ReviewHydrogen deuterium exchange mass spectrometry in biopharmaceutical discovery and development - A review

•The pharmaceuticals industry is increasingly shifting to protein therapeutics.•Hydrogen deuterium exchange mass spectrometry is uniquely well suited to support biopharmaceutical development.•Applications for hydrogen deuterium exchange span drug discovery, development and manufacturing.•Future developments will allow improved sensitivity, structural resolution and a broader range of dynamics to be monitored.

Protein therapeutics have emerged as a major class of biopharmaceuticals over the past several decades, a trend that has motivated the advancement of bioanalytical technologies for protein therapeutic characterization. Hydrogen deuterium exchange mass spectrometry (HDX-MS) is a powerful and sensitive technique that can probe the higher order structure of proteins and has been used in the assessment and development of monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs) and biosimilar antibodies. It has also been used to quantify protein-ligand, protein-receptor and other protein-protein interactions involved in signaling pathways. In manufacturing and development, HDX-MS can validate storage formulations and manufacturing processes for various biotherapeutics. Currently, HDX-MS is being refined to provide additional coverage, sensitivity and structural specificity and implemented on the millisecond timescale to reveal residual structure and dynamics in disordered domains and intrinsically disordered proteins.

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