학술논문
Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells
Document Type
article
Author
Hua-Long Xiong; Yang-Tao Wu; Jia-Li Cao; Ren Yang; Ying-Xia Liu; Jian Ma; Xiao-Yang Qiao; Xiang-Yang Yao; Bao-Hui Zhang; Ya-Li Zhang; Wang-Heng Hou; Yang Shi; Jing-Jing Xu; Liang Zhang; Shao-Juan Wang; Bao-Rong Fu; Ting Yang; Sheng-Xiang Ge; Jun Zhang; Quan Yuan; Bao-Ying Huang; Zhi-Yong Li; Tian-Ying Zhang; Ning-Shao Xia
Source
Emerging Microbes and Infections, Vol 9, Iss 1, Pp 2105-2113 (2020)
Subject
Language
English
ISSN
22221751
2222-1751
2222-1751
Abstract
ABSTRACTThe 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.