Real-Time Cellular Thermal Shift Assay to Monitor Target Engagement.

ABSTRACT: Determining a molecule's mechanism of action is paramount during chemical probe development and drug discovery. The cellular thermal shift assay (CETSA) is a valuable tool to confirm target engagement in cells for a small molecule that demonstrates a pharmacological effect. CETSA directly detects biophysical interactions between ligands and protein targets, which can alter a protein's unfolding and aggregation properties in response to thermal challenge. In traditional CETSA experiments, each temperature requires an individual sample, which restricts throughput and requires substantial optimization. To capture the full aggregation profile of a protein from a single sample, we developed a prototype real-time CETSA (RT-CETSA) platform by coupling a real-time PCR instrument with a CCD camera

SUBMITTER: Sanchez TW 

PROVIDER: S-EPMC9486815 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature