Quantification of SARS-CoV-2 neutralizing antibody by a pseudotyped virus-based assay
Quantification of SARS-CoV-2 neutralizing antibody by a pseudotyped virus-based assay
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DOI：
10.1038/s41596-020-0394-5
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发表时间：
2020-09-25
期刊：
影响因子：
14.8
通讯作者：
Wang, Youchun
中科院分区：
文献类型：
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作者：
Nie, Jianhui;Li, Qianqian;Wang, Youchun
假型病毒因其安全性和多功能性而成为有用的病毒学工具。在水泡性口炎病毒（VSV）假型病毒生产系统的基础上，我们在生物安全二级设施中开发了针对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的基于假型病毒的中和测定。与结合抗体测试相比，中和测定可以区分抗体家族中的保护剂。该协议包括 SARS-CoV-2 S 假型病毒的生产和滴定以及基于它的中和测定。可以评估针对病毒附着和进入的各种类型的样品的效力，包括来自动物和人类的血清样品、单克隆抗体和融合抑制剂（肽或小分子）。如果提前制备了假型病毒原液，则需要2天才能获得候选样本的效价数据。在实施本协议之前，需要具备处理细胞的经验。本协议中使用基于 VSV 的假病毒生产系统生产和滴定 SARS-CoV-2 S 假病毒，使其能够在生物安全 2 级条件下以及在中和测定中使用，以评估样品中中和抗体或分子抑制剂的水平。
Pseudotyped viruses are useful virological tools because of their safety and versatility. On the basis of a vesicular stomatitis virus (VSV) pseudotyped virus production system, we developed a pseudotyped virus-based neutralization assay against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in biosafety level 2 facilities. Compared with the binding antibody test, the neutralization assay could discriminate the protective agents from the antibody family. This protocol includes production and titration of the SARS-CoV-2 S pseudotyped virus and the neutralization assay based on it. Various types of samples targeting virus attachment and entry could be evaluated for their potency, including serum samples derived from animals and humans, monoclonal antibodies and fusion inhibitors (peptides or small molecules). If the pseudotyped virus stock has been prepared in advance, it will take 2 days to get the potency data for the candidate samples. Experience in handling cells is needed before implementing this protocol.The production and titration of the SARS-CoV-2 S pseudotyped virus using a VSV-based pseudovirus production system in this protocol enable its use under biosafety level 2 conditions as well as in a neutralization assay to assess the level of neutralizing antibodies or molecular inhibitors in a sample.