Abstract

NanoBRET™ Target Engagement Assay technology offers a sensitive, specific method to measure the interaction of small-molecule drugs with their target protein in live cells. This live-cell assay method facilitates quantitative determination of compound affinity and target protein occupancy in live cells in addition to kinetic parameters such as residence time under physiological conditions. Recent developments in the field have led to the generation of a pan-KRAS tracer for the investigation of engagement at this challenging target. The presentation will highlight a suite of tools available for KRAS target engagement in addition to other tools to address target engagement at downstream effector proteins in the RAS-RAF-MEK-ERK pathway. Data will also be presented to show how the recently introduced Lumit™ technology can be used to detect activation of ERK by detection of phospho-ERK using a simple, homogeneous no-wash assay format. Targeted protein degradation based approaches in drug discovery are increasingly important. Data will also be presented to show how luminescent tagging of endogenous KRAS (G12C) using CRISPR gene editing with HiBiT tag can be used to detect targeted degradation of KRAS using a VHL-based PROTAC molecule.