgtccca ggtgcag 27
<210> 31
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 31
caaggagtct gttccgaggt gcag 24
<210> 32
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 32
atgaggstcc cygctcagct gctgg 25
<210> 33
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 33
ctcttcctcc tgctactctg gctcccag 28
<210> 34
<211> 25
<212> DNA
<213> Artificial Sequence
<400> 34
atttctctgt tgctctggat ctctg 25
<210> 35
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 35
ggtcctgggc ccagtctgtg ctg 23
<210> 36
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 36
ggtcctgggc ccagtctgcc ctg 23
<210> 37
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 37
gctctgtgac ctcctatgag ctg 23
<210> 38
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 38
ggtctctctc scagcytgtg ctg 23
<210> 39
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 39
gttcttgggc caattttatg ctg 23
<210> 40
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 40
ggtccaattc ycaggctgtg gtg 23
<210> 41
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 41
gagtggattc tcagactgtg gtg 23
<210> 42
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 42
gtgctgtcct tgctgtcctg ct 22
<210> 43
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 43
caccagtgtg gccttgttgg cttg 24
<210> 44
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 44
tgctcgggga tccgaattct 20
<210> 45
<211> 20
<212> DNA
<213> Artificial Sequence
<400> 45
tcgagtgcgg ccgcaagctt 20
Claims (15)
1.一种抗SARS-CoV-2抗体或其抗原结合片段,所述抗体特异性地结合所述SARS-CoV-2表位,其中,所述SARS-CoV-2表位包含如SEQ ID NO:23所示的序列;
所述抗体或其抗原结合片段包含重链可变区VH和轻链可变区VL,所述VH包含VHCDR1、VHCDR2和VHCDR3,以及所述VL包含VLCDR1、VLCDR2和VLCDR3,其中,
VHCDR1的氨基酸序列如SEQ ID NO:11所示,VHCDR2的氨基酸序列如SEQ ID NO:12所示,和VHCDR3的氨基酸序列如SEQ ID NO:13所示;所述VL由如下氨基酸编码:VLCDR1的氨基酸序列如SEQ ID NO:14所示,VLCDR2的氨基酸序列如SEQ ID NO:15所示,和VLCDR3的氨基酸序列如SEQ ID NO:16所示;或者,
VHCDR1的氨基酸序列如SEQ ID NO:17所示,VHCDR2的氨基酸序列如SEQ ID NO:18所示,和VHCDR3的氨基酸序列如SEQ ID NO:19所示;所述VL由如下氨基酸编码:VLCDR1的氨基酸序列如SEQ ID NO:20所示,VLCDR2的氨基酸序列如SEQ ID NO:21所示,和VLCDR3的氨基酸序列如SEQ ID NO:22所示。
2.根据权利要求1所述的抗体或其抗原结合片段,其包含连接子,其中,编码所述连接子的序列包含如SEQ ID NO:3所示的序列。
3.根据权利要求1所述的抗体或其抗原结合片段,其中,编码所述抗体或其抗原结合片段包含如下所示序列中的一种或多种:
(1)VH的氨基酸序列如SEQ ID NO:1所示,和VL的氨基酸序列如SEQ ID NO:5所示;
(2)VH的氨基酸序列如SEQ ID NO:7所示,和VL的氨基酸序列如SEQ ID NO:9所示;
(3)如SEQ ID NO:24或26所示的氨基酸序列。
4.一种多核苷酸,其中,所述多核苷酸选自(5)-(6)中的任一项:
(5)如SEQ ID NO:2和SEQ ID NO:6所示的核苷酸序列,或如SEQ ID NO:8和SEQ ID NO:10所示的核苷酸序列;
(6)如(5)所示的核苷酸序列的反向互补序列的核苷酸序列。
5.根据权利要求4所述的多核苷酸,其中,所述多核苷酸选自(9)-(10)中的任一项:
(9)如SEQ ID NO:25或27任一序列所示的核苷酸序列;
(10)如SEQ ID NO:25,27任一序列所示的序列的反向互补序列的核苷酸序列。
6.一种载体,其中,所述载体包含根据权利要求4-5任一项所述的多核苷酸。
7.一种分离的宿主细胞,其中,所述宿主细胞包含如权利要求6所述的载体。
8.一种制备稳定表达目标蛋白的宿主细胞的方法,其中,所述方法包含利用权利要求6所述的载体,转化初始宿主细胞的步骤。
9.根据权利要求8所述的方法,其中,所述宿主细胞为中国仓鼠卵巢细胞。
10.一种制备目标蛋白的方法,所述方法包含利用权利要求7所述的宿主细胞、或通过权利要求8所述的方法,制备所述目标蛋白。
11.一种抗体或其抗原结合片段,其中,所述抗体或其抗原结合片段为权利要求10所述的方法制备。
12.一种试剂盒,其中,所述试剂盒包含根据权利要求1-3任一项或权利要求11所述的抗体或其抗原结合片段。
13.权利要求12所述的试剂盒在制备用于检测COVID-19的试剂盒中的应用。
14.一种药物组合物或疫苗,其中,所述药物组合物或疫苗中含有根据权利要求1-3任一项或权利要求11所述的抗体或其抗原结合片段。
15.权利要求1-3任一项或权利要求11所述的抗体或其抗原结合片段,或权利要求14所述的药物组合物或疫苗在制备用于治疗或预防COVID-19的药物的应用。
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AU2021209282B2 (en) | 2020-09-04 | 2022-06-02 | Newsoara Biopharma Co., Ltd. | Anti-Sars-Cov-2 Neutralizing Antibodies |
AU2021209287B1 (en) | 2021-01-19 | 2022-03-24 | Newsoara Biopharma Co., Ltd. | Expression Vector for Anti-Sars-Cov-2 Neutralizing Antibodies |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1566143A (zh) * | 2003-06-25 | 2005-01-19 | 中国科学院上海生命科学研究院 | Sars冠状病毒核衣壳蛋白的抗原决定簇及其应用 |
CN111333722A (zh) * | 2020-03-03 | 2020-06-26 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | SARS-CoV-2抑制剂及其应用 |
CN111454354A (zh) * | 2020-04-14 | 2020-07-28 | 吉林大学 | 抗2019-nCoV的抗体、制剂及其制备方法和应用 |
-
2020
- 2020-08-10 CN CN202010794796.8A patent/CN112300274B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1566143A (zh) * | 2003-06-25 | 2005-01-19 | 中国科学院上海生命科学研究院 | Sars冠状病毒核衣壳蛋白的抗原决定簇及其应用 |
CN111333722A (zh) * | 2020-03-03 | 2020-06-26 | 江苏省疾病预防控制中心(江苏省公共卫生研究院) | SARS-CoV-2抑制剂及其应用 |
CN111454354A (zh) * | 2020-04-14 | 2020-07-28 | 吉林大学 | 抗2019-nCoV的抗体、制剂及其制备方法和应用 |
Non-Patent Citations (1)
Title |
---|
Safety, tolerability, and immunogenicity of a recombinant adenovirus type-5 vectored COVID-19 vaccine: a dose-escalation, open-label, non-randomised, first-in-human trial;Feng-Cai Zhu等;《lancet》;20200522;第395卷;第1845-1854页 * |
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