靶向SARS-CoV-2受体结合结构域的嵌合抗体表达及功能研究  

Expression and functional study of chimeric antibodies targeting SARS-CoV-2 receptor binding domain

作　　者：石慧春 焦凡珂 徐巍 陈永康 陆路[2,3] 邹鹏 SHI Huichun;JIAO Fanke;XU Wei;CHEN Yongkang;LU Lu;ZOU Peng(Shanghai Public Health Clinical Center,Fudan University,Shanghai 201508,China;Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS),School of Basic Medical Sciences,Shanghai Medical College,Fudan University,Shanghai 200032,China;Shanghai Institute of Infectious Disease and Biosecurity,Shanghai 200032,China)

机构地区：[1]复旦大学附属公共卫生临床中心,上海201508 [2]复旦大学上海医学院基础医学院教育部/卫健委/医科院医学分子病毒学重点实验室,上海200032 [3]上海市重大传染病和生物安全研究院,上海200032

出　　处：《微生物与感染》2023年第1期2-12,共11页Journal of Microbes and Infections

摘　　要：严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)感染所致的2019冠状病毒病(coronavirus disease 2019,COVID-19)给人民的生命健康造成了严重的威胁,目前该病毒仍然在广泛传播,针对SARS-CoV-2有效的治疗方法仍有限,因此寻找广谱中和抗体阻断SARS-CoV-2感染具有重要的意义。本文成功表达并纯化了3株靶向SARS-CoV-2的受体结合结构域(receptor-binding domain,RBD)的人鼠嵌合抗体,并且检测了这几株抗体对多种SARS-CoV-2假病毒和活病毒的中和活性。本研究表明这3株抗体能特异性地中和SARS-CoV-2野生型假病毒,IC 50分别为0.03μg/mL、0.06μg/mL、0.03μg/mL。此外,这3株抗体对Alpha株假病毒、Delta株假病毒和Lambda株假病毒也具有广谱中和活性,并可以抑制SARS-CoV-2 Delta株活病毒的复制。机制研究表明,这些抗体可以阻断SARS-CoV-2的S蛋白介导的细胞-细胞融合,且RBD上的K417、E484和N501位点可能是其与这3株抗体结合的关键位点。Coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection has a serious impact on people’s health and lives.At present,SARS-CoV-2 is still widely spread,but effective treatments for SARS-CoV-2 are still limited.Therefore,it is of great significance to find broad-spectrum neutralizing antibodies to block SARS-CoV-2 infection.In this study,we successfully expressed and purified three human-mouse chimeric antibodies targeting receptor-binding domain(RBD)of SARS-CoV-2,and then the neutralizing activities of these antibodies against a variety of SARS-CoV-2 pseudoviruses and authentic virus were tested.The results showed that these three antibodies could specifically neutralize SARS-CoV-2 wild-type pseudovirus,and the IC 50 values were 0.03μg/mL,0.06μg/mL and 0.03μg/mL,respectively.Furthermore,these antibodies exhibited broad-spectrum neutralizing activity against Alpha,Delta and Lambda pseudoviruses,and could also inhibit the replication of SARS-CoV-2 Delta authentic virus.Mechanism studies showed that these antibodies could block cell-cell fusion mediated by SARS-CoV-2 S protein,and K417,E484 and N501 on receptor-binding domain(RBD)may be the key sites for binding with these three antibodies.

关 键 词：严重急性呼吸综合征冠状病毒2 受体结合结构域 嵌合抗体 中和抗体 

分 类 号：R373.1[医药卫生—病原生物学]