Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2

Jie Hu; Qingzhu Gao; Changlong He; Ailong Huang; Ni Tang; Kai Wang

doi:10.1016/j.gendis.2020.07.006

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Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2

更新时间：2022-06-24

- Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2
- Genes & Diseases Vol. 7, Issue 4, (2020)
- 作者机构：
  
  Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Department of Infectious Diseases, The Second Affiliated Hospital, Chongqing Medical University,Chongqing,China,400010
- 作者简介：
- 基金信息：
- DOI：10.1016/j.gendis.2020.07.006
  CLC：
- Published：2020，
  
  Published Online：1 December 2020，
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Jie Hu, Qingzhu Gao, Changlong He, et al. Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2. [J]. Genes & Diseases 7(4).(2020)
DOI：

Jie Hu, Qingzhu Gao, Changlong He, et al. Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2. [J]. Genes & Diseases 7(4).(2020) DOI： 10.1016/j.gendis.2020.07.006.

摘要

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative virus of the coronavirus disease 2019 (COVID-19) pandemic. To establish a safe and convenient assay system for studying entry inhibitors and neutralizing antibodies against SARS-CoV-2

we constructed a codon-optimized

full-length C-terminal mutant spike (S) gene of SARS-CoV-2. We generated a luciferase (Luc)-expressing pseudovirus containing the wild-type or mutant S protein of SARS-CoV-2 in the envelope-defective HIV-1 backbone. The key parameters for this pseudovirus-based assay

including the S mutants and virus incubation time

were optimized. This pseudovirus contains a

Luc

reporter gene that enabled us to easily quantify virus entry into angiotensin-converting enzyme 2 (ACE2)-expressing 293T cells. Cathepsin (Cat)B/L inhibitor E−64d could significantly block SARS-CoV-2 pseudovirus infection in 293T-ACE2 cells. Furthermore

the SARS-CoV-2 spike pseudotyped virus could be neutralized by sera from convalescent COVID-19 patients or recombinant ACE2 with the fused Fc region of human IgG1. Thus

we developed a pseudovirus-based assay for SARS-CoV-2

which will be valuable for evaluating viral entry inhibitors and neutralizing antibodies against this highly pathogenic virus.

关键词

Keywords

Antiviral therapeuticsCoronavirusNeutralizing antibodiesPseudovirusSARS-Cov-2Spike protein

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