Overcoming Challenging Targets: What To Do When Midpoint CTE Returns Inconclusive Results
Midpoint CETSA is a common method for assessing target engagement by measuring protein stability at a single temperature, but it can obscure important mechanistic differences in complex proteins. This article explains how multi-temperature CETSA profiles reveal conformational state-specific binding, improving compound ranking and selectivity assessment. It highlights when to move beyond midpoint CETSA, especially for covalent mechanisms, mutant selectivity, and structurally diverse compounds, providing a practical decision framework for drug discovery.
