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A protease-activatable luminescent biosensor and reporter cell line for authentic SARS-CoV-2 infection.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Gerber, Pehuén Pereyra 
Duncan, Lidia M 
Greenwood, Edward Jd  ORCID logo  https://orcid.org/0000-0002-5224-0263
Marelli, Sara 
Naamati, Adi 

Abstract

Efforts to define serological correlates of protection against COVID-19 have been hampered by the lack of a simple, scalable, standardised assay for SARS-CoV-2 infection and antibody neutralisation. Plaque assays remain the gold standard, but are impractical for high-throughput screening. In this study, we show that expression of viral proteases may be used to quantitate infected cells. Our assays exploit the cleavage of specific oligopeptide linkers, leading to the activation of cell-based optical biosensors. First, we characterise these biosensors using recombinant SARS-CoV-2 proteases. Next, we confirm their ability to detect viral protease expression during replication of authentic virus. Finally, we generate reporter cells stably expressing an optimised luciferase-based biosensor, enabling viral infection to be measured within 24 h in a 96- or 384-well plate format, including variants of concern. We have therefore developed a luminescent SARS-CoV-2 reporter cell line, and demonstrated its utility for the relative quantitation of infectious virus and titration of neutralising antibodies.

Description

Funder: NIHR Cambridge BRC

Keywords

Biosensing Techniques, COVID-19, COVID-19 Testing, Cell Line, Humans, Luminescent Measurements, Peptide Hydrolases, SARS-CoV-2, Viral Proteins, Virus Replication

Journal Title

PLoS Pathog

Conference Name

Journal ISSN

1553-7366
1553-7374

Volume Title

18

Publisher

Public Library of Science (PLoS)
Sponsorship
Medical Research Council (MR/P008801/1)