Assay Method Information

Assay Name:  Enzyme Inhibition Assay and Ki Values Determination
Description:  To determine the Ki values of active compounds, 25 nM Mpro was mixed with increasing concentrations of compounds (from 4 nM to 4,000 nM with twofold dilutions) and hydrolysis of 15 µM fluorescent peptide was monitored. Initial hydrolysis rates of fluorescent peptide were plotted as a function of compound concentrations and Ki values were obtained by fitting the data into the Morrison equation (42) with SE from triplicates. The Michealis Menten constant of enzyme (Km) value used for Ki calculations is 17 µM.
Affinity data for this assay
 

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