Presentation: P01016

Identification of enzyme inhibitors by screening compound libraries is a common procedure in drug discovery. HTS enzymatic assays should have a fine-tuned detection limit of inhibitory activity (iadl) in order to detect potentially useful compounds that may be weak and/or very diluted inhibitors. Traditionally, activity measurements are performed under initial rate conditions. In this study we show that improved detectability and robustness are obtained at substrate conversion levels beyond the initial rate region. These results were obtained with a mathematical model for iadl. The model is a function of the substrate conversion level of enzyme activity control and assay parameters that account for the experimental noise. The substrate conversion level where detectability and its robustness are improved was traced down for each member of a series of simulations with random combinations of plausible assay parameters values. This point is in all cases beyond the initial velocity range.

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