CETSA as a tool for assessing selectivity of warheads and PROTACS and for detecting binders that remain intact
Poster
Enhancing PROTAC research: CETSA as a tool for assessing selectivity of warheads and PROTACS and for detecting binders that remain intact
CETSA® (Cellular Thermal Shift Assay) is an effective method for studying interactions between small molecules and their protein targets, particularly useful in targeted protein degradation (TPD) as it improves insights from degradation profiles. CETSA can be conducted in cell lysates and intact cells without modifying compounds, proteins, or cellular environments.
By using mass spectrometric readouts, CETSA assesses binding selectivity for warheads targeting specific proteins and evaluates selectivity across various PROTAC scaffolds. The impact of effective PROTAC interactions can be monitored through protein degradation assays. However, parallel assessment of target engagement by CETSA, shows that not all proteins that bind the PROTAC warhead undergo degradation. Thus, cellular effects from PROTACs may arise from both primary target degradation and traditional protein-ligand interactions, potentially leading to off-target effects similar like those seen in traditional small molecule drug development projects.