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Article

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

Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo namesti 2, 166 10 Prague 6, Czech Republic
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Takuya Terai
Molecules 2021, 26(13), 3792; https://doi.org/10.3390/molecules26133792
Received: 10 May 2021 / Revised: 11 June 2021 / Accepted: 14 June 2021 / Published: 22 June 2021
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. View Full-Text
Keywords: high-throughput screening; virus; drug; discovery; papain-like; protease; SARS-CoV-2; flavivirus; TBEV high-throughput screening; virus; drug; discovery; papain-like; protease; SARS-CoV-2; flavivirus; TBEV
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MDPI and ACS Style

Cihlova, B.; Huskova, A.; Böserle, J.; Nencka, R.; Boura, E.; Silhan, J. High-Throughput Fluorescent Assay for Inhibitor Screening of Proteases from RNA Viruses. Molecules 2021, 26, 3792. https://doi.org/10.3390/molecules26133792

AMA Style

Cihlova B, Huskova A, Böserle J, Nencka R, Boura E, Silhan J. High-Throughput Fluorescent Assay for Inhibitor Screening of Proteases from RNA Viruses. Molecules. 2021; 26(13):3792. https://doi.org/10.3390/molecules26133792

Chicago/Turabian Style

Cihlova, Bara, Andrea Huskova, Jiri Böserle, Radim Nencka, Evzen Boura, and Jan Silhan. 2021. "High-Throughput Fluorescent Assay for Inhibitor Screening of Proteases from RNA Viruses" Molecules 26, no. 13: 3792. https://doi.org/10.3390/molecules26133792

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