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Infectious SARS-CoV-2 neutralization assay

DS Deyong Song
WW Wenbo Wang
CD Chuangchuang Dong
ZN Zhenfei Ning
XL Xiu Liu
CL Chuan Liu
GD Guangying Du
CS Chunjie Sha
KW Kailin Wang
JL Jun Lu
BS Baiping Sun
YZ Yanyan Zhao
QW Qiaoping Wang
HX Hongguang Xu
YL Ying Li
ZS Zhenduo Shen
JJ Jie Jiao
RW Ruiying Wang
JT Jingwei Tian
WL Wanhui Liu
LW Lan Wang
YD Yong-Qiang Deng
CD Changlin Dou
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Neutralizing activity of mAbs was measured using a standard plaque reduction neutralization with Vero cells. Briefly, fivefold serial dilutions of mAbs were added to approximately 100 PFU of SARS-CoV-2 and incubated for 1 h at 37 °C. Then, the mixture was added to Vero cell monolayers in a 24-well plate in duplicate and incubated for 1 h at 37 °C. The mixture was removed, and 1 ml of 1.0% (w/v) LMP agarose (Promega) in DMEM plus 4% (v/v) FBS was layered onto the infected cells. After further incubation at 37 °C for 2 days, the wells were stained with 1% (w/v) crystal violet dissolved in 4% (v/v) formaldehyde to visualize the plaques. PRNT50 values were determined using non-linear regression analysis. All experiments were performed followed the standard operating procedures of the approved Biosafety Level-3 facility.

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