CN114316040B - 一种抗新型冠状病毒的全人源单克隆抗体及其用途 - Google Patents
一种抗新型冠状病毒的全人源单克隆抗体及其用途 Download PDFInfo
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Abstract
本发明涉及一种抗新型冠状病毒的全人源单克隆抗体及其用途,属于生物工程技术领域。所述的全人源单克隆抗体或其片段包括重链可变区和轻链可变区,重链可变区的氨基酸序列如SEQ ID NO:11、13、15、17或19中任一项所示;轻链可变区的氨基酸序列如SEQ ID NO:12、14、16、18或20中任一项所示。本发明还提供该抗体的编码基因、表达载体、用途和组合物。本发明的结合分子能阻止新型冠状病毒侵染易感细胞,且是全人源的,与其它动物源性(如鼠源性)抗新型冠状病毒的分子相比,免疫原性低,且亲和性良好,不仅治疗效果好,而且副作用低,同时可靠的有效性及安全性也为此类抗体的标准化生产成药提供了很大的保障。
Description
技术领域
本发明涉及生物工程技术领域,具体地涉及一种高亲和力抗新型冠状病毒SARS-CoV-2的全人源单克隆抗体及其用途。
背景技术
单克隆抗体药物产业现已成为全球生物制品行业中占比最大的子行业,该类药物具有特异性强、疗效快、半衰期短和副作用小等临床治疗优势,其代表了药品治疗领域的最新发展方向。单抗药在全球生物制品行业中的市场占比已由1997年2.5%上升到2015年的34.7%,到2015年,全球市场规模已达到916亿美元。2010-2015年也是我国单抗药物行业的高速发展期,据中投顾问预测,到2020年,我国单克隆抗体药物产业市场规模将达到280亿元,2016-2020年年均复合增长率达30%,远超Research and Markets预测的未来5年全球单克隆抗体药物产业9.84%的增速水平。单克隆抗体技术在近年内得到了迅猛的发展,从最初的动物抗体、嵌合性抗体到部分人源化,乃至最终完全的人源化抗体,单抗药物的免疫原性反应与副作用已经大幅降低,其疗效不佳也得到了显著的改善。考虑到异体蛋白/抗体的免疫原性和治疗的效果,治疗性单克隆抗体正朝着人源化和全人源单克隆抗体的方向发展,尤其是全人源单克隆抗体的开发将成为抗体药物研发领域的主要方向。此外,治疗性全人源单克隆抗体药物研发的主要技术包括: 1) 由人源单克隆抗体转基因鼠生产的完全抗体;2) 由Phage Ribosome/mRNA and Yeast cell displays技术生产的部分重组抗体如scFv, Fab, Diabody, dAb等;3) 由人杂交瘤或人B淋巴细胞系生产的完全抗体。目前人源单克隆抗体的生产主要来源于转基因鼠和从噬菌体等体系表达生产。然而,由于由重组技术生产的不完全单克隆抗体对靶组织/细胞的特异性和亲和力较弱,其应用前景不佳。
自2019年底新冠疫情爆发以来,新冠病毒的大面积传播对人类公共健康造成了严重的威胁。至今新冠病毒的不断变异和再次感染爆发提醒我们,冠状病毒仍然是严重危害共同安全的高危致病病原体,虽然目前新冠疫苗已开始在人群中普遍接种,但是随着气候和生态环境的变化以及人与动物互动的增加,未来似乎不可避免地出现新的冠状病毒感染爆发,尽快开发出有效的抗冠状病毒药物,已成为临床治疗的迫切需要。
目前已有 20 多种针对新冠抗体产品投入使用,并一度将该病原体的刺突蛋白推入十大靶点名单。其中部分已经获得了FDA的紧急使用授权,但这些药物并没有获得完全批准。抗体药物用于病毒感染性疾病的治疗已有报道,如抗血清用于治疗SARS和重症H5N1丙型肝炎病毒感染者的案例已经证明抗体在治疗病毒感染中起重要作用。全人源单克隆抗体不仅可以阻断病毒与靶细胞的结合,通过补体和T细胞、NK 细胞等效应细胞发挥作用可将病毒杀灭或将被病毒感染的细胞清除,此外,单抗药物还具有免疫原性低、特异性强、疗效快和副作用小等明显优势。冠状病毒主要分为α,β,γ和δ冠状病毒四个亚家族,其中α冠状病毒和β冠状病毒可以感染哺乳动物,γ冠状病毒和δ冠状病毒可以感染鸟类。目前发现的可以感染人类的冠状病毒有7个,其中SARS-CoV-2属于β亚型冠状病毒。SARS-CoV-2的Spike蛋白(S蛋白)是该病毒最重要的表面蛋白,与病毒的传染密切相关。S蛋白含有S1和S2两个亚基,其中S1亚基包含的受体结合区域(S-RBD),主要负责识别细胞受体;而S2蛋白则含有膜融合过程所需的基本元件。我们的生物信息学研究发现,SARS-CoV-2的S1-RBD区域与SARS和MERS等相关冠状病毒有一定的相似性,而S2区域与其它6种致病性冠状病毒高度相似性保守,且到目前为止市面上还没有针对S2蛋白所开发的单克隆抗体药物。因此,开发效果好、可针对多数冠状病毒、在临床上能大规模使用的特异性治疗冠状病毒感染的生物药物具有十分重要的意义。
发明内容
本发明人经过深入的研究,选取SARS-CoV-2表面具有良好免疫原性的S2蛋白作为抗原表位,通过抗原蛋白序列构建质粒后表达和纯化抗原蛋白,之后检测SARS-CoV-2感染后康复人群的血清抗体水平,并从中筛选出的含有较高血清抗体水平的多个个体,然后采集这些个体的抗凝全血,分离其外周血液的单核细胞,建立噬菌体文库,利用S2蛋白(高度保守,可针对包括SARS-CoV-2在内的所有致病性冠状病毒及今后可能出现的变异病毒株)为抗原蛋白,经过ELISA逐级筛选得到目的抗体,再通过转染HEK 293T细胞,获取含有目的抗体的细胞培养上清,最后进行中和效价实验并确定其抗病毒效果,从而筛选出可表达针对新型冠状病毒(SARS-CoV-2)及其它致病性冠状病毒具有较强中和活性的抗体细胞株。本发明的单克隆抗体是全人源的,其重链、轻链可变区以及恒定区均来源于人的基因,因此具有免疫原性低、安全性高的特点,试验结果证明,本发明的单克隆抗体具有高亲和力的特点,本发明提供的单克隆抗体能具有显著抗新型冠状病毒(SARS-CoV-2),为此类抗体的标准化生产成药提供了很大的保障。
