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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
by
Hou, Wang-Heng
, Wang, Shao-Juan
, Zhang, Liang
, Yuan, Quan
, Wu, Yang-Tao
, Ma, Jian
, Li, Zhi-Yong
, Zhang, Tian-Ying
, Xiong, Hua-Long
, Qiao, Xiao-Yang
, Fu, Bao-Rong
, Yang, Ren
, Zhang, Bao-Hui
, Huang, Bao-Ying
, Zhang, Ya-Li
, Shi, Yang
, Xu, Jing-Jing
, Ge, Sheng-Xiang
, Xia, Ning-Shao
, Yao, Xiang-Yang
, Yang, Ting
, Liu, Ying-Xia
, Cao, Jia-Li
, Zhang, Jun
in
Animals
/ Antibodies, Monoclonal - immunology
/ Antibodies, Neutralizing - blood
/ Antibodies, Neutralizing - immunology
/ Antibodies, Viral - blood
/ Antibodies, Viral - immunology
/ Betacoronavirus - immunology
/ Cell Line
/ Chlorocebus aethiops
/ Coronavirus Infections - diagnosis
/ Coronaviruses
/ COVID-19
/ Cricetinae
/ neutralization assay
/ Neutralization Tests - methods
/ Pandemics
/ Pneumonia, Viral - diagnosis
/ Proteins
/ pseudovirus
/ SARS-CoV-2
/ Severe acute respiratory syndrome coronavirus 2
/ Spike Glycoprotein, Coronavirus - genetics
/ Spike Glycoprotein, Coronavirus - immunology
/ Vaccines
/ Vero Cells
/ Vesicular stomatitis Indiana virus - genetics
/ Vesicular stomatitis Indiana virus - immunology
/ vesicular stomatitis virus
/ VSV-SARS-CoV-2
2020
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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
by
Hou, Wang-Heng
, Wang, Shao-Juan
, Zhang, Liang
, Yuan, Quan
, Wu, Yang-Tao
, Ma, Jian
, Li, Zhi-Yong
, Zhang, Tian-Ying
, Xiong, Hua-Long
, Qiao, Xiao-Yang
, Fu, Bao-Rong
, Yang, Ren
, Zhang, Bao-Hui
, Huang, Bao-Ying
, Zhang, Ya-Li
, Shi, Yang
, Xu, Jing-Jing
, Ge, Sheng-Xiang
, Xia, Ning-Shao
, Yao, Xiang-Yang
, Yang, Ting
, Liu, Ying-Xia
, Cao, Jia-Li
, Zhang, Jun
in
Animals
/ Antibodies, Monoclonal - immunology
/ Antibodies, Neutralizing - blood
/ Antibodies, Neutralizing - immunology
/ Antibodies, Viral - blood
/ Antibodies, Viral - immunology
/ Betacoronavirus - immunology
/ Cell Line
/ Chlorocebus aethiops
/ Coronavirus Infections - diagnosis
/ Coronaviruses
/ COVID-19
/ Cricetinae
/ neutralization assay
/ Neutralization Tests - methods
/ Pandemics
/ Pneumonia, Viral - diagnosis
/ Proteins
/ pseudovirus
/ SARS-CoV-2
/ Severe acute respiratory syndrome coronavirus 2
/ Spike Glycoprotein, Coronavirus - genetics
/ Spike Glycoprotein, Coronavirus - immunology
/ Vaccines
/ Vero Cells
/ Vesicular stomatitis Indiana virus - genetics
/ Vesicular stomatitis Indiana virus - immunology
/ vesicular stomatitis virus
/ VSV-SARS-CoV-2
2020
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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
by
Hou, Wang-Heng
, Wang, Shao-Juan
, Zhang, Liang
, Yuan, Quan
, Wu, Yang-Tao
, Ma, Jian
, Li, Zhi-Yong
, Zhang, Tian-Ying
, Xiong, Hua-Long
, Qiao, Xiao-Yang
, Fu, Bao-Rong
, Yang, Ren
, Zhang, Bao-Hui
, Huang, Bao-Ying
, Zhang, Ya-Li
, Shi, Yang
, Xu, Jing-Jing
, Ge, Sheng-Xiang
, Xia, Ning-Shao
, Yao, Xiang-Yang
, Yang, Ting
, Liu, Ying-Xia
, Cao, Jia-Li
, Zhang, Jun
in
Animals
/ Antibodies, Monoclonal - immunology
/ Antibodies, Neutralizing - blood
/ Antibodies, Neutralizing - immunology
/ Antibodies, Viral - blood
/ Antibodies, Viral - immunology
/ Betacoronavirus - immunology
/ Cell Line
/ Chlorocebus aethiops
/ Coronavirus Infections - diagnosis
/ Coronaviruses
/ COVID-19
/ Cricetinae
/ neutralization assay
/ Neutralization Tests - methods
/ Pandemics
/ Pneumonia, Viral - diagnosis
/ Proteins
/ pseudovirus
/ SARS-CoV-2
/ Severe acute respiratory syndrome coronavirus 2
/ Spike Glycoprotein, Coronavirus - genetics
/ Spike Glycoprotein, Coronavirus - immunology
/ Vaccines
/ Vero Cells
/ Vesicular stomatitis Indiana virus - genetics
/ Vesicular stomatitis Indiana virus - immunology
/ vesicular stomatitis virus
/ VSV-SARS-CoV-2
2020
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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
Journal Article
Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells
2020
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Overview
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.
Publisher
Taylor & Francis,Taylor & Francis Ltd,Taylor & Francis Group
Subject
/ Antibodies, Monoclonal - immunology
/ Antibodies, Neutralizing - blood
/ Antibodies, Neutralizing - immunology
/ Antibodies, Viral - immunology
/ Betacoronavirus - immunology
/ Coronavirus Infections - diagnosis
/ COVID-19
/ Neutralization Tests - methods
/ Pneumonia, Viral - diagnosis
/ Proteins
/ Severe acute respiratory syndrome coronavirus 2
/ Spike Glycoprotein, Coronavirus - genetics
/ Spike Glycoprotein, Coronavirus - immunology
/ Vaccines
/ Vesicular stomatitis Indiana virus - genetics
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