In:
PLOS Pathogens, Public Library of Science (PLoS), Vol. 18, No. 2 ( 2022-2-10), p. e1010265-
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.
Type of Medium:
Online Resource
ISSN:
1553-7374
DOI:
10.1371/journal.ppat.1010265
DOI:
10.1371/journal.ppat.1010265.g001
DOI:
10.1371/journal.ppat.1010265.g002
DOI:
10.1371/journal.ppat.1010265.g003
DOI:
10.1371/journal.ppat.1010265.g004
DOI:
10.1371/journal.ppat.1010265.g005
DOI:
10.1371/journal.ppat.1010265.s001
DOI:
10.1371/journal.ppat.1010265.s002
DOI:
10.1371/journal.ppat.1010265.s003
DOI:
10.1371/journal.ppat.1010265.s004
DOI:
10.1371/journal.ppat.1010265.s005
DOI:
10.1371/journal.ppat.1010265.s006
DOI:
10.1371/journal.ppat.1010265.s007
DOI:
10.1371/journal.ppat.1010265.s008
DOI:
10.1371/journal.ppat.1010265.s009
DOI:
10.1371/journal.ppat.1010265.s010
DOI:
10.1371/journal.ppat.1010265.s011
DOI:
10.1371/journal.ppat.1010265.s012
DOI:
10.1371/journal.ppat.1010265.s013
DOI:
10.1371/journal.ppat.1010265.s014
DOI:
10.1371/journal.ppat.1010265.r001
DOI:
10.1371/journal.ppat.1010265.r002
DOI:
10.1371/journal.ppat.1010265.r003
DOI:
10.1371/journal.ppat.1010265.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2022
detail.hit.zdb_id:
2205412-1
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