前期生物信息学研究表明,新型冠状病毒(SARS-CoV-2)S1蛋白的RBD区域序列与SARS和MERS等相关冠状病毒有一定的相似性,而其S2区域与已发现的其他致病性冠状病毒具有高度相似性,提示S2蛋白的保守程度高,由于S2在不同冠状病毒中均具有高度的相似性,因此这也为今后可能发生的冠状病毒感染疫情的防治提供一定的战略储备。因此在抗原靶点的选择时,我们选择S2蛋白作为抗原表位,最终获得了具有高安全性,高亲和力的抗新型冠状病毒(SARS-CoV-2)的全人源单克隆抗体。
本发明一方面提供一种抗新型冠状病毒的全人源单克隆抗体,其具有重链可变区和轻链可变区:
所述重链可变区的氨基酸序列如SEQ ID NO:11、13、15、17或19中任一项所示;或具有如SEQ ID NO:11、13、15、17或19中任一项所示氨基酸序列经替换、缺失或增加一个或几个氨基酸形成的具有同等功能的氨基酸序列;
所述轻链可变区的氨基酸序列如EQ ID NO:12、14、16、18或20中任一项所示;或具有EQ ID NO:12、14、16、18或20中任一项所示氨基酸序列经替换、缺失或增加一个或几个氨基酸形成的具有同等功能的氨基酸序列。
优选地,所述全人源单克隆抗体具有如下任一组重链可变区和轻链可变区:
(i)重链可变区的氨基酸序列如SEQ ID NO:11所示(R2P1-D8-H);轻链可变区的氨基酸序列如SEQ ID NO:12所示(R2P1-D8-L);
(ii)重链可变区的氨基酸序列如SEQ ID NO:13所示(R2P2-D10-H);轻链可变区的氨基酸序列如SEQ ID NO:14所示(R2P2-D10-L);
(iii)重链可变区的氨基酸序列如SEQ ID NO:15所示(R2P2-G5-H);轻链可变区的氨基酸序列如SEQ ID NO:16所示(R2P2-G5-L);
(iv)重链可变区的氨基酸序列如SEQ ID NO:17所示(R2P2-D7-H);轻链可变区的氨基酸序列如SEQ ID NO:18所示(R2P2-D7-L);
(v)重链可变区的氨基酸序列如SEQ ID NO:19所示(R2P2-F8-H);轻链可变区的氨基酸序列如SEQ ID NO:20所示(R2P2-F8-L)。
上述重链可变区和轻链可变区组成的全人源单克隆抗体能特异性结合新型冠状病毒(SARS-CoV-2)S2蛋白。
在本发明的另一方面,提供一种编码上述全人源单克隆抗体的核酸分子。
优选地,所述核酸分子具有如SEQ ID NO:1、3、5、7或9中任一项所示的重链可变区的核苷酸序列;和如SEQ ID NO:2、4、6、8或10中任一项所示的轻链可变区的核苷酸序列。
更优选地,所述核酸分子具有如下任一组重链可变区和轻链可变区:
(i)重链可变区的核苷酸序列如SEQ ID NO:1所示(R2P1-D8-H);轻链可变区的核苷酸序列如SEQ ID NO:2所示(R2P1-D8-L);
(ii)重链可变区的核苷酸序列如SEQ ID NO:3所示(R2P2-D10-H);轻链可变区的核苷酸序列如SEQ ID NO:4所示(R2P2-D10-L);
(iii)重链可变区的核苷酸序列如SEQ ID NO:5所示(R2P2-G5-H);轻链可变区的核苷酸序列如SEQ ID NO:6所示(R2P2-G5-L);
(iv)重链可变区的核苷酸序列如SEQ ID NO:7所示(R2P2-D7-H);轻链可变区的核苷酸序列如SEQ ID NO:8所示(R2P2-D7-L);
(v)重链可变区的核苷酸序列如SEQ ID NO:9所示(R2P2-F8-H);轻链可变区的核苷酸序列如SEQ ID NO:10所示(R2P2-F8-L)。
在本发明的另一方面,提供一种表达载体,所述的表达载体包括上述核酸分子。
该表达载体除包含上述的核酸分子,还可以包含适当启动子或者控制序列。该表达载体可以用于转化适当的宿主细胞,以使其能够表达蛋白质。
在本发明的另一方面,提供一种宿主细胞,所述的宿主细胞包括上述表达载体。
宿主细胞可以是原核细胞,如细菌细胞;或是低等真核细胞,如酵母细胞;或是高等真核细胞,如哺乳动物细胞。代表性例子有:细菌细胞如大肠杆菌,链霉菌属;鼠伤寒沙门氏菌;真菌细胞如酵母;植物细胞;昆虫细胞如果蝇 S2 或Sf9;动物细胞如CHO、COS7、NSO或Bowes 黑素瘤细胞等。特别适用于本发明的宿主细胞是真核宿主细胞,尤其是哺乳动物细胞,如 293T 细胞。
在本发明的另一方面,提供上述全人源单克隆抗体的制备方法,包括以下步骤:
S1.抗原蛋白序列的表达与纯化:从大肠杆菌系统中表达出纯化后的可以用于抗新型冠状病毒SARS-CoV-2单克隆抗体药物筛选的S2蛋白;
S2.噬菌体展示,获得特异性抗体序列:对多名康复者的血液样本进行建库,进行4-5轮的噬菌体生物淘选并利用ELISA进行抗体验证,获得特异性的抗体序列,并且进行重组抗体的表达与纯化,得到与S2蛋白具有高亲和性的抗体;
S3.新型冠状病毒的假病毒中和实验:选取步骤S2中与S2蛋白亲和性较高的抗体进行梯度稀释,与新型冠状病毒假病毒共孵育后一起加入到HEK293T-ACE2细胞中,6小时后更换新鲜完全培养基继续培养细胞48小时,通过检测细胞荧光素酶的活性判定抗体中和效率。
本发明通过人外周血单核细胞特异筛选法筛选到的全人源单克隆抗体,是由活病毒蛋白(抗原)在人体内激活自身免疫系统产生的抗体,与杂交瘤技术和转基因小鼠等技术制备的抗体相比优势明显,具有免疫源性低、亲和力高、特异性强、抗体产生多样性高、安全性高、药代动力学好、用量低、功效高、疗效快等优点,而且无资源及规模化生产限制,更容易标准化生产及质量控制,同时也解决了血源制品难以避免的潜在污染源问题,该抗体药物具有重要的应用价值。
