Kinase Screening and Profiling

Volume 1360 of the series Methods in Molecular Biology pp 97-106

A High-Content Assay to Screen for Modulators of EGFR Function

  • Christophe AntczakAffiliated withHTS Core Facility, Memorial Sloan Kettering Cancer CenterCenter for Proteomic Chemistry, Novartis Institutes for Biomedical Research
  • , Hakim DjaballahAffiliated withHTS Core Facility, Memorial Sloan Kettering Cancer CenterInstitut Pasteur Korea Email author 

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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 purified 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 trafficking, and internalization are imaged and quantified 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.

Key words

RNAi siRNA Iressa Kinase Inhibitor High-content assay EGFR Cancer Drug discovery INCA2000 INCA3000 Microscopy HTS