Work

Protocol for covalent ligand discovery via library-versus-proteome screening

Year: 2026

Type: article

Abstract: Here, we present a dynamic combinatorial library-versus-proteome activity-based protein profiling (DCL-ABPP) workflow for high-throughput covalent ligand discovery. We describe steps for integrating a dynamic combinatorial library with proteome-wide mass spectrometry to generate and screen hundreds of ligands in situ , eliminating the need for pre-synthesis. We detail procedures for two complementary modes: competitive screening, which identifies enzyme inhibitors (e.g., serine hydrolases) through family-wide probe competition; and direct screening, which maps covalent ligand binding sites on cysteines. For complete details on the use and execution of this protocol, please refer to Huang et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104786

Authors: Yuchen Huang, Lexuan Hou, Lin Zhu, Xi Yang, Gang Li

Institution: Shenzhen Bay Laboratory

Language: English

Topic: Click Chemistry and Applications

Subfield: Organic Chemistry

Field: Chemistry

Domain: Physical Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: National Natural Science Foundation of China, Shenzhen Bay Laboratory, Guangdong Special Support Plan