Why is there a significant difference between IC50 in enzymatic assay and KD of ITC?

Debomita Bhattacharya @Debomita-Bhattacharya

18 July 2023 1 3K Report

When checked in enzymatic assay, a compound with IC50 37 nM showed a KD value of 20 µM against ITC. How can this discrepancy be explained?

Adam B Shapiro

It is possible for a binding interaction to generate no heat at a given temperature. The result may be different if you change the temperature.

The observation of a 20 µM Kd may represent a secondary weak binding mode, or some non-binding phenomenon involving the compound by itself.

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