在本发明的另一方面,提供所述的全人源单克隆抗体或其结合片段在用于制备检测新型冠状病毒感染的试剂或试剂盒,以及用于制备治疗、预防新型冠状病毒(SARS-CoV-2)感染的药物中的用途。
本发明旨在以S2蛋白作为表位抗原,利用我们已建立的单克隆抗体筛选平台快速研发针对SARS-CoV-2以及其它致病性冠状病毒包括今后可能出现的变异病毒株的特异性全人源单克隆抗体,以期为冠状病毒感染的防治提供有效的药物。按我国《药品注册管理办法》规定,本研究产品属于治疗用生物制品第1类,为原药研发品种,具有重大的社会意义和应用价值。
在本发明的另一方面,提供一种组合物,包括治疗有效量的所述的全人源单克隆抗体中的一种或多种抗体组成的抗体混合物,以及药学上可接受的载体。
本文所用的术语“药学上可接受的”是指当分子本体和组合物适当地给予动物或人时,它们不会产生不利的、过敏的或其它不良反应。本文所用的“药学上可接受的载体”应当与本发明的单克隆抗体或其片段相容,即能与其共混而不会在通常情况下大幅度降低组合物的效果。
可作为药学上可接受的载体或其组分的一些物质的具体例子是糖类,如乳糖、葡萄糖和蔗糖;淀粉,如玉米淀粉和土豆淀粉;纤维素及其衍生物,如羧甲基纤维素钠、乙基纤维素和甲基纤维素;西黄蓍胶粉末;麦芽;明胶;滑石;固体润滑剂,如硬脂酸和硬脂酸镁;硫酸钙;植物油,如花生油、棉籽油、芝麻油、橄榄油、玉米油和可可油;多元醇,如丙二醇、甘油、山梨糖醇、甘露糖醇和聚乙二醇;海藻酸;乳化剂,如 Tween;润湿剂,如月桂基硫酸钠;着色剂;调味剂;压片剂、稳定剂;抗氧化剂;防腐剂;无热原水;等渗盐溶液;和磷酸盐缓冲液等。
本发明的组合物可根据需要制成各种剂型,并可由医师根据患者种类、年龄、体重和大致疾病状况、给药方式等因素确定对病人有益的剂量进行施用。给药方式例如可以采用注射或其它治疗方式。
上述药物组合物可包含两或多个对于新型冠状病毒具有中和活性的单克隆抗体或其片段。
在本发明的另一方面,提供一种检测新型冠状病毒(SARS-CoV-2)的试剂盒,其包括所述的单克隆抗体或其片段。
以所述的单克隆抗体或其片段为基础,可制备方便、快速且准确地检测新型冠状病毒(SARS-CoV-2)的试剂盒。作为本发明的一种检测方式,采用间接 ELISA法,将待测的抗原包被于固相载体上,利用本发明的单克隆抗体或其片段进行检测。
作为本发明的一种优选方式,所述的单克隆抗体或其片段是抗体,可根据双抗夹心法的原理来检测。双抗夹心法常规的做法是将一抗( 如本发明的单克隆抗体) 固定于载体,然后使一抗与抗原反应,洗涤后再与二抗反应(所述的二抗携带可检测信号,或可与携带可检测信号的物质结合),最后进行化学发光或酶联显色反应检测信号。双抗夹心法特别适用于具有两个或两个以上表位的抗原的检测。
为了在检测时更方便,所述试剂盒中除了含有本发明的单克隆抗体或其片段以外,还可以包含其它检测试剂或辅助试剂,所述的辅助试剂例如是 ELISA 试剂盒中常规使用的一些试剂,这些试剂的特性以及它们的配制方法均是本领域技术人员所熟知的,如显色剂、标记物、二抗、抗抗体、增敏剂等。本领域人员应理解,各种变化形式的检测试剂盒均是包含在本发明中的,只要在其中利用了本发明的单克隆抗体或其片段作为识别新型冠状病毒(SARS-CoV-2)的试剂。
此外,在所述试剂盒中还可包含使用说明书,用于说明其中装载的试剂的使用方法。
在获得了本发明提供的单克隆抗体或其片段后,可以利用多种免疫学相关方法来检测样品中SARS-CoV-2的S2蛋白,从而得知待测样品的供体是否感染新型冠状病毒(SARS-CoV-2),这些方法均被包含在本发明中。较佳地,所述的方法是以非疾病诊断为目的的。
在本发明的另一方面,提供一种非治疗性地抑制新型冠状病毒(SARS-CoV-2)的方法,所述的方法包括给予患者有效量的所述单克隆抗体或其片段。
在本发明的另一方面,提供一种非治疗性检测新型冠状病毒(SARS-CoV-2)的方法,所述的单克隆抗体或其片段与待测样品进行接触,通过检测所述的单克隆抗体或其片段与受试样品的结合情况,获得新型冠状病毒(SARS-CoV-2)的存在情况以及存在量。
如本文所用,术语“待测样品”涵盖了多种样品类型,包括生物学来源的血液及其它体液样品,实体组织样品如活检组织样品或者组织培养物,或者衍生自其中的细胞或者其后代。该术语还包括在获得后已经通过任何方式处理的样品,例如用试剂处理、溶解、或者富集某些成分如蛋白质或者多核苷酸。该术语涵盖了得自任何物种的各种临床样品,也包括培养的细胞、细胞上清和细胞溶解产物。
本发明与现有技术相比,其有益效果有:
本发明成功制备了抗新型冠状病毒SARS-CoV-2的全人源单克隆抗体,该单克隆抗体具有高亲和力的特点,所述单克隆抗体能特异性结合新型冠状病毒SARS-CoV-2,能显著抗新型冠状病毒SARS-CoV-2。
本发明所述抗体为全人源性单克隆抗体,相比鼠源抗体,全人源抗体的基因完全来源于人的基因,没有其他种属的成分,在人体内不发生抗鼠抗抗体等毒副作用,具有更好的生物相容性,更适合和更有潜力成为治疗新型冠状病毒SARS-CoV-2的大分子药物,为此类抗体的标准化生产成药提供了很大的保障。
附图说明
图1为本发明技术路线图;
图2为本发明五种抗体蛋白表达及纯化图谱,其中,MW为分子量标记,IN为阴性对照,FT为流穿,W为洗涤液,E为洗脱液;
图3为本发明五种抗体蛋白纯化图谱,其中,MW为分子量标记。
具体实施方式
以下实施例进一步说明本发明的内容,但不应理解为对本发明的限制。在不背离本发明精神和实质的情况下,对本发明方法、步骤或条件所作的修改或替换,均属于本发明的范围。
