| Assay Method Information | |
| | SAR-CoV-2 MPro Enzyme inhibition IC50 determination |
| Description: | The inhibition of the SAR-CoV-2 MPro Enzyme was measured by the method of Mesecar et al.20: The fluorescence of the cleaved FAM peptide (excitation 480 nm/emission 520 nm) was measured using a fluorescence intensity protocol on a Pherastar FX (BMG). The assay reaction buffer contained 20 mM HEPES (pH 7.3), 50 mM NaCl, 10% Glycerol, 1 mM TCEP, 0.01 % Tween-20. Compounds were pre-incubated with 5 nM SARS-CoV-23CL protease for 15 minutes at 23 °C. Enzyme reactions were initiated with the addition of 375 nM [5-FAM]-AVLQSGFR-[Lys(Dabcyl)-K-amide substrate and allowed to proceed for 30 minutes at 23 °C. Percent inhibition or activity was calculated based on control wells containing no compound (0% inhibition/100% activity). IC5o values were generated using a 4-parameter hill slope fit model using Vault software (CDD). |
| Affinity data for this assay | |
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