Assay Method Information

Assay Name:  SARS-CoV-2 Protease FRET Assay
Description:  Pure SARS-CoV-2 3CLP was obtained as previously described in detail, and the enzymatic activity was confirmed according to an established protocol; see ref 42. The FRET substrate was synthesized according to the methods previously described in ref 42. The IC50 values for each inhibitor tested were determined using methods described previously in ref 42 via a FRET-based kinetic assay with a 30 min inhibitor incubation period prior to the addition of the substrate. GraphPad Prism software (GraphPad 8.3.1) was used to determine the IC50. Compounds were tested at eight different concentrations (100 to 0.00001 μM). Triplicate experiments were performed for each data point, the appropriate Hill slopes were generated, and the value was presented as a mean standard error.
Affinity data for this assay
 

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