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Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors.
Ihssen, Julien; Faccio, Greta; Yao, Chunyan; Sirec, Teja; Spitz, Urs.
  • Ihssen J; Biosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland.
  • Faccio G; Biosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland.
  • Yao C; Biosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland.
  • Sirec T; Biosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland.
  • Spitz U; Biosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland.
STAR Protoc ; 2(3): 100793, 2021 09 17.
Article in English | MEDLINE | ID: covidwho-1368796
ABSTRACT
This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of Mpro, the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of Mpro mainly rely on in silico molecular dynamics simulations or on FRET (Fluorescence Resonance Energy Transfer) substrates. The protocol is based on an aminomethyl coumarin substrate. High sensitivity, specificity, and an easily detectable fluorescent read-out are the advantages offered by this rapid assay, which allows high throughput screening of new Mpro inhibitors.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Antiviral Agents / Protease Inhibitors / Viral Proteins / Fluorescence Resonance Energy Transfer / Coronavirus 3C Proteases / SARS-CoV-2 / COVID-19 Drug Treatment Limits: Humans Language: English Journal: STAR Protoc Year: 2021 Document Type: Article Affiliation country: J.xpro.2021.100793

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Antiviral Agents / Protease Inhibitors / Viral Proteins / Fluorescence Resonance Energy Transfer / Coronavirus 3C Proteases / SARS-CoV-2 / COVID-19 Drug Treatment Limits: Humans Language: English Journal: STAR Protoc Year: 2021 Document Type: Article Affiliation country: J.xpro.2021.100793