Abstract
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.
Highlights
A novel severe acute respiratory syndrome coronavirus (SARS-CoV-2) has caused pandemic disease after its emergence at the end of 2019
We developed a novel cell fusion assay based upon spikehACE2 interaction, which demonstrated excellent correlation with standard spike pseudotyping for inhibition by convalescent sera, cloned antibodies, and biologics, after
Cell fusion assay to quantify SARS-Cov-2 spike interaction with human Angiotensin-Converting Enzyme 2 (hACE2) following competing interests: An invention disclosure has been filed with Yale University and the Department of Veteran’s Affairs regarding the cell fusion assay
Summary
A novel severe acute respiratory syndrome coronavirus (SARS-CoV-2) has caused pandemic disease after its emergence at the end of 2019. It was declared as a major global public health issue by WHO in March 2020 [1]. As of May 18th 2021, SARS-CoV-2 has infected nearly 164 million people and has caused close to 3.4 million deaths world-wide Remdesivir is the only FDA-approved drug for the treatment for COVID-19 patients (https://www.fda.gov/drugs/drug-safety-and-availability/ fdas-approval-veklury-remdesivir-treatment-covid-19-science-safety-and-effectiveness), and three vaccines have Emergency Used Authorization (EUA) from the FDA Gov/emergency-preparedness-and-response/coronavirus-disease-2019-covid-19/covid19-vaccines), and four vaccines have been formally approved by the EMA
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