In this study, we present a live-cell ebBRET-based biosensor platform which includes twelve distinct biosensors for monitoring SH2-domain containing proteins-mediated downstream signaling of RTKs. These biosensors are designed to measure receptor proximal events that are engaged upon receptor activation at the plasma membrane and early endosomal compartments. We demonstrate here the applicability of our biosensor platform to perform in-depth characterization of RTK biology and pharmacology. This technology revealed different modes of action of ligands and inhibitors, extensive RTK signal profiling of EGFR mutants, and trafficking of RTK effectors. It represents a powerful tool for the analysis of RTK mutations and for the identification of new generation of TKIs and antibodies directed against RTKs.