Hydrogen Deuterium Exchange (HDX) Mass Spectrometry Service

Hydrogen-deuterium exchange mass spectrometry (HDX-MS) is a mass spectrometry technique that uses the proton instability mass present on the amide of the protein backbone to study the protein structure. The principle is that when a protein is immersed in a solution of deuterium water, the amide hydrogens in the main chain of the protein molecule will exchange with the deuterium in solution at different rates, depending to some extent on the conformation. The hydrogen on the surface of the protein in close contact with the deuterium water has a faster rate of hydrogen-deuterium exchange than that formed by the hydrogen located inside the protein. The rate of hydrogen-deuterium exchange for different sequence fragments of the protein is determined by mass spectrometry to derive information about the protein spatial structure and to infer the degree of activity at different sites.

The benefits of HDX-MS include: independence from the size and complexity of the analytical system; low sample requirements; high sensitivity; and the ability to detect coexisting protein conformations.

Creative Proteomics offers a wide range of solutions for hydrogen-deuterium exchange mass spectrometry (HDX-MS) projects. We are equipped with the nanoACQUITY UPLC HD-Exchange System analytical system with sub-dimensional nano-chromatographic particle packing for ultra-high separation. The on-line automated pre-treatment is used inline with the high-resolution mass spectrometer to improve experimental efficiency and fully guarantee the effectiveness of pre-treatment and reliability of identification results.

Creative Protomics's HDX-MS service can provide:

• Epitope mapping
• Identify drug binding sites and drug-induced conformational changes
• Analyze protein-protein interactions
• Provide information on protein aggregation, conformational changes and regions of intermediate structures formed during aggregation
• Study the effects of mutations on protein structure and dynamics
• Study the effect of agents on the structure or stability of biological drugs
• Analyze information on conformational differences between different states of proteins or protein complexes and elucidate structured versus unstructured regions of proteins
• Study the effects of phosphorylation and other post-translational modifications on protein structure and dynamics

Workflow of HDX-MS

General workflow for HDX-MS experiments (Masson et al., 2013)

Sample Delivery Requirements

• Sample type: purified protein or molecule/antibody of known sequence and known binding conditions
• Total sample volume: ≥ 0.10mg; > 0.05mg (antibody/antigen Epitope mapping)
• Sample concentration: ≥ 0.5mg/mL
• Sample purity: ≥ 90%

If you would like to learn more about HDX-MS services or have other needs, please contact us. We look forward to cooperating with you.