Elsevier

Virology

Volume 556, April 2021, Pages 73-78
Virology

Detecting SARS-CoV-2 3CLpro expression and activity using a polyclonal antiserum and a luciferase-based biosensor

https://doi.org/10.1016/j.virol.2021.01.010Get rights and content
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Highlights

  • Coronavirus 3CLpro activity is efficiently measured using a luciferase-based biosensor.

  • Small molecule inhibitors of 3CLpro block activation of the biosensor.

  • Polyclonal antiserum raised against SARS-CoV 3CLpro recognizes SARS-CoV-2 3CLpro in WB and IF assays.

Abstract

The need to stem the current outbreak of SARS-CoV-2 responsible for COVID-19 is driving the search for inhibitors that will block coronavirus replication and pathogenesis. The coronavirus 3C-like protease (3CLpro) encoded in the replicase polyprotein is an attractive target for antiviral drug development because protease activity is required for generating a functional replication complex. Reagents that can be used to screen for protease inhibitors and for identifying the replicase products of SARS-CoV-2 are urgently needed. Here we describe a luminescence-based biosensor assay for evaluating small molecule inhibitors of SARS-CoV-2 3CLpro/main protease. We also document that a polyclonal rabbit antiserum developed against SARS-CoV 3CLpro cross reacts with the highly conserved 3CLpro of SARS-CoV-2. These reagents will facilitate the pre-clinical evaluation of SARS-CoV-2 protease inhibitors.

Keywords

SARS-CoV-2
3CLpro
Main protease
Mpro
nsp5
Protease inhibitors
pGlo biosensor

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