A high-content assay to screen for modulators of EGFR function

Abstract

Cell-based assays have the potential and advantage to identify cell-permeable modulators of kinase function, and hence provide an alternative to the conventional enzymatic activity-driven discovery approaches that rely on purifi ed recombinant kinase catalytic domains. Here, we describe a domain-based high- content biosensor approach to study endogenous EGFR activity whereby EGF-induced receptor activation, subsequent traffi cking, and internalization are imaged and quantifi ed using time-dependent granule formation in cells. This method can readily be used to search for EGFR modulators in both chemical and RNAi screening; with potential applicability to other receptor tyrosine kinases.