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Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells

2020; Taylor & Francis; Volume: 9; Issue: 1 Linguagem: Inglês

10.1080/22221751.2020.1815589

2222-1751

Hualong Xiong, Yangtao Wu, Jiali Cao, Ren Yang, Yingxia Liu, Jian Ma, Xiaoyang Qiao, Xiangyang Yao, Baohui Zhang, Yali Zhang, Wangheng Hou, Yang Shi, Jingjing Xu, Liang Zhang, Shaojuan Wang, Baorong Fu, Ting Yang, Shengxiang Ge, Jun Zhang, Quan Yuan, Baoying Huang, Zhiyong Li, Tianying Zhang, Ningshao Xia,

Animal Virus Infections Studies

The global pandemic of coronavirus disease 2019 (COVID-19) is a disaster for human society. A convenient and reliable neutralization assay is very important for the development of vaccines and novel drugs. In this study, a G protein-deficient vesicular stomatitis virus (VSVdG) bearing a truncated spike protein (S with C-terminal 18 amino acid truncation) was compared to that bearing the full-length spike protein of SARS-CoV-2 and showed much higher efficiency. A neutralization assay was established based on VSV-SARS-CoV-2-Sdel18 pseudovirus and hACE2-overexpressing BHK21 cells (BHK21-hACE2 cells). The experimental results can be obtained by automatically counting the number of EGFP-positive cells at 12 h after infection, making the assay convenient and high-throughput. The serum neutralizing titer measured by the VSV-SARS-CoV-2-Sdel18 pseudovirus assay has a good correlation with that measured by the wild type SARS-CoV-2 assay. Seven neutralizing monoclonal antibodies targeting the receptor binding domain (RBD) of the SARS-CoV-2 S protein were obtained. This efficient and reliable pseudovirus assay model could facilitate the development of new drugs and vaccines.