实施例中未注明具体条件的实验方法,通常按照常规条件,例如Sambrook等人,分子克隆:实验室手册(New York: Cold Spring Harbor Laboratory Press ,1989)中所述的条件,或按照制造厂商所建议的条件。
图1为本发明技术路线图,包括抗原蛋白序列载体的构建、原核系统的表达,纯化及验证,得到S2抗原蛋白、噬菌体展示,多轮淘选,筛选噬菌体、每轮过后用Elisa进行多克隆抗体检测、单克隆抗体筛选,逐步检测、特异性抗体序列分析,重组抗体的表达与纯化、新冠假病毒-抗体中和实验、获得抗SARS-CoV-2全人源单克隆抗体。
实施例:新型冠状病毒(SARS-CoV-2)的全人源单克隆抗体的制备
1.S2噬菌体文库的构建
1.1 外周血单核细胞的获取
从已感染新型冠状病毒的志愿者体内抽取新鲜血液3.5mL,置于抗凝管中。将新鲜抗凝全血与PBS等体积稀释后,缓慢加入到淋巴细胞分离液(国产Solarbio,Cat.NO.P8610)中,离心之后将其小心拿出,吸第二层云雾状的单核细胞层于装有PBS的离心管中洗涤,离心弃上清,再用PBS洗涤1次,收集到的细胞即为外周血单核细胞(periphery bloodmononuclear cell,PBMC)。
1.2 提取单核细胞总RNA,构建噬菌体文库
细胞中提取总RNA, 反转录合成cDNA, 用直接PCR及半巢式PCR 扩增人抗体重链(VH)及轻链(V和V)可变区基因。采用改进的重叠延伸PCR法将VH基因和VL基因连接成人单链抗体(scFv)基因, 并将 scFv文库基因克隆到噬菌体载体。
2.利用S2噬菌体文库展示技术筛选单克隆抗体
以S2蛋白为固定相,以噬菌体展示文库为流动相,利用体外生物淘洗法来进行筛选。具体操作过程为:首先设立1mL 50μg S2抗原蛋白组(Ag组),500μL 10μg抗原对照组(NC组),4℃孵育12h,之后用0.05% PBST冲洗三次,在试管中加入5%的牛奶,37℃孵育2h,封闭过后用0.05%PBST冲洗三次,用1%milk-PBST稀释噬菌体库,将文库噬菌体输入管中,32℃孵育2小时,用0.05% PBST洗3次,PBS洗2次。用1mL甘氨酸- HC洗脱粘合剂,然后用Tris-HCl中和,直到pH为7.0。在试管上涂5 mL 5%牛奶/PBST, 37℃孵育2h,随后用0.05%PBST冲洗3次,将噬菌体转移到试管中,32℃孵育1小时,然后将噬菌体转移到EP管中。随后检测洗脱噬菌体的滴度,培养大肠杆菌TG1至OD600=0.4-0.6,稀释洗脱液,将10μL稀释液与180μL大肠杆菌TG1混合,37℃培养30分钟,倒于2×YT-A (Amp 100μg/mL)平板上,将盘子倒置放置在37℃过夜。
扩增洗脱的噬菌体:(1)将10μL的大肠杆菌TG1加入800μL的2YT培养液中,37℃培养至OD600=0.4-0.6。将培养到对数期的TG1转移到10mL 2YT-G(最终浓度2%葡萄糖)培养液中,37℃摇床培养至OD600=0.4-0.6,加入洗脱产物,37℃孵育30分钟,37℃摇匀30分钟,加入30mL 2YT-AG培养液(终浓度0.1% Amp,2%葡萄糖),37℃摇瓶培养1h,加入M13KO7(M13KO7:TG1=20:1),37℃孵育30分钟,37℃摇匀30分钟,菌液5000rpm离心10分钟,重悬于40mL 2YT-AK中,在30℃的摇床上孵育过夜,8000rpm离心10分钟,弃去上清,重悬1mL PBS, 12000rpm离心5分钟,将上清转移到新的1.5mL离心管中。扩增噬菌体滴度的检测:培养大肠杆菌TG1至OD600=0.4-0.6。稀释洗脱液,将10μL稀释液与180μL大肠杆菌TG1混合。37℃培养30分钟,倒于2×YT-A (Amp 100μg/mL)平板上。将盘子倒置放置在37℃过夜,其具体的生物筛选结果如表1所示。
表1 生物筛选结果
根据表1的筛选结果,经过四轮生物淘选,最后得到1.2×107(pfu)目的噬菌体文库。
3.利用ELISA技术对文库进行逐级筛选
(1)多克隆噬菌体ELISA检测:用抗原包覆平板(试验组:4μg/mL蛋白S2;对照组:0μg抗原)4℃过夜,随后用0.05% PBST洗涤2次,PBS洗涤1次,用300μL 5%的milk在37℃下封闭2小时,用0.05% PBS冲洗3次,将100μL稀释的扩增噬菌体加入孔中,32℃孵育2小时,用0.05% PBST冲洗3次,在阻断缓冲液中稀释抗m13-hrp抗体(1:400),每孔加入100μL, 32℃孵育1小时,用0.05% PBST冲洗3次,每孔加入100μL TMB,室温孵育,每孔加入100μL 2MHCl。使用设定在450-630nm的酶标仪读数,筛选结果如表2所示。
表2 多克隆噬菌体ELISA检测结果
根据表2的检测结果,经过四轮多克隆噬菌体筛选,确定以第二轮噬菌体进行单克隆筛选。
(2)单克隆噬菌体ELISA检测:
1)根据以上实验结果,选择R2噬菌体进行单克隆筛选,从检测洗脱效价的平板中挑选192个克隆。37℃振荡250 rpm培养这些噬菌体,直到OD600=0.4-0.6,M13KO7感染培养物(MOI=20:1) 37℃孵育30min, 37℃摇匀30min。2×YT-AK (Amp 100μg/mL, Kan 100μg/mL)等体积悬浮沉淀,在30℃培养过夜,离心,培养物和上清可用于ELISA,用抗原包覆平板(试验组:4μg/mL蛋白S2;对照组:0μg抗原)过夜,4℃,洗孔3次。
用300μL 5%的milk在37℃下封闭2小时,把管子洗三遍,孔中加入100μL的噬菌体上清,32℃孵育2小时,把管子洗三遍。在阻断缓冲液中稀释抗m13-hrp抗体(1:400),每孔加入100μL, 32℃孵育1小时,把管子洗三遍,TMB每孔加入100μL室温孵育,每孔加入100μL2MHCl。使用设定在450-620nm的酶标仪读取板,然后对高特异性克隆进行测序。
