tetano
Editor, Senior Moderator
Virology
. 2021 Jan 26;556:73-78.
doi: 10.1016/j.virol.2021.01.010. Online ahead of print.
Detecting SARS-CoV-2 3CLpro expression and activity using a polyclonal antiserum and a luciferase-based biosensor
Amornrat O'Brien[SUP] 1 [/SUP], Da-Yuan Chen[SUP] 2 [/SUP], Matthew Hackbart[SUP] 1 [/SUP], Brianna J Close[SUP] 3 [/SUP], Timothy E O'Brien[SUP] 4 [/SUP], Mohsan Saeed[SUP] 5 [/SUP], Susan C Baker[SUP] 6 [/SUP]
Affiliations
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: 3CLpro; Main protease; Mpro; Protease inhibitors; SARS-CoV-2; nsp5; pGlo biosensor.
. 2021 Jan 26;556:73-78.
doi: 10.1016/j.virol.2021.01.010. Online ahead of print.
Detecting SARS-CoV-2 3CLpro expression and activity using a polyclonal antiserum and a luciferase-based biosensor
Amornrat O'Brien[SUP] 1 [/SUP], Da-Yuan Chen[SUP] 2 [/SUP], Matthew Hackbart[SUP] 1 [/SUP], Brianna J Close[SUP] 3 [/SUP], Timothy E O'Brien[SUP] 4 [/SUP], Mohsan Saeed[SUP] 5 [/SUP], Susan C Baker[SUP] 6 [/SUP]
Affiliations
- PMID: 33548599
- DOI: 10.1016/j.virol.2021.01.010
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: 3CLpro; Main protease; Mpro; Protease inhibitors; SARS-CoV-2; nsp5; pGlo biosensor.