Article
Development of an efficient reproducible cell-cell transmission assay for rapid quantification of SARS-CoV-2 spike interaction with hACE2

https://doi.org/10.1016/j.crmeth.2022.100252Get rights and content
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open access

Highlights

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    An efficient cell-cell transmission assay for SARS-CoV-2 spike-hACE2 interaction

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    Uses two stable cell lines and a single, helper-dependent adenovirus vector

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    Reliably tests effects of small-molecule drugs, monoclonal antibodies, and vaccines

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    Rapid, reproducible assay, easily modifiable to encode spike variants of concern

Motivation

Reproducible and facile methods of measuring viral replication are indispensable in the development of vaccines and therapeutic medications against SARS-CoV-2. We sought to develop a rapid, time-saving, cell-cell transmission assay that can be used to measure and quantify SARS-CoV-2 virus-cell binding and entry to circumvent the challenges faced by the assays currently used as gold standards for SARS-CoV-2 neutralization studies.

Summary

Efficient quantitative assays for measurement of viral replication and infectivity are indispensable for future endeavors to develop prophylactic or therapeutic antiviral drugs or vaccines against SARS-CoV-2. We developed a SARS-CoV-2 cell-cell transmission assay that provides a rapid and quantitative readout to assess SARS-CoV-2 spike hACE2 interaction in the absence of pseudotyped particles or live virus. We established two well-behaved stable cell lines, which demonstrated a remarkable correlation with standard cell-free viral pseudotyping for inhibition by convalescent sera, small-molecule drugs, and murine anti-spike monoclonal antibodies. The assay is rapid, reliable, and highly reproducible, without a requirement for any specialized research reagents or laboratory equipment and should be easy to adapt for use in most investigative and clinical settings. It can be effectively used or modified for high-throughput screening for compounds and biologics that interfere with virus-cell binding and entry to complement other neutralization assays currently in use.

Keywords

SARS-CoV-2
cell-cell transmission
pseudotyping
quantitative assay
neutralization
stable cell line
infectivity assay
convalescent sera
post-vaccination sera

Data and code availability

  • All the Raw datasets generated during this study have been deposited at Mendeley Data and are publicly available as of the date of publication. DOIs are listed in the key resources table.

  • This paper does not report original code.

  • Any additional information required to reanalyze the data reported in this paper is available from the lead contact upon request.

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Lead contact