How HDX-MS Epitope Mapping Works
HDX-MS Epitope Mapping Workflow
Hydrogen-Deuterium-eXchange Mass Spectrometry (HDX-MS) measures the exposure levels of amide hydrogen atoms on the surface of a protein or protein complex. The first step causes the exposed hydrogen atoms to be readily exchanged with deuterium. Hydrogen atoms that are buried within the protein structure – for instance at the binding site of an antibody-antigen complex – are not exchanged as readily. By examining the peptides using Bottom-up mass spectrometry, and comparing them with the amino acid sequence, it is then possible to determine which regions are the binding locations between the interacting partners.
- Labeling – incubate different fractions with D2O solution in a time series
- Quench – chill and control temperature to prevent further exchange or back-exchange
- Digest – prepare peptides for analysis
- LC-MS/MS
- Data processing – determine deuterium uptake of digested peptides acid and identify binding site(s)
- Comparison – compare similar samples – e.g. antibodies with similar sequences
Applications of our HDX-MS Service
While our HDX-MS service is commonly used for epitope mapping in an antibody-antigen interaction, it can also be used to study other protein-protein interactions, protein conformation changes, multi-protein complexes, small molecule interactions with proteins, disulfide linked proteins, glycosylation sites, protein interactions with DNA or RNA, disordered proteins, etc.
The Rapid HDX Advantage.
High Resolution Even for Large Ab:Ag Complexes.
Quality and Expertise in Epitope Mapping.
With over 7 years of experience in HDX-MS, we are capable of working with a variety of analytes and ligands, and novel projects. No two projects are ever the same and so experimental conditions are always individually optimized for overall project success.
Part of a Complete Epitope Characterization and Antibody Sequencing Platform.
Our HDX-MS service is one part of our antibody solution platform that also offers isolation, protein sequencing, recombinant expression, kinetic analysis and epitope mapping. Combining more than one service into a single workflow gives convenient access to richer, higher quality results.
HDX-MS Epitope Mapping Pricing.
HDX-MS Epitope Mapping
- Detailed epitope mapping analysis for 1 antibody-antigen pair.
- Sample requirements: Antigen 200 μg, Antibody 200 μg
- Turn around time: 4 weeks
HDX-MS Epitope Screening
- Fast analysis of binding location of 10+ antibodies to one antigen.
- Sample requirements: Antigen 500 μg, Antibodies 100 μg each
- Turn around time: 4 weeks
Custom HDX-MS Project
- Custom analysis of protein complexes, conformation changes, ligands and other structural questions.
- Please inquire about experiment design, and sample requirements.
Getting started.
Talk to Our Scientists.
We Have Sequenced 8000+ Antibodies and We Are Eager to Help You.
Through next generation protein sequencing, Rapid Novor enables reliable discovery and development of novel reagents, diagnostics, and therapeutics. Thanks to our Next Generation Protein Sequencing and antibody discovery services, researchers have furthered thousands of projects, patented antibody therapeutics, and developed the first recombinant polyclonal antibody diagnostics.
Talk to Our Scientists.
We Have Sequenced 8000+ Antibodies and We Are Eager to Help You.
Through next generation protein sequencing, Rapid Novor enables timely and reliable discovery and development of novel reagents, diagnostics, and therapeutics. Thanks to our Next Generation Protein Sequencing and antibody discovery services, researchers have furthered thousands of projects, patented antibody therapeutics, and ran the first recombinant polyclonal antibody diagnostics
Talk to our scientists. We have sequenced over 8000 antibodies and we are eager to help you.
The Science of Epitope Mapping.
Antibody Sequencing & Discovery Services.
Other Services.
Antibody Sequencing & Discovery Services.
REmAb®
Monoclonal antibody sequencing from small antibodysamples, no need for hybridoma or DNA information. Full sequence in record time.
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REpAb®
Polyclonal antibody sequencing and antibody discovery service. Sequence antibodies from blood or a polyclonal mixture.
Explore REpAb
MATCHmAb™
Antibody verification by real-time peptide mapping. Antibody sequence confirmation service for reproducibility.
Explore MATCHmAb
HDX-MS
Mass spec based epitope mapping. Epitope mapping for identify the binding site of an antibody to its corresponding antigen with the highest confidence and resolution.
Explore Epitope Mapping
SPR Analysis
Label-free evaluation of antibody-antigen binding relies on a Surface Plasmon Resonance technology. Combined with our expertise in antibody interactions, this results in high sensitivity and the lowest noise even in complex analytes.
Explore SPR Analysis
NovorIg™
Quantify the immune response by accurately profiling the vast antibody-protein repertoire and its different isotypes, and subtypes in serum over time.
Explore Immune Response Profiling