tetano
Editor, Senior Moderator
Emerg Microbes Infect
. 2020 Sep 7;1-38.
doi: 10.1080/22221751.2020.1815589. Online ahead of print.
Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells
Hua-Long Xiong[SUP] 1 [/SUP], Yang-Tao Wu[SUP] 1 [/SUP], Jia-Li Cao[SUP] 1 [/SUP], Ren Yang[SUP] 2 [/SUP], Ying-Xia Liu[SUP] 3 [/SUP], Jian Ma[SUP] 1 [/SUP], Xiao-Yang Qiao[SUP] 1 [/SUP], Xiang-Yang Yao[SUP] 4 [/SUP], Bao-Hui Zhang[SUP] 1 [/SUP], Ya-Li Zhang[SUP] 1 [/SUP], Wang-Heng Hou[SUP] 1 [/SUP], Yang Shi[SUP] 1 [/SUP], Jing-Jing Xu[SUP] 5 [/SUP], Liang Zhang[SUP] 1 [/SUP], Shao-Juan Wang[SUP] 1 [/SUP], Bao-Rong Fu[SUP] 1 [/SUP], Ting Yang[SUP] 5 [/SUP], Sheng-Xiang Ge[SUP] 1 [/SUP], Jun Zhang[SUP] 1 [/SUP], Quan Yuan[SUP] 1 [/SUP], Bao-Ying Huang[SUP] 2 [/SUP], Zhi-Yong Li[SUP] 4 [/SUP], Tian-Ying Zhang[SUP] 1 [/SUP], Ning-Shao Xia[SUP] 1 [/SUP]
Affiliations
Abstract
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.
Keywords: SARS-CoV-2; VSV-SARS-CoV-2; neutralization assay; pseudovirus; vesicular stomatitis virus.
. 2020 Sep 7;1-38.
doi: 10.1080/22221751.2020.1815589. Online ahead of print.
Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells
Hua-Long Xiong[SUP] 1 [/SUP], Yang-Tao Wu[SUP] 1 [/SUP], Jia-Li Cao[SUP] 1 [/SUP], Ren Yang[SUP] 2 [/SUP], Ying-Xia Liu[SUP] 3 [/SUP], Jian Ma[SUP] 1 [/SUP], Xiao-Yang Qiao[SUP] 1 [/SUP], Xiang-Yang Yao[SUP] 4 [/SUP], Bao-Hui Zhang[SUP] 1 [/SUP], Ya-Li Zhang[SUP] 1 [/SUP], Wang-Heng Hou[SUP] 1 [/SUP], Yang Shi[SUP] 1 [/SUP], Jing-Jing Xu[SUP] 5 [/SUP], Liang Zhang[SUP] 1 [/SUP], Shao-Juan Wang[SUP] 1 [/SUP], Bao-Rong Fu[SUP] 1 [/SUP], Ting Yang[SUP] 5 [/SUP], Sheng-Xiang Ge[SUP] 1 [/SUP], Jun Zhang[SUP] 1 [/SUP], Quan Yuan[SUP] 1 [/SUP], Bao-Ying Huang[SUP] 2 [/SUP], Zhi-Yong Li[SUP] 4 [/SUP], Tian-Ying Zhang[SUP] 1 [/SUP], Ning-Shao Xia[SUP] 1 [/SUP]
Affiliations
- PMID: 32893735
- DOI: 10.1080/22221751.2020.1815589
Abstract
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.
Keywords: SARS-CoV-2; VSV-SARS-CoV-2; neutralization assay; pseudovirus; vesicular stomatitis virus.