PLoS Pathogens (Jun 2021)

Rapid, reliable, and reproducible cell fusion assay to quantify SARS-Cov-2 spike interaction with hACE2.

  • Min Zhao,
  • Pei-Yi Su,
  • Danielle A Castro,
  • Therese N Tripler,
  • Yingxia Hu,
  • Matthew Cook,
  • Albert I Ko,
  • Shelli F Farhadian,
  • Benjamin Israelow,
  • Charles S Dela Cruz,
  • Yong Xiong,
  • Richard E Sutton,
  • Yale IMPACT Research Team

DOI
https://doi.org/10.1371/journal.ppat.1009683
Journal volume & issue
Vol. 17, no. 6
p. e1009683

Abstract

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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.