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Detecting SARS-CoV-2 3CLpro expression and activity using a polyclonal antiserum and a luciferase-based biosensor.
O'Brien, Amornrat; Chen, Da-Yuan; Hackbart, Matthew; Close, Brianna J; O'Brien, Timothy E; Saeed, Mohsan; Baker, Susan C.
  • O'Brien A; Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL, USA.
  • Chen DY; Department of Biochemistry, Boston University School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, MA, USA.
  • Hackbart M; Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL, USA.
  • Close BJ; National Emerging Infectious Diseases Laboratories, Boston University, MA, USA; Department of Microbiology, Boston University School of Medicine, Boston, MA, USA.
  • O'Brien TE; Department of Mathematics and Statistics, Loyola University Chicago, Chicago, IL, USA.
  • Saeed M; Department of Biochemistry, Boston University School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, MA, USA. Electronic address: msaeed1@bu.edu.
  • Baker SC; Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL, USA. Electronic address: sbaker1@luc.edu.
Virology ; 556: 73-78, 2021 04.
Article in English | MEDLINE | ID: covidwho-1049897
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / Coronavirus 3C Proteases / SARS-CoV-2 / Immune Sera / Luciferases Type of study: Experimental Studies / Prognostic study / Randomized controlled trials Limits: Animals Language: English Journal: Virology Year: 2021 Document Type: Article Affiliation country: J.virol.2021.01.010

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / Coronavirus 3C Proteases / SARS-CoV-2 / Immune Sera / Luciferases Type of study: Experimental Studies / Prognostic study / Randomized controlled trials Limits: Animals Language: English Journal: Virology Year: 2021 Document Type: Article Affiliation country: J.virol.2021.01.010