Work

Developments in hydrogen/deuterium exchange mass spectrometry at sub-zero temperatures

Year: 2025

Type: article

Abstract: HDX-MS has become a popular and established method for studying the dynamic conformational landscape and interactions of proteins in solution. Over the years, HDX-MS has been used to analyze increasingly complex protein systems in terms of size, purity, and biologically relevant conditions. For example, HDX-MS has been employed to study membrane proteins in their native lipid membranes from bacteria, intact viruses, and individual proteins within living cells. However, these exciting and complex new applications pose significant challenges to traditional HDX-MS experiments, particularly the need to perform sample pretreatment and LC separation on a short timescale at zero degrees to prevent inadvertent deuterium labeling information loss (back-exchange). A promising development in HDX-MS is the use of sub-zero temperature separation to reduce back-exchange and enable extended chromatographic separation. In this review, we aim to provide an overview of the current state of sub-zero temperature HDX-MS and the challenges associated with its use.

Source: TrAC Trends in Analytical Chemistry

DOI: 10.1016/j.trac.2025.118539

Authors: Anton Berg Hansen, Omid Rouhi, Kasper D. Rand

Institution: University of Copenhagen

Language: English

Topic: Mass Spectrometry Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: hybrid

Estimated APC paid: $5,150

Funder: European Research Council