Rapid Evaluation of Small Molecule Cellular Target Engagement with a Luminescent Thermal Shift Assay

Determination of target engagement for candidate drug molecules in the native cellular environment is a significant challenge for drug discovery programs. The cellular thermal shift assay (CETSA) has emerged as a powerful tool for determining compound target engagement through measurement of changes...

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Bibliographic Details
Published inACS medicinal chemistry letters Vol. 12; no. 8; pp. 1288 - 1294
Main Authors Mortison, Jonathan D, Cornella-Taracido, Ivan, Venkatchalam, Gireedhar, Partridge, Anthony W, Siriwardana, Nirodhini, Bushell, Simon M
Format Journal Article
LanguageEnglish
Published American Chemical Society 12.08.2021
Subjects
Letter
BiTSA
KRAS
HiBiT
cellular target engagement
TP53
Thermal shift assay
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Summary:Determination of target engagement for candidate drug molecules in the native cellular environment is a significant challenge for drug discovery programs. The cellular thermal shift assay (CETSA) has emerged as a powerful tool for determining compound target engagement through measurement of changes to a protein's thermal stability upon ligand binding. Here, we present a HiBiT thermal shift assay (BiTSA) that deploys a quantitative peptide tag for determination of compound target engagement in the native cellular environment using a high throughput, plate-based luminescence readout. We demonstrate that BiTSA can rapidly assess cellular target engagement of small molecule ligands against their cognate targets and highlight two applications of BiTSA for differentiating small molecules targeting mutant KRAS and TP53.
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ISSN:1948-5875
1948-5875
DOI:10.1021/acsmedchemlett.1c00276