Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a technique that provides critical information on both protein conformation and dynamics. Proteins contain hydrogen atoms at the backbone amide positions that exchange with hydrogen present in the surrounding solvent. This exchange process can be monitored experimentally by incubating proteins in deuterium and monitoring the resulting exchange using high resolution mass spectrometry. Proteins will increase in mass as deuteration proceeds, and the rate of hydrogen exchange reflects aspects of the protein structure. The technique can be used in areas such as epitope mapping, protein-ligand binding, protein complexes and comparability studies. HDX-MS has been made more routinely accessible with Waters commercialized integrated systems with lower sample requirements, automated sample handling, data collection and data analysis. Advancements in the technology continues to improve the systematic capabilities of HDX and efficiencies of HDX workflows.
This presentation gives an overview and theory of the HDX-MS technique, explains the Waters approach to HDX-MS workflows and describes application examples of using HDX-MS in probing protein conformation, conformational changes, dynamics, protein folding, and the effects of binding.