Open AccessRapid, reliable, and reproducible cell fusion assay to quantify SARS-Cov-2 spike interaction with hACE2
Author(s) -
Min Zhao,
Pei-Yi Su,
Danielle Castro,
Therese Tripler,
Yingxia Hu,
M. Katherine Cook,
Albert I. Ko,
Shelli Farhadian,
Benjamin Israelow,
Charles S. Dela Cruz,
Yong Xiong,
Richard E. Sutton
Publication year - 2021
Publication title -
plos pathogens
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.719
H-Index - 206
eISSN - 1553-7374
pISSN - 1553-7366
DOI - 10.1371/journal.ppat.1009683
Subject(s) - hek 293 cells , neutralization , cell culture , cell fusion , virus , virology , cell , computational biology , microbiology and biotechnology , chemistry , biology , biochemistry , genetics
COVID-19 is a global crisis of unimagined dimensions. Currently, Remedesivir is only fully licensed FDA therapeutic. A major target of the vaccine effort is the SARS-CoV-2 spike-hACE2 interaction, and assessment of efficacy relies on time consuming neutralization assay. Here, we developed a cell fusion assay based upon spike-hACE2 interaction. The system was tested by transient co-transfection of 293T cells, which demonstrated good correlation with standard spike pseudotyping for inhibition by sera and biologics. Then established stable cell lines were very well behaved and gave even better correlation with pseudotyping results, after a short, overnight co-incubation. Results with the stable cell fusion assay also correlated well with those of a live virus assay. In summary we have established a rapid, reliable, and reproducible cell fusion assay that will serve to complement the other neutralization assays currently in use, is easy to implement in most laboratories, and may serve as the basis for high throughput screens to identify inhibitors of SARS-CoV-2 virus-cell binding and entry.