Limited Proteolysis-Coupled Mass Spectrometry (LiP-MS): A Cutting-Edge Tool for De Novo Drug Target Discovery

The identification of drug targets is fundamental to drug development. Traditional affinity-based target screening methods, including chemical biology probes and biotin labeling techniques, rely on the presumption of pre-existing knowledge of targets, thereby limiting their ability to uncover novel mechanisms of action. Recently, limited proteolysis combined with mass spectrometry (LiP-MS) has emerged as a hypothesis-free approach. By detecting drug-induced conformational alterations in proteins and integrating these observations with high-throughput mass spectrometry analysis, LiP-MS enables target identification without prior chemical modification. This article presents a comprehensive review of the underlying principles and workflow of LiP-MS, focusing on recent advancements, existing challenges, and strategies for its integration with complementary technologies. Furthermore, it delineates the advantages of LiP-MS relative to conventional proteomic methods and summarizes drug targets identified through LiP-MS in recent studies.