Biochemical assay development for drug discovery: a sequential optimization from protein expression to enzymatic activity

Background The drug discovery process based on high-throughput screening (HTS) requires highly demanding efforts to comply with the strict criteria of homogeneity, sensitivity, processivity, reproducibility and miniaturization. For enzymatic targets, the ability to screen in parallel multiple parameters is a prerequisite to identify the permissive conditions to develop functional assays and to upscale the protein production to entirely support an HTS campaign, which generally requires several milligrams of pure protein.