摘要
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‐overexpressingHEK‐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 showeddifferent 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.
基金
This work was supported by grants 2020‐SKT‐14 and 2021‐YKT‐01 from the Shanghai Institute for Food and Drug Control
LX‐2021‐06 from the Shanghai Drug Administration
19DZ2294600 from the Shanghai Science and Technology Committee.
