Hydrogen Deuterium Exchange - Mass Spectrometry

Hydrogen deuterium exchange (HDX) MS works in many applications, including understanding how small-molecule therapeutics bind to protein targets, and performing epitope mapping. With this technology, deuterium in a solution replaces the amide hydrogen in a biomolecule’s backbone, and MS measures these changes.

Amide hydrogen on the backbone of a biomolecule exchanges with deuterium in solution at different rates partly dependent on conformation. MS can measure these changes and therefore infer the degree of activity at various sites. These locations can be identified and mapped to the primary sequence, which allows for easier visualization and reveals information about protein structure. Relative folding and dynamics can be determined from the different rates of uptake at different locations.

With UPLC-MS^E, your lab can generate a comprehensive dataset, measuring all peptides without bias by peptide signal intensity. This is an important feature for HDX studies, where key information might be found in low-intensity peptides.

HDX-MS opens new windows into biomolecule dynamics by providing information on the relative deuterium uptake of different conformations of a protein, or locations within a protein. Recent developments have made HDX with mass spectrometry a more accessible tool to study the dynamics of higher order structure (HOS) of biotherapeutics. Industry and regulatory trends have clearly moved towards an emphasis on conformation and the relationships between biomolecules, making HDX a key tool being rapidly adopted in more biotherapeutic analysis areas.