| Assay Method Information | |
| | SARS2 Coronavirus 3CL Protease Assay |
| Description: | The enzymatic activity of SARS2 coronavirus 3CL protease was determined in a FRET (fluorescence resonance energy transfer)-based assay measuring the cleavage of a peptide substrate by recombinantly expressed and purified enzyme. Cleavage of the peptide SEQ ID NO:1 (CPC Scientific) by SARS2 3CL protease was measured in reaction buffer (50 mM Hepes pH 7.5, 0.01% Triton X-100, 0.01% BSA, 2 mM DTT). SARS2 3CL protease (5 nM final concentration) was pre-incubated with compound for 30 minutes before reaction initiation with peptide substrate (15 uM final concentration). Room temperature reactions (4 h) were quenched by addition of a high dose of inhibitor and read on an appropriate plate reader (excitation wavelength=495 nm, emission wavelength=520 nm). Data were analyzed by a standard 4 parameter fit to determine IC50 values. |
| Affinity data for this assay | |
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