2)阳性克隆验证ELISA:将50μL的阳性克隆加入2mL 2YT-AG培养基中,培养至OD600=0.4-0.6。M13KO7感染培养物(MOI=20:1) 37℃孵育30min,37℃摇匀30min。2×YT-AK(Amp100μg/mL, Kan 100μg/mL)等体积悬浮沉淀。在30℃培养过夜,离心,培养物和上清可用于ELISA,用抗原包覆平板(试验组:4μg/mL蛋白S2;对照组:0μg抗原)过夜,4℃,洗孔3次。
用300μL 5%的milk在37℃下堵孔2小时,把管子洗三遍,孔中加入100μL的噬菌体上清,32℃孵育2小时,把管子洗三遍,在阻断缓冲液中稀释抗m13-hrp抗体(1:400),每孔加入100μL, 32℃孵育1小时,把管子洗三遍,TMB每孔加入100μL室温孵育,每孔加入100μL 2MHCl。使用设定在450-620nm的酶标仪读取板,然后对高特异性克隆进行测序,验证阳性克隆ELISA(IgG)表达,将抗原(试验组:4μg/mL protein S2)涂于平板上过夜,4℃,洗孔3次。用300μL 5%的milk在37℃下封闭2小时,把管子洗三遍,加入100μL稀释后的IgG,32℃孵育2小时,把管子洗三遍,在阻断缓冲液中稀释羊抗人IgG HRP(1:700),每孔加入100μL, 32℃孵育1小时,把管子洗三遍。TMB每孔加入100 μL,室温孵育,每孔加入100 μL 2M HCl,使用设定在450-620nm的酶标仪读取板,检测结果如表3所示。
表3 单克隆噬菌体ELISA检测结果
根据表3的检测结果,得到8个与S2结合能力强的噬菌体,并将表中R2P1-E1、R2P1-H1、R2P1-F2、R2P1-D8、R2P2-G5、R2P2-D7、R2P2-D10和R2P2-F8克隆送去测序。通过对测序结果的分析,我们最终得到了五条(R2P1-D8、R2P2-G5、R2P2-D7、R2P2-D10和R2P2-F8)唯一且正确的序列。随后进行五种抗体的表达与纯化用XtenCHO细胞通过Xten转染方案转染五种抗体,收集培养基和细胞,直到存活率下降到50%以下(转染后14天),其中,上述五种抗体蛋白表达及纯化图谱如图2-3所示。
4.新冠假病毒抗体中和实验
实验前一天,将待感染HEK293T-ACE2 细胞(Cat.GM-C09233)细胞接种于96孔细胞培养板中,接种量约为 1×104个细胞/孔,次日进行病毒感染时,细胞密度在30%左右为佳。本实验选取之前制备的五种中结合能力较好的G5、F8抗体进行假病毒中和实验,根据抗体母液的浓度,将其按3倍梯度连续稀释。两种抗体原始浓度皆为 3mg/mL,第一孔S1稀释浓度是60μg/mL,中和孵育的终浓度是30μg/mL。取0.37μL/孔假病毒:取 597.78μL完全培养基,加入2.2μL病毒原液,混匀,55μL分装10个EP管(预留5μL)。 按连续稀释的抗体G5、F8分别与上述假病毒液等体积混合,并在室温孵育1小时。从培养箱取出提前铺好HEK293T-ACE2细胞的96孔板,在镜下观察细胞是否污染,密度,状态,用排枪将上层培养基吸去(注意枪头不要碰到96孔板底部的细胞,可将96孔板倾斜后吸去)。 吸取100μL稀释好的假病毒抗体混合液加入铺好HEK293T-ACE2细胞的96孔板中(96孔板抗体布局如表4所示),注意沿孔壁加入避免冲起细胞。处理6小时更换新鲜完全培养基继续培养48小时。感染假病毒换液后48小时,通过荧光素酶检测试剂盒(Promega/ E1910)在酶标仪上检测荧光素酶的活性判定抗体中和效率,结果如表5所示。
表4 96孔板抗体布局
表5 抗体中和效率
从表5的检测结果可以看出,抗体从高浓度到低浓度,样品OD值存在一定的上升趋势,说明本发明抗体具有一定的中和保护作用。上述抗体作为高亲和力抗新型冠状病毒SARS-CoV-2的全人源单克隆抗体,可用于制备检测、治疗或者预防新型冠状病毒SARS-CoV-2感染的药物中,同时可为此类抗体的标准化生产成药提供了很大的保障。
最后需要强调的是,以上所述仅为本发明的优选实施例,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种变化和更改,凡在本发明的精神和原则之内,所做的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
SEQUENCE LISTING
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<120> 一种抗新型冠状病毒的全人源单克隆抗体及其用途
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<211> 120
<212> PRT
<213> R2P1-D8-H重链可变区核苷酸序列
<400> 11
Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
5 10 15
Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
20 25 30
Phe Thr Ser Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln
35 40 45
Gly Leu Glu Trp Met Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr
50 55 60
Ser Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg Asp
65 70 75
Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser
80 85 90
Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Leu Leu Arg Asp
95 100 105
Pro Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
110 115 120
<210> 12
<211> 113
<212> PRT
<213> R2P1-D8-L轻链可变区核苷酸序列
<400> 12
Gln Ser Val Val Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly
5 10 15
Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly
20 25 30
Asn Asn Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro
50 55 60
Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly
65 70 75
Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly
80 85 90
Thr Trp Asp Ser Ser Leu Ser Ala Val Val Phe Gly Gly Gly Thr
95 100 105
Lys Val Thr Val Leu Ala Ala Ala
110
<210> 13
<211> 120
<212> PRT
<213> R2P2-D10-H重链可变区核苷酸序列
<400> 13
Met Ala Gln Val Gln Leu Gln Gln Ser Gly Gly Gly Leu Val Gln
5 10 15
Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
20 25 30
Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys
35 40 45
Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr
50 55 60
Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
65 70 75
Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala
80 85 90
Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Gly Ala Gln His Arg
95 100 105
Pro Gly Thr Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
110 115 120
<210> 14
<211> 112
<212> PRT
<213> R2P2-D10-L轻链可变区核苷酸序列
<400> 14
Gln Pro Val Leu Thr Gln Pro Pro Ser Ala Ser Glu Thr Pro Gly
5 10 15
Gln Arg Leu Ser Ile Ser Cys Ser Gly Asn Arg Ser Asn Ile Gly
20 25 30
Ser His Pro Val Asn Trp Tyr Gln Gln Val Pro Gly Thr Ala Pro
35 40 45
Lys Leu Leu Ile His Gly Asp Asn Lys Arg Pro Ser Gly Val Pro
50 55 60
Asp Arg Phe Ser Ala Ser Lys Ser Gly Thr Ser Ala Ser Leu Val
65 70 75
Ile Ser Gly Leu Gln Pro Glu Asp Glu Ala Asp Tyr Tyr Cys Ala
80 85 90
Ser Trp Asp Asp Arg Leu Ile Gly Val Phe Gly Gly Gly Thr Gln
95 100 105
Leu Thr Val Leu Ala Ala Ala
110
<210> 15
<211> 120
<212> PRT
<213> R2P2-G5-H重链可变区核苷酸序列
<400> 15
Met Ala Gln Val Gln Leu Gln Glu Ser Gly Ala Glu Val Lys Lys
5 10 15
Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
20 25 30
Phe Thr Ser Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln
35 40 45
Gly Leu Glu Trp Met Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr
50 55 60
Ser Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg Asp
65 70 75
Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser
80 85 90
Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Leu Leu Arg Asp
