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

Sheng Wang; Lizhen Liu; Can Wang; Ziqiang Wang; Xuhua Duan; Gang Chen; Hu Zhou; Hong Shao

doi:10.1016/j.bsheal.2021.12.006

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. Epub 2022 Jan 3.

Authors

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 Zhou⁵, Hong Shao^{1

2}

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.

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.