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. 2025:2905:171-179.
doi: 10.1007/978-1-0716-4418-8_11.

Click Chemistry for Target Engagement Studies

Silvia Castelli  1 , Sonia Troiani  2 , Ulisse Cucchi  2
Affiliations

Click Chemistry for Target Engagement Studies

Silvia Castelli et al. Methods Mol Biol. 2025.

Abstract

The expanding number of targeted covalent small molecule inhibitors in clinical development increases the importance of identification and quantitation of on- and off-target engagement for these molecules in order to better understand the level of target binding and inhibition and potential side effects. Here we describe the optimization of a click chemistry-based chemoproteomic approach, to study the target engagement of covalent molecules. Using the BTK inhibitor ibrutinib as a casa study, we aim to characterize its mechanistic profile in cell-systems.

Keywords: Chemoproteomic; Click chemistry; Covalent inhibitors; Gel-based analysis; Target engagement.

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