95 100 105
Pro Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
110 115 120
<210> 16
<211> 113
<212> PRT
<213> R2P2-G5-L轻链可变区核苷酸序列
<400> 16
Gln Ser Val Leu Thr Gln Pro Ser Ser Val Ser Ala Ala Pro Gly
5 10 15
Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly
20 25 30
Lys Asn Tyr Val Ser Trp Tyr Gln His Leu Pro Gly Thr Ala Pro
35 40 45
Lys Leu Leu Thr Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro
50 55 60
Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Ala
65 70 75
Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly
80 85 90
Thr Trp Asp Ser Ser Leu Ser Glu Val Val Phe Gly Gly Gly Thr
95 100 105
Lys Leu Thr Val Leu Ala Ala Ala
110
<210> 17
<211> 120
<212> PRT
<213> R2P2-D7-H重链可变区核苷酸序列
<400> 17
Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
5 10 15
Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
20 25 30
Phe Thr Ser Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln
35 40 45
Gly Leu Glu Trp Met Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr
50 55 60
Ser Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg Asp
65 70 75
Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser
80 85 90
Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Leu Leu Arg Asp
95 100 105
Pro Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
110 115 120
<210> 18
<211> 113
<212> PRT
<213> R2P2-D7-L轻链可变区核苷酸序列
<400> 18
Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly
5 10 15
Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly
20 25 30
Asn Asn Tyr Val Ser Trp Tyr Gln Gln Val Pro Gly Thr Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Val Ile Pro
50 55 60
Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly
65 70 75
Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly
80 85 90
Thr Trp Asp Ser Ser Leu Ser Ala Val Val Phe Gly Gly Gly Thr
95 100 105
Gln Leu Thr Val Leu Ala Ala Ala
110
<210> 19
<211> 120
<212> PRT
<213> R2P2-F8-H重链可变区核苷酸序列
<400> 19
Met Ala Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
5 10 15
Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
20 25 30
Phe Thr Ser Tyr Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln
35 40 45
Gly Leu Glu Trp Met Gly Ile Ile Asn Pro Ser Gly Gly Asn Thr
50 55 60
Ile Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg Asp
65 70 75
Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser
80 85 90
Glu Asp Thr Ala Val Tyr Tyr Cys Ala Ser Met Gly Glu Arg His
95 100 105
Ala Phe Asp Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
