tetano
Editor, Senior Moderator
Biosaf Health
. 2022 Jan 3.
doi: 10.1016/j.bsheal.2021.12.006. Online ahead of print.
Establishment of a pseudovirus neutralization assay based on SARS-CoV-2 S protein incorporated into lentiviral particles
Sheng Wang[SUP] 1 2 [/SUP], Lizhen Liu[SUP] 1 2 3 [/SUP], Can Wang[SUP] 1 2 [/SUP], Ziqiang Wang[SUP] 1 2 [/SUP], Xuhua Duan[SUP] 1 2 4 [/SUP], Gang Chen[SUP] 1 2 [/SUP], Hu Zhou[SUP] 5 [/SUP], Hong Shao[SUP] 1 2 [/SUP]
Affiliations
Abstract
The Coronavirus Disease 2019 (COVID-19) is still causing a wide range of infections and deaths due to the high variability of the SARS-CoV-2 virus. 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.
. 2022 Jan 3.
doi: 10.1016/j.bsheal.2021.12.006. Online ahead of print.
Establishment of a pseudovirus neutralization assay based on SARS-CoV-2 S protein incorporated into lentiviral particles
Sheng Wang[SUP] 1 2 [/SUP], Lizhen Liu[SUP] 1 2 3 [/SUP], Can Wang[SUP] 1 2 [/SUP], Ziqiang Wang[SUP] 1 2 [/SUP], Xuhua Duan[SUP] 1 2 4 [/SUP], Gang Chen[SUP] 1 2 [/SUP], Hu Zhou[SUP] 5 [/SUP], Hong Shao[SUP] 1 2 [/SUP]
Affiliations
- PMID: 35005601
- PMCID: PMC8721934
- 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 SARS-CoV-2 virus. 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.