HDX

Hydrogen-deuterium exchange mass spectrometry (HDX-MS) is a very powerful technique for probing a protein’s higher order structure: protein conformation, dynamics, and interaction. The technique is based on the intrinsic exchange of backbone amide hydrogen to deuterium when the sample is incubated in D2O. The incorporation of deuterium results in a mass increase that can be measured by MS. In the bio-pharmaceutical industry, HDX-MS is widely utilized to ensure batch-to-batch reproducibility, stability, comparability, and to provide epitope mapping analysis.

The BioMS HDX-MS platform currently includes a LEAP-PAL3 autosampler custom built for HDX sample preparation, and an Ultimate 3000 micro-LC system coupled to an Orbitrap MS. The automated sample handling is essential to provide the throughput, reproducibility, and measurement precision needed for analysis of large sample series.

Proteins have very heterogeneous chemical and physical characteristics and therefore the scientific hypothesis will define the design of the HDX experiment.

Please feel free to contact us with questions or discussion of your HDX-MS needs.

HDX-MS at BioMS comprises:

  • Bottom-up, continuous uptake, proteolytic fragmentation HDX-MS experiments to map;
    • Folding, Identifying disordered regions
    • Interaction analysis
    • Epitopes
  • HDX data analysis and visualization with specialized software, to provide both an easily interpreted overview and detailed information.

A typical HDX-MS experiment entails H/D exchange by incubating the sample in D2O during defined time-points, proteolytic digestion at low pH, LC-MS separation of peptides, and data analysis.