110 115 120
<210> 20
<211> 113
<212> PRT
<213> R2P2-F8-L轻链可变区核苷酸序列
<400> 20
Gln Ser Val Val Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly
5 10 15
Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly
20 25 30
Asn Asn Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
35 40 45
Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro
50 55 60
Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly
65 70 75
Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly
80 85 90
Thr Trp Asp Ser Ser Leu Ser Ala Val Val Phe Gly Gly Gly Thr
95 100 105
Lys Leu Thr Val Leu Ala Ala Ala
110
Claims (6)
1.一种抗新型冠状病毒的全人源单克隆抗体,其特征在于,所述全人源单克隆抗体具有重链可变区和轻链可变区;
重链可变区的氨基酸序列如SEQ ID NO:11所示;轻链可变区的氨基酸序列如SEQ IDNO:12所示。
2.一种编码如权利要求1所述全人源单克隆抗体的核酸分子,其特征在于,具有如下重链可变区和轻链可变区:
重链可变区的核苷酸序列如SEQ ID NO:1所示;轻链可变区的核苷酸序列如SEQ IDNO:2所示。
3.一种表达载体,其特征在于,包括权利要求2所述的核酸分子。
4.一种宿主细胞,其特征在于,包括权利要求3所述的表达载体,且所述宿主细胞不是植物细胞。
5.一种如权利要求1所述的全人源单克隆抗体在制备检测新型冠状病毒感染的试剂或试剂盒,以及制备治疗或者预防新型冠状病毒感染的药物中的用途。
6.一种组合物,其特征在于,包括治疗有效量的权利要求1所述的全人源单克隆抗体,以及药学上可接受的载体。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111592594A (zh) * | 2020-03-13 | 2020-08-28 | 北京大学 | 一种抗新型冠状病毒的单克隆抗体及其应用 |
CN113150129A (zh) * | 2021-01-28 | 2021-07-23 | 四川大学华西医院 | 抗新冠病毒SARS-CoV-2表面S2蛋白的单链抗体及其应用 |
CN113264998A (zh) * | 2021-01-28 | 2021-08-17 | 四川大学华西医院 | 抗新冠病毒SARS-CoV-2表面S1蛋白的单链抗体及其应用 |
CN113563464A (zh) * | 2021-08-01 | 2021-10-29 | 中国疾病预防控制中心性病艾滋病预防控制中心 | 人源化高中和活性抗新型冠状病毒单克隆抗体及应用 |
CN113929774A (zh) * | 2021-10-15 | 2022-01-14 | 中国科学院微生物研究所 | 一种新型冠状病毒及其突变体的单克隆抗体及其应用 |
CN113943368A (zh) * | 2021-10-15 | 2022-01-18 | 中国科学院微生物研究所 | 一种新型冠状病毒及其突变体的单克隆抗体及其应用 |
CN114106164A (zh) * | 2021-12-09 | 2022-03-01 | 杭州旭科生物技术有限公司 | 抗新型冠状病毒s蛋白的单克隆抗体及其应用 |
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Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111592594A (zh) * | 2020-03-13 | 2020-08-28 | 北京大学 | 一种抗新型冠状病毒的单克隆抗体及其应用 |
CN113150129A (zh) * | 2021-01-28 | 2021-07-23 | 四川大学华西医院 | 抗新冠病毒SARS-CoV-2表面S2蛋白的单链抗体及其应用 |
CN113264998A (zh) * | 2021-01-28 | 2021-08-17 | 四川大学华西医院 | 抗新冠病毒SARS-CoV-2表面S1蛋白的单链抗体及其应用 |
CN113563464A (zh) * | 2021-08-01 | 2021-10-29 | 中国疾病预防控制中心性病艾滋病预防控制中心 | 人源化高中和活性抗新型冠状病毒单克隆抗体及应用 |
CN113929774A (zh) * | 2021-10-15 | 2022-01-14 | 中国科学院微生物研究所 | 一种新型冠状病毒及其突变体的单克隆抗体及其应用 |
CN113943368A (zh) * | 2021-10-15 | 2022-01-18 | 中国科学院微生物研究所 | 一种新型冠状病毒及其突变体的单克隆抗体及其应用 |
CN114106164A (zh) * | 2021-12-09 | 2022-03-01 | 杭州旭科生物技术有限公司 | 抗新型冠状病毒s蛋白的单克隆抗体及其应用 |
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