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

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 st...

Full description

Saved in:

Bibliographic Details
Published in:	PLoS pathogens Vol. 18; no. 2; p. e1010265
Main Authors:	Gerber, Pehuén Pereyra, Duncan, Lidia M, Greenwood, Edward Jd, Marelli, Sara, Naamati, Adi, Teixeira-Silva, Ana, Crozier, Thomas Wm, Gabaev, Ildar, Zhan, Jun R, Mulroney, Thomas E, Horner, Emily C, Doffinger, Rainer, Willis, Anne E, Thaventhiran, James Ed, Protasio, Anna V, Matheson, Nicholas J
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 10-02-2022 Public Library of Science (PLoS)
Subjects:	Antibodies Assaying Biology and life sciences Biosensing Techniques - methods Biosensors Cell activation Cell Line Coronaviruses COVID-19 COVID-19 - diagnosis COVID-19 - virology COVID-19 Testing - methods Engineering and Technology Experiments Flow cytometry Health aspects High-throughput screening HIV Human immunodeficiency virus Humans Infections Luminescent Measurements - methods Medicine and health sciences Microscopy Peptide Hydrolases - analysis Peptide Hydrolases - genetics Peptide Hydrolases - metabolism Physical Sciences Physiological aspects Protease Proteases Proteinase Proteins Quantitation Research and Analysis Methods SARS-CoV-2 - enzymology SARS-CoV-2 - genetics SARS-CoV-2 - physiology Severe acute respiratory syndrome Severe acute respiratory syndrome coronavirus 2 Titration Viral diseases Viral infections Viral Proteins - analysis Viral Proteins - genetics Viral Proteins - metabolism Virus Replication Viruses United Kingdom
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	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.
Bibliography:	new_version The authors have declared that no competing interests exist.
ISSN:	1553-7374 1553-7366 1553-7374
DOI:	10.1371/journal.ppat.1010265