2. Academic Validation
  3. Establishment of a pseudovirus neutralization assay based on SARS-CoV-2 S protein incorporated into lentiviral particles

Establishment of a pseudovirus neutralization assay based on SARS-CoV-2 S protein incorporated into lentiviral particles

  • Biosaf Health. 2022 Feb;4(1):38-44. doi: 10.1016/j.bsheal.2021.12.006.
Sheng Wang 1 2,  Lizhen Liu 1 2 3,  Can Wang 1 2,  Ziqiang Wang 1 2,  Xuhua Duan 1 2 4,  Gang Chen 1 2,  Hu Zhou 5,  Hong Shao 1 2
Affiliations

Affiliations

  • 1 Department of Biochemical Drugs and Biological Products, Shanghai Institute for Food and Drug Control, Shanghai 201203, China.
  • 2 NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai Institute for Food and Drug Control, Shanghai 201203, China.
  • 3 School of Pharmacy, Fudan University, Shanghai 201203, China.
  • 4 Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.
  • 5 Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
PMID: 35005601 DOI: 10.1016/j.bsheal.2021.12.006
Abstract

The coronavirus disease 2019 (COVID-19) is still causing a wide range of infections and deaths due to the high variability of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Therefore, it is necessary to establish a reliable and convenient pseudovirus-based neutralization assay to develop drug targeted variants of SARS-CoV-2. Based on the HIV-1 backbone, we generated a high titer luciferase (Luc)-expressing pseudovirus packaging system. Three dominant S mutant substitution pseudovirus were also established and identified compared to wide type in hACE2-overexpressing HEK-293T cells (293T-ACE2 cells). Compared to Serine Protease Inhibitor camostat mesylate, the cysteine protease inhibitor E-64d could significantly block all SARS-CoV-2 mutant S pseudovirus Infection in 293T-ACE2 cells. Furthermore, the neutralization ability of two antibodies targeted receptor-binding domain (RBD) of SARS-CoV-2 spike protein (S) was evaluated, which showed different inhibition dose-effect curves among four types of S pseudovirus. Overall, we developed a pseudovirus-based neutralization assay for SARS-CoV-2, which would be readily adapted to SARS-CoV-2 variants for evaluating antibodies.

Keywords

COVID-19; Neutralizing antibody; Pseudovirus; RBD; SARS-CoV-2 variants.

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