Integrated Platform for Mass-Spectrometric Studies of Protein Structure

Summary
Technology for studying the higher order structure (HOS) and interactions of biomolecules is a
highlighted NIGMS SBIR/STTR research topic. Mass-spectrometric methods for studying
HOS include hydroxyl radical footprinting as well as chemical methods to covalently label
amino acid side-chain functional groups, and also hydrogen/deuterium exchange (HDX) on
protein backbone amides. These methods require only small amounts of not-necessarily-pure
material, and can study conformation and dynamics of proteins in solution with or without
ligands. Mass spectrometric methods are especially well suited to biopharma comparability
and epitope mapping studies. Currently, there is no software that can analyze mass
spectrometric data from both covalent labeling and HDX experiments, even though the two
methods both probe solvent accessibility and, if combined, would give finer resolution and
greater confidence. The outcome will be vendor-neutral commercial software with advanced
analytical and reporting capabilities that can handle both HDX and covalent labeling
experiments.Narrative
The higher order structure (HOS) of biomolecules determines function, binding partners,
immune response, and safety and efficacy of therapeutics. Mass-spectrometric
methods for studying HOS include hydrogen/deuterium exchange and covalent labeling
experiments such as oxidative footprinting. The proposed project will result in unique
software to analyze all types of mass spectrometric data for HOS studies, and will find
use in both academic research and the biopharmaceutical industry.