Delaware Valley Mass Spectrometry Discussion Group

Abstract: HotSpot Therapeutics, Inc. is pioneering the discovery and development of a new class of allosteric drugs that target certain naturally occurring pockets on proteins called “natural hotspots”. These pockets are decisive in controlling a protein’s cellular function, and we believe they have significant potential for new drug discovery by enabling the systematic design of potent and selective small molecules with novel pharmacology.

We present our implementation of microflow LC, Q-TOF-MS and Zeno MS/MS for routine intact protein analysis, label-free PTM characterization, and isobaric tag-based proteomic profiling. To maximize throughput and to minimize downtime, all protein and proteomic workflows are carried out on the same trap-elute LC system and with a single ionization source. LC-MS complements our suite of biophysical, biochemical and modeling tools for the in vitro characterization of protein-ligand binding. Furthermore, when coupled with structure-based design of chemical probes, microflow LC and Zeno-MS/MS enabled the characterization of target engagement in live cells with performance comparable to nanoLC-MS3 and produced physiologically relevant information that expedited hit-to-lead progression in our drug discovery efforts.