Hydrogen-Deuterium Exchange Mass Spectrometry (HDX MS)

Hydrogen deuterium exchange mass spectrometry (HDX MS) is a mass spectrometry technique for studying the spatial conformation of proteins. The main principle of this technique is to place the protein in a heavy water solution, where the hydrogen atoms on the protein surface exchange with the deuterium atoms in the heavy water. The exchanged protein is then digested by enzymes to produce peptide fragments, and the mass of the peptide segments is identified by mass spectrometry: peptides located on the protein surface are more likely to undergo hydrogen-deuterium exchange compared to those located inside the protein, thus increasing in mass, which can be used to infer the epitope conformation of the protein. Additionally, HDX MS can be used to study dynamic changes in protein structure, protein interaction sites, and identification of active sites on protein surfaces, among other applications.

Hydrogen deuterium exchange mass spectrometry can quickly study the amino acid sequences located on the surface of natural proteins in solution, making it widely applicable in studying dynamic epitope active sites of proteins. Since HDX MS relies on the principle of hydrogen-deuterium exchange, back-exchange of hydrogen and deuterium atoms can greatly affect the accuracy of experimental results, making it crucial to avoid such back-exchange. Currently, controlling hydrogen-deuterium back-exchange mainly involves shortening the liquid chromatography analysis time and maintaining the temperature and pH within the range of the lowest back-exchange reaction coefficient.

BiotechPack uses the advanced Waters nanoACQUITY UPLC HD-Exchange System, which employs sub-2-micron chromatographic particles. Compared to ordinary larger particles, this allows for a significant reduction in liquid phase analysis time without compromising chromatographic separation performance, offering ultra-high resolution. Furthermore, BiotechPack integrates automated online pretreatment with high-resolution mass spectrometers, ensuring the reliability of experimental data and improving experimental efficiency, fully guaranteeing the reliability of pretreatment effects and identification results. You only need to inform us of your experimental objectives and send us your samples, and we will handle all subsequent project matters, including sample processing, enzymatic digestion, mass spectrometry detection, and raw data analysis.

Hydrogen Deuterium Exchange Mass Spectrometry HDX MS Analysis Process

Hydrogen Deuterium Exchange Mass Spectrometry HDX MS - BiotechPack Co., Ltd.

Features of Hydrogen Deuterium Exchange Mass Spectrometry

• Low sample requirement and relatively low purity requirement
• Direct study of native spatial conformation of proteins in solution without needing protein crystals
• Study of active sites and spatial epitopes in dynamic changes

BiotechPack Platform Advantages

• UPLC-HDX-MS system with precise temperature control at 0±1℃ to minimize hydrogen-deuterium back-exchange reactions
• Automated pretreatment and sample injection analysis with precise control of hydrogen-deuterium exchange reaction time
• High reproducibility spectra obtained using the nanoACQUITY UPLC system
• Comprehensive, timely, and interactive result management using MassLynx®, DynamX HDX data analysis, IDENTITYE, and ProteinLynx Global SERVER™

Application Areas of Hydrogen Deuterium Exchange Mass Spectrometry

• Study of protein spatial conformation and active sites
• Dynamic changes in protein spatial epitopes
• Dynamic study of interactions between proteins and drugs
• Study of interaction sites between proteins
• Effects of protein mutations or other treatments on protein conformation and dynamics

Sample Requirements

• Protein-ligand and protein samples
• Sample concentration >1µg/µl
• Sample amount >10µg
• Note: Samples should preferably be free of detergents.

Bilingual Project Report

In the final technical report, BiotechPack will provide you with a detailed bilingual version (Chinese/English) of the technical report, including:

1. Experimental steps (Chinese and English)
2. Relevant mass spectrometry parameters (Chinese and English)
3. Information on identified substances
4. Raw data
5. Analysis results of protein spatial epitopes, conformation, etc.

One-stop Service of HDX MS Analysis

You only need to place an order and send your samples, and BiotechPack will provide one-stop service: sample processing, instrumental analysis, data analysis, and project reporting.

Related Services

Protein Interaction Analysis
Protein Structure Identification
Protein Circular Dichroism Analysis
Native Mass Spectrometry Analysis