Engaged but not degraded

Target engagement profiling of degrader compounds with CETSA CETSA® (Cellular Thermal Shift Assay) is a label-free method for studying small molecule-protein interactions and is especially useful in targeted protein degradation (TPD) to complement degradation profiling.Antibody-based detection with AlphaLISA was used to investigate degradation kinetics for three target proteins: CDK4, CDK9, and BRD4. CETSA combined with mass spectrometry readouts enables selectivity profiling of full PROTACs and corresponding warheads. While degradation assays measure outcome, CETSA also detects target engagement, revealing that not all PROTAC-bound proteins are degraded. Thus, PROTACs may drive cellular effects through both degradation and conventional ligand interactions, including potential off-target effe...