High-Throughput Fluorescent Assay for Inhibitor Screening of Proteases from RNA Viruses

0544070 - ÚOCHB 2022 RIV CH eng J - Journal Article
Cihlová, Bára - Hušková, Andrea - Böserle, Jiří - Nencka, Radim - Bouřa, Evžen - Šilhán, Jan
High-Throughput Fluorescent Assay for Inhibitor Screening of Proteases from RNA Viruses.
Molecules. Roč. 26, č. 13 (2021), č. článku 3792. E-ISSN 1420-3049
R&D Projects: GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : high-throughput screening * virus * drug * discovery * papain-like * protease * SARS-CoV-2 * flavivirus * TBEV
OECD category: Biochemistry and molecular biology
Impact factor: 4.927, year: 2021
Method of publishing: Open access
https://doi.org/10.3390/molecules26133792

Spanish flu, polio epidemics, and the ongoing COVID-19 pandemic are the most profound examples of severe widespread diseases caused by RNA viruses. The coronavirus pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demands affordable and reliable assays for testing antivirals. To test inhibitors of viral proteases, we have developed an inexpensive high-throughput assay based on fluorescent energy transfer (FRET). We assayed an array of inhibitors for papain-like protease from SARS-CoV-2 and validated it on protease from the tick-borne encephalitis virus to emphasize its versatility. The reaction progress is monitored as loss of FRET signal of the substrate. This robust and reproducible assay can be used for testing the inhibitors in 96- or 384-well plates.
Permanent Link: http://hdl.handle.net/11104/0321123