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HDX-MS with Millisecond Time Resolution
Applied Photophysics collaborated with AstraZeneca, CPI, Living Systems Institute (LSI) - University of Exeter to build a prototype HDX-MS system that allows measuring HDX labelling times from 50 ms to over 300 seconds with single millisecond precision in a variety of biomolecules in a fully automated online manner.
Our research director, Dr Lindsay Cole, led the Applied Photophysics team that built the system’s hardware and software, leveraging APP's experience in fast kinetics, and is now part of the study published in Analytical Chemistry.
The research "Online Fully Automated System for Hydrogen/Deuterium-Exchange Mass Spectrometry with Millisecond Time Resolution" presents a novel system. This system can analyse the kinetics of protein interactions using hydrogen/deuterium exchange mass spectrometry (HDX-MS).
HDX-MS is a powerful technique for researchers to study protein interactions and conformational changes. The technique involves exposing proteins to deuterium oxide (D2O), which replaces the hydrogen atoms in the protein backbone, with exposure times varying between 50 ms and many minutes. The rate at which the deuterium atoms exchange with the hydrogen atoms in the protein backbone provides information about the protein's structure and dynamics.
The authors of this study developed a fully automated system that allows for HDX-MS analysis with millisecond time resolution. The system consists of fully automated sample loading, a fluidic mixing device, and liquid chromatography with a mass spectrometer system.
One of the key advantages of this system is its ability to analyse protein structural change and dynamics at the peptide and residue level of highly disordered protein regions, like intrinsically disordered proteins/regions. An important aspect of protein conformational dynamics that is difficult to study due to the lack of structural biology tools can provide insight here.
Overall, the Online Fully Automated System for Hydrogen/Deuterium-Exchange Mass Spectrometry with Millisecond Time Resolution is a significant development in the field of HDX-MS. The system has multiple beneficial features for researchers. It can analyse protein interactions in real-time and is fully automated. This makes it a valuable tool for studying protein structure and dynamics.
Read this publication here: https://pubs.acs.org/doi/full/10.1021/acs.analchem.2c05310