| Assay Method Information | |
| | In Vitro Inhibitory Activity of Compounds Against SARS-CoV-2 Papain-Like Protease |
| Description: | Test compounds were assayed at 10 concentrations from 10 μM, in duplicate for the IC50 determination. The assay buffer contained 50 mM HEPES (pH 7.5), 0.01% Triton-X 100, 0.1 mg/ml BSA and 5 mM DTT. The final concentrations of the PLpro protein and substrate in the assay were 6.25 nM and 25 μM, respectively.Compounds were 3 folds serially diluted to 10 concentrations and added to an assay plate (384 w format) using ECHO, in duplicate wells. The final concentrations are 10 μM, 3.33 μM, 1.11 μM, 0.37 μM, 0.123 μM, 0.041 μM, 0.014 μM, 0.0046 μM, 0.0015 UM and 0.00051 μM.20 μL of 7.8 nM of PLpro protein were added to an assay plate containing compounds using a Multidrop. The compounds and PLpro protein were pre-incubated at room temperature for 30 min. Then 5 μL of 125 μM of substrate were added to an assay plate using a Multidrop. The final concentrations of PLpro and substrate were 6.25 nM and 25 μM, respectively. |
| Affinity data for this